It is a little chilly in here The pointy nipples get hard when it's cold Protection from what the cold how does you're you're Tightening up everything so that more heat from the body is not leaving Is that actually what it is?   No, no, no That's why why do you think everything shrivels up because if it's all the everything's going to the core mind usually expands not in the cold Anyways today We're doing it. Oh, I should open my laptop. Yeah trailer dropped last night And it was about midnight our time and it's good thing We didn't react to it because we were sleeping because it didn't have subs. Oh that would have sucked Because we were Considering staying up and watching it at about what 1115 I messaged you instead of fading. Yeah So yeah, that would have really sucked if we had pushed and you would have sucked too because we would have just been like sitting here going but it's apparently only a minute 30 and I mean team didn't notice it didn't have subs till much later. So I got what they say And so we'll have what they're saying like goddess like we did before Yeah, they didn't they noticed too late that it wasn't subbed got it So we know what they're saying, but we don't know it as they're saying a big productions Just please I'm begging you as a idiot who only speaks English Put English on your subs right away, please Anyways, but so yeah, this is a His new film attack. We've seen the teaser for it look like a big action-packed. Yep style of film. Yeah expecting Booming bombing bloodshed and bone breaking Boner all of the boners you need from John Abraham wait what okay? Okay, we'll read it after it wake me up when you're done Hama is gonna be a don't go to bed.  Exactly I don't know Super combat combo Very soon So tell me I don't know Okay Resistance going down The hero That's what we expected. That's how an action film That's how an action trailer should be. Yep short tight show us a bunch of action that will get us pumped. Yep Good visuals as well Go to our actual What they said? So in the beginning they said two months into our life in our life are important two moments in our life are important First one is when we are born and second is when we find out why did we take birth? when In other words when we were born and the second is when we find out why we were born ah And then ending terrorist is this guy going to save y'all India's superhero. Gotcha Yeah, so I didn't really actually even think we needed this I know I kind of got that just yeah, it's gonna be kind of action kind of lines like that But badass kind of stunt work and action and all that I thought it was really cool I didn't I love the cut to the music. I did too And a lot of it looks as is not a surprise A lot of the visuals remind me of gaming and it probably appeals a lot to the visual aesthetic that gamers like with Wargames, so What do you mean game? Gaming like it looks like the like a man in a video game. Yeah. Yeah a lot um Have you ever played any video games like in terms of like the newer kind to that? Yeah, once or twice But that's it with like I'm with Micah. Yeah, once or twice But none of them have ever really I mean I actually in all sincerity have enjoyed more just sitting and watching Micah play when he plays mm-hmm. I it just doesn't Intrigued me at all Maybe that's who that was Ilham Because it for a second.  Ah, he's a big action director. So he did this Eek the tiger That's I believe a big salmon con. I remember the title Did he know a special thing some bad luck and bang bang bang bang bang another lady. Nope Ad second unit ad on bang bang. He was bunty. I know you said a long time ago that you'd like when trailers Do stuff to music yes, I do like that because you like doing stuff to music. Yep Exactly, right? They kind of go together. Yeah, it's true. Although. I don't coordinate my bangs with the bangs Sometimes that depends on the song depends on the song. I mean there are some songs that are just designed for that. Toto's Africa You're going for Toto if I hear that blasting from somebody's house, I know they're getting jiggy with it I especially synchronized to that part. Yeah, so we're Leland got come yeah from there. Yeah. Yeah, we got it Anyways, yeah really cool Really cool trailer Please let us know what you thought about the trailer and also if you're a studio watching because we know all of you big studios watch Please put subs on your videos immediately. Yeah, I would help. I really appreciate that. I mean, I'm just a dummy We know we realize we're not your target audience Exactly, we know that you don't need an audience outside of India But I know you'd like to have that audience outside of India And why not when there's a huge population outside of India that speak English and it would just be really helpful Yeah, especially if it's like in the Hindi language because obviously Hindi's I think them Is it the most spoken or is Dhamma still the most spoken? It's a good question that I don't know the answer to I know obviously been a lot of people to speak Bengali, too But I know the second most English-speaking nation in the world is India. Yep, so I'm just I know most of your population Also might speak English Well, and when you consider the fact that regardless of whatever language you put that in a Lot of people for example, they speak Dhammel But they don't speak Hindi and they speak English They speak Bengali in English, but they may not speak Hindi Well, a lot of Bengali speakers speak Hindi But my point being is if they have their mother tongue in a second language typically the second language is is English I think even more than it is Hindi and Johnny How many people in India would you say on average speak more than their native tongue? Most all have at least two languages. Oh, yeah, for sure But at least three right would you say more speak Hindi as a second language or more speak English as a second? I know we're being subjective Like if somebody speaks to lugo are they more likely to have also learned Hindi or English Yeah, yeah, so a lot of the South which is if they had any films with English subs That would be very helpful for the non Hindi speaking Indians Yeah, see how many it's kind of crazy because obviously in America we will speak American We only speak American And if they speak another language it will be Spanish usually obviously that makes sense. That's the closest That's the number one here But still that's not like a common thing. No It's very common for Spanish speakers to speak English because everyone else in the world speaks more than one language It's just us Americans. We just people do stuff and many many Americans here are like well if you move into here You ought to speak American We know it's called English. We're making fun of them people here.  Yeah, that would help Anyways, let us know what you told about the trailer down below


Oh, let's go live. And it appears we are officially on. So hello everybody and welcome back to another session of our Sussex Vision Seminar Series, as always within the Worldwide Neuroinitiative. I'm George Caffetzis, a former master's student in Thomas Euler's lab, and currently a PhD student with Tom Baden. And as your host for today, I would like once again to begin by thanking Tim Vogels and Panos Bozellos for putting forward this ever-growing and developing initiative towards a greener and much more accessible seminar world. Having said that, please allow me of course to get back to the reason we all gathered here for today and introduce our guest from Ljubljana University, Professor Gregor Belusic. Following his bachelor's studies in biology, Gregor went on and obtained in 2003 his PhD in biological sciences from the same university. The topic, TRG in transgenic Drosophila, with some work conducted in Germany and some in the Netherlands. In 2013, he was appointed as an assistant professor in Ljubljana University and he has been located there ever since, nowadays holding the title of Associate Professor of Animal Physiology. At the forefront of technological development, including among others a multispectral light synthesizer in his lab, they use a plethora of techniques ranging from ERG to single cell IFIS or spectrophotometry and to behavioral assays in attempts to understand how insects see the world. And today we will be hearing about their latest findings on butterfly vision with a phylogenetic aspect as a bonus as they have studied directly of about 10 different butterfly species. So without any further ado from my side, please all welcome Professor Belusic for his token title opponent processing in the expanded retinal mosaic of Nymphaline butterflies. Gregor, this stage is all yours officially. Thank you, George. Hello, good people of the internet. So let me share my screen with you and yeah, can you see it? You can see the power point and the laser pointer. Okay, so today I will tell you something about the processing of color by means of direct interphotoreceptor of ponency in the largest family of the urinal butterflies, which is the Nymphaline butterfly or brush footed butterflies. The welcome slide shows you the reflection from the eye of the blue morpho which belongs to this great butterfly family and the colorful pattern is called the eye shine phenomenon that can be readily observed in many of the butterflies, virtually all Nymphalids and some other families of butterflies and that can be exploited to infer about the function of the retinal photoreceptors that make this eye shine so colorful. We will hear today about the redomatidia and the non-redomatidia which are apparently green, yellow, blue and all colors due to all kinds of screening and visual pigments in theomatidia and we, as we'll hear, have managed to decipher a bit of this phenomenon and now we can more or less say what's inside hidden below this beautiful mosaic. So first a few words on color vision. Color vision based on chromatic sets of spectral photoreceptors was first I think proposed by from Helmholtz and he just suggested that human vision should be based on three types of spectral photoreceptors but that is definitely true and valid in nowadays but this alone cannot explain some more complex phenomenon in color vision like the after effects, the non-existing colors like combination of red and green, et cetera. So that's why hearing German psychologists proposed in the same century the theory of opponent processing of color where he suggested that inputs from different spectral photoreceptors, spectral channels are compared in some kind of interneurons that weigh the different inputs with different polarities and then report to the brain the outcome which is usually presentable in as a family of curves of sensitivity that oscillate between the positive and negative parts and tell us that our neurons that convey signals from the retina process color in the opponent manner. So a neuron can weigh blue versus yellow color which is a sum of green and yellow or it can weigh red against green. Bushbaum and Gottschalk in 83 proposed and proved that the opponent system should be accompanied by an acromatic signal channel reporting the absolute luminance and thereby the information transferred to the brain is optimal so two opponent axis and one non-opponent or acromatic axis. What is new is that now we can measure the opponent signals not at the level of the interneurons but in the retina, in the photoreceptors themselves. The first reports in the literature date from like 83 where Tomislav Matić has published an article in comparative physiology on spectral sensitivity in Papilla butterfly where you can see that the spectral sensitivity curves go to negative parts and he showed that this is due to electrical interactions in the retina. The problem with this was that it is quite difficult to discern this hyperpolarizing responses of like bioreceptors hyperpolarizing in the green or blue receptors hyperpolarizing in the green orange from field potentials from ERG from extracellular artifacts and usually this has been attributed to bad preparation, poor electro penetration but now we know that it's a true phenomenon that can be treated and even exploited to measure the sensitivity of opponent units not only on the primary units. Subsequently, it has been also recorded in other aspects of animal vision like polarization vision. Roger Harley has published an article on dorsal rim receptors in the flies where you can see that the responses from polarization sensitive receptors can have negative going tails so which are from the antagonistic polarization detectors oriented at 90 degrees. This has been conveniently shown with calcium imaging in receptor terminals in the dorsal rim by Peter Weir and Dickinson in 2016. Now, in the recent years, we have a number of key publications that convince us that the opponent processing is indeed taking place in the receptors themselves without the intervention of interneurons such as the case like in Drosophila where color vision is conveyed by the central photoreceptors, UV blue or UV green sensitive and in their terminals, you can measure with calcium imaging negative signals in the spectrally opponent parts of their sensitivity. We have published also an article together with the lab of Kentaro Aricava on opponent signals in the accents of photoreceptors in Papillio where spectrally antagonistic receptors project synapses and utilize histamineergic chloride channels to convey inhibitory inputs into the spectrally opponent photoreceptors. Another class of histamineergic channels is used to convey signals from the photoreceptor to the LMC interneurons for downstream processing. So the same neurotransmitter histamine opens two different kinds of histamineergic channels. One are fast with the sharp dependence in the LMCs and the other are slightly less sensitive in the photoreceptor accents. Another striking example are the photoreceptors of zebrafish as published by my host lab today where you can see signals with opposed polarity in different parts of the spectrum. So like blue plus, red minus, UV plus, I think it's also green minus receptors at a certain level. I think this is mediated by interneurons. So it's not totally direct between the photoreceptors but nevertheless the signal is present in the photoreceptors. So the opponent processing is taking place at the very first stage of vision. We focused in our research on diurnal butterflies, specifically nymphalic butterflies. So diurnal butterflies have nothing but excellent color vision which can be shown beautifully with behavioral experiments. So they are able to discriminate colors from grace which means that they possess true color vision which is the ability to discriminate spectral composition independent of the intensity and they can be nicely conditioned and demonstrated that they are able to show that to discriminate colors as far as, or as little as one nanometer apart from each other, wavelengths one nanometer apart from each other. This is all based on either simple sets of photoreceptors like in the monarch where you have three different options, UV blue and green options or expanded sets of obscenes like in Papillio. And this together with lots of color filters, results usually in expanded sets of photoreceptors where receptors and non-visual pigments mutually produce a very complicated spectral sense of detectors in the retina. Brush food butterflies have been traditionally thought to be all trichromatic insects similar to the honeybees which possess UV blue and green photoreceptors. And this has notoriously been showing up in all physiological studies so far. Parantica, the night related to the monarch or Polygonia or Sasakia, they all appear to have trichromatic sense of photoreceptors. But behavioral evidence has been accumulated in the years that these butterflies, although they possess only three spectral channels, presumably or at least three options, they are able to distinguish parts in the red part of the spectrum. So like red from orange or red from deep red. This has been demonstrated for the monarch and for Heliconius by Adriana Grisco and Collab. So even though there is no extra option here, they still can see colors in the red part. So to identify the red receptors, we have studied a number of these beautiful butterflies and recorded from the retina with single cell electrode technique with sharp microelectrodes, recorded their eye shine. We studied their anatomy and we found the corresponding substrate for color vision in the red part of the spectrum in some of them, but not all of them. So we collected some at the faculty, some on the beach of the Adriatic Sea and some were from Costa Rica, a butterfly supplier. So we could do experiments all year long. To study them, we lean very much on our own produce from our garage, which is the so-called net synth, a spectral combiner that utilizes the diffraction rating to combine the output of the LEDs into a tunable spectral sequence of stimuli, with which you can very quickly record the spectral sensitivity of impaled photoreceptors. So as expected in Apatura Ilya, which is one of the infallid butterflies that we studied, we have found the green photoreceptors that are actually very broad bed sensitive, the UV receptors and the blue receptors, which are showing signs of opponent input from the green photoreceptors. So actually the UV receptors can be called UV plus, green minus and blue plus, green minus for the blue receptors. We are very sure that this is due to the direct interphotoreceptor-opponent interactions via histamine-ergic chloride channels. The green receptors are not opponent in this case. So Apatura possesses a basic infallid retina. We studied a number of animals found many, many cells and they all belong to three well-definable spectral classes, UV, blue or green sensitive. The UV and blue receptors always have a maximal sensitivity to vertically polarized lights so doors are ventrally oriented. And we know that these are the massive receptors R1 and R2 at the distal part of the butterfly raptom that are long visual fibers that convey color vision directly to the medulla to the second optical ganglion. The rest is the green receptors which are either oriented horizontally or diagonally. So this can be told by measuring their polarization sensitivity. And these are mostly dedicated to acromatic motion vision but of course they also contribute to the discrimination of colors in the green part of the spectrum. Now this opponents in the UV and blue receptors is of course dependent on light intensity used but somehow in the blue receptors, for example, you can see that the positive and negative going part are well matched so that the gain in the blue and the green receptors is optimized that the opponent system works at a huge range of light intensity and allows for color discrimination and deem to very bright light. It is also surprisingly not very dependent on the aperture of the stimulus. So either if you illuminate light, only one or a number of matidia, the effect is very slight, more matidia creates slightly bigger negative response but this suggests that the opponent process is taking place in a single photoreceptor. The opponent response is of course slightly delayed in the blue receptor because there is a synapse between the green and the blue receptor but for example if you look in the ultraviolet receptor the latency to UV response is slightly longer than the latency to a green receptor which is probably due to the very high transduction gain in the UV receptor which then becomes slightly slower than the massive green opponent counterparts which can strike the opponent signal even before the UV receptor gets depolarized. Mapping of receptive fields show that everything comes indeed from a single omatidium and these receptors like blue and green receptors combine into combinations with either high polarization sensitivity in the main part and low or high polarization sensitivity in the opponent part. So omatidia can either contain all green receptors converging onto single UV or blue receptors or pairs of horizontal converging onto vertical and one type of omatidia is suitable for color vision the other is supposedly suitable for polarization vision because you have also polarization of poignancy between the blue and green photoreceptors. Apatura contains the basic mosaic of nymphalids which is based on the long visual fibers that are either UV or blue sensitive and short visual fibers that are green sensitive and always opponent to the long visual fibers. The opposite is never true or can never be observed in the short visual fibers so that the long visual fibers would feed opponent synapses onto the short visual fibers. Together these are one and two blue or UV sensitive form three types of omatidia that are randomly distributed across the retina combinations blue, blue, blue, UV or UV, UV. This forms the retinal mosaic that has a predictable pupil sensitivity. So we also measured the pupillary response of these receptors of these omatidia by illuminating them with monochromatic iso-quantal stimuli and we observed how is the shine from the reflected light from the omatidia decreased upon stimulation with monochromic light. Thereby we could record the action spectra of distinct omatidia. And you can see that a dark adapted eye has a uniform eye shine. The UV or blue adapted eye has a different pattern of attenuation of pupil excitation. And in Vanessa, we noticed that indeed we could confirm that the omatidia best responded to green stimuli, but some omatidia responded strongly to UV and blue, some to UV and not to blue and some to blue and not to UV, which corresponds to the three omatidial types randomly distributed across the retina, like shown here in this patch of omatidia that has been studied with pupillometry or optical retinography. Then in some nymphalates, we nevertheless found some additional spectral classes, a novel spectral class that we found particularly exciting because these are receptors that we found in, for example, in arginis, the fritillary that are depolarized in green and hyperpolarized upon red flashes. The question, of course, is whether the hyperpolarization in the red is an ERG artifact or true synaptic interaction. Therefore, we injected current through the electrode. And first, with positive current, we depolarized the cell, which led to the decrease of the depolarizing responses in the green. And it also increased the hyperpolarizing responses in the red. So we then turned to negative current, which hyperpolarized the cell, increased the responses in the green, and reversed the hyperpolarizing responses in the red. So the responses in the red are due to the current through histamineurgic chloride channels. And this chloride current can be reversed if you are injecting sufficient current close to the synaptic site, which is in the axons in the lamina. In the retina, this is quite difficult to do. But if you are close to the synaptic site and it works, you can show that your current can be reversed and it's a chloride inhibitory current. And then we used selective adaptation with the monochromatic light. First, if we adapted the cell, which was green plus, red minus with red light, we killed the response in the red and we isolated the green unit. Then we also shown monochromatic green light. We silenced the green part and we isolated the red unit. And therefore, we showed that two spectral sensitivities of the units in the opponent pair could be measured independently using selective chromatic adaptation.  And we could measure for a longer period in those receptors. We found those green plus, red minus receptors. And we could measure the spectral sensitivity of the red unit using selective adaptation. And it appeared to be a beautiful, spiking, very sharply tuned red receptor with a maximum at around 620 nanometers. Of course, we also measured the sensitivity of the green unit with red adaptation. And these are the green phases. A receptor maximum is sensitive to green light at 520 nanometers. So this type of receptor has been found in all of these butterflies, nine species, the NAUS, prepona, heliconeus, Haraxes, the two-tailed Pasha, fritillaries, arginis, and spaharia, melitea, and morpho. They all show these negative-going responses in the red, but not in Apatura or Vanessa. The telltale sign for the discrimination between those two butterflies versus the rest is their eye shine. So if you look at the reflected light, you can see that in Apatura and Vanessa, the eye shine is uniform with no red color to matidia, while in Haraxes, the eye shine has non-redo matidia, which are yellow to green, even blue sometimes, and red or matidia. It has been noticed also before that species with mosaic eye shine with red or matidia, like heliconeus, are indeed able to discriminate red from orange or red from deep red, but not Vanessa, which has a uniform eye shine. It can only discriminate like red from blue, but not red from orange. So it's not in terms of the independent color vision. Of course, we took heliconeus from the butterfly supplier, and we immediately shown in the same species the existence of this red minus unit. The cellular identity of this green plus red minus cell was surprisingly R1 or R2. We showed it with Lucifer yellow dye injection. So in the location of the usual UV or blue receptor, in the long visual fiber, we found the green sensitive cell, which means that these cells express long wavelength opsin, and this has been sitting in the bioarchive for a while now, found by McCulloch and Adriana Briscoe, that in heliconeus, a class of R1 and 2 receptors express a green opsin. Indeed, they even co-express a green and blue opsin. So we could confirm the co-expression, also in heliconeus, because this green cell is actually picking in the blue with a tail in the green. So a more difficult question was the cellular identity of the red unit. We were never able to directly record from the red unit. We found many hundreds of cells. Our number is something like 500 recorded cells, and we don't have a single recording from the red unit directly. We made multiple correlative experiments to show that this unit is actually probably the basal cell R9 sitting in every butterfly or matidium with a so far unknown role. This unit has a very small dynamic range. It operates a bright light, because it's at the base of the retina shaded with visual and screening pigments, and it's a small cell with a few microvillage. So therefore, its sensitivity is not very high. If you make an anatomical section to the eye, to her axisi, which I've shown before, half a millimeter of eye, and only 12 microns at the bottom is this biloved R9 receptor heavily pigmented. But the lucky coincidence is that it's sitting directly on top of a mirror made from tracheoles that actually doubles its optical path. So its optical length is about 25 microns. So it's still very small, but we know from mosquitoes that cells with such size can operate and can convey vision or color vision. Second, the latency of the response was in the red unit very, very long, compared to the latency of opponent response of green units in the UV plus and blue plus, green minus receptor. Here, the difference is on the order of one or two milliseconds, but the red unit becomes very slow at low light intensity, and even at high light intensity, the delay is long, which means, again, that it's a small cell with a very high transduction gain that makes everything very slow. And finally, we measured its polarization sensitivity with rotating polarizer. And we found that both the green unit and the red unit best responded to a vertical polarizer. So the excitation of the green unit and the inhibition by the red unit were maximal at vertical polarizer. And the only candidate cells with vertical microvilla in butterfly retina are R1 and 2, which is the green unit, and the basal R9, which is the red unit. Finally, we mapped the receptive fields of both units, and we found that there is no pooling across the retina. All signals emerge from a single omatidium. So the green and the red unit sensitivity spatially overlap. We created then a very simple optical model. We projected supercontinuum white light through the random, and we calculated light absorption by each micrometer slab of the random. And we know from published articles that R9 expresses a green opsin. But due to the filtering with overlaid random years of green sensitives for the receptors, most of the green light is absorbed by them, and the sensitivity of the red unit is shifted to red part. But then the subclass of the material contains this red pigment besides the random. And these are the omatidia that are shining red in the eye shine. And this pigment suppresses the sensitivity in the green part, thereby making our selective adaptation experiments at least possible. Because if you project green light, it will not excite the red unit because of this red screen pigment. Surprisingly, very, very similar sensitivities of cells, of basal cells, have been published for wild honeybees in the article of Page and Menzel from 1992. So some wild honeybees, which have mine receptors, probably contain red-sensitive basal receptors, which, of course, opens new avenues for honeybee research, at least for the wild honeybees. Then we analyzed not only electrophysiological, but also eye shine data across the new phallid phylogeny. And we found that across new phallids, the red omatidia with red receptors occur really sporadically. So closely related tribes of subfamilists can have either only green receptors, no red receptors, or red receptors. These all have only UV blue-green sets of receptors. These have red receptors. So it's surprisingly spread along the phylogeny, which means that all these traits, the green sensitive R1 and 2, the basal R9, the screening pigment, they all occur upon a simple genetic switch that can be switched on or off. If you look carefully in the tribe Arginini, the fritillaries, the retina is even sexually dimorphic. So only the males have the red receptors. Now I'll explain this a bit later. Back to the page in Menzel article, many hymenoptera show red receptors, which is most likely made possible by the basal R9. So again, this can be switched on or off among across the species. So together, these omatidia that are red shining create an expanded new phallid retina, expanded mosaic with expanded receptor sets. We decided to present the sensitivity of such animals like the two-tailed pasha with basic receptors, which are UV plus, green minus, blue plus, green minus, and green non-opponent receptors. Along with the expanded set, which is UV plus, yellow minus, blue plus, yellow minus, then green plus, red minus, and red receptors, plus the yellow plus, which is due to the polarization sensitivity, probably the diagonal receptors 5 to 8 in the proximal tier of the retina in the red omatidia. So receptors actually function as broadband detectors, serving quite possibly motion vision, expressing very likely a green opsin, but with sensitivity slightly deformed due to the red screen pigment and the filtering in the raptor. What is very important is that the polarization sensitivity of these green plus cells is super small. So they are almost insensitive. It can be measured that it's vertical, but it's very low. This will be important later in our pupillometry. Again, in this expanded retina, you can see blue and UV receptors combining with opponent units that are usually with very low polarization sensitivity. In a few cases, there are units with polarization sensitivity in the opponent part. But the blue receptors can be either blue plus, green minus or blue plus, yellow minus. And to prove that this is formed by the green or yellow receptors in non-red or red omatidia, we made a simple simulation where we subtracted the sensitivity of a green-adapted blue receptor, which is the steel curve in both cases, from the sensitivity of a green receptor or a yellow receptor, weighed by a certain linear factor. And the result is the dashed curve, which perfectly fits to the recorded sensitivity of a blue plus, green minus, and blue plus, yellow minus receptor. So we see that these opponent combinations can occur in both types of omatidia and perhaps contribute to color vision if this negative going tail has any significance in the subsequent processing of color in the medulla. Whether it has or not is a very difficult question, but still relevant, as we will see later. The spectral sets of receptors in the expanded retina are highly conserved across Nifalidae. We could find identical sets of receptors in Charaxes, the Puteil Pasha, also in the Monarch. Note these orange receptors, the red receptor, the blue plus receptor. Then in Prepona, again, an orange receptor, red receptor, also morpho, orange receptor, red receptor. So this is quite a surprising find that is so conserved, despite the eyeshines being so differently looking. Probably the eyeshines is more influenced by the eye size and by the dynamics of the pigment than by the spectral sets of the photoreceptors. Similar sensitivities have been, by the way, published also by McCulloch and Adriana Briscoe in Helikonium. So they are sitting in the article elsewhere. Now, accidentally we came across the same sister species of Arginis in a study together with the lab of Kentaroa Ricava. We recorded from Arginis Pafia, a polyarctic species, and Kentaroa and Peru Chen have recorded from a few years ago from Arginis Sagana. Their main aim was to show the putative sexual dimorphism in the retina because females have such black, prominent wings. Pafia has orange wings in both sexes. There are some black moths also around in Europe, but I think more importantly, the males have very, very large entreconia, which are structures for dispersing the pheromones, and therefore the female search for males may be more dependent on olfaction than on vision. When we looked into the retina with the eye shine with the oftalmoscope, we have seen immediately that the females of both species have a uniform non-mosaic eye shine, which is the panel A and C, while the males have dark omatidia, if shown with monochromatic light, that appear red if shown with white light. So the eye shine in the males is mosaic with red omatidia and in the females, non-mosaic. We made histological analysis and we found this red screening pigments that I've shown before in the male eyes and not in the female eyes. And these are the screens that are able to screen to filter incident light into the basal photoreceptor if the omatidium is shining red. Again, electrophysiological analysis has indicated that the basic set is present in the females, UV blue-green receptors, while the males have, in addition to the basic set, UV blue-green, also blue plus, yellow minus, green plus, red minus, orange receptors and red receptors, all due to the special filtering pigments in the red shining omatidia and the basal red-sensitive R9 cell. Such an expanded retinal mosaic has been found already in Heliconius, which belongs to the same subfamily as Arginis. So it's both are Heliconini, but Arginis is a tribe of this. So in Heliconius, the sexual dimorphism is of two types. One is due to the application of the UV pigment, which has co-evolved with the UV pigmentation on the wings, and two UV pigments then create six types of omatidia because the combination can be blue-blue, UV-1 blue or UV-2 blue and UV-1, UV-1 and UV-2, UV-2, et cetera. Six types and similar in Heliconius with red omatidia and red receptors, the opsin in R1 and 2, which is now green, co-expressed with blue, leads to six omatidial types, UV-blue, blue-blue, UV-UV, green-UV, green-blue and green-green. So the whole retinal mosaic is like duplicated in order to get another extra spectral channel for seeing in the red. That is quite a high price to pay for an extra channel, but this is apparently the developmental way to go in the butterflies. So to summarize the opulent processing in infallid butterflies, the basic mosaic is based on UV and blue long visual fibers, being fed opulent signals from the long, from the short visual fibers sensitive to green. The red omatidia additionally contain this green sensitive R1 and 2 and having opulent signal fed from the basal R9, which has light filtered with red screen pigments. It takes all these factors to create a red receptor. An additional factor that is required to create red receptor is also a shift of the main green opsin from 520 to about 540, 45 nanometers, which has been observed in all these species that we have studied. So if you want to put red sunglasses to create a red receptor, which expresses the green opsin, it's not sufficient to have a simple short wavelength green, 520, because then the red filter will be effectively an infrared filter. You must go to a longer wavelength green receptor in order to exploit the property of the red filter. The predicted pupillary response of these cells is now shown below. So there will be high response to blue and to green in a blue, blue omatidia. Response to UV and blue in blue, UV omatidia, response to UV light and green in UV, UV omatidia, and the new red omatidia will be presumably sensitive to blue and green, UV and green, or green and green. So no UV response. And those wavelengths symbolize the response to a rotating polarizer. We've seen before that these green receptors are very, very insensitive to polarized light. So the pupillary response to polarized light will be very independent from the polarizer orientation in the green receptor, which is not true for the blue or UV receptors. So we then illuminated the eye of the two-tailed Pasha with monochromic light and the principal component analysis of the pupillary response yielded six classes that perfectly corresponds to the prediction. Blue-blue receptors had strong response in the blue and weak in the green. UV blue, strong response in the UV, and in the blue, UV UV, strong response in the UV. And the three, this rectangle symbolize the size of oscillation upon polarizer rotation. Those three types in the lower row have very small response to polarizer in the green. So we believe that these are the blue-green omatidia, green-green omatidia, and green-UV omatidia. So we found six types of omatidia that were then mapped to the original eye shine. So all the top row omatidia, ABC, which are BB, UB, and UU, were mapped to the non-red omatidia, while ESG, the omatidia with the green receptors were mapped to the red omatidia. So together, the non-red and the red created this matrix, which corresponds to the slightly boosted image of the original eye shine. So we have proven that the green receptors, together with the red receptors, reside in red-colored omatidia. This is now how we understand the eye shine, which is, again, shown on the right-hand side. We record this from the central part, but as usual, the eye is regionalized as in all other butterflies the dorsal part contains very few red omatidia and the ventral part is enriched because the ambient in the ventral hemisphere is much richer in the long-wavelength colors than the dorsal hemisphere. Similarly, convincing results was found in the paper kite, which belongs to the danaïne. Again, pupillary response was strong in the green in three types of omatidia and the response to polarized light was very independent of polarizer orientation, which is not true for the bottom row, where you have UV blue and blue blue omatidia, which strongly responds to polarized light at different orientations. So again, six omatidial types in a species with a mosaic eye shine with red omatidia. We can now consider the butterfly eye shine explained, but of course it's slightly more complicated than just saying that it has non-red and red omatidia because you can see that Harax eye shine in a dark-adapted state has non-red omatidia, which can even appear blue and light-adapted appear green or yellow. This is due to the massive conversion of the main visual pigment from rhodopsin to metrodopsin. This can be triggered with white light and this photoisomerization creates a large differential spectrum and then a sum of rhodopsin and metrodopsin can be fitted to this differential spectrum and you can immediately get the main rhodopsin, which is here, 544 nanometer rhodopsin, the green rhodopsin in this species, which has red receptors. Apatura with a non-mosaic eye shine has a 520 nanometer opsin, so a convincing shift of the opsin to create red receptors. So meanwhile, we have contributed a bit to the study of Papillio lamina conducted by the lab of Kentaro-Rikawa, Kentaro and the co-workers have painstakingly analyzed the synaptic connectome of the lamina. They have traced all the cells in these atomatidia and reconstructed all the synapses, nine photoreceptors in each matidium, four LMC neurons and we have contributed a bit of modeling of the responses based on the synaptic strain. What we can learn from this matrix of connections in Papillio is first that spectrally identical cells, R3, R4, or R5, R2, R9 are not opponent to spectrally identical counterparts, which makes perfect sense, except in some parts, R5 and R6, which are probably spectrally identical, but with microvilli oriented at different angles, which forms then polarization analyzers. Second, in Papillio, long visual fibers, R1 and R2 can be presynaptic to the short visual fibers, R5 to R8 and R9, which is very different from lymphalates, where this never happens, at least as we could observe with electrophysiology. The result is then a stunning complexity of the retina that is underlying this fascinating ability of the animal to discriminate one nanometer apart, monochromic light, but we could also explain the actual physiological measurements, for example, the process dominating the spectral sensitivity of the LMC neuronesis convergence. So the spectral sensitivity can be explained just by a linear sum of the spectral sensitivities of the overlaying photoreceptors, and this can be explained either without knowing or with knowing the connectome. So the connectome perfectly explains that you have three types of formatidia and three types of spectral LMCs that differ in the UV and the red part slightly and possibly could convey the long wavelength information to the medulla, although the sensitivity is relatively broad and all the spectral richness of the photoreceptors is somehow lost. So the question remains, how is then the color information conveyed into the medulla here in Papillio? One possible answer is that it goes through the opponent tail of the long visual fibers that are red, UV plus, green minus, blue plus, green minus, violet plus, broadband minus, or blue plus, red minus. And these sensitivities are again, nicely explained by the linear weights of the synapses and by the physiological results. So this is again showing the previous measurements in Papillio lamina. To conclude, we have now seen the opponent processing phallus and in Papillio, and we can of course ask ourselves, what is the adaptive significance of this spectral design and cellular design of early opponent processing circuitry in butterflies? The first answer that has been proposed by many authors before us is that the spectral opponents leads to a de-correlation of spectral channels and it narrows down the spectral sensitivity of the photoreceptors because they are log amplifiers. And even if the opsin has only one per mile or even less absorbance in a very remote spectral part, the log amplification machinery in the cell will create a large response, for instance, to red light in a green sensitive cell. And with the opponent, this is then reduced, it's narrowed down. Second, the opponent circuitry, as shown now, appears to form an achromatic channel and two opponent channels which conform to the theory proposed by Bushbaum and Gottschall for optimal information transfer. This has been suggested, for example, by Rudi Behnia, he just last year in Drosophila. Another finding by the host lab is that the spectral tuning of opponent channels actually maximizes the variance of information across spectral channels. So Tom and co-workers have went to the nature, measured natural spectra in the natural habitat of the zebrafish and found that the principal component analysis of this spectra nicely matches the spectral characteristics of the achromatic brightness channel and the two opponent channels. We can add a little bit of our insight into this mosaic. So the question here posed is, how is the transduction gain in the photoreceptors adjusted so that the strength of the opponent is just right, which means that the opponent's responses do not kill the main cell or vice versa. So first in Ivalida, the short visual fibers are always opponent to the long visual fibers. Never the opposite happens. And the long visual fibers are short wavelength receptors in principle, UV and blue. And short visual fibers are a long wavelength receptors, green or further. Under the natural illuminance, for example, the skylight, the daylight spectrum, the quantum catch is very low in the UV receptors, slightly higher in the blue receptors and the highest in the green receptors. Therefore, the transduction gain is very high in UV receptors, high in blue receptors, and low in green receptors. This has been noted, for example, by von Helversen, who behaviorally measured the absolute sensitivity in the honeybee, a very high sensitivity in the UV, smaller in the blue, and smaller in the green. We have measured sensitivity in the mouse, for example. In UV receptors are one and a half log units more sensitive than blue, and blue are more sensitive than green receptors. So their gain is adjusted to the composition of the daylight so that all receptors get equally depolarized by exposing themselves to the average light, which brings them into the optimal range of depolarization for optimal signal-to-noise ratio. Now, a tendency from UV and blue receptors to green receptors would, presumably, very much negatively, detrimental influence the signal in the green receptors, because these cells, upon slight exposure to UV or blue patches, would strongly suppress the signal in the green receptors, which is detrimental for motion vision. Therefore, we've seen that in the expanded retina, the green versus red channel has found its own bypass through the motion vision in the means of the long visual fiber, which is sensitive to green, receiving opponents from the red. This is an exception where R9 is a point to a long visual fiber, but R9 is heavily shaded, it has low photon catch, and the green cell is not a part of the motion vision circuitry. So an obvious answer to the hypothesis is, of course, opponent circuitry has evolved to function under natural illumination. Not surprising. And perhaps another hypothesis stemming from our finding is whether the option co-expression and filters and all the sophisticated tricks that the butterflies use in the retina evolved to perhaps adjust finally the gain in opponent pairs to detect colors in some particular parts of the spectrum, where, for example, the signals from the wings of conspecifics or from the flowers are the strongest. So aside from this generalistic opponent circuitry, which has been found across species, butterflies apparently have also more specifically evolved channels where these optical tricks are perhaps used for getting the game just right so that the opponents works perfect. I shall conclude this lecture with small advertising for the articles that describe all the stories that I've told right now. One has been published in the proceedings of the Royal Society B on the finding of the red receptor and the green receptor. The retinal mosaic explained is now the review in philosophical transactions and the sexual dimorphism in fritillaries in the same journal. The article on Papillot is in preparation and should be submitted quite soon. So you will be able to read all about it in detail probably next year. I would like to thank you for your attention and to my co-workers from the lab, especially Primoz Spiri, who developed all the fancy optical measuring devices, the pro-pillometry. Marco Illich has contributed a lot to the analysis, to the modeling. Andrei is our anatomical expert. Aleš has contributed to the electrophysiology. Marco to the optical measurement. The butterflies were supplied by Costa Rica entomological supplies and by a colleague Mark Jals from the UK who supplied Apatura. They were determined also by our colleagues from the zoology department here in biology and the graduate students that have worked on the butterflies at Lucia and Iost. All done in collaboration with Kintaro Aikawa and Dukas Teringa, financed mostly by the Air Force Office of Scientific Research and a little bit by the Slovenian Research Agency. Thank you very much. I have not been able to see your questions because I'm actually in quite a Zoom and your questions are on YouTube. So this will be moderated by George if there are questions. Yes, here is where I come in again. And thank you very much Gregor for this impressive work like both in wealth and in breadth. And I really enjoyed personally how your work in non-model species can inform work in others like as the Hanebis as you mentioned already. There are already a number of questions appearing in the chat so I will keep mine for later on. I will restrict myself to being the host and the moderator of this discussion. And I would like to remind our audience that they can either post your question in the chat or join us in 10 minutes from now in the Zoom room that we are currently sitting in to ask in person or continue in a more informal get together. So given that you touched on it towards the end of your talk, I will start with the last question that appears on the chat currently from Charlotte Lewis. Why do they bother making the red receptor in the first place? And you already mentioned something on it. It seems very energy and cost inefficient. So if you could design it, would you design it differently, let's say? Yes, the solution to the red receptor in most other butterflies is very different. So the piered butterflies and papillionate butterflies, they utilize their diagonal proximal tier of receptors R5 to 8, which are equipped with a long wavelength sensitive opsin and a bit of filters. And they create a very powerful, sensitive system of red receptors. And this solution in infallid butterflies appears indeed slightly clumsy, energy consuming, but filters are involved all over the place because red opsin is seemingly not trivial to evolve. So you need some kind of long wavelength opsin plus the filters to create a red receptor. And of course the selective pressure to design this receptor must be enormous because as we have seen the phylogeny, these receptors occur and appear and disappear all across the infallids, sorry for clicking this. But yeah, they can be switched off among, on and off among the sexes and present in very closely related sister species or absent in those species. So if I would design them, I would definitely go the papillionate and piered way because it's so much stronger system. But then maybe those receptors, the diagonal red receptors can influence the motion vision system because as we have seen in the publications from Papillio, motion system in Papillio motion detection system is not independent from color contrast. So it can be triggered with color contrast only. And maybe that's the price to pay that your motion detection becomes sensitive to color variation. And I think there will be like some really nice insights from Kit Longden early next January, like when we continue the series about this specific topic that we just mentioned. Next question is from Luisa Ramirez. Do you think that a reason for having an interphotoreceptor mechanism instead of an interneural mechanism such as horizontal cell connections could be again in the image resolution? And if not, do you have other explanation for such a difference across species? So the main difference that I've shown here was for example, the difference between Drosophila and the butterflies. In Drosophila, you can only see the opponents in the receptor terminals. That's why they have never been recorded in the receptor somata except for the dorsal rim. And we hypothesized that the Drosophila laminar has been secondarily reduced due to the development of the neural superposition. So this very complicated developmental sequence that leads to the convergence of different material to common interneurons. Then somehow abolishes any possibility to create this fancy connectome that allows for color opponent circuitry. But direct opponents between the photoreceptors is probably the most economical way to do it because avoiding one interneuron is a good way to save on energy and resources. So if you have the laminar dedicated to other tasks like motion vision, high sensitivity motion vision, this is the path that the flies have taken, then color vision comes second. And this is very nicely seen in all behavioral tests of color vision in flies which are just off the scale if you compare them to the butterflies. So performing quite poorly. And but of course the gain is that they have colonized all the worlds very successfully and they can torment us in any kind of any time of the year while butterflies are much more sensitive, maybe. And yeah, if an animal can avoid an interneuron then this of course also comes at the price of flexibility but of course our findings do not exclude the existence of interneurs in the medalla. Of course there will be inter-o-material opponents weighing from different types ofomatidia, different spectral sets in adjacentomatidia, then interneurons can compare the signals from them, of course at the price of the spatial resolution because then the precision, single-matidium precision is lost. But that's not something very new because we also do it. We also compare spatially different spectral receptors and our resolution of color vision is also inferior from the acromatic vision. So the last question or I should say questions appearing are from Tom. I will start with the first part. How well do the spectral zero crossings tend to match any difference in a respective spectra in cone-specific wings? We have not systematically measured the spectra of the wings and I can immediately tell that those points where the spectra crossed zero are not fixed because those receptors tend to have different moods of opponents. You can have a sleeping beauty where there is no opponents in the butterfly and it's very difficult to get any opponent signals in the receptors. They can be eventually enhanced with receptor, depolarization with heart injection or you can have an opponent with very strong flavor and so what I'm suggesting here is that the opponent may be state dependent. So the next thing we are going to test is of course the effects of neuromodulators on this opponent and this will then affect also the crossing zone. So it's not totally fixed. But the wing spectra are well measurable and this is of course an attempting thing to measure because we all speculate that the butterfly vision is sophisticated also to see their cone specifics. So yeah. The continuation of this question again from Thomas, do you think that this is a major driving force in selection or is it possibly more about wider statistics across the scene that kind of thing as well? Yeah, I mean we have seen highly conserved sensitivity across species. So you can see that this is quite a generalistic visual system not very particularly tuned to the wing spectra. Of course it would be probably good to measure exactly these crossing points and correlate them but yeah, a blue butterfly with the red receptor apparently uses the red receptor to maybe evaluate the host plant. Maybe that's a major driving force, the quality of the greenery where the position takes place because the spectrum of the non-quality leaves is very much changed in the red part or the far red part. So I think lots of this is anyway devoted to the generalistic vision not so specifically tuned to the wings. But of course then you have this fancy broadband and violet receptor simpapelio for example which are in any color vision model excluded because they do not seem to contribute to generalistic color vision. So the violet and broadband receptors in papelio but they can of course be used for other tasks like wing recognition or broadband receptors could measure the general illumination, et cetera. So there are exceptions to it of course. All right, thank you very much. So as there are no more questions appearing in the chat I would like to thank you once again Gregor and I would like to remind the audience that they can join us in the post-talk informal get-together even as bystanders if they want to continue following this discussion because soon I will be terminating the live broadcast. So you can either follow the link that you will find posted there or we will see you in one of our next talks. And given that there are no questions appearing for now like I have a couple of them that I would like to ask you Gregor. So one of them is like you saw it towards the end of your presentation that the retinal mosaic changes like based on the adaptation and it appeared that there was like a dose of dental gradient. Is this like something consistent or it was like just one? This has been noted basically in all butterflies with measured eyeshine. This is very predictive, predictable. So the top part is enriched in shortwave interceptors and the bottom part is enriched in long-wagel interceptors. So this is very usual. Okay and the second maybe a little bit crazy. Is there any circadian adaptation of the mosaic that we know of? Whoa, this is cool. I mean, as I said, this may be all stay dependent but the mosaic as such is fixed and it cannot change but of course the opponent connections may change somehow but what we have seen is actually only intra-o-material connections. So no inter-o-material stuff going on. Thank you. And one last question that is of general nature. So you mentioned that in some butterflies we don't see these opponents with that green opponents. Do we know if these butterflies like displayed different behavioral or ethological components that separates them from the rest or this is not unknown? I would like to be able to answer this. There's been theories put forward in the lab that maybe those with simplified mosaics, simplified visual system to chromatic are more towards generalists being present earlier around migratory species like Vanessa but the monarch is also migratory and has beautiful red receptors. So there is no clear answer to this of course. And even in the coloration as I said, morpho with the red receptor is like very counterintuitive. Yeah, I guess it's quite a complex niche to dissect and like find the cause and effect relation between behavior and expression patterns. Okay, so at this stage I would conclude I think the live broadcast given that it's also the first talk that the baby is attending and that you go into like an interview mode and ask you how you feel Gregor, how you feel baby and so on. But yeah, I would like to thank everyone for tuning in. And again, as I already said, if you want to tag along, please do make sure you click on the link that is available in the chat. Thank you very much. Thank you folks for listening and thank you for inviting me and to see you just around the corner. Okay, and we are officially off air.


It's a massive store network that puts 70% of the US population within 3 miles of one of its locations. It's CBS, the nation's second largest drug store chain, and it was beaten to the medication's home delivery punch by Amazon. The online retail giant, in fact, delivered a knockout. Take a look at its stock on the leak of its approval for business as a pharmaceutical distributor in more than a dozen states. 972.43 on its October 26 closing. On the news, bam, up to 1195 on the 27th. Now it's resting at 1,128.61. Highest we've seen. Joining me to discuss CBS's next move is Melissa Armo, the founder of The Stock Swoosh. Last we left CBS, hey, good to have you. The last we left CBS, it was absolutely terrified. It went from a high in October of 76, October high, that is, 76.46 on the 20th, on the 27th. After the announcement, it took a dive to 68.99 today, 98.70. A recovery or a holding pattern, we spoke when the news broke and you were bearish on CBS short term, buy, sell, or hold. What do you think investors and CBS shareholders should do right now? Well, right now, the chart is actually in a downtrend. Why? Because it broke major support. So anytime a chart breaks major support, it switches trends. So for CBS right now, if you're still long it, then you're considering it for as a long-term hold. Right now, in the short term, I'm talking in the next few months, CBS could continue to trade lower. Next target is 65, 60 bucks. It certainly can trade down quite a while until it flips. Now the one thing I will say that is a rumor, it hasn't happened yet, but there's a rumor out that CBS may buy Etna and that would be really big for the stock and it could explode if that happens. However, no one knows if it's going to happen and they had their earnings report, CBS had their earnings report Monday morning, November 6th, the stock dropped on the report and they didn't mention anything about purchasing Etna. So we don't really know if that's going to happen and if it does, will it happen in 2017 or not until 2018? Okay, well let's talk about Jeffrey's cut the stock price target for CBS health shares from 86, 76. It was due according to it to uncertainty over Amazon's potential competitive threat here. Do you think that was a wise move? Is it too early to really know the impact Amazon's going to have on this? Well I think it's going to have an impact on it and that's one of the reasons why the best thing that CBS could do is have a vertical move, a vertical integration into trying to capture some of that pharmaceutical business that clearly Amazon is going to go after. Now Amazon hasn't started selling pharmaceuticals yet. So again we don't know when that's going to start happening. But they just came out this week, they announced on Monday that CBS is going to start to do same day delivering in certain areas and next day delivery. That's going to help, however that's not going to be enough. What they really need to do is more acquisitions. They acquired Caremark, that was a good acquisition for them.  So what they can do is make it more like engaging for people. In other words, say they end up buying Aetna and you end up having insurance then with CBS. Well you'll get reductions in co-pays, you'll be incentivized to think of CBS and you'll think, oh CBS, I'll go to get my drugs at CBS, I'll order my prescriptions at CBS. CBS is a very loyal customer base, they're not going anywhere. Let's just say for example they don't even buy Aetna, CBS isn't going to go out of business anytime soon, like you just said 70% of Americans live within three miles of a CBS. Unfortunately though, it's going to impact the pharmaceutical business. It's not just CBS, other companies too, Amazon's going to step in and whenever that happens, it's definitely going to affect companies in the healthcare industry. Do you think that CBS can ramp up in time though? We don't know what the timeline is on Amazon actually rolling this out but it's doing operations in Manhattan for deliveries, it's going to roll out, what is it, one or two days shipping here nationwide trying to get in there. Is it going to speed up enough for people to say have the automatic click that says I want to do this because Amazon is so streamlined across all these sort of sectors. Can CBS catch up for shareholders to respond and really have confidence this is even going to work? It seems like a bit of a gamble. Well that's what I'm saying right now, CBS is in a downtrend, that's what I'm saying right now. CBS is not a buy, it's not a buy at all, I mean that's why some of these different companies have cut what they've said were the targets. I even think 76 is kind of unrealistic right now because the stock like I said is downtrending so they could drop before they flip around and what's going to make the company flip around. I don't think doing the same day or the next day delivery is going to make this flip around, that's what I'm saying. They need to take a foothold and do something else, more acquisitions. They need to engage customers better and that's something that Amazon does good very well but if CBS ends up buying Etna, Etna's one of the third largest health insurance companies so that would just be huge for them to think of it as the one stop place when you think of doing your insurance or your drugs or any of that kind of stuff. I think for now the stock is down but like I said if they end up merging and buying Etna and that comes out in December which it is rumored that that could happen yet this year the stock could explode. It could go over 76, it could go over 80. The major support of the stock was at 80 and when it broke that it started trending down and the way that stocks act when they start trending down people get very very anxious and they sell so if you're still in it long you really truly believe it's something big is going to happen and they're going to flip it around but as of right now that's not the thing. I will tell you one quick thing like I said the last time I was here the market made another new high again today. Any stock that is not making new highs with the market is not a good buy it's not even a good hold. The nice thing about investing and trading is guess what if you're in something and you're up you could sell it and you can always buy it back again. Well that's true so you know this is the CVS topic is unfolding continually. We got to bring you back in October to see if if your guesses turn out right on this. Thank you so much Melissa Armo founder of Stock Swoosh LLC. Thank you. Thanks thanks for having me.


Coach, how would you summarize this when we're a well-fought game and slipped in the end? Yeah, disappointed. I thought we had, you know, especially in the regulation, we had a chance to close it out.   So disappointed because I thought we competed. We gave ourselves a chance, but I do think games like this, we're going to play more of these games as we close out the season. So, you know, tighten it up our free throw shooting number one. I think number two, just a couple of mental mistakes there that we'll have to clean up and film, but use it as a learning experience and use it to get better. On the unfortunate end, you've seen what it can do to your team when you have your own player go down with a tough injury. What is it like when it's the opposing team? I mean, that's what our guys are talking about in the locker room now. They're not talking about the game or the loss or talking about, you know, a player was having a great game and a heck of a player. And it's just, you know, you just pray that it's not a long-term thing and he can bounce back, but just takes the wind out of yourself. So it's just not the same. And like I said, you know, we've been through it. So tough, tough, tough for the crowd, tough for everybody. The free throw shooting, that was kind of a night-long thing, but you had some key turnovers again in the last, I mean, it didn't have two minutes, really two in the last minute that really cost you. Was that just mental mistakes, physical mistakes? Listen, I think part of it was their activity on defense. I give them credit. I do think just as a general thing, like we are kind of good to really good is making that extra pass, you know, and that's collective. I can look at all five of our guys that were out there. They could have made maybe a good to great pass. And that's how you close out against a playoff team, an excellent Western conference team. But I thought we didn't do that enough and got to do a better job. How tough was it that you lost to both of your Senators fouling out? Yeah, we lost three guys fouling out. We played JD at the fight with thought about Ronde, but Ronde hadn't played all game, and I was a little hesitant in there. JD had his rhythm, and JD can guard fives, but we can't cry over spilt milk. It happened, we had guys fouled out. I will give our guys credit. I thought our physicality, you know, we battled with them. We were in the box and rang with them. We didn't, you know, we took a few punches but didn't go down. And I'm proud of our guys. I thought they tussled and fought with a, you know, big, strong team. How big is that, especially when in the rebounding battle, headed into these physical games you have coming up? Yeah, no, it's our kind of next step, I think, as a team. And we're capable. You know, I thought Ed was a monster tonight. I thought he was fantastic. You know, really, he kind of sets the tone. J.A., listen, that's where he's got to get better. He knows it, we all know it. Has to, you know, do a better job rebounding the ball. But again, I thought we fought with them. We didn't back down and, you know, that gave us a chance at the fact that we're a little more physical tonight. When you look at these past four games starting with the Clippers game, has that aspect of the game changed for you throughout those four games? So you think they're kind of learning the toughness they need for playoff? That's a good point. I think the only way you learn about it is to go through it. And we're going through it right now. And you understand where these teams are, too. You know, they're top teams in the Western Conference. And you learn a lot from these games. I'm going to watch the table and watch how they screen. I'm going to watch how they rebound, how many guys they send to the board. I think, you know, you learn things from really good teams. So definitely, definitely is going to help us down the road. Thanks, everybody.


Good afternoon, everyone. My thanks to you for waiting. I just had some other matters that I had to attend to. I understand that the purpose of this media conference is to discuss the Government's response to the discipline review. I just start off by outlining that there were in fact two reviews. The initial review was conducted by the CMC in terms of the police discipline process, and from memory the CMC came up with 11 recommendations One of those recommendations, the third one, recommended further work, and the Government committed to that further work by establishing the independent panel that's referred to in today's media release, and that work commenced in March this year. That group came up with their own review, which had 57 recommendations, as indicated in the Government's media release today. In respect of both reviews, the CMC review and the independent panel's review, the police service made a submission to the Government in relation to that, and I'm sure that was considered by Cabinet. And of course, this media release today indicates that the Government's position has been identified, but I haven't had time yet to read the Government's position in respect of what I assume, of course, is both reviews. I'll try and do that later today. Other matters, of course, understandably, I'm sure you would appreciate overtaken this for me at the moment. But can I say that obviously it's an important matter, and we totally support that the police discipline system does indeed need to be transparent. It needs to be straightforward as it possibly can be, and what is sometimes an adversarial legislative process. It needs to have public confidence, and it needs, certainly, to be something that occurs far quicker than is often the case now. Some of the things, of course, we'll have to look at will be the resourcing implications for the service. And obviously the Government, if they set policy, we're duty bound, of course, to comply with Government policy, which we will. There's no argument about that. I believe that some of these changes may well require legislative changes, and of course until that legislative change is in place that couldn't be proceeded with. But just with those few words, I'm happy to take questions that you have. Can you expand on resourcing implications? Do you have concerns that resourcing will be hit? I think it's fairly clear that one of the recommendations that's been supported is the idea of regional investigative teams. And that's probably, for me at the moment, the one that I need to turn my mind to the most. We have eight police regions, so how large those teams are likely to be, how many officers would be required. And clearly in our larger regions, which are the South East region, the north side of Brisbane particularly, but also the North Coast region, we have something like 1,500 police, compared to other smaller regions numerically, where we might have six or 700. So the numbers presumably will vary. But then what you have to factor in distance and travel as well. So we'll need to do an assessment of the numbers, and then work out where those numbers are going to come from, how we're going to provide them, and how soon we can do that. I think you said it may result in police being pulled from the bigger, the two bigger regions, where's the need to sell them? No, I'm sorry if I misled you. Now what I was saying was that there are eight police regions. My understanding is that the government have supported the concept of regional investigative teams to be based, you know, at regional headquarters. Officers of the eight police regions, and my point was only that some of the regions are much larger than others. So for example, the North Coast region, which takes in six districts, or Redcliffe, Kabalsha, the Sunshine Coast, Gimping, Meribur and Bundaberg, and has 1,500 police, may need more investigators than a region such as the Northern region, which takes in primarily Townsville and Mount Islay, and has about six or seven hundred officers here. What do you think of the reform to allow the CNC to override the decision-making and assistant commissioner? Look, I think that's, was that from the Sunday Mail story this morning? Yeah. Yeah. Look, with the greatest respect, I think that I'd rather not comment on the Sunday Mail story. I'd rather see the actual detail in terms of the government response, but certainly that was part of the recommendations of the independent review, that there'd be an ability by the CNC, as I understood it, to seek to have a substitute sanction or result put into place, but I'd need to get a full appreciation of the detail of about how it is proposed that will occur and work, what legislative framework would be put into place to enable it, and really what the practical implications of it are. And I'm sorry that I'm not in a position to comment on at this stage, but I am happy to do so in the future, when we've had a bit of chance to fully consider these matters and advise you of any implications for us. Just in relation to the Slaks Creek House Fire, if we can, where are the investigations currently up to there, and what is the plan for police over the next few days? I'm sorry, could you, I just missed the first part of it. The Slaks Creek House Fire. The Slaks Creek Fire. Obviously, those investigations are ongoing, and as is the case with any fatal fire, there will be a report to the coroner, and I would imagine that this is going to take a long, long time to put together.  So I would think this will take months and months to finalise the investigation. So you're saying that the cause is unknown? I'm not aware that the cause has been definitively established, but even if we were, the proper process is that we would not disclose the cause. We would complete our investigation in its totality, and the judicial entity, if you like, that has responsibility for determining this matter, ultimately, as the coroner. So the steps would be that we'd prepare a report for the coroner, and it would be the coroner who would ultimately make the determination about causal factors. Do you have a preliminary sort of idea of what may have caused the fire? I think my understanding is that the police involved in this do not believe, do not believe that there are any suspicious circumstances at all. I think that's a really important point to make, and that's been restated, and I feel confident, absolutely, in their judgment in that regard. That being the case, then, the cause is some cause of an accidental unintentional nature, presumably, but the precise nature of that is still, in my view, yet to be fully determined. Some states, they don't, like, fire would go at some electrical fault, and maybe you get told that, or whatever it is. Yes. Like something like that, or is that not going to happen this time? No, that possibility is within scope in terms of the causal factors. So it could be months before that house is removed, it's not like they should be... Yeah, good question. It's such a sad scene, as an insight. The investigation will be complete soon in terms of the house, so we won't need access to the house again in the very near future. And I would imagine that the aspect of what happens then to the house would be the determination of the owner of the residence and the Logan Council. You're aware if police are being able to identify into body or how long that process will take? My understanding is that that would not have been completed as yet. That is a very difficult task, in generally speaking, that could involve dental examination and or DNA. Yes. Do you have any idea how long that process will take before the bodies can be returned to the family? Look, it will be done as soon as we possibly can and can I say we're extremely grateful to the family and the broader Pacific Island community for their understanding of the processes that we have to go through. But it is absolutely essential that before a body is released for a service, that we have the identification of that person correct. And I would ask and seek the further understanding and patience of the family in that regard. I think everyone would appreciate that, that you simply cannot second and guess at this and we have to be certain when we release people for a service that we have identification positively confirmed. Okay, positive identification on the end. Is it three people? Look, I'm sorry, I don't know. That may be so. If the information is able to be released after the family are advised and the coroner gives approval, we will make that information available. I thank you for your time today.


I welcome you all once again to my channel and today's topic will be item analysis. And those who are new, I am Dr. Rashmi Singh, Assistant Professor of the Department of Education at S.Hanna Girls degree college, University of Allahabad. Join my telegram group, explore education to share your feedback, ask your query and certain other things which we cannot do here. My lecture will be bilingual that is covered English as well as Hindi and useful for teaching examinations and do subscribe my channel if you like my video content and delivery. Okay? So let's start. First of all, item analysis means Aekang, Vishlesh and Hindi. When we study educational measurement and evaluation, research methodology, achievement test, standardization achievement test, then it is mentioned that the item we will choose in our test, its analysis is necessary. Why is it necessary? First of all, let's talk about the examination. Why is it necessary? Why is it necessary? It is because we take the examinations of the students, they do the research. So there is a very good article. Assessment the bridge between teaching and learning. See, what a good definition. In assessment, the bridge between teaching and learning, dial in William's days if our students learned what we taught, we would never need to assist. It is only through assessment that we can discover whether the instructional activities in which we engage our students resulted in the intended learning. We can learn whether the teaching was changed in learning. And what is the assessment? It is really the bridge between teaching and learning. So what is the assessment? It is really the bridge between teaching and learning. And what are the exams? Exams as a window into student learning gaps. So the gaps that are coming in student learning. We can see them there, we can see them there, we can see them through exams. And consequently, a way to show student learning. And we can see how much student learning has been done. So you can understand how important the examination is. The teaching, the theoretical portion, when the learning is being changed in the practical portion, we are able to understand it through the examination. So why did we bring this here? Because the items that will be kept in the item analysis are very important. Why? Because the items that will be kept in the item analysis are very important. Why? Because they can really be assessed. They can be evaluated. They can be examined. They can be examined properly by their students. That is why the item analysis is important. One item that you will keep in your question paper, you have to do its analysis. Then it can be understood in the right way. Whose study was right. Sorry. Let's start with the item analysis. What is the item analysis? It is an investigation. It investigates the performance of items considered individually either in relation to some external criteria or in relation to the remaining items on the test. The items that we are going to do, the items that we are going to keep in the question paper, the items that we are going to keep in a test, what it does? It investigates the performance. Either it investigates the external criteria or it investigates all items on the test with each other. What is more? It is a statistical technique. It is a statistical technique which is used for selecting and rejecting the items of the test. What should we do first? We write down many questions. But we ask questions in the examination. This is mostly about your objective test. So what do we do in it? First, we make a lot of questions. For example, if you want to make a set of 50 questions, then make more than 60, 75, 100. And then we will do its item analysis. Then we will come to know which item is suitable for it, which will be our blueprint and which one will be removed from it. Rejected because it is not the right item. So only through this item analysis comes to be known. It is a statistical technique which is used for selecting and rejecting the items of the test. On the basis of which basis will you either keep it or remove it? There is a difficulty value, especially which we do not have. Discriminative power and item distractors. That is, how much is its difficulty value? Discrimination power is not in that item. And is there an item distractor or not? What is the problem? We are creating the right answer. We have the right answer to reach the right answer. We see these three things and we keep them or remove them. The difficulty value and discriminative power are very important. And the question is asked. Subjective, objective, even formula is asked. It will be given to you. It will be given to you. You will have to remove it. How much is the difficulty value? How much is the discrimination power? Okay. And what is this? It is an important tool to uphold test effectiveness and fairness. This is a very good tool for us to make our test effective and fair. And it can increase the efficacy of examination. That is, the test we are going to take will increase its efficiency. If we do item analysis, then by testing knowledge accurately. Why? Because the knowledge, the knowledge that we want in our students, we will be able to accurately test it. Why? Because of item analysis. If we do it properly, then it is more beneficial. When you were reading reliability, validity, then it was said that if you do item analysis, then your test can be more reliable and more valid. Why? Because it will do the same thing that you have kept there. Then functions, if we look at it, then three important things. Collecting the appropriate items for test. That is, we will keep the most important, most perfect item in our test. Rejecting inappropriate, which will remove inappropriate items. And modification in the structure of the items.  The special thing is that we do not have two points. Difficulty value, discriminative power and distractor can also be talked about verbally. So, difficulty value is the difficulty. What is the difficulty? This is what comes to mind from Hindi. What is this? It is the percentage of people. That is, what is the subject of those people? This is the subject of your test administering. Who answers an item correctly? That is, the number of people who correct that item. They stick to the correct answer. That is the difficulty value. So, what happens is that we feel that there is a little resistance here. You can understand it like this. Do not call it item difficulty. Call it item facility. Then you will understand it correctly. Because many scientists say that it should not be called item difficulty. It should be called item facility. Why? Because what is the definition? Percentage of people who answer an item correctly. That is, the difficulty value of an item is 0.9. What does this mean? 90% of people correct the question. So, 90% of people, 90% of students correct the question. What does this mean? It is very easy. So, the difficulty value is less. So, it is a reverse relationship. The more the difficulty value is, the easier the question will be. The less the difficulty value is, the harder the question will be. You can understand it like this. It is reverse. So, first of all, the percentage of people who answer an item correctly. Then it is the relative frequency with which the examiners choose the correct response. It is similar. Then, item difficulty has a powerful effect on both the variability of test scores and the position with which test scores discriminate among group of examinees. That is, there are two effects. One is variability of test scores. How does test score vary? It is also affected by this. And the position with which test scores discriminate among group of examinees. And how do test scores behave in group of examinees? This also tells us that upon item difficulty. It also has a formula. If you have a descriptive examination, then you will have to write it. Otherwise, you can ask questions based on this formula in the objective. You can ask the formula. What is dv? Difficulty value is equal to rh plus rl upon nh plus nl. Now, what is rh? Rightly answered in highest group. Now, you will know if this highest group is the lowest group. So, the questions that people have asked in the highest group and the questions that people have asked in the lowest group means the total questions that have been asked. This means that if we have taken everyone in the whole group. And nh plus nl means that the number of examinees was in the highest group or the number of examinees was in the lowest group. So, rh plus rl upon nh plus nl. Okay. And when we take its percentage into 100, then we will get the corner. Similarly, value will be 0.9. So, what will be its value? Its value will be 0 to 1. You can also ask how much is the range of difficulty value? So, it is 0 to 1. So, the highest difficulty value is 100%. 100% means that everyone has answered all the questions. So, there is no point in asking questions. Then you will get to know which difficulty value range is the one that you have to keep the item in your test or discard it. Okay. This is its formula. Then, in this case, non-response examinees are available. If there are examinees in your test, students who have not applied tic, then the formula will change. What will happen? db is equal to rh plus rl upon nh plus nl minus nr. Those people will be eliminated who have not applied tic. What is nr? The number of non-response examinees. Okay. Now, let's see how difficult the value will be. So, what will we do with the evaluation? What is it? So, it is good to keep our test item and we have to keep it or reject it. We should also know that our main goal is to keep the right items in our test. And those who are not right, those who are not good, those who are not doing research, those who do not have tic, then remove them. So, this is the criteria. If there is a difficulty value between 10-9-2 and 10-9-3, this means that this is the most difficult. How difficult is it? Because only 20-30% of people can correct that question. Similarly, it is getting harder. If 30-40% is correct, then it is also difficult. If 40-60% is correct, then it means that it is a little difficult. If 60-70% is correct, then easy questions and 70-80% is correct. So, it is a very easy question. So, we have to keep this. That is, we will keep around 10-4-10-6 so that it is moderate and difficult. That is, those who will be able to do the examination after studying will be able to do it. No, it will be written well. If it is a difficult question, then they will not be able to do it. So, it should be a little difficult. Why? Because we are going to take the examination. We are not going to ask. We are not going to examine the children, the students, how much they have done. So, we will give them some difficulty value. Okay? Then, discrimination index. How will our item not be able to differentiate? So, who is able to differentiate between the high achievers and the low achievers? Between the superiors and the inferiors?  Only then, someone will be able to do the examination. Otherwise, if we take the examination and all the children have passed 100% mark, then we will not be able to solve any problem. Okay? So, how will we define discrimination index? Index of discrimination is that ability of an item. Our aim is that on the basis of which discrimination is made between superiors and inferiors. That is, high achievers, sorry, the spelling is wrong. High achievers and low achievers have the ability to differentiate between the high achievers and the low achievers. Whose aim is called discrimination index or discrimination index? And its formula is DI is equal to Rh minus RL. That is, the highest group that the people did the right thing and the lowest group that the people did the right thing will be minus RL. And whose group will we give the run? Sorry, NH and NL. That means the highest group has the number or the lowest group has the number. So, which one will we choose? Which one is larger? The one whose name will be more. The number of the highest group's examinees or the number of the lowest group's examinees whose number is more, we will give the run to the difference between the two. So, let's take out the discrimination index. Now you will understand the highest group and the lowest group. First of all, the test is scored. We have taken the test. Scores are rank ordered. We have read a crumb that the nominal-ordinal-interval ratio is the ordinal, where we know the rank. So, what we have said is that rank ordered is the lowest number that we have established. That is how we rank in the class. Then what we will do? 27% of the highest and lowest scores are selected. We will select the topmost 27% and the bottom 27% Now you will say why 27%? Brother, 50% do it, 20% do it, 30% it looks good to cut. If you run it, it will be cut. Why did you take 27%? So, there is a logic behind it. Who told us that we don't use it like this. Kelly Mahode told us we told it in 1940. Okay. Sorry. Then, number of sorry, number of correct answers is the highest 27% then the highest 27% is the top one. What do we do in that? We minus the bottom 27% and then who runs the result? Number of people in the larger of the two groups. This is the formula. R H minus R L upon N H and N L what is it? Which one is larger? Who is more whose mind is more we will run away from it. Okay. Now, let's see why did you take 27%? Percentage of 27% is used because this value will maximise differences in normal distributions while providing enough cases for analysis. That means there was a lot of research on this. They did a lot of research. You must have taken arbitrarily which group was taken how many percent was taken how many percent was taken but when they took 27% the statistics involved they found that the normal distribution maximises differences. We are getting most of the difference here and most of the cases for analysis if we take 27% then this is the 27% of Kali Mahudaya which is famous and it is done with 27% only. But some students consider it to be a bit different which is a student J.C. Nandini who said taking 25% would be better but the 27% of Kali is more you can say famous and it is used as a thumb rule. So if we take a sample which is 24% then it will be easier to make a group. Okay Okay Discrimination which is index there are three ways zero discrimination positive discrimination and negative discrimination.  So zero oblique none or zero oblique no that that item is not discriminating. So we don't have to keep this item in our test and it is of no use. Then there is positive discrimination which means that the high achiever the superior if he corrects the question and the low achiever does the wrong thing which means the right discrimination which we were talking about which we want in our test or in our item is positive discrimination and the items in which the positive discrimination comes out we keep it in our test. Then negative discrimination is completely wrong. What happens is the achievers do the wrong thing and the inferior achievers do the right thing. So you will say how this can happen this can happen that it is very poorly worded or poorly worded or something like that that the low achiever guessed it and the high achiever or the superior is getting very confused so we don't have to keep the questions of negative discrimination in our test okay and above our regarded as high discrimination used so if our item comes out then we keep it in our final test. So this is about discrimination index and difficulty value. This should be DI discrimination index then relationship between discrimination index and not contradictory to each other. No one is opposing each other but both are complete. Both should consider in selecting good items if we want that our item analysis is good if we want our test to be good then we have to take care of both of them and what to do if an item is negatively discriminated or zero discrimination is to be rejected whatever the difficulty value difficulty value we said if 10 4 6 10 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 20 21 22 22 23 24 25 26 27 28 29 29 30 31 31 31 31 31 31 31 31 31 31 31 31 35 30 35 28 31 37 running 39 39 39 40 40 40 40 41 41 80 Ee 42 halfway I have covered a very important topic today, item analysis. Theoretical question is also asked if you are doing MA, MAID, BA, BAID and if you are in an objective question. Thank you all and don't forget to like and subscribe my channel. If you like it and to find this channel in telegram group scan to join this and the name of my telegram channel is explore education and join my telegram group to achieve all this. So done from my side.