I'm Marcia Joyner and we are Navigating the Journey and today the Journey takes us into the land of the Hawaiian Homes Act, the Hawaiian Homes Everything and our guest is known to all of you that have watched us over the years and this is my dear friend Bill Isla who is now the director of Hawaiian Homes. Is that the correct title? Yes. And the first time Bill was with us was when he was director of Department of Land and Natural Resources which has been a long time ago. Yes. Yes. So welcome, welcome, welcome Bill. Thank you so much for always coming when we call. Thank you for inviting. First tell us about the Hawaiian Homes Act. What is it? Sure. What, no it is, it is still in effect. It is still in effect, it is a congressional act passed in 1920, implemented, started implementing in 1921 by, mainly driven by Prince Kohio who was a non-voting member of Congress. Correct at the time, but had a large, had a large following. Prince Kohio because of his responsibility to Hawaiians and the people of Hawaii saw the degrading of conditions for native Hawaiians as they were dispossessed from land. So he went to Congress, took him several years to get Congress to pass the Hawaiian Homes Commission Act whose primary purpose is to put Hawaiians back on lands and secondarily rehabilitate them. Rehabilitate meaning a lot of things because it is pretty broadly described in the act. Rehabilitate people? Yes. The conditions of native Hawaiians at that time in urban areas, they were in tenements. Is this the plantation era? This is before the plantation era. So this is, well, right about when the plantations were at their height. So basically Hawaiians were dispossessed from land, they moved to the urban areas to find jobs, cramped conditions, low wages, disease, lots of bad social indicators if we were to measure them today. So he saw all of this and went to Congress to argue for the opportunity for Hawaiians to get back on the land to become farmers and ranchers and even aquaculturists. So aquaculture is actually written into the act. That was kind of interesting. So one of the primary reasons for our being is to get people on the land. There are three different ways. We can do it in a residential lease, an agricultural lease or a pastorial lease. So the department has... What's a pastorial? Pastorial is like a ranch lease because back then there were very few neighborhood stores and people got their meat from either they raised their own or they went to the ranches and purchased cattle. So over the last almost 100 years, I think, depending on when you count, either 1920 or 1921, it's either the 99th year or the 100th year next year. And we're still discussing about what the proper time is to honor Prince Kohio and everyone that assisted. So we have 20, a little over 27,000 people that are still waiting for one of these leases. What takes so long? What takes so long is the cost of developing land in Hawaii. The type of land that we have, some of it is flat, much of it is not. Some of it has water resources, much of it does not. So we spend a lot of time trying to bring water to... So you have to put in... Your office has to put in the infrastructure. That is correct. We basically pay for infrastructure. In the modern world, it's road, sewers, water, electricity. Shouldn't it be solar instead of electricity? We have a program where we allow beneficiaries and leases to install solar to help reduce their costs. Yeah, but I think some people still like that comfort of knowing that in case something happens, a storm or something, that they have that ability to access point electric. So in the first, I want to say, 40, 50 years of the act, it was a federal act. And very little federal funds were applied. So the department at that time was... Its main source of funding was revenue off of leases that were still in effect. So much of the lands that came to the HHL at that time were under leased to sugar companies and big ranches. So as part of the act, it said that we would honor those leases until they ran out. However, payment for these leases was really what paid for the operations of the department. So those payments came to you? Those payments came to the precursor of the Department of Wine Homelands. So I'm not sure what it was called under the territory, but basically this managing agency. It did the best it could with the resources that it had. It did not have a lot of money allocated for improvements, infrastructure, water and those kinds of things. We fast forward to 1959. Hawaii becomes a state. One of the requirements for Hawaii becoming a state, the federal government says, is you will take responsibility for this program. So since 1959, the department was created as a state agency. And a lot of folks, a lot of our beneficiaries like to think of themselves as quasi-independent. However, because we're a state agency, I have to operate under the state rules, regulations, and the federal oversight that has rules and regulations. So that makes me unpopular with some of our beneficiaries who are on the path to nation within a nation or independent status. However, I keep reminding them that I have these rules and regulations that I have. And then we go to the legislature every year with the governor's assistance to request monies. The last two years we've been fairly well-blessed. We've got about $25 million in our capital improvement budget for each year. The capital improvement is for the infrastructure. That'll help us build infrastructure. So roads, sewers, it could go to the development of a well, the infrastructure to take the transport that water from that well either to another county system or perhaps to our system, because we manage three water systems. We lose a lot of money on those systems, but it's a necessary benefit to our homesteaders, especially on Moloka'i, where a lot of the larger aglots are. Well, now tell me this, you have land on each island? Yes, including Lanai, which we just received, I want to say, in the last 15 years. Yes. Ni'ao? No, no land on Ni'ao, it's private, the only one. Yes. Okay, now Moloka'i and that Moloka'i Ranch for sale, all of that land is just sitting there. So do you get to any part of that? You get a part of their lease, Moloka'i Ranch? No, we do not. That's private. I mean down the road, if they decide to sell in the state and the state were to purchase some of those lands, and it's possible that we could get additional lands, but we have plenty of lands on Moloka'i. What we're missing is water to develop those lands. Water on Moloka'i is a big deal. So what's needed on Moloka'i is another well drilled outside of the current aquifer, because right now we have three straws in that aquifer, and it's beyond sustainable yield right now. So if we had either Moloka'i Ranch or the county dig another well, then we could utilize the excess allocation to increase water to our beneficiary. Now I understand that Moloka'i Ranch wants to sell the whole thing, and they don't want to divvy it up. Is that true? I've heard that. I have not been in any of the negotiations. Because that's a lot of land to sell at one time. It is, but I think the foreign company wants to get out of that and invest that money elsewhere. So they need the whole thing? They want to sell it all at once, including their operations of their water system attached to Moloka'i Ranch. That's really a big... You could put a lot of people on that land. We could, and we could put a lot more people on our land if we just had more water. Developing more water is a priority on the island of Moloka'i for us. One of the biggest challenges for me as Chair is that we have a certain segment of the beneficiary class that wants the department to emphasize more economic development on their behalf. And so we have opposition to leasing of lands that are not required for homesteading. That has always been part of our history. So all of these lands that were leased to the sugar companies in the ranches prior to is what basically paid for the operations, the improvements on the homelands prior to becoming a state agency. Now that they're gone and the land is... Do you still get any residuals from those companies? On state lands that were formally leased to sugar and had sugar operations, we're supposed to be getting 30% of the revenue. So let's say somebody built a hotel on lands that were formally used for sugar. We're supposed to get 30% of those revenues. Are people building hotels on the land? I guess on Maui, huh? It's totally possible. There's actually a hotel on Kauai that came into our inventory as part of the Act 14 settlement. So we're basically paying ourselves with the lease rent. And there are other leases that were formally sugar. We're also supposed to get 30% off of any water licenses. And you're saying, suppose... Yes. So... Is that an operative word? Yes, yes. So all the water licenses... You're trying to tell me... All the water licenses have expired. There's been several court cases about how the new process should occur. It hasn't occurred yet. So no new water licenses have been issued. As long as that's the case, we're not receiving 30% of the revenue off of those water licenses. And then the challenge becomes... In Hawaii, where water is a public trust resource, how do you value that water? You have to put a dollar? For the value of the license, if we're going to collect... No, no, I meant on the water. We would have to put a value on the license, right? And then collect 30% off of that. Okay. That's being discussed right now. Yes, nobody knows. Well, it's in court. It's in the legislature. It's in many avenues right now, and we haven't answered that question. Okay, now, how does OHA fit with your office, Hawaiian Homes? So the Department of Hawaiian Homes and the Hawaiian Homeland program was created in 1921. OHA was created in 1978. Right. So how do they work together or do they? We have the same beneficiary class. Although OHA expands their beneficiary class to those who have less than 50%. They receive, it's supposed to be 20%. The wording is 20% of the revenue off of seeded lands. However, they receive a formula. So they're basically getting about 15.1, 15.2 million per year. And then additional allocations from the legislature in addition to the revenue that they generate off of their, let's call it an endowment for now, money that they have for investing. And so that's how OHA is funded. Now can they build, can they put people on the land? That's not your land, but some of their land. They could. And I understand that the trustees are looking for it. We've partnered because we share beneficiary classes, right? So we've partnered. OHA has committed to funding some of the bonds that we have put out there that have led to houses being built, roads being built, infrastructure being built. So OHA continues to pay $3 million a year to help us pay for those bonds. Oh, great. Now we have to take a break and we'll be back in just one minute. And then we're going to talk about the elephant in the room. Sure. Mauna Kea. We'll be right back. Hello, I'm Mufi Haname. I want to tell you about a great show that appears on Think Tech Hawaii. It's all about tourism. In fact, we call it Tourism 101 where we talk about the issues and challenges that faces our number one industry throughout the state. We'll have some interesting guests from a very informative dialogue and allow you an opportunity to maybe learn a little bit more about why this industry is so important for our state. It's been great for us in the past. We need it today and especially going forward. That's Tourism 101 on Think Tech Hawaii. Mahalo. Aloha. My name is Victoria and I'm a host at The Adventures in Small Business. This is a collaboration between U.S. Small Business Administration, Hawaii District Office, and its partners where we showcase the stories of local entrepreneurs and small businesses. Talk about how to start a business. Talk about great tips for small business owners. Please join us every Thursday, 11 a.m. at Think Tech Hawaii. See you soon. Mahalo. I'm Marcia and we're back and we are talking with my dear friend and all of you know I only talk to dear friends with Bill Isla and Bill is the director of Hawaiian Homes and he was the director of land and natural resources and a whole lot of other stuff. Yes.  So let's start there and let's see what that impact on the Hawaiians is or has been or will be. Let's start there. Sure. So the conditional use permit to construct in a conservation district, the telescope, came before the Board of Land and Natural Resources while I was the chair. After undergoing environmental impact statement, many different management plans, many different studies and the board voted to approve the CDUP permit at the time. There was a question of whether the board, excuse me, could make that decision without taking into account a request for a contested case hearing prior to making the decision. That was challenged. Ultimately, that was taken all the way to the Hawaii Supreme Court and they ruled against basically the board's decision. Subsequently, under new chair, the whole process started all over again. That decision was affirmed and through many different appeals, the permit process again went all the way up to the Supreme Court and Supreme Court ruled that yes, indeed, the University of Hawaii TMT followed the law and therefore TMT should be allowed to proceed and that brings us to the present day situation where there are some folks on the road blocking the movement of equipment up to the mountain to begin the construction. Well, it seems to me in listening that most people don't know that it went all the way to the Supreme Court. Most people don't have a sense of the background of what has gone on these 10 years prior to this and listening to what's going on, there doesn't seem to be anyone that the mountain, the management of the mountain or anything, will come to the microphone and explain all of this. So people around the world have no concept. It's almost like one day there's a telescope and now these people are protesting and saying, you know, and well, you know, I come from a long line of protesters, so that's not an issue. But if we don't know the full background, then what they're asking for does not have any meaning. But we need to know. So I think there's been a change in how people gather information and with the, you know, with the prevalence, if you will, of social media. There are many, many thoughts out there on social media about things and that's very difficult to, as you and I would do in the past, do our homework and study something very intently before making a decision, right? So many folks make up their minds based on what they see on social media without doing the full sort of analysis. And that's where we are today. Particularly for me, what I've been accused of lately as the chair of the Department of Homeland Homelands is not practicing my fiduciary duty as a fiduciary for the, advocating for the road, Mauna Kea Access Road.  We are obtaining advice, of course, the Attorney General's Office. And back in 1994, 1995, the legislature, as part of a larger settlement with the past bad practices of removing and using without commission of Homeland Homelands, created the settlement in Act 14. Act 14 gave the department a little more than $600 million for past unauthorized use of Department of Homeland Homelands. It required DLNR to put back another 16,000 plus acres into our inventory, our land inventory to make it whole. And then there's a small settlement on Kauai issues, and then there was some money that was charged for a school in Nanakuli being on Homeland and not compensating for it, and a couple of other small things. So with that act in 1994, 1995, it settled all of these claims. The legislature knew that it was going to take a while to get through the compensation part. So they basically said that by initiating this settlement, the Department of Transportation has operational authority, jurisdiction over the road, Mauna Kea Access Road, actually. So I have beneficiaries who don't like that, and they're promoting another view. However, as I'm advised by, I'm a stated agency, advised by state attorneys, Act 14 settled this. So this is something that's being actively debated, I think, on the mountain by beneficiaries and non-beneficiaries who don't agree that the Department of Transportation has the road and has the authority to close the road. And so this is actually causing a lot of discomfort in my community and actually showing his face in some of the Hawaiian Homes Commission meetings and taking up a lot of time away from actually trying to get more people on the land. So it's just for your department, it's the road. But as big as the mountain is, there are sacred places, I'm sure. Of course. But all of these other telescopes are there. What about that? Because they're not on Hawaiian homelands? I don't have a say on that. But in terms of the protest, it seems to be only geared to one telescope and not the others. I think depending on who you talk to that are occupying the road right now on the mountain, they all have feelings about whether it's one telescope or all the rest of the telescope. Yeah. I would just point out that, you know, Monarchy Access Road represents about 67 acres of more than 300 acres of roads and highways that either the counties or the state use without our permission. And that's part of the settlement. So it's kind of a larger discussion about all the roads across Hawaii that either the state or the county use without our permission. Oh boy. So is there, I know this is asking a lot, a way to settle this amicably so that the people that are protesting, the people that want a sacred place on the mountain can have a sacred place without this doing away with the telescope. Is as big as the mountain is, can't there be sacred places? I believe the answer is yes. And the mayor of Hawaii Island, Mayor Harry Kim and the governor are open to any types of suggestions or negotiations on how to achieve just that. Because that seems to me, I can't see that the telescope is either or. I just can't believe as big as that mountain is that it's an either or. That's something that's being discussed as time goes by, right? The mayor has held probably about three or four meetings and I don't know how many individual meetings to try to accomplish just that. So I don't know that he's been successful as of yet, but he's still trying. Oh bless him. My goodness, can you imagine last year with the volcano and now this, he has been through so much. Well, Mayor Kim has a lot of experience, so. But how much can his heart take? You know what? He still rides his bike six miles a day. Yes, I know. He's such a treasure, a real treasure. And I just have to know that this is going to settle. There's got to be a settlement for both sides. I mean, it can't be either or. I have to believe everybody is praying for that. Yeah, it just can't be either or. And my view, of course, and when I met you, a man of the sea, a man of the ocean, the Hawaiians were traveling around the world guided by the stars when the Europeans thought they'd fall off dead. They have been so far out in advance of everybody else on the planet. And so for me, the telescope is just further that knowledge. It just furthers that knowledge. So I can't see that it has to go away. I just think that they belong together. It's more knowledge. It takes the Hawaiians even further ahead of the rest. They should be ahead of the world. The Hawaiian should be at the top of the world. Polygyne Voyaging Society, but just less than a year ago, got back from a worldwide voyage, right? Yes. Carrying the message of we need to protect our canoe, the planet Earth. And so I like to think that there are cooler heads that are thinking about how everybody benefits and how the children of Hawai'i benefits going forward. Yes. Well, Bill, thank you. Thank you for coming. Thank you for spending this time with us, and you will be back. Sure. Aloha. Thank you all for coming and spending this time with us, and we'll see you next time.


Quiz, verbs like sing, which is correct? Number one, Simon with deep feeling, saying, sung, both are correct. Saying, too, they, into action, to save her life. Spraying, sprung, both are correct, both are correct. I, enough money to pay the bill. Have brung, have brought, both are correct. Have brought, for, ouch, I, buy a bee. Sting, was stung, both are correct, was stung. Five, Marty, his clothes on a rack. Hung, was hung, both are correct. Hung. To learn more about this topic, click on the link above. Or, check out our websites for more free English learning resources. ESLgold.com and freeenglishstudy.com.


Thank you very much. I'm glad it worked out. Finally, so the title of my talk is genome and transcriptome dynamics in cancer cells. To start the most important slides, the member of my laboratories who in different capacities contributed to what I'm going to talk about, we have active collaborations. The University of Lübeck, the Karolinska Institute in Stockholm, and the University of Göttingen, which is part of a clinical research unit which is funded by the German research community. So what I will do is, at the first third or half of my talk, discuss with you what we know about chromosomal changes and patterns of aneuploidy in carcinomas, essentially as part, as kind of an introduction to the second topics. When I discuss briefly what the consequences of these cytogenetic abnormalities are on the transcriptome, on the gene expression in cancer cells, and then I will exploit what we can do in order to translate what we have learned from these more basic research topics to translational medicine, both in terms of genomic aberrations and alterations of the transcriptome. What you see here are chromosomes from a patient with chronic myelogenous leukemia. And you can see there is a single chromosomal translocation between chromosome 9 and 22. And at this point, I have to pause and acknowledge Janet Rowley, who actually discovered the balanced nature of this chromosomal translocation. In fact, this slide was performed in a collaboration with her. She unfortunately passed away just two months ago. She was a staunch supporter of the Intramural Research Program in general and the Genome Institute in particular, where she served as a scientific advisor since its inception. And, as you know, this translocation has profound effects, not only in terms of diagnosis, but for treatment, because if you target the genetic event, which is the fusion of BCR and ABL, which results in aberrant tyrosine kinase signal with a drug, you can get these patients into remission. And the paradigm of chromosomal translocation not only applies to CML, but to many other hematological malignancies as well, which some of them are listed here. But I chose to show you this slide for another reason as well, which is clearly there is one genetic event, which we now cause transformation, but you can also appreciate that the rest of the genome remains normal. So, what it is that there is a single genetic event, which is sufficient to transform these hematological cells. If you look at solid tumors, carcinomas in particular, tumors of epithelial origins, the picture changes dramatically. Here is a normal kerotype, and here are the metaphases from a breast cancer cell in SKBr3, and you can see a profound degree of chromosomal instability, not only numerically, but also structurally. Then you can see giant marker chromosomes, which are a reflection of oncogene amplification, in this case, the CMIC and the HER2NU oncogene. Then you have other features, this is probably a little dark, cancer cells, which we call anibloid, which is reflected in enormous degree of variability in the DNA content from one cell to another. You can also appreciate apolamytosis, which clearly will lead to a different chromosome count in these cells. You can see the phenomenon of anaphase, which results in lost chromosome and is caused by telomere attrition. Some pictures like this and this led to the perception that if you look at most solid tumors in adults, it looks like someone set off a bomb in the nucleus. This perception led to the interpretation that in solid tumors different from the hematological malignancies, we have a cytogenetic chaos, which is induced by catastrophic mitosis, you have seen one, keratobic complexity, heterogeneity of the tumor cell population and ongoing chromosomal instability. The perception of a cytogenetic chaos also led to the perception that chromosomal aberrations in solid tumors different from the hematological malignancies would rather be a consequence rather than the cause of the disease. What I will show you now is, I will discuss with you now whether this is indeed the case. Arguably, we had to develop methodologies to dissect the complex aberrations in solid tumors. Most important was comparative genomic hybridization. I will show more details later, but in short, it allows you to map genomic imbalances in the entire tumor cell population. We have spectral keratiming, which we developed years ago, which allows you to paint all chromosomes in different colors. And then we can use specific probes to enumerate chromosomal copy numbers directly in non-dividing cells, cytological preparations or tissue sections, and it can also be used to identify translocation because you see a fusion of different colors. And now, with the advent of global gene expression profiling or sequencing, we can interpret and query how these aberrations affect the gene expression levels in a global sense. So, having had these tools, we then try to understand the dynamics of changes in the transition from normal epithelium to invasive disease. And initially, we turned to two model systems which are cervical carcinogenesis, called rectal carcinogenesis, mainly for the reason because the morphological changes are very well described and they are accessible. So, you know the sequence. It goes from low-grade dysplasia to high-grade dysplasia and eventually to invasive disease. And while it looks a little different in the colon, it's the same. You have primalictan lesions that if they would be removed, you would cure the patients. So, we used a combination of molecular cytogenetic tissue microdissection techniques, gene expression profiling, and they asked the question, is there a non-random distribution of chromosomal changes in one of those different tumor entities? Is there a stage-specific sequence of events? In other words, some changes early, others late. And which additional genetic or epigenetic changes do occur? And it is now completely clear that the distribution of chromosomal gains and losses is non-random. In red, you see the aberrations for the cervix. Let's just look at chromosome three. On the left side are the gains. On the left side are the losses. Chromosome five has gained. But the rest of the genome is relatively unaffected. In the colon, which is displayed in green, you can see invariably gains of chromosome seven, extra copies of the long arm of chromosome eight, chromosome 20, and very specific and unique for colorectal tumor genesis gains of chromosome 13. And I just want to point out, we and others have now studied more than 500 cases of cervical carcinomas here. The blood is normalized for 10. And you can appreciate that all cervical carcinomas have extra copies of the long arm of chromosome three. In other words, this aberration is as common as is the Philadelphia chromosome in CML. We do know that we have risk factors, high risk HPV, most commonly 16 and 18, which is required for cervical cancer to occur, but it's not sufficient. And in colorectal tumors, you know that inflammation, colitis, ulcerative Crohn's disease, greatly increases the risk for colorectal tumors to occur. But the distribution in these conditions is the same as in sporadic tumors. Conclusion, we do not have a cytogenetic chaos, but we have a stability on a different plateau of copy number changes. It's like a speciation. So, but we do have the problem that we have an enormous degree of variability. So the question comes up, how can we reconcile, excuse me, anibloidy and ongoing chromosomal instability, which is a fact with a strictly conserved pattern of genomic imbalances that we observe. And in order to answer this question, we engage in a collaboration with the Navy at that time across straight, across two streets from here, and collected patient samples where we have ductal carcinoma in situ and invasive ductal carcinoma on the same slide. For that, we use tissue microdisection, and we've prepared from both of these lesions independent slides where we had these single-cell suspension here, you see, for one case, the histology. And then we chose, we looked at the distribution of genomic imbalances that is specific for breast cancer. You can again appreciate that it's specific, but it's different from both colon and cervix. And we chose probes, fish probes, fluorescence in cytohibrosis to enumerate a copy number of these regions together with control probes directly on these cells that we prepared from DCIS and IDC, and that was done, this is a technical detail in repeat hybridization, but at the end of the day, we could enumerate 10 independent loci on many of these cells that were on the slide DCIS and synchronous IDC. And we don't have to go through here in detail, but that allowed us to enumerate in the entire population of the cancer copy number gains for a certain chromosome and copy number losses for a certain chromosome. And here you can see there are many cells that have the same pattern, and this is a DCIS and this is a synchronous IDC. You can appreciate that here we have a major pattern that is recapitulated in the invasive component, but you can also appreciate that we have an enormous degree of heterogeneity. In some instances, the clone that was predominant in the DCIS actually disappeared in the IDC. And together with our colleagues from NCBI and in the collaboration, very fruitful and pleasant collaboration with Russell Schwartz, Kahn-Eggie-Millen, we then could reconstruct the dynamics that occurred from the transition from DCIS to invasive disease. And two pathways occurred. One was clonal stability. Here you can see the DCIS on the left and the IDC on the right. You can see the major clone, which is defined by these copy number orations, prevailed and was also found in this component. And here as well, and in this case, it was the same. But much to our surprise, we also noted that in some cases the entire population of clones that were present in the pre-invasive disease disappeared. Only those that acquired extra copy of the MIG oncogene made it to invasive disease. And that again told us that the transition from pre-invasive disease, in many cases, is determined by the acquisition of extra copies of the C-MIG oncogene. But you can also appreciate that in many cases the ductal carcinoma in situ are already governed by a enormous degree of chromosomal instability. Therefore, I'm not a clinician, but I just cannot see that any other intervention than surgery would remove these clones. But anyway, despite this chromosomal instability, if we then look at all these clones that we have analyzed in copy number changes, we can conclude that chromosome 1Q, which is frequently gained, is very rarely lost. And chromosome 8Q, MIG oncogene, which is frequently gained, is very rarely lost. And those chromosomes here, P53, for instance, which is frequently lost, is rarely gained, despite this enormous degree of chromosomal instability. And this makes it then consistent with the genomic imbalances that are specific for copy number changes. So despite chromosomal instability, what is the pressure for Darwinian selection is the maintenance of the genomic copy number changes that are the defining features for any one of those carcinomas. This now, of course, triggers the questions. We have these changes, mostly whole chromosome arms or, in other instances, whole chromosomes in the colon gain of entire chromosome 7. This now triggers, I think, a fundamental question. How does it affect the transcriptome? You could ask the question, but you could test the hypothesis. The expression of all or most genes located on a chromosome is affected by chromosomal gain or loss. Or the expression of only a few genes whose reduce or increased expression is critical for tumor genesis is the target of chromosomal aneuploidy during tumor genesis, and this was not clear. And in order to identify...  The distribution of genomic imbalances is cancer-specific to an extent that you don't have to have any other information, but the distribution of genomic imbalances is a cervical carcinoma or a colorectal carcinoma. Specific genomic imbalances occur before the transition to invasive disease, which is very important if you want to convert that to a diagnostic test, which I will show you later. And, as I said before, there is no cytogenetic chaos but a stability on a different plateau of genomic copy number changes. So, as I said, we then ask, what are the consequences of chromosomal aneuploidy of the transcriptome, which I will discuss briefly in the next few slides, and then I will turn to how we can use this knowledge to improve the diagnosis of cancer and premalignant lesions. So in order to address the question what are the consequences on the transcriptome, we use the...an old technique which is called microcell mediated chromosome transfer to generate artificial chromosomal trisomies and this is how it looks. Here we have a normal kerotype and when we do this manipulation, we can put in an extra chromosome, in that case chromosome 3, but you can see, which is now present in 3 copies, but you can see that the rest of the kerotype is unaffected. Then we collected RNA from the control and RNA from the plus 3 cell and performed global gene expression profiling. Here again the control, some genes are up, some genes are down, which is what you would expect, but if you look at the plus 3, you can appreciate that most, if not all genes, go up in expression levels. And this, by the way, is the same if you look at the constitutional chromosomal trisomies 13, 18, 21, where this increase is also not restricted to a few genes but affects most of the genes and in a resting on a site, it's tolerated for chromosome 13, 18, 21, because these are the gene poorest chromosomes despite they have a larger size, 13, of course it's larger than chromosome 20. So aneuploidy results in the significant increase of average message levels of genes on the affected chromosomes. The degree of increase closely follows genomic copy number. Therefore, chromosomal aneuploidy is not only target a few specific genes, but result in a massive and complex deregulation of the cancer transcriptome. And just contrast that to what we see as a consequence of the Philadelphia chromosome where you have one aberration. Now you have a thousand genes that go up and you also have to appreciate of course there are numerous transcription factors that affect genes and other chromosomes. So it's really a massive deregulation which we have not yet understood in its complexity. This is not only the case in our model system but also in real tumors. Here a control, a normal DNA copy number and this is the gene expression level which is normal as well. But if you have copy number decreases or copy number increases, the gene expression goes down in this region, goes up in this region. It's completely clear. Function follows form. And this is established not only from our laboratory but now accepted in the scientific community. To summarize we have a normal cell carried typically stable. Then we have chronic inflammation for instance in the colon, HPV infection. We have for whatever reasons in the breast hyperproliferation we have just a cell cycle accident. What happens then? That we have extra copies of a given chromosome and this chromosome is tissue specific that can occur with or without additional mutation. Then what I call nuclear aneuploidy is very low so that there is not much difference in the DNA content from one cell to another. The trisomies affect global gene expression levels and as you will see later these events are the basis for global expansion of these early lesions. If this is not being treated then we arrive at a cancer cell where we have additional mutations. As we know P53 occurs relatively late in colorectal tumor genesis. We have an enormous degree of variability in the DNA content from one cell to another. But the genomic aneuploidy persists that why CGH was successful. As I have shown you before what is the selective pressure is the maintenance of the specific genomic imbalances. So now after this introduction I want to show you how we convert that to translational applications. Three topics I hope I can cover them all. First I will discuss with you the role of genomic instability in the prognosis of breast cancer and then we look at cervical carcinoma and if time how we use transcriptional profiling to predict treatment response in patients with rectal cancer. So breast cancers usually present in two flavors. One have a relatively stable genome which are called diploid and others are genomically instable have high degree of aneuploidy use if you take home lessons this is the Washington Monument and this is the Manhattan Skyline and many years ago our collaborator Greta Auer at Kowalinska I was not on that paper at the time discovered that patients who have a genomically stable tumor have a far better prognosis than patients who have an aneuploid tumor. Profound differences. And then in 2002 and as I said many other papers followed Wanderweber published a paper in the New England Journal who showed that there is by gene expression a good prognosis signature and a poor prognosis signature. They even came to the same conclusion and if you look at these curves you cannot argue that they are very similar. So we try to understand the nature of the similarity and perform gene expression profiling of a set of diploid tumors and a set of aneuploid tumors. And we could easily separate the aneuploid ones from the diploid ones and this separation was based on a signature of 12 genes which perfectly separated almost perfectly separated the stable from the instable breast cancers. And then we asked the question can we use this aneuploidy specific gene expression signature to predict to recapitulate the published data sets and that worked in all instances and the numbers are very high with very high statistical significance. And then the next question was whether we can ask the prognostic signatures that were derived from these data sets to predict the degree of genomic instability in our cases. And that worked perfectly well. The oncotype DX which is used in the clinic as you know and the good prognosis signature of the oncotype DX predicted that the tumors were diploid and the poor prognosis signatures predicted that the tumors are aneuploid and you could see that works very well. And the same occurred for the mama print. The good prognosis signature predicted genomic stability. The poor prognosis signature predicted aneuploidy. And just on the side luminal area is another gene expression signature which is now which you know is associated with the good prognosis predicted genomic stability and the basal signature under her two new signature associated with the poor prognosis predicted genomic instability. In summary, the degree of genomic instability and gene expression signature of poor prognosis are linked. The degree of genomic instability is the major biologically determinant of poor prognosis. And it is not only the degree of biological genomic instability if you look at the tumor cell but it's the fact that you have many multiple clones which provide the tumor with the nimbleness to react to environmental challenges including therapy. The next application which we developed for over more than 10 years is the identification of individual progression risk in cervical dysplasia. And here you remember what I showed you that there is invariably a gain of chromosome 3 the long arm of chromosome 3 in cervical carcinomas. And this gain never occurs in normal cells. So we developed in collaboration with Abbott a probe set that targets these regions that just control probes which help us to just control probes which help us to in normal rate you have the same scenario if you test for her to do amplification you also use a centromere for chromosome 17 as a control. And then we applied this to routinely collected PAP cervical smears. And I want to really make the point in cervical cytology the challenge is not to diagnose cancer. Because we all actually do not want to diagnose cancer because it's too late. We want to diagnose early lesions which can be cured by surgery. The challenge in cervical cytology is that there are two challenges. I mean everybody can say well this looks different than this but you have a normal cervical cell which is defined by a small nucleus and a large cytoplasm and you can easily discern that from these cancer cells but the distinction from here to here is much harder. Even the distinction from here to here is essentially impossible. What is even more important is that only 15% of the low grade dysplastic lesions would progress. Therefore it would require treatment despite the fact that about 90% of these low grade lesions are already positive for HPV. So HPV does not really help. It only helps if it's negative because then you know there won't be a cervical carcinoma but most of them are positive so it doesn't really help. So we argued that if all these carcinomas have a gain of a chromosome 3Q but none of the normal have it. And there is only a fraction of the low grade lesions would progress and would require treatment that those that progress already have the aberration that defines the invasive disease and that those 85% that would spontaneously regress are cytogenetically normal. And we then explored that and you can see here with this probe set for three copies 2, 2, 2, 2, 2, 2. This is normal. Cervical interaberythal neoblasia grade 2 which is a moderate dysplasia. You can see here 1, 2, 3 copies of chromosome 3Q. The rest of the genome is still normal. And in some of the carcinomas you can see clouds of 3Q of the turc oncogene which we choose as a probe that is reminiscent of the amplification of her 2 in breast cancer. And then this is unfortunately very difficult to see. I come back to the what I said it is the basis for clonal expansion. Here you can see a pap smear which was stained. We then destained it. Here a normal cell 2 copies. And here we have a nest of cells which were collected. They all have 3 copies of chromosome 3Q. But they are next to each other. Meaning they originated from each other. So once a cell has acquired 3 copies that is the basis for clonal expansion. So not only do we have a diagnostic test but we also can visualize the emergence of cancer by looking at the copy number changes of that particular chromosome. We confirmed that in many different studies this was one in collaboration with women who had pap smear randomly and those that had a suspicious pap smear then underwent colposcopy and biopsy. So we could correlate the findings under pap smear with the histology with a gold standard and the conclusion is that the gain of 3Q the human telomerase gene which is on there has the highest combined sensitivity and specificity for the detection of histologically confirmed high grade lesions. And we are now conducting an even larger study to extend that to low grade lesions and to ascus. We conducted another study which was aimed at validating 3Q as a molecular marker of progression which as I showed you at the beginning is the most important and this is how it was done. We collected a group of patients who are now much higher which were diagnosed with severe dysplasia and before they had a low grade dysplasia. Then we had another group of women which had the same low grade dysplasia but returned to normal. We had another group of women which were diagnosed despite being enrolled in an active surveillance program with severe dysplasia or even carcinoma but the pap smear before was normal so that should not happen. So we then hypothesized that those we know already these are positive these are all negative, these are all positive but we hypothesized that those low grade lesions that show progression are positive for 3Q and those that show regression are negative for 3Q and this worked essentially with the sensitivity of 100% and the specificity of 95% and we can discuss that later it's a particular feature of HPV but you can see that the point of no return in the progression risk for individual lesion is the acquisition of the specific cytogenetic abnormality that then defines the cancer entity. We also were expected a little bit shocked that in some cases where the pap smear was assessed as being normal and after a relatively short latency the woman was diagnosed with a carcinoma we could detect extra cubbies in a third of the cases in the cases that were cytologically assessed as being normal and I'm just showing you some examples here here there is a syn-1 lesion which regressed two copies, two copies so there would not have been any other treatment required here we have a syn-2 lesions and you can see here for instance these two cells and they have both three copies this lesion is progressed to high grade dysplasia and carcinoma and this is the case that I mentioned to you before which was assessed as being normal but you can see you have four copies of chromosome 3Q in these set of cells and again they are next to each other indicating that the acquisition of this operation is the basis for clonal expansion I shall also emphasize so I talked to cytologists at Hopkins and they come in the training program twice a year and every time at the beginning the wrong diagnosis goes up and then they learn a little and it goes down because it's obviously relatively difficult to unambiguously assess the morphology and when I go to cytology meetings there are whole days where they discuss on how to best repeat the reading of a pap smear to avoid false negatives and the fact that it's relatively often repeated is an acknowledgement of the fact that an individual pap smear is relatively ambiguous but you cannot argue that there are four copies so it's a binary you can ask your six year old to count to four and make the statement this is not true therefore they cannot even say make the statement as cervical carcinoma they can even say there is a woman whose low grade dysplastic lesion will eventually be an invasive carcinoma so it took us a long time to convince gynecologists and pathologists to embrace that because the procedure for the collection of this material would not even have to be changed because in particular with liquid based cytology there is always leftover but now finally quest diagnostic has embraced the test and is offering it in its portfolio so now I want to switch gear a little bit I don't know what the time is I'm fine and just talk a little bit how we not only use genomic operations but aberrations of the transcriptome and inject genomic information into a problem of treatment response in patients with rectal cancer and you know that better than I do there are many treatment options in solitumis it can be surgery alone surgery plus adjuvant chemo radiation or chemotherapy in colon carcinomas or near adjuvant chemo radiotherapy followed by surgery or other approaches or in some instances dependent on the mobility of the patient chemo radiation alone and the problem in rectal cancer is a real clinical problem because based on a large study which was conducted from the term on rectal cancer study group and has been adopted now in most of Europe I know it's not quite completely adopted in the United States the standard treatment of locally advanced rectal cancer is near adjuvant chemo radiotherapy mostly based on five of you in radiation followed by surgery and then chemo therapy and the reason for that is that they found out that if pre-surgery chemo radiotherapy is administered there is a higher rate of R0 there was a higher rate of sphincter preserving resections and there was a reduction of local recurrences significant from I believe 14 to 6% the profound clinical problem is that the response to near adjuvant chemo radiotherapy is very heterogeneous you can have complete pathological response in that case where after surgery there is not a single tumor cell left or you have essentially complete resistance where the tumor just doesn't bother being treated with 5FU and radiations so we generated together with a former postdoc of mine Michael Godimi who is now the chair of surgery at the university in Goettingen Germany a clinical research unit you can look it up in order to address that problem and we aimed at identifying predictors of response and identify targets of that and understand mechanisms that could explain this profound difference in response versus resistance and in order to do so we initially designed a pilot study with 30 patients they all had pre-treatment clinical stage based on rectal ultrasound we then collected tumor biopsies and performed gene expression profiling they obviously I can barely see from here but then they went to this neo adjuvant scheme which is chemotherapy followed by surgery and four rounds of 5FU and then we had a long term follow-up the pathological staging is shown here and as I said we performed gene expression profiling and then evaluated local and distant recurrences after medium follow-up times of 44 months and the gene expression profiling worked actually fairly well here we had I forgot how many, there were 30 cases approximately half of them had non-responding tumors a little fewer had tumors that show complete response and the gene expression profiling could actually fairly well discern these two groups but then we looked at the genes that were up-regulated in the tumors that were resistant and among those what a transcription factor TCF4 and this obviously rings a bell because transcription factor TCF4 is the major effect of wind signaling of the major effect of wind signaling of course not only in colorectal tumor genesis and wind signaling is intricately involved in colorectal tumor genesis affect mutations in the adenomatosis which actually poses coli gene resulted in stability of catenin which then actually increases the expression of the transcription factor TCF4 which in turn turns on cyclin D1, CMIC and other notorious oncogenes and therefore transform these cells so that was a very interesting lead so we hypothesized that silencing of a gene that is over expressed in resistant tumors would increase sensitivity to chemo-radio therapy and we did this silencing with the approach of using RNA interference and I don't go into details here but if you inject small interfering RNAs into cells you can reduce either protein synthesis you can cleave the transcript which results in a loss of function and you can do that essentially for any gene of interest so we used small interfering RNA against the transcription factor TCF4 and asked the question whether silencing of TCF4 would increase sensitivity of colorectal cancer cells to chemo-radio therapy here you can see the results it was a very effective silencing in the reduction of protein we repeated that with many different constructs and when the cells were then subjected to radiation not shown here but also to chemo-radiation this is a large scale you can see here the control and you can see here a profound sensitization after silencing the transcription factor TCF4 which in primary tumor samples was over-expressed in those that were resistant to treatment conclusion reduced expression of TCF4 leads to a sensitization to radiation we are now exploring whether we can recapitulate this effect using small molecule inhibitors of wind signaling and together with this clinical research unit in Göttingen we are about to design clinical trials to test that in patients so in summary we have performed gene expression profiling which I have shown you in rectal carcinomas that are resistant or sensitive to near adjuvant chemo-radiotherapy we have also varied many different levels of the genome using CGH to see whether we have specific genomic imbalances that could explain the sensitivity we have performed microRNA expression profiling we explored KRS and BVF mutation which by the way did not explain the different sensitivity to radiation we looked at the methylation status and in collaboration with Steven Chanock the Illumina platform to look for single nucleotide polymorphisms in the group of patients that respond differentially to explore whether certain haplotypes for instance in genes that are involved in drug metabolism could explain the profound differences in the clinical course we do all this in order to address the problem that all drives us which is that patients come into the clinic with different clinical features they have a different genetic makeup and a different tumor biology and in the case of rectal carcinomas they also have a different treatment toxicity and this is not only true for rectal carcinomas for instance primalignant lesions in the cervix could be summarized here however despite these acknowledged differences which we do not understand to the extent we should all these patients receive identical therapy the goal of all the stuff of course is to learn enough from all these different techniques I have not listed next generation sequencing here to learn enough about the genetic makeup so that we can assign patients to therapies from which they would benefit the most with that I'm at the end and I thank you for your attention I'm happy to take this I have one a question that all biology is only understandable by natural selection and evolution is it possible that by using chemotherapy in a way we encourage resistant tumors to grow out? I think it is I think it is I mean tumor heterogeneity was already I don't have it on this slide unfortunately but what we are now doing we look at these rectal carcinomas that are complete responders and those that are resistant and ask the question whether in those that are then resistant there are minor clones that expand during chemotherapy completely passable but I can also say it's now being more and more acknowledged that we cannot think if we look at the bulk of the tumor we have understood the tumor so this is a signal to noise question really what are the comments and questions? in breast and colon cancer is there a normal tissue in the same patient like those described in the series? the question was let's just look at cervical carcinomas or colorectal carcinomas whether those cytogenetic abnormalities that are the early events in tumor genesis can already be detected in the adjacent normal epithelium not on the chromosomal level but we are now exploring the possibility that on those chromosomes that are early gained in any one of those diseases that there are genes which are already higher more highly transcribed so that the initial impetus to gain these chromosomes would be a physiological one but on the chromosomal normal tissue has no chromosomal approvals is there a normal person without cancer? no so the question was what is the role of non-coding RNAs whether they are short or long in the maintenance of genomic instability and the answer is I don't know the answer is nobody knows obviously through the genome institute and other initiatives we will learn much more about it hopefully through other initiatives as well but the role of non-coding RNAs in the maintenance of the genome is not yet established is that an indicator that you are getting increased risk genetic defects or do you look at that or it has something totally different? so the question is to which extent established serum markers would reflect the risk of developing specific aberrations I cannot, there are no studies I cannot answer that and the serum markers are very likely but there are obviously problems and many people have looked for colon and other tissue pancreas but there is no direct correlation it could be also factors that are unrelated to the presence of certain genomic imbalances which is whether you have a blood vessel through the tumor and you can shed cells so what I believe will become a possibility in the future for earlier detection is the detection of circulating tumor cells the reason I ask that is I have a lady with metastatic ovarian cancer who is in total remission and her scans are normal and she just has an increase in her CA-125 so as a non-oncologist how am I supposed to deal with that or just let the oncologist handle it which comes to me and asks me the question I'm not going to make it such a subject so can you comment on the tempo of translation of this exciting science to the clinic just a question I would have in my head is this something I'm going to have to face tomorrow a year from now or 10 years from now it seems like it's pretty soon for the cervical casinomas it's being implemented now there are several laboratories around the world who actually use it for samples samples that are ambiguous the goal would be to all the cases that are HPV positive should get this genetic test because then you know what's going to happen and I usually restrain from such statements in biology or genetics but this is black and white it's you have it, you progress you don't, you do not, period so this is probably the most advanced we are now, as I mentioned to you before we work with the Collins Institute where we have 9000 archived breast cancer cases from all of which we have the clinical follow up the suites are very good with that there is no population not so much mobility and we have the clinical follow up and we have the DNA content so we are now in the process of correlating in a large definitive data set the degree of an employee with prognostication so that's going to be next we have just received funding and getting out of trials where we can try where we can evaluate in larger numbers the value of gene expression profiling for prediction of response I recall reading a note from Ben Stavida who was a former director of the Cancer Institute that we were entering a phase where in arbitrary cancer you need a very skillful doctor and this stuff is really beginning to bring that home we need more other comments or questions? it's only done as a reflex when you start seeing abnormal cells is that likely going to change? I know with it there were several studies I believe they suffered from budget cuts and sequestration exactly addressed the question whether HPV should be the first test because it essentially would make sense because those that are negative don't get cervical cancer I completely agree but I have not seen the result of any there was one out of Seattle, there was one that was initiated by Mark Schiffman at the NCI and others but there is no I don't want to step on anyone's toes but maybe there is also a priori resistance by the cytopathologists to change or by the gynecology to change something which has been successful and I mean one should be very careful to change something that is successful into something where you first have to validate it on a large basis but from a biological point and a genetic point of view there would be no reason not to change the practice to what you just suggested It is a fact that we clinicians stumble along behind investigators and it takes a while to translate I think Other comments or questions? If you have them please come down Thank you very much


Hi everyone. I'm Jocelyn. I'm president of the Gustavus Choir. On behalf of the choir, I just want to thank you so much for all of your wonderful gifts to Gustavus. Some of those gifts include our special singing mask that we're using this year. It allows us to sing safely and together. We are currently in Beck Hall rehearsing for Christmas in Christ Chapel and we want to share a little bit of that music with you right now.


Good morning everybody, little frosty out here this morning. We had a fog overnight and that's what happens. So we're getting a bunch of work done to the host right now. And we have our friends from Browns plumbing and heating over right now. And they're installing a new high efficiency furnace, a new hot water tank, and a new air conditioner. But the air conditioner is actually going in next week. There's no real rush for that, as you can tell. They're gonna do that next week. But we got it all on a deal, so we bought it all together. Our furnace was over 30 years old and was long past its life expectancy. We had a little bit of trouble with the motherboard in there as well. It was a little bit of a short, nothing life-threatening or anything, but it was an issue that needed to be resolved. And we decided it's pretty old furnace, we better replace it. So the furnace we got now is a two stage high efficiency furnace. It's gonna have a 26% better efficiency. So our gas bills should go down 26%, up to 26%. So they're working in the basement on that right now. We got the new hot water tank installed as well right away because our hot water tank was also over 30 years old. They have a life expectancy of what? Like 15 years, 10 years, 10, 15 years. But that replaced all at the same time. So now this house is gonna be good to go for the next 20 years. The furnace will be good for like 20, 25 years. And the hot water tank, like I said, 10, 15 years. And then we'll have the AC installed so it'll be ready to go in spring. Now, we're in the back here. I believe we're gonna install our AC.  Right now we have it installed. The current AC is on the other side of the house there, outside the fence. And all of the lines are running up in the roof of the basement, which is all plastered in with drywall. And you'd have to rip out the drywall or cut out the drywall to get to the lines. So what we did was we just did a delete on that air conditioner over there.  And then we're gonna, because this is our furnace room and our laundry room right down here with that little window. Now the line's coming out right here. We're gonna have them go underneath the door here to there. I'm gonna build a little step here to cover them up so that we don't accidentally kick it when we come in and out of the door. We were gonna put it right in this area here, but there's already quite a bit going on here. You can see those white pipes over there. Those are the new intake and exhaust. The old furnace, which was in high efficiency, went up through the chimney there. Now you can see there's still emission error. You can still see that there's some exhaust coming out of there now. That's because the hot water tanks will still use the chimney. They have the new hot water tank installed and it's heating up the water right now and that's what you're seeing. The new high efficiency furnace, this is how they do it. They come out the side of the house here. Now the intake and the exhaust right there. They're gonna clean that up yet. Knock that down. On the air conditioner, the lines will come out of the house somewhere around here. We couldn't put it here because these two exhaust vents there, one's for the stove hood exhaust, one's for the dryer. You've got your water lines over there. We don't want the AC right beside the deck there either. So, probably one of the lines. I'll put it somewhere over here. And we'll have a brand new AC. So that one spring hits and the sun decides to come up a little higher. Warm us up a bit. It'll keep ourselves nice and cool. What do you guys think, Chevy? Diesel? They heard something. I think there's a dog over there, mate. Probably. So now we're on the side of the yard here on the side of the house. This is where our older air conditioner is. Quick, it's pretty old. So that's the unit and that's original as far as we know. That's about 30 years old. So this guy's gonna be going. They're busy working down there. I'm just staying out of their way. So now the big reveal. I'm excited. I've already seen it. I'm gonna show you guys. I don't know. I know I'm old now because I get excited about getting a new furnace and hot water tank installed. So here it is. I'm gonna turn you around here. It's a Lennox installed by Browns. Plumbing and heating. We get all of our work done through them. They're phenomenal, amazing. Highly recommend them if you're in the area and you don't know who to do. You're plumbing, heating, anything like that. Give them a call. We've gone through them for years now and they've always been on point. You know, we got everything inspected after we moved in here. And the inspection didn't go that great. It was Browns that came in and did the inspection where on their MVP program where we pay a monthly fee and then they come out and inspect your furnace once a year, inspect your plumbing once a year and inspect your air conditioning once a year for free. And they do all of your plumbing as well too. So they came out and did the inspection as soon as we moved in here, we booked an appointment with them. They came and they checked out that furnace. Remember I showed you that furnace in there? That thing was built in 1991. Installed in this house in 1994. So it was over 30 years old. It was past its life expectancy. And the fan in there, it sounded like a jet engine when it started up because it was taking so much energy it just didn't wanna turn over the fan anymore. And there was also a problem with our circuit board in there as well where it could become a safety issue if it evolves into anything further. So we got them to check out everything else. The hot water tank that was in here before as well. Also original, 30 years old. How many of you have heard of a hot water tank lasting 30 years? Our last hot water tank that was in here was 30 years old. So that was where that needed to be replaced. The inside of it was all gummed up. They flushed it out for us during that inspection. Which improved it a lot. But they could tell that it was... They didn't know how it was still working. So that needs to be replaced. And as well, I showed you our air conditioner outside. Also 30 years old and needs to be replaced. So we got it all done. All in one shebang. And we saved a lot of money by doing that. We saved about $3,500 by doing it all at once. And here we are. So this house is practically brand new now. I mean, it's still very well put together. The structure and the frame and the bones of the house is practically like brand new. But don't gotta worry about that. And now we have the furnace that's brand new. It's a high efficiency two stage furnace. So it has two levels depending on how much power it needs. So it's supposed to save us power that way. It's also a high efficiency furnace. Remember I showed you that last furnace and it vented the exhaust out the chimney. I think I explained this to you earlier in the vlog. It used to go out here, right? Now just the hot water tank goes out here and they put a sleeve up there all the way up to the top of the chimney to ensure nothing leaks at all. The furnace however, no longer goes up the chimney. This is a high efficiency one. So this one's a little different. It has the intake right here and the exhaust there. Goes around here, up there and out the side of the house. Think I also explained this to you before. Sorry, I'm trying to remember what I already told you. So everything in here is good to go, brand new. We're gonna have to move the location of the air conditioner like I was saying because these are the lines for the old air conditioner here, right? They come around here, they go up here and then they go through here and into the dry walled ceiling of the basement. So I don't know about you guys but if you are building a house in the near future and you're gonna build a basement, don't dry wall the ceiling of the basement. Okay, take it from Trucker Josh, it's a pain. I haven't had the biggest pain yet because we haven't had any big problems other than this but if you dry wall the ceiling of the basement, imagine all of this, like all of these pipes and everything, all of that is in there, in the whole basement. And if you need to access any of it, you gotta actually cut out the dry wall and re-dry wall it. It's a big, big pain. If you're doing a basement, put tile ceiling in the basement, way easier to access stuff. But anyways, because this goes through the dry walled, we can't get to it. Unless we rip all the dry walled, we don't wanna do that. And then you gotta get paint matched and paint everything, redo it. They would take care of it. Browns would take care of all of this for us but we don't wanna go that route.  Just leave that in there. The new air conditioner is going to come out of the same area here. But instead of going all the way down to the other end of the house, like way out over there, it's just gonna zip and go right outside the house right here. And then that way, the air conditioner will be there. The garage is just over here, right? So it'll just be over there way shorter, way easier to keep track of and easier to maintain as well if anything ever goes wrong. Easier to get at, because what if something went wrong with one of these lines in there, right? Again, we have to rip out the whole dry wall ceiling and the whole basement is like that. It's all drywall, right? Even in here. So don't get me wrong, I really like the look of the drywall. We didn't do that. We bought the house like this. I think it looks great. I'm glad they did it for the looks. I'm not complaining about it at all. But when you do come into circumstances like this where you need to access all of the lines and the pipes and anything else that's up there, it's tile ceilings the way to go. Remove the tiles and get up there. Anyway, so that was our endeavor. It was pretty expensive. Yes, it was. If you're wondering, yes. Yes, it was expensive. But the only thing we have in here now that hasn't been replaced that might need replacing in the next 15 years is the water softener system. But that one's still doing pretty good. And I don't think we'll need to replace that anytime soon. But that is the oldest thing in our house right now. Like check this thing out. Brand new hot water tank. See, they got all the gas lines going up here.  That is code now to have that here. Up there, you got the little gooseneck up there. The drain, the exhaust, they completely redid. Goes around the same route as the other one. And this furnace is in the same spot as the other one. Comes up, this is all new up to here. And this is where they connected the new to the old. And that's the old ducting there. Lennox is a great brand. We went with the Lennox air conditioner as well. But the air conditioner is just a one stage. Remember at our old house, we had that super, super fancy air conditioner. We bought a variable speed air conditioner there. It costs us a lot of money. We wanted the best and we got the best. We didn't need it. We shouldn't have spent that money. We did. That air conditioner alone was more expensive than this furnace and that air conditioner combined. But we did get that deal because we bought both Thorol three at the same time, right? It's hard to tell. This is an expense that should only come up about every 20, 25 years. So it should be good until about 2050. Now it's 2023, the end of 2023 right now. And close to 2050 probably is when we'll be good till the hot water tank might need to be replaced before that. But the rest of it, hopefully anyways. So that's my big show and tell. Woo. I'm an adult now. These things excite me. I should find out what that model number is because you guys probably want to know the model of that furnace. I'm sorry, I don't have it right now. And we're waiting on the paperwork, all the invoices to be finalized because this just got done. But if you want to know, I can share it in a future vlog. Two stage furnace, that's all. That was all I really had to show you today guys. I'm still waiting to go back on the road. And I'm enjoying my time at home. So we've got a couple more things to do. I got to do some work to Old Blue in the shop yet. Needs a little bit more work done to it. We'll do that tomorrow. And then we'll be packing our bags, loading her up and headed down to Texas. I got word back from my friend who drives down in Texas every now and then. You know him, Moses. I met him on the road a couple of months ago. He drives to Texas every now and then. And he confirmed what I thought. He's like, yeah, don't sleep anywhere near the border. Don't go. When you got to go down to Laredo and you got to go that close to Mexico, the goal, what you do is you stay several hundred miles away for night into the US. And then when you have to deliver, all you do is you go down there to be on time, you get in, you get out. Get in there, drop your freight, get out. He said, don't stay there. My delivery's gonna be later in the evening too. We'll talk about this more in the future. But I won't be staying anywhere around there. We're down in Laredo, Texas. So you'll hear more about that cause I'm excited. Thanks for joining me anyways guys, for my little show and tell. I'm sorry I don't got more to show you but I'm on holiday, okay? So if you want my honest opinion, you're lucky you're getting any vlog. I just love doing this so much. Take care everybody, see you tomorrow.


Padre, Padre, Padre, I like the thought that you had to be vibrating at a certain frequency to receive that frequency. Yeah, that was super cool. You can't receive an FM frequency on an AM dial, so you have to be tuned in to be able to receive a certain signal. Super simple concept. But bang on, that was really cool actually Padre. That was super cool. That's why people say, are you listening to me? Are you listening to me? Can you hear me? Are you tuned in to me? Maybe if that doesn't switch that. Are you listening to me? Can you hear me too? Are you tuned in to me? Are you vibing with me? Are we at the same frequency? Maybe the hippies at it, I don't know. But that's pretty cool. I thought that was pretty good, right? That's why maybe when people do, as Diem Turner mentioned, when you do ecstasy with a small group of like-minded people, close friends, it amplifies the feeling. Same goes with mushrooms, right? And if you're especially mushrooms, if you're on a mushroom trip in an environment where you're not vibing with people, oh wow, wow, that's nastiness, right? Salute evil to hope, hope you're doing well, hope you're doing well. That was fun. That was fun. Those damn hippies were onto something, they were onto something, but they lost it. They lost it, right? There was a handful of them that remained true to the calling, but there weren't that many to begin with, I don't think, right? Like people talk about the, I've read a little bit about this. People talk about the 60s, you know, everyone vibing, blah, blah, blah. But that was only in pockets. Majority of the United States and Canada was still not the hippie culture, right? They weren't really, they sold out, bro, they sold out. They sold out. Again, there wasn't that many of them, right? There weren't pockets, but if, and one of the main pockets was San Francisco, but look at the shit coming out of San Francisco, right? Is that from the hippie culture or is that hijacked, right? I don't know. So to, I tripped on shrooms with people who weren't having a good vibe. It went from one of the best to the worst experiences real fast. Yeah. That's what mushrooms does on a flip of a dime or was it a flip of a dime or whatever it is, it all, it could turn on you so fast. Same with LSD, turn on you so fast.  I would do it again. What about Woodstock? Woodstock was, there were people that died in Woodstock, right? The hails angels with the bodyguards, there's some, there's some people that died in Woodstock. You know, there's lots of lovey-dovey things happening, but there were some nasty stuff going on too, weren't there a lot of people there? Yeah, but that's, you're talking all of the United States, take it all to one music festival and the hardcore's would have been migrating to that, right? How big was Woodstock relative to the entire United States? Not that everybody went there, but there was a lot of people there, right? Look at Woodstock 99, clearest case of selling out of everything. Yeah, I didn't, that was like vomit inducing, right? That was the wacko. And look at that. Look at the people, some of those musicians. Look at, what do you call it, Neil Young?  Man, I used to like Neil Young.