[Federal Register Volume 76, Number 45 (Tuesday, March 8, 2011)]
[Rules and Regulations]
[Pages 12558-12559]
[FR Doc No: 2011-5246]

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DEPARTMENT OF TRANSPORTATION

Federal Aviation Administration

14 CFR Part 73

[Docket No. FAA-2011-0055; Airspace Docket No. 11-AAL-2]

Amendment to Special Use Airspace Restricted Areas R-2203, and R-
2205; Alaska

AGENCY: Federal Aviation Administration (FAA), DOT.

ACTION: Final rule; technical amendment.

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SUMMARY: This action amends the using agency of Restricted Areas R-2203
A, B, & C; Eagle River, AK, and R-2205, Stuart Creek, AK. These changes
reflect the U.S. Army's current organization in Alaska. There are no
changes to the boundaries, designated altitudes, time of designation,
or activities conducted within the affected restricted areas.

DATES: Effective Date 0901 UTC, May 5, 2011.

FOR FURTHER INFORMATION CONTACT: Ken McElroy, Airspace, Regulations and
ATC Procedures Group, Office of Mission Support Services, Federal
Aviation Administration, 800 Independence Avenue, SW., Washington, DC
20591; telephone:<PHONE_NUMBER>.

SUPPLEMENTARY INFORMATION:

The Rule

    This action amends Title 14 Code of Federal Regulations (14 CFR)
part 73 by changing the R-2203 and R-2205 using agency of Special Use
Airspace to, ``U.S. Army, AK (USARAK), Commanding General, Fort
Richardson, AK''. This is an administrative change and does not affect
the boundaries, or operating requirements of the airspace, therefore,
notice and public procedures under 5 U.S.C. 533(b) is unnecessary.
    Section 73.22 of Title 14 CFR part 73 was republished in FAA Order
7400.8S, effective February 16, 2010.
    The FAA has determined that this regulation only involves an
established body of technical regulations for which frequent and
routine amendments are necessary to keep them operationally current.
Therefore, this regulation: (1) Is not a ``significant regulatory
action'' under Executive Order 12866; (2) is not a ``significant rule''
under Department of Transportation (DOT) Regulatory Policies and
Procedures (44 FR 11034; February 26, 1979); and (3) does not warrant
preparation of a regulatory evaluation as the anticipated impact is so
minimal. Since this is a routine matter that will only affect air
traffic procedures and air navigation, it is certified that this rule,
when promulgated, will not have a significant economic impact on a
substantial number of small entities under the criteria of the
Regulatory Flexibility Act.
    The FAA's authority to issue rules regarding aviation safety is
found in Title 49 of the United States Code. Subtitle I, section 106
describes the authority of the FAA Administrator. Subtitle VII,
Aviation Programs, describes in more detail the scope of the agency's
authority.
    This rulemaking is promulgated under the authority described in
subtitle VII, part A, subpart I, section 40103. Under that section, the
FAA is charged with prescribing regulations to assign the use of the
airspace necessary to ensure the safety of aircraft and the efficient
use of airspace. This regulation is within the scope of that authority
as it amends Restricted Areas in Alaska.

Environmental Review

    The FAA has determined that this action qualifies for categorical
exclusion under the National Environmental Policy Act in accordance
with 311d, FAA Order 1050.1E, ``Environmental Impacts: Policies and
Procedures.'' This airspace action is not expected to cause any
potentially significant environmental impacts, and no extraordinary
circumstances exist that warrant preparation of an environmental
assessment.

List of Subjects in 14 CFR Part 73

    Airspace, Prohibited areas, Restricted areas.

The Amendment

    In consideration of the foregoing, the Federal Aviation
Administration amends 14 CFR part 73 as follows:

PART 73--SPECIAL USE AIRSPACE

0
1. The authority citation for part 73 continues to read as follows:

    Authority:  49 U.S.C. 106(g), 40103, 40113, 40120; E.O. 10854,
24 FR 9565, 3 CFR, 1959-1963 Comp., p. 389.

Sec.  73.22  [Amended]

0
2. Section 73.22 is amended as follows:
* * * * *

R-2203A Eagle River, AK [Amended]

    By removing the existing Using Agency information and
substituting the following:
    Using agency. U.S. Army, AK (USARAK), Commanding General, Fort
Richardson, AK.

R-2203B Eagle River, AK [Amended]

    By removing the existing Using Agency information and
substituting the following:
    Using agency. U.S. Army, AK (USARAK), Commanding General, Fort
Richardson, AK.

R-2203C Eagle River, AK [Amended]

    By removing the existing Using Agency information and
substituting the following:
    Using agency. U.S. Army, AK (USARAK), Commanding General, Fort
Richardson, AK.
* * * * *

R-2205 Stuart Creek, AK [Amended]

    By removing the existing Using Agency information and
substituting the following:
    Using agency. U.S. Army, AK (USARAK), Commanding General, Fort
Richardson, AK.

    Issued in Washington, DC, on March 2, 2011.
Rodger A. Dean,
Acting Manager, Airspace, Regulations and ATC Procedures Group.
[FR Doc. 2011-5246 Filed 3-7-11; 8:45 am]
BILLING CODE 4910-13-P


EDVMS MEETING DEC 10 12, 2003 PUBERTAL MALE AND FEMALE RAT ASSAYS EDSTAC TIER 1 Endocrine Disrupter Screening and Battery 1998 In Vitro ER Binding and/ or Transcriptional Activation AR Binding and/ or Transcriptional Activation Steroid Hormone Synthesis In Vivo Uterotropic Assay in Ovariectomized Adult Rat ( 3d)

Pubertal Female Rat Assay including Thyroid ( 20d)

Hershberger Assay with Castrate immature male rat ( 10d)

Frog Metamorphosis Assay Short term Assessment of the Fish Reproductive System EDSTAC Screening Battery:

Detects Estrogens by sc and oral routes in vivo and in vitro Detects Androgens and Anti in a senstive assay using simple endpoints ( organ weights) and in vitro.

Detects HPG/ CNS alterations related to FSH, LH,

prolactin, dopamine, GH with simple, precise developmental landmark without RIAs.

Detects Inhibition of Steroidogenesis in vivo ( in female)

with simple, precise endpoints and in vitro Detects Thyroid effects in intact Female Rat ( RIAs)

and Frog Metamorphosis Uses extensively utlized in vivo endpoints/ assays Questions?

Given the number of chemicals run in more than one lab, is a formal interlab study still needed?

Is the pubertal male assay sensitive enough to replace the Hershberger assay in T1S?

Should we evaluate the adrenal or other endpoints in more detail?

What mechanisms of action need further evaluation to establish the sensitivity of the assay?

Should we be running concurrent dose range finding studies?

Can we reduce the sample size in these assays? Comments Data analyses presented used SAS Proc GLM with BW21 as a covariate.

Post hoc t tests were used to compare means to control, by block, when F value significant.

We had several a priori hypotheses for many of the chemicals because they had been tested prior to the RTI and TI 2003 studies Analyses done by LEG at EPA Use of relative organ weights is not appropriate for EDC screening as the results can be very misleading. ANCOVA not much better when treatment affects body weight.

Need MTD of 10% Importance of Strain Effects in Screening Assays???

The OECD program to validate the rat uterotrophic bioassay: Phase Two  Dose Response Studies.

Jun Kanno, Lesley Onyon, Shyamal Peddada , John Ashby, Elard Jacob and William Owens.

doi: 10.1289/ ehp. 5780 ( available at http:// dx. doi. org/). Online 23 January 2003 Abstract:

The Organisation for Economic Co operation and Development has completed Phase 2 of an international validation program for the rodent uterotrophic bioassay. The purpose of the validation program was to demonstrate the performance of two versions of the uterotrophic bioassay, the immature female rat and the adult ovariectomized rat, in four standardized protocols.

. ........ It is noteworthy that these results were reproducible under a variety of different experimental conditions ( e. g., animal strain, diet,

housing, bedding, vehicle, animal age, and so on), indicating that the bioassay's performance as a screen is robust.

Pubertal Assessment of the LE versus the SD rats TI 2000. For each sex there were 12 rats/ sex/ strain, run in two complete blocks to determine intralab variability using 6 treatment groups ( a control and 5 chemicals at a single dose).

Factorial design  2 Strains x 6 Treatments x 2 Blocks for each sex Endocrine Disrupting Chemicals: Prepubertal Exposures and Effects on Sexual Maturation and Thyroid Activity in the Female Rat. A Focus on the EDSTAC Recommendations Jerome M. Goldman, Susan C.

Laws, Sharon K. Balchak, Ralph L. Cooper, and Robert J. Kavlock.

Critical Reviews in Toxicology, 30( 2): 135 196 ( 2000)

Modified Research Protocol for Assessment of Pubertal Development and Thyroid Function in Juvenile Female Rats.

Purpose and Applicability The purpose of this protocol is to outline procedures to quantify the effects of environmental compounds on pubertal development and thyroid function in the intact juvenile female rat. This assay detects agents that display antithyroid, estrogenic, antiestrogenic ( estrogen receptor [ ER]

or steroid enzyme mediated) activity, or alter puberty via changes in luteinizing hormone ( LH),

follicle stimulating hormone ( FSH), prolactin ( PRL) and growth hormone ( GH) secretion, or via alterations in hypothalamic function.

Required Endpoints:

Growth ( body weight). Liver, kidney, pituitary, and adrenal weights Age and weight at vaginal opening Serum thyroxine ( T4), thyroid histology and thyroid stimulating hormone ( TSH)

Uterine and ovarian weights and histology Vaginal cytology Optional Endpoints:

Serum tri iodothyronine ( T3), estradiol ( E2), and prolactin Thyroid weight Liver, kidney, pituitary, adrenal and vaginal histology Ex vivo ovarian and pituitary hormone production Hypothalamic neurotransmitter concentrations Estrous cycle length ( requires extension of dosing) 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 Assign to Treatments based upon body weight.

15/ group Dose Female Necropsy Our Immature ( 21  43 Days of Age) Intact Female Rat Protocol to Evaluate Pubertal Development and Thyroid Function.

Detects inhibition of steroidogenesis, antithyroid and ( anti) estrogenic activities and altered HPG maturation.

Wean Days of age Endpoints:

Growth Age and weight at vaginal opening ( VO)

Vaginal cytology Serum thyroxine and thyroid stimulating hormones Ovarian and uterine weights and histology Liver, kidney, pituitary and adrenal weights Dose daily Daily examination for VO and lavage LEGJR Published studies that have used the pubertal female assay Atrazine  2 labs Nonyphenol DES  2 labs Methoxychlor  VO and cycles Estradiol Octylphenol  VO and cycles Nonylphenol  VO and cycles Tamoxifen Fadrazole Ketoconazole Finasteride Testolactone Flutamide Perchlorate  in prep PBDE  submitted PTU MBC  VO and cycles DBP  VO and cycles Chemicals studied this year in the pubertal female assay. New data RTI  completed Atrazine 75 or 150 mg/ kg/ d Fenarimol 50 or 250 mg/ kg/ d Methoxychlor 25 and 50 mg/ kg/ d Bisphenol A 400 or 600 mg/ kg/ d Ketoconazole 50 and 100 mg/ kg/ d PTU 2 and 25 mg/ kg/ d TherImmune 2000 ( 2x2x6 factorial.

Blocks, LE versus SD and chemicals.

n= 6/ strain/ block)  completed Ethynyl estradiol .005 mg/ kg/ d Tamoxifen 10 mg/ kg/ d PTU 240 mg/ kg/ d Ketoconazole 100 mg/ kg/ d Pimozide 30 mg/ kg/ d Methoxychlor 100 mg/ kg/ d TherImmune 2003  completed Ethynyl estradiol 0.0025 and 0.005 mg/ kg/ d Methoxychlor 12.5, 25 and 50 mg/ kg/ d Phenobarbital 50 and 100 mg/ kg/ d Relationship between reduction in body weight due to restricted feeding and delayed age at VO 70 80 90 100 Percent of control body weight  6  5  4  3  2  1 0 1 2 3 4 5 6 7 Delay in VO in Days FR PTU2 PTU 25 BPA400 BPA600 MET25 MET50 KETO50 KETO100 FEN50 FEN250 ATR75 ATR150 Estrogens in the pubertal female assay Easily detected by accelerated VO if they are effective with oral administration in the rat Effect of Ethynyl Estradiol on puberty in the SD female rat TI SD 2003 31.9 31.1 28.5 Oil EE0025 EE005 25 30 35 AGE AT VO ** **

33.4 32.5 28.7 Oil EE0025 EE005 AGE AT Estrus 80 67 20 Oil EE0025 EE005 % Cycling Normally ** ** Effect of Ethynyl estradiol on AGE AT VO in the SD female rat TI SD 2003. Note: Ovarian and Uterine histological changes noted in almost all females in TI SD/ LE 2000 ( SD more affected) but no affects noted in TI SD 2003 study.

31.9 31.1 28.5 Oil EE0025 EE005 26 28 30 32 34 36 38 TI SD 2003 ** 34.9 25.9 36.3 26 Oil EE005 24 26 28 30 32 34 36 38 SD LE TI SD/ LE 2001 ** Effects of Methoxychlor in the RTI SD 2003 Pubertal Female SD rat assay 32.9 29.8 27.4 Con A MET 25 MET 50 22 24 26 28 30 32 34 36 Age VO 128 105 89.5 CON A MET 25 MET 50 80 90 100 110 120 130 140 Wt at VO * * *

35.1 31.1 28.8 Con A MET 25 MET 50 25 30 35 40 Age 1st P/ E * * * Effects of Methoxychlor ON AGE AT VO in the Female RTI 1994 LE, EPA 1989 and RTI SD 2003 rat studies 32.9 29.8 27.4 Con A MET 25 MET 50 22 24 26 28 30 32 34 36 RTI SD 2003 *

* 32 26.3 25.4 Con A MET 25 MET 50 EPA LE 1989 29 25.6 Con A MET 50 RTI LE 1994 *

* * Effects of Methoxychlor in the Pubertal Female SD rat assay 94 85 126 106 87 91 107 94 89 91 100 100 95 90 111 102 77 91 102 98 92 92 100 100 BWT OVWT UT WET UT DRY THYROID LIVER ADRENAL KID PIT BW GAIN Normal Ovary Histo Normal thyroid Histo 80 90 100 110 120 Con A MET 25 MET 50 Lack of effect of BPA in the RTI SD 2003 Pubertal Female SD rat assay on age and weight at VO. Octylphenol and Nonylphenol accelerate VO in this assay. BPA is positive in the in vitro and uterotropic assays but negative here 32.3 33 32.3 CON B BPA 400 BPA 600 30 31 32 33 34 35 36 Age VO 123 116 108 CON B BPA 400 BPA 600 100 110 120 130 140 Wt at VO * Effects of BPA in the RTI SD 2003 Pubertal Female SD rat assay 86 66 74 68 94 81 86 88 86 80 100 100 89 73 82 83 88 83 87 90 86 85 100 100 BWT OVWT UT WET UT DRY THYROID LIVER ADRENAL KID PIT BW GAIN Normal Ovary Histo Normal thyroid Histo 60 70 80 90 100 110 Con A BPA 400 BPA 600 Relationship between reduction in body weight due to restricted feeding versus ovary and uterine weights 80 85 90 95 100 Percent of control body weight 60 70 80 90 100 110 Percent OF CONTROL weight FR BPA400 BPA600 Ovarian weight 80 85 90 95 100 Percent of control body weight 60 70 80 90 100 110 Percent OF CON weight FR BPA400 BPA600 Uterine Weight Daily oral ( mg/ kg/ d) Octylphenol treatment induces pseudoprecocious puberty in the 21 day old female rat.

Data are expressed as the number of days that VO was accelerated ( Gray and Ostby, 1998). Also see Stoker et al.

and Goldman et al., 2001.

0 50 100 150 200 0 1 2 3 4 5 6 78 Days VO is accelerated Octylphenol " Antiestrogens"

and Inhibitors of Steroidogenesis Tamoxifen mixed agonist/ antagonist ICI and ZM  ER antagonists Antralex  GnRH antagonist Fadrazole  Aromatase inhibitor Fenarimol  fungicide that inhibits aromatase in vivo and in vitro Ketoconazole  like fenarimol, but less specific for P450 aromatase Results as expected, or better, except Fenarimol Effects of TAMOXIFEN 10 MG/ KG in the SD AND LE Pubertal Female rat from TI 2000 26.7 26.5 34.9 36.3 SD LE 20 25 30 35 40 CON TAM AGE VO 64.2 66.2 111 130 SD LE 50 60 70 80 90 100 110 120 130 140 CON TAM Weight at VO Effects of TAMOXIFEN 10 MG/ KG in the SD and LE Pubertal Female rat from TI 2000 36.8 44.2 64.7 81.7 SD LE 20 30 40 50 60 70 80 90 CON TAM OVARIAN WT 111 102 292 275 SD LE 50 100 150 200 250 300 CON TAM Uterine Weight Effect of "( anti) estrogens" on age at VO in the Female rat. Data from Ashby et al., 2002; Marty et al., 1999; and Kim et al., 2002. ZM 189,154 and ICI 182,780 are ER antagonists. Tamoxifen is a mixed ER agonist and antagonist. Antralex is a GnRH antagonist.

 14  12  10  8  6  4  2 0 2 4 6 8 FAD .6 FAD 1.2 FAD 6 ANTARELIX DES 5 TAM 50 TAM 200 Deviation in days from Control 80 0 10 0 0 6 12 18 24 30 36 42 48 54 60 66 72 78 84 90 96 Control ZM oral ICI oral ANTERELIX sc % VO day 33 Effects of FENARIMOL in the Pubertal Female SD rat assay RTI SD 2003 34.2 33.7 32.9 Age VO 30 31 32 33 34 35 36 Con A Fenarimol 50 Fenarimol 250 115 133 128 Wt at VO 100 110 120 130 140 Con A Fenarimol 50 Fenarimol 250 * Lack of effect of FENARIMOL on VO in the Female rat from RTI SD 2003 and EPA LE 1990 studies 34.2 33.7 32.9 26 28 30 32 34 36 Con A Fenarimol 50 Fenarimol 250 RTI SD 2003 29.3 31.6 30.4 26 28 30 32 34 36 Con Fenarimol 35 Fenarimol 70 EPA LE 1990 Effects of FENARIMOL in the Pubertal Female SD rat assay RTI SDS 2003 90 88 84 81 78 146 102 95 82 86 100 100 101 109 101 103 80 118 121 106 100 101 100 100 BWT OVWT UT WET UT DRY THYROID LIVER ADRENAL KID PIT BW GAIN Normal Ovary H Normal thyroid 70 80 90 100 110 Con A Fenarimol 50 Fenarimol 250 Relationship between reduction in body weight due to restricted feeding versus ovary and uterine weights 80 85 90 95 100 Percent of control body weight 60 70 80 90 100 110 120 Percent OF CONTROL weight FR FEN50 FEN250 Ovarian weight 80 85 90 95 100 Percent of control body weight 60 70 80 90 100 110 Percent OF CON weight FR FEN50 FEN250 Uterine Weight Effects of KETOCONAZOLE in the RTI SD 2003 Pubertal Female SD rat assay. OVARIAN HISTOPATH IS KEY FOR THIS CHEMICAL 32.3 33 33.3 Con A KETO 50 KETO 100 30 31 32 33 34 35 36 Age VO 123 127 123 CON B KETO 50 KETO 100 100 110 120 130 140 Wt at VO Effects of KETOCONAZOLE on age at VO in the Female rat RTI SD 2003, TI SD/ LE 2000 and MSM 1999 32.3 33 33.3 Con A KETO 50 KETO 100 30 31 32 33 34 35 36 RTI 2003 32.3 33 33.3 Con A KETO 50 KETO 100 TI 2000 33.5 36 39.7 Con A KETO 50 KETO 100 MSM 1999 * Effects of KETOCONAZOLE in the RTI SD 2003 and TI SD/ LE 2000 Pubertal Female SD rat assay.

Percent of females with ovarian histologic lesions 100 80 0 0 20 40 60 80 100 CON KETO 50 KETO 100 0 100 0 100 0 20 40 60 80 100 SD LE RTI SD 2003 TI SD/ LE 2000 Effects of KETOCONAZOLE in the RTI SD 2003 Pubertal Female SD rat assay 92 84 73 73 100 108 109 91 89 0 100 99 96 91 89.3 103 110 167 108 100 98 10 100 BWT OVWT UT WET UT DRY THYROID LIVER ADRENAL KID PIT BW GAIN Normal Ovary Histo Normal thyroid Hist 70 80 90 100 110 CON KETO 50 KETO 100 Relationship between reduction in body weight due to restricted feeding versus ovary and uterine weights 80 85 90 95 100 Percent of control body weight 65 70 75 80 85 90 95 100 105 Percent OF CONTROL weight FR KETO50 KETO100 80 85 90 95 100 Percent of control body weight 65 70 75 80 85 90 95 100 105 Percent OF CON weight FR KETO50 KETO100 Effects of CNS active chemicals on puberty in the female Atrazine  alters CNS dopamine,

prolactin and LH surges, also may affect aromatase and etc. Results reproducible Phenobarbital  alters CNS, pituitary and liver function, enhancing hormone metabolism and secretion.

Results robust and reproducible Effects of ATRAZINE in the Pubertal RTI SD 2003 Female rat age and weight at VO and age at first estrus. Note: EPA needs to provide more guidance on analysis of estrous cycle data 32.9 34.2 36.4 Con A ATR 75 ATR 150 30 31 32 33 34 35 36 37 38 39 Age VO 128 128 130 CON A ATR 75 ATR 150 100 110 120 130 140 Wt at VO * 35.1 35.8 38.1 Con A ATR 75 ATR 150 32 33 34 35 36 37 38 39 40 41 Age 1st P/ E ** Effects of ATRAZINE at 100 150 mg in the Pubertal Female rat assay. RTI ( SD) versus Laws ( Wistar)

and Ashby et al ( SD and AP strains).

32.9 34.2 36.4 Con A ATR 75 ATR 150 30 31 32 33 34 35 36 37 38 39 Age VO RTI * 32.5 35.9 37.1 32.8 34.3 36.3 Con  Laws ATR 50 ATR 100 30 31 32 33 34 35 36 37 38 39 Age VO EPA 1 2 *

* 38 41 39 42.2 Con ATR 100 35 36 37 38 39 40 41 42 43 44 45 SD AP *

* Effects of ATRAZINE in the RTI SD 2003 Pubertal Female SD rat assay 86 81 87 86 84 90 96 93 78 80 100 100 92 95 97 96 84 93 111 97 94 89 100 100 BWT OVWT UT WET UT DRY THYROID LIVER ADRENAL KID PIT BW GAIN Normal Ovary Histo Normal thyroid Hist 80 90 100 110 Con A ATR 75 ATR 150 Relationship between reduction in body weight due to restricted feeding versus ovary and uterine weight 80 85 90 95 100 Percent of control body weight 65 70 75 80 85 90 95 100 105 Percent OF CONTROL weight FR ATR75 ATR150 Ovarian Weight 80 85 90 95 100 Percent of control body weight 65 70 75 80 85 90 95 100 105 Percent OF CON weight FR ATR75 ATR150 Uterine Weight Effects of Phenobarbital on female rat puberty TI SD 2003 31.9 33.1 32.8 34.5 Oil PB25 PB50 PB100 29 30 31 32 33 34 35 36 37 AGE AT VO * ** 118 123 121 128 Oil PB25 PB50 PB100 115 120 125 130 135 Wt at VO * Effects of Phenobarbital on female rat puberty.

TI SD 2003 8.67 9.79 10.76 11.32 Oil PB25 PB50 PB100 8 9 10 11 12 13 LIVER WT 41.4 52 52.6 51.8 Oil PB25 PB50 PB100 30 35 40 45 50 55 60 ADRENAL WT **

** **

** ** ** Antithyroid effects on puberty in the female PTU and the corresponding growth retardation have surprisingly little affect on pubertal landmarks Effects of PTU in the RTI SD 2003 Pubertal Female SD rat assay 32.3 33.3 33.1 CON B PTU 2 PTU 25 30 31 32 33 34 35 36 Age VO 123 127 118 CON B PTU 2 PTU 25 100 110 120 130 140 Wt at VO Effects of PTU in the RTI SD 2003 Pubertal Female SD rat assay 72 75 104 99 433 60 58 69 86 60 100 97 88 105 104 268 93 87 95 97 96 100 BWT OVWT UT WET UT DRY THYROID LIVER ADRENAL KID PIT BW GAIN Normal Ovary Histo Normal thyroid Hist 10 100 1000 Con A PTU 2 PTU 25 OBSERVATIONS ON THE PUBERTAL FEMALE ASSAY Responds very well to estrogens Responds very well to GnRH antagonist and potent aromatase inhibitors Responds well to ketoconazole, but effects not the pubertal landmarks Reproducible responses to Atrazine Reproducible responses to Phenobarbital Negative for Fenarimol at doses tested.

Need a study at the MTD with doses from dose range finding, not guessing EDVMS MEETING DEC 10 12, 2003 PUBERTAL MALE AND FEMALE RAT ASSAYS Endocrine Disrupting Chemicals: Prepubertal Exposures and Effects on Sexual Maturation and Thyroid Function in the Male Rat. A Focus on the EDSTAC Recommendations. Tammy E. Stoker, Louise G. Parks, L. Earl Gray, and Ralph L. Cooper. Critical Reviews in Toxicology, 30( 2): 197 252 ( 2000)

Research Protocol for Assessment of Pubertal Development and Thyroid Function in Immature ( 23 to 53 Day Old) Male Rats Purpose and applicability The purpose of this protocol is to outline procedures to quantify the effects of environmental compounds on pubertal development and thyroid function in the intact juvenile/ peripubertal male rat. This assay detects compounds that display antithryoid, estrogenic, androgenic,

antiandrogenic [ androgen receptor ( AR) or steroid enzyme mediated] activity, or alters puberty via changes in follicle stimulating hormone ( FSH), luteinizing hormone ( LH),

prolactin, growth hormone ( GH) or hypothalamic function.

Required endpoints Growth ( body weight)

Age and weight at preputial separation Serum thyroxin ( T4), thyroid histology and TSH Seminal vesicle plus coagulating gland weight ( with and without fluid), Ventral prostate weight, Levator ani plus bulbocavernosus weight, Epididymal and testis weights and histology Optional measures Serum testosterone, estradiol, LH, prolactin and tri iodothyronine ( T3)

Liver, kidney, adrenal and pituitary weights and histology ex vivo testis and pituitary hormone production, Hypothalamic neurotransmitter levels Alternative Assays for Possible Inclusion in T1S the Pubertal Male assay was an Alternative assay that EDSTAC wanted developed and evaluated for screening If standardized and validated and found to be functionally equivalent, or better,

then an alternative assay could be included in T1S. 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 Assign to Treatments based upon body weight.

15/ group Dose Male Necropsy Our Immature ( 21  52 Days of Age) Intact Male Rat Protocol to Our Immature ( 21  52 Days of Age) Intact Male Rat Protocol to Evaluate Pubertal Development and Thyroid Function. Detects Evaluate Pubertal Development and Thyroid Function. Detects inhibition of steroidogenesis, antithyroid and ( anti) androgenic inhibition of steroidogenesis, antithyroid and ( anti) androgenic activities and altered HPG maturation.

activities and altered HPG maturation.

Wean Days of age Required endpoints:

Growth Age and weight at preputial separation ( PPS)

Serum thyroxine and thyroid stimulating hormones Thyroid Histology Seminal vesicle plus coagulating gland weight ( with fluid)

Ventral prostate weight, Levator ani/ bulbocavernosus muscle weight Testis and epididymal weights and histology Liver, kidney, adrenal and pituitary weights Optional endpoints Serum hormones and ex vivo testis and pituitary hormone production Dose daily Daily examination for PPS LEGJR 20 30 40 50 Age in days 0 1 2 3 4 ng/

ml Serum T Serum DHT Serum Androstanediol The serum androgen profile The serum androgen profile changes dramatically during changes dramatically during puberty puberty 20 30 40 50 Age in days 10 100 1000 10000 mg Epididymis Testes Ventral Prostate Seminal Vesicle Reproductive organ weights grow Reproductive organ weights grow rapidly during puberty rapidly during puberty 0 10 20 30 40 50 60 70 80 90 Time ( min)

0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 Serum LH 829 837 965 855 0 10 20 30 40 50 60 70 80 90 Time ( min)

0.0 0.5 1.0 1.5 2.0 Serum Testosterone 829 837 965 855 Basal Serum LH and Testosterone levels ( ng/ ml) in Control LE Adult Male rats measured repeatedly at ten minute intervals.

Large CVs require large effects of large sample sizes ( Fail et al., 1996) Modified USEPA Endocrine Disrupter Screening and Testing Battery with the pubertal male assay replacing the Hershberger assay In Vitro ER Binding and/ or Transcriptional Activation AR Binding and/ or Transcriptional Activation Steroid Hormone Synthesis In Vivo Uterotropic Assay in Ovariectomized Adult Rat ( 3d)

Pubertal Female Rat Assay including Thyroid ( 20d)

Pubertal Male Rat Assay including Thyroid ( 30d)

Frog Metamorphosis Assay Short term Assessment of the Fish Reproductive System EDSTAC Screening Battery:

Detects Estrogens by sc and oral routes and in vitro Detects Androgens and Anti in the pubertal male assay using simple endpoints ( PPS and organ weights)

and in vitro.

Detects HPG/ CNS alterations related to FSH, LH, prolactin,

dopamine, GH with simple, precise developmental landmark without RIAs in pubertal female and male.

Detects Inhibition of Steroidogenesis in vivo ( in male and female) with simple, precise endpoints and in vitro Detects Thyroid effects in intact pubertal Male and Female Rat using RIAs and Frog Metamorphosis Uses extensively utlized in vivo endpoints/ assays Chemicals investigated in the pubertal male Chemicals investigated in the pubertal male assay at RTI and TI this year assay at RTI and TI this year RTI  completed RTI  completed Atrazine 75 and 150 mg/ kg/ d Atrazine 75 and 150 mg/ kg/ d p, p' DDE 50 and 100 mg/ kg/ d p, p' DDE 50 and 100 mg/ kg/ d Vinclozolin 30 and 100 Vinclozolin 30 and 100 mg/ kg/ d mg/ kg/ d Methoxychlor 25 and 50 Methoxychlor 25 and 50 mg/ kg/ d mg/ kg/ d PTU 2 and 25 mg/ kg/ d PTU 2 and 25 mg/ kg/ d Ketoconazole 50 and 100 Ketoconazole 50 and 100 mg/ kg/ d mg/ kg/ d Linuron 50 and 100 mg/ kg/ d Linuron 50 and 100 mg/ kg/ d Phenobarbital 50 and 100 Phenobarbital 50 and 100 mg/ kg/ d mg/ kg/ d TherImmune 2000 ( 2x2x6 factorial.

TherImmune 2000 ( 2x2x6 factorial.

Blocks, LE versus SD and chemicals,

Blocks, LE versus SD and chemicals,

n= 6/ strain/ block/ chemical)

n= 6/ strain/ block/ chemical)

completed completed Flutamide 50 mg/ kg/ d Flutamide 50 mg/ kg/ d Methyl Testosterone 80 mg/ kg/ d Methyl Testosterone 80 mg/ kg/ d PTU 240 mg/ kg/ d PTU 240 mg/ kg/ d Ketoconazole 100 mg/ kg/ d Ketoconazole 100 mg/ kg/ d Pimozide 30 mg/ kg/ d Pimozide 30 mg/ kg/ d DBP 1000 mg/ kg/ d DBP 1000 mg/ kg/ d TherImmune 2003  completed TherImmune 2003  completed Phenobarbital 25, 50 and 100 Phenobarbital 25, 50 and 100 mg/ kg/ d mg/ kg/ d Vinclozolin 10, 30 and 100 mg/ kg/ d Vinclozolin 10, 30 and 100 mg/ kg/ d Flutamide 25 and 50 mg/ kg/ d Flutamide 25 and 50 mg/ kg/ d Published studies using the pubertal male assay p, p' DDE  several labs Vinclozolin  2 labs Linuron  PPS Cyproterone acetate Flutamide  3 labs Finasteride  2 labs Ketoconazole  2 labs Testolactone TP BBP DBP  2 labs Methoxychlor  2 labs BPA DES PTU Perchlorate  in prep PBDE  submitted Phenobarbital Fenitrothion MBC  PPS Atrazine  2 labs Relationship between reduction in body weight due to restricted feeding versus and delay in age at PPS 80 85 90 95 100 Percent of control body weight  10  5 0 5 10 Delay in PPS in Days Feeding Vinclozolin Linuron DDE1 DDE2 DBP DBP2 KETO TH MT TH FLUT TH FR SMEC AR antagonists Flutamide Vinclozolin p, p' DDE Linuron Results

all chemicals produced expected robust responses,

easily detected all of the above. Reproducible 41.1 53.3 54 Oil F25 F50 35 40 45 50 55 60 AGE AT PPS DAYS Effects of flutamide at 25 and 50 mg/ kg on puberty in the male rat TI SD 2003 ** ** 214 319 311 Oil F25 F50 150 200 250 300 350 400 WT AT PPS DAYS ** ** 698 175 135 Oil F25 F50 0 200 400 600 800 1000 SD TI 2003 SV WT Effects of flutamide at 25 and 50 mg/ kg In the male rat TI SD 2003 ( RED) and TI SD/ LE 2000 ** ** 2.71 3 3.5 Oil F25 F50 2.6 2.8 3 3.2 3.4 3.6 TESTES WT ** **

371 80 477 76 Oil F50 0 100 200 300 400 500 SD LE SV WT ** 2.67 3.12 3.09 3.52 Oil F50 2.6 2.8 3 3.2 3.4 3.6 SD LE TESTES WT ** Effects of Vinclozolin in male pubertal SD rat assay RTI 2003 41.4 43.8 46.8 0 30 100 40 40.5 41 41.5 42 42.5 43 43.5 44 44.5 45 45.5 46 46.5 47 47.5 48 Age at PPS 219 243 260 0 30 100 180 200 220 240 260 280 Weight at PPS **

**

**

** Effects of Vinclozolin on AGE AT PPS in male SD and LE rats 41.4 43.8 46.8 0 30 100 40 41 42 43 44 45 46 47 48 49 50 RTI SD 2003 **

**

**

** 40.7 44.4 47.8 0 25 100 RTI LE 1994 **

**

41.1 42.7 45.5 0 30 100 EPA LE 1996 ** ** 204.9 232 270 0 25 100 RTI LE 1997 Effects of Vinclozolin in Weight at PPS in male LE and SD rats 219 243 260 0 30 100 180 200 220 240 260 280 300 RTI SD 2003 **

** ** 204 208 232 0 30 100 EPA LE 1996 ** Effects of Vinclozolin in male RTI 2003 pubertal rat assay 96 55 77 77 82 107 102 84 97 103 94 108 BWT SV LABC PROSTATE EPIDIDYMIS TESTIS 50 60 70 80 90 100 110 120 30 100 % OF CONTROL WEIGHTS ** **

**

**

**

**

* Effects of Vinclozolin on male reproductive organ weights in the RTI SD 2003 study compared to EPA LE 1996 pubertal rat assay at 100 mg/ kg/ d 96 71 100 80 86 96 55 107 77 82 BWT SV TESTIS PROSTATE EPIDIDYMIS 50 60 70 80 90 100 110 120 RTI EPA % OF CONTROL WEIGHTS ** **

**

**

**

** The intact male assay is much less sensitive to the effects of vinclozolin on ASO than are seminal vesicle weights in our Hershberger or pubertal male assays.

0 50 100 150 200 250 300 30 40 50 60 70 80 90 100 Percent of Control Intact Male Hershberger Pubertal 2 labs The Adult Intact Male assay is much less sensitive to the Effects of vinclozolin on ventral prostate weight. Males treated with TP in the Hershberger Assay.

0 50 100 150 200 250 300 0 20 40 60 80 100 %

Control Intact Male Hershberger Pubertal 2 Labs It is critical that the T1 Screening assay be able to detect we ak antiandrogens like DDE and linuron because they cause reproductive tract malformation in utero. Exposure to 100 mg p, p' DDE/ kg dam's body weight on gestational days 14 18 alters development of the External Genitalia. Gray et al, 1999.

Control SD Male Rat Treated SD Male with Cleft Phallus, Exposed Os, Hypospadias and Slight Vaginal Pouch Effects of p, p' DDT and p, p' DDE treatment during sexual Effects of p, p' DDT and p, p' DDE treatment during sexual differentiation in Dutch Belted rabbits. ( Veeramachaneni et differentiation in Dutch Belted rabbits. ( Veeramachaneni et al., 1996; 2000; Palmer et al., 2000)

al., 1996; 2000; Palmer et al., 2000)

Does exposed weekly to p, p' DDT at 25 mg/ kg Does exposed weekly to p, p' DDT at 25 mg/ kg from GD 1 through 6 weeks postpartum,

from GD 1 through 6 weeks postpartum,

followed by 10 mg/ kg directly to male up to followed by 10 mg/ kg directly to male up to 12 weeks of age.

12 weeks of age.

Effects Effects Cryptorchidism Cryptorchidism Atypical germ cells in undescended testes inclduing Atypical germ cells in undescended testes inclduing Carcinoma in situ like cells Carcinoma in situ like cells At 8 and 12 weeks At 8 and 12 weeks serum p, p' DDT at 231 and 38 ppb serum p, p' DDT at 231 and 38 ppb serum p, p' DDE at 187 and 37 ppb serum p, p' DDE at 187 and 37 ppb Effects of p, p' DDE in male pubertal rat assay RTI SD 2003 41.4 44.9 45.7 0 50 100 40 40.5 41 41.5 42 42.5 43 43.5 44 44.5 45 45.5 46 46.5 47 47.5 48 Age at PPS 219 251 259 0 50 100 180 200 220 240 260 280 Weight at PPS ** ** **

** Effects of p, p' DDE in male pubertal rat assay 41.4 44.9 45.7 0 50 100 40 41 42 43 44 45 46 47 48 49 50 RTI SD 2003 ** ** **

** 43.3 43.5 48 0 30 100 40 41 42 43 44 45 46 47 48 49 50 EPA LE 1994 ** Effects of p, p' DDE in male pubertal rat assay RTI SD 2003 98 88 88 97 90 101 100 102 96 109 96 102 BWT SV LABC PROSTATE EPIDIDYMIS TESTIS 80 90 100 110 50 100 % OF CONTROL WEIGHTS * ** 100 97 101 105 100 100 70 37 34 100 102 88 0 50 100 150 200 250 300 350 30 40 50 60 70 80 90 100 110 Intact Male Assay Hershberger Pubertal M % control The Adult intact male assay is insensitive to the Effects of p, p' DDE at 100 mg/ kg/ d on SV versus ASO weight. Males treated with TP in the Hershberger Assay. 100 88 115 97 99 0 50 100 150 200 250 300 350 30 40 50 60 70 80 90 100 110 120 IM H Pubertal % Control The intact male assay is insensitive to the Effects of p, p' DDE at 100 mg/ kg/ d on ventral prostate weight.

Males treated with TP in the Hershberger Assay. Effects of gestational day 14 18 treatment with linuron ( 75 mg/ kg/ d) and/ or benzyl butyl phthalate ( BBP) on fetal male rat testicular testosterone production in 3 h, fetal testis and whole body testosterone levels, and nipple/ areola induction. All values differ significantly from control ( Hotchkiss in prep)

CON LIN BBP LIN+ BBP 0 2 4 6 8 10 Numbers of areolas/ nipples CON LIN BBP LIN+ BBP 0 1 2 3 4 5 TESTICULAR T PRODUCTION ( ng/ testis/ 3 CON LIN BBP LIN+ BBP 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 TESTICULAR T ( ng/ testis)

CON LIN BBP LIN+ BBP 0 0.05 0.1 0.15 0.2 0.25 WHOLE BODY T ( ng/ fetus)

Low dose Effects of Linuron on Sexual Differentiation of the male rat ( McIntyre et al, 2000, Toxicol Appl Pharm 167, 87 99). Dams were dosed with linuron at 12.5, 25 or 50 mg/ kg/ d from GD 12 to 21. No direct exposure to pups. This pesticide is negative in published teratology and multigenerational tests. Could the current tests detect these malformations in only 20 F1 males/ group?

0 0 3.6 1.8 11.6 11.6 11.4 4.6 0 12.5 25 50 Dose of Linuron mg/ kg/ d 0 2 4 6 8 10 12 14 Hypoplastic Testis Hypoplastic Epididymal Percent of Male Offspring Malformed Effects of Linuron in male pubertal rat assay RTI SD 2003 39.6 43.6 45.5 0 50 100 38 38.5 39 39.5 40 40.5 41 41.5 42 42.5 43 43.5 44 44.5 45 45.5 46 46.5 47 Age at PPS 207 227 224 0 50 100 190 200 210 220 230 Weight at PPS **

**

** * Effects of Linuron on PPS in the EPA LE 1987 and RTI SD 2003 male rats 39.6 43.6 0 50 38 39 40 41 42 43 44 45 46 47 RTI SD 2003 43 45.5 0 40 38 39 40 41 42 43 44 45 46 47 EPA LE 1987 ** * Effects of Linuron in male pubertal rat assay RTI SD 2003 84 56 71 70 84 93 93 73 85 74 92 97 BWT SV LABC PROSTATE EPIDIDYMIS TESTIS 50 60 70 80 90 100 50 100 % OF CONTROL WEIGHTS * * **

** ** ** ** ** ** 100 94 104 93 81 0 50 100 150 60 70 80 90 100 110 Intact Male Hershberger Pubertal M % Control The Adult intact male assay is much less sensitive to the Effects of Linuron at 100 mg/ kg/ d on ventral prostate weight. Males treated with TP in the Hershberger Assay. 100 105 104 96 89 0 50 100 150 200 50 60 70 80 90 100 110 Intact Male Hershberger Pubertal % Control The Adult intact male assay is much less sensitive to the Effects of Linuron at 100 mg/ kg/ d on SV versus ASO weight. Males treated with TP in the Hershberger Assay.

MTD The Phthalate Esters Inhibitors of Fetal Male Rat Testicular Hormone Synthesis. Not an AR antagonist Negative in Developmental Toxicology/ Teratology Studies Negative in some Multigenerational Studies Results in Transgenerational Studies DEHP ++

BBP ++

DBP ++

DINP +

DEP negative DMP negative DOTP negative Effects of DBP in male rat TI 2000 SD/ LE 43 44.2 0 1000 40 41 42 43 44 45 46 47 48 49 50 SD AGE at PPS ** **

47.5 51.8 0 1000 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 LE AGE at PPS ** ** Effects of DBP in male rat EPA LE 1985, 1986 42.4 48.4 0 1000 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 LE EPA 1985 AGE at PPS ** ** **

39.5 42.6 43.4 44.4 0 250 500 1000 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 LE EPA 1986 AGE at PPS * ** ** 3.09 2.44 OIL DBP 2.00 2.50 3.00 Testes weights 477 382 OIL DBP 350 400 450 500 Seminal Vesicle weights 623 514 OIL DBP 500 550 600 650 LABC weights 5.9 21.1 20 17.5 BWT TESTES SV LABC 0 5 10 15 20 25 % REDUCTION INDUCED BY DBP DBP reduces reproductive tissue weights in the pubertal male assay using SD rats, the least affected strain. TI 2000 0.62 1.01 1.76 1.33 Testis SV 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 0 1000 LE EPA 1985 Nec at 82 days Effects of 1000 mg/ kg DBP on testis weight in the LE and SD male rat from studies TI SD/ LE 2002, EPA LE 1985 and EPA SD 1981 0.45 0.35 1.44 0.75 Testis SV SD EPA 1981 Nec at 52 days **

** * 0.37 0.174 1.34 0.37 Testis SV LE TI 2000 Nec at 53 54 days ** 1.22 0.381 1.55 0.477 Testis SV SD TI 2000 Nec at 53 54 days *

**

**

**

Strain* Treatment Interaction p< 0.01 SD less affected SD less affected SD not less affected 100 101 102 101 105 100 86 65 48 0 200 400 600 800 1000 40 50 60 70 80 90 100 Percent of Control Intact Male Assay Hershberger Pubertal Male The intact male assay is less sensitive to the effects of the phthalate ester DBP at the limit dose of 1 g/ kg/ d on ASO weight than is seminal vesicle weights in our Hershberger or pubertal male assays. EFFECTS OF AN ESTROGENIC AND POTENTIALLY ANTIANDROGENIC PESTICIDE NO EFFECT ON PPS, AS EXPECTED FROM EARLIER WORK REDUCED ANDROGEN DEPENDENT TISSUE WEIGHTS POSITIVE IN THE PUBERTAL MALE Effects of Methoxychlor in male pubertal rat assay RTI SD 2003 41.4 41.8 41.8 0 25 50 40 40.5 41 41.5 42 42.5 43 43.5 44 44.5 45 Age at PPS 219 219 220 0 25 50 170 180 190 200 210 220 230 240 Weight at PPS Effects of Methoxychlor on PPS in the male rat from EPA LE 1989 study verus RTI SD 2003 41.4 41.8 41.8 0 25 50 40 40.5 41 41.5 42 42.5 43 43.5 44 44.5 45 RTI SD 2003 42.4 40.6 43.4 0 25 50 40 40.5 41 41.5 42 42.5 43 43.5 44 44.5 45 EPA LE 1989 NO EFFECT ON PPS AT THESE DOSES IN EITHER STUDY Effects of Methoxychlor in male pubertal rat assay 96 73 86 95 96 100 96 91 97 94 101 101 BWT SV LABC PROSTATE EPIDIDYMIS TESTIS 70 80 90 100 25 50 % OF CONTROL WEIGHTS ** * ANDROGENIC EFFECTS IN THE PUBERTAL MALE ASSAY VERY POSITIVE RESPONSES EXACTLY AS EXPECTED ON PPS, TESTIS AND SEX ACCESSORY TISSUES 47.5 32.9 43 36.8 CON MT 25 30 35 40 45 50 SD LE AGE AT PPS DAYS Effects of METHYLTESTOSTERONE at 80 mg/ kg on puberty in the male rat TI SD/ LE 2000 ** 185 144 238 119 CON MT 100 150 200 250 WT AT PPS ** **

** 477 710 371 729 CON MT 200 300 400 500 600 700 800 900 1000 SD LE SV WT Effects of METHYLTESTOSTERONE at 80 mg/ kg In the male rat TI SD/ LE 2000 ** 2.67 0.6 3.09 0.89 CON MT 0 1 2 3 TESTES WT **

221 335 182 374 CON MT 0 100 200 300 400 500 VP WT ** AN INHIBITOR OF P450 ENZYMES, INCLUDING STEROIDOGENESIS VERY POSITIVE REPRODUCIBLE REPRODUCTIVE AND OTHER ENDOCRINE ( ADRENAL) AND ORGAN ( LIVER)

EFFECTS Effects of Ketoconazole in male pubertal rat assay RTI SD 2003 39.6 42.3 44.1 0 50 100 38 38.5 39 39.5 40 40.5 41 41.5 42 42.5 43 43.5 44 44.5 45 Age at PPS 207 227 235 0 50 100 190 200 210 220 230 240 250 260 Weight at PPS **

**

**

** Effects of Ketoconazole ON THE AGE AT PPS 39.6 42.3 44.1 0 50 100 38 38.5 39 39.5 40 40.5 41 41.5 42 42.5 43 43.5 44 44.5 45 RTI SD 2003 **

** 47.5 48.9 43 46.3 0 100 38 39 40 41 42 43 44 45 46 47 48 49 50 SD LE TI SD/ LE 2000 ** Effects of Ketoconazole on reproductive organs in TI SD/ LE 2000 and RTI SD 2003. Also, consistent increases in adrenal and liver weights at both doses.

**

** 638 546 567 448 623 471 0 100 400 450 500 550 600 650 700 **

639 423 367 223 477 261 0 100 150 200 250 300 350 400 450 500 550 600 650 700 SD TI LE TI SD RTI 266 208 181 123 221 137 0 100 100 150 200 250 300 **

** VP SV LABC ** * Relationship between reduction in body weight due to restricted feeding versus ventral prostate weight 80 85 90 95 100 Percent of control body weight 60 70 80 90 100 110 120 Percent OF CONTROLventral prostate weigh Feeding Vinclozolin KETO TH FR SMEC R square = 0.819 # pts = 5 y =  36.6 + 1.42x Effects of Atrazine in male pubertal rat assay 41.4 42 42.9 0 75 150 40 40.5 41 41.5 42 42.5 43 43.5 44 Age at PPS 219 209 200 0 75 150 170 180 190 200 210 220 230 240 Weight at PPS * ** ATRAZINE A CNS ACTIVE CHEMICAL DELAYS PPS IN ALL STUDIES VARIABLE EFFECTS ON BODY WEIGHT Effects of ATRAZINE in the Pubertal Male rat assay RTI versus Stoker al.

41.4 42 42.9 Con A ATR 75 ATR 150 40 41 42 43 44 45 Age PPS RTI 42.1 43.8 43.6 43.4 Con  Stoker ATR 50 ATR 100 ATR 150 40 41 42 43 44 45 46 Age PPS EPA *

*

* *

1.5 d 0.6 d 1.7 d 1.3 d 1.5 d Effects of Atrazine in male pubertal rat assay. RTI versus Stoker data at 150 mg/ kg/ d 81 76 79 81 87 103 90 84 75 94 BWT SV LABC PROSTATE EPIDIDYMIS TESTIS 70 80 90 100 Stoker RTI % OF CONTROL WEIGHTS PHENOBARBITAL AFFECTS CNS, LIVER METABOLISM AND ENDOCRINE ORGANS DIRECTLY ROBUST, REPRODUCIBLE RESPONSES ON MALE PPS AND REPRODUCTIVE ORGAN WEIGHTS Effects of Phenobarbital in male pubertal rat assay 39.6 41.3 43 0 50 100 38 38.5 39 39.5 40 40.5 41 41.5 42 42.5 43 43.5 44 44.5 45 Age at PPS 207 220 224 0 50 100 190 200 210 220 230 Weight at PPS ** ** *

* Effects of Phenobarbital in male pubertal rat assay 39.6 41.3 43 0 50 100 38 38.5 39 39.5 40 40.5 41 41.5 42 42.5 43 43.5 44 44.5 45 RTI SD 2003 ** ** 41.1 41.1 43.6 Oil PB50 PB100 38 39 40 41 42 43 44 45 TI SD 2003 ** Effects of Phenobarbital in male pubertal rat assay 92 76 82 88 88 92 119 97 100 95 97 99 97 117 BWT SV LABC PROSTATE EPIDIDYMIS TESTIS Thyroid 70 80 90 100 110 120 130 50 100 % OF CONTROL WEIGHTS * ** ** ** * * OBSERVATIONS ON THE PUBERTAL MALE ASSAY Produced expected results for all chemicals Both SD and LE strains responded significantly Appears to detect all the activities as expected More sensitive than the Adult Intact Male Assay More sensitive than the Hershberger to DBP but not to the AR antagonists PRIORITIZATION FOR SCREENING BASED UPON QSAR, HPTS, VOLUME, ETC TIER 1 SCREENING About 100 rats per chemical ( about 10,000 Rats/ 100 chemicals)

HOLD 95% Negative No Testing 90% () 10%(+) True + ( 5%)

False + ( 5%)

( Type II Error)

( 10 chemicals per 100 screened)

Replicate Tier 1 Repeat False positives 5%

( about 50 rats per repeat or 500 rats per 100 chemicals screened) Positives 5% Tier 2 Testing ( about 1,500 rats per chemical,

or 7,500 rats per 100 screened)

Estimate of Animal Use in EDSP ( Assuming 5% of Chemicals are endocrine active)

18,000 Rats per Hundred Chemicals Evaluated Gray et al., in press. Toxicology Letters. ICT IX LEGJR


[Federal Register Volume 79, Number 169 (Tuesday, September 2, 2014)]
[Notices]
[Pages 52097-52098]
[FR Doc No: 2014-20889]

-----------------------------------------------------------------------

SECURITIES AND EXCHANGE COMMISSION

[File No. 500-1]

In the Matter of QSGI Inc.; Order of Suspension of Trading

August 28, 2014.
    It appears to the Securities and Exchange Commission
(``Commission'') that there is a lack of current and accurate
information concerning the

securities of QSGI Inc. (``QSGI'') because QSGI has not filed any
periodic reports for any reporting period subsequent to June 30, 2011.
QSGI is a Florida corporation with its principal place of business
listed as West Palm Beach, Florida. Its stock is quoted on OTC Link,
operated by OTC Markets Group Inc., under the ticker: QSGI. The
Commission is of the opinion that the public interest and the
protection of investors require a suspension of trading in the
securities of QSGI.
    Therefore, it is ordered, pursuant to Section 12(k) of the
Securities Exchange Act of 1934, that trading in the securities of the
above-listed company is suspended for the period from 9:30 a.m. EDT,
August 28, 2014, through 11:59 p.m. EDT, on September 11, 2014.

    By the Commission.
Jill M. Peterson,
Assistant Secretary.
[FR Doc. 2014-20889 Filed 8-28-14; 4:15 pm]
BILLING CODE 8011-01-P


[Federal Register Volume 81, Number 91 (Wednesday, May 11, 2016)]
[Proposed Rules]
[Pages 29202-29206]
[FR Doc No: 2016-11095]

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DEPARTMENT OF TRANSPORTATION

Federal Aviation Administration

14 CFR Part 39

[Docket No. FAA-2016-6430; Directorate Identifier 2015-NM-176-AD;]
RIN 2120-AA64

Airworthiness Directives; The Boeing Company Airplanes

AGENCY: Federal Aviation Administration (FAA), DOT.

ACTION: Notice of proposed rulemaking (NPRM).

-----------------------------------------------------------------------

SUMMARY: We propose to supersede Airworthiness Directive (AD) 2005-13-
30, which applies to all Boeing Model 737-100, -200, and -200C series
airplanes. AD 2005-13-30 currently requires repetitive inspections to
detect discrepancies of certain fuselage skin panels located just aft
of the wheel well, and repair if necessary. Since we issued AD 2005-13-
30, an evaluation by the design approval holder (DAH) indicates that
the fuselage skin is subject to widespread fatigue damage (WFD), and we
have received reports of cracks at the chem-milled steps in the
fuselage skin. This proposed AD would add new fuselage skin inspections
for cracking, inspections to detect missing or loose fasteners and any
disbonding or cracking of bonded doublers, permanent repairs of time-
limited repairs, related investigative and corrective actions if
necessary, and skin panel replacement. We are proposing this AD to
detect and correct fatigue cracking of the fuselage skin panels, which
could cause rapid decompression of the airplane.

DATES: We must receive comments on this proposed AD by June 27, 2016.

ADDRESSES: You may send comments, using the procedures found in 14 CFR
11.43 and 11.45, by any of the following methods:
     Federal eRulemaking Portal: Go to http://www.regulations.gov. Follow the instructions for submitting comments.
     Fax:<PHONE_NUMBER>.
     Mail: U.S. Department of Transportation, Docket
Operations, M-30, West Building Ground Floor, Room W12-140, 1200 New
Jersey Avenue SE., Washington, DC 20590.
     Hand Delivery: Deliver to Mail address above between 9
a.m. and 5 p.m., Monday through Friday, except Federal holidays.
    For service information identified in this NPRM, contact Boeing
Commercial Airplanes, Attention: Data & Services Management, P.O. Box
3707, MC 2H-65, Seattle, WA 98124-2207; telephone<PHONE_NUMBER>,
extension 1; fax<PHONE_NUMBER>; Internet https://www.myboeingfleet.com.
You may view this referenced service information at the FAA, Transport
Airplane Directorate, 1601 Lind Avenue SW., Renton, WA. For information
on the availability of this material at the FAA, call<PHONE_NUMBER>. It
is also available on the Internet at http://www.regulations.gov by
searching for and locating Docket No. FAA-2016-6430.

Examining the AD Docket

    You may examine the AD docket on the Internet at http://www.regulations.gov by searching for and locating Docket No. FAA-2016-
6430; or in person at the Docket Management Facility between 9 a.m. and
5 p.m., Monday through Friday, except Federal holidays. The AD docket
contains this proposed AD, the regulatory evaluation, any comments
received, and other information. The street address for the Docket
Office (phone:<PHONE_NUMBER>) is in the ADDRESSES section. Comments will
be available in the AD docket shortly after receipt.

FOR FURTHER INFORMATION CONTACT: Wade Sullivan, Aerospace Engineer,
Airframe Branch, ANM-120S, FAA, Seattle Aircraft Certification Office
(ACO), 1601 Lind Avenue SW., Renton, WA 98057-3356; phone: 425-917-
6430; fax:<PHONE_NUMBER>; email<EMAIL_ADDRESS>
SUPPLEMENTARY INFORMATION:

Comments Invited

    We invite you to send any written relevant data, views, or
arguments about this proposed AD. Send your comments to an address
listed under the ADDRESSES section. Include ``Docket No. FAA-2016-6430;
Directorate Identifier 2015-NM-176-AD'' at the beginning of your
comments. We specifically invite comments on the overall regulatory,
economic, environmental, and energy aspects of this proposed AD. We
will consider all comments received by the closing date and may amend
this proposed AD because of those comments.
    We will post all comments we receive, without change, to http://

www.regulations.gov, including any personal information you provide. We
will also post a report summarizing each substantive verbal contact we
receive about this proposed AD.

Discussion

    Fatigue damage can occur locally, in small areas or structural
design details, or globally, in widespread areas. Multiple-site damage
is widespread damage that occurs in a large structural element such as
a single rivet line of a lap splice joining two large skin panels.
Widespread damage can also occur in multiple elements such as adjacent
frames or stringers. Multiple-site damage and multiple-element damage
cracks are typically too small initially to be reliably detected with
normal inspection methods. Without intervention, these cracks will
grow, and eventually compromise the structural integrity of the
airplane. This condition is known as widespread fatigue damage. It is
associated with general degradation of large areas of structure with
similar structural details and stress levels. As an airplane ages, WFD
will likely occur, and will certainly occur if the airplane is operated
long enough without any intervention.
    The FAA's WFD final rule (75 FR 69746, November 15, 2010) became
effective on January 14, 2011. The WFD rule requires certain actions to
prevent structural failure due to WFD throughout the operational life
of certain existing transport category airplanes and all of these
airplanes that will be certificated in the future. For existing and
future airplanes subject to the WFD rule, the rule requires that DAHs
establish a limit of validity (LOV) of the engineering data that
support the structural maintenance program. Operators affected by the
WFD rule may not fly an airplane beyond its LOV, unless an extended LOV
is approved.
    The WFD rule (75 FR 69746, November 15, 2010) does not require
identifying and developing maintenance actions if the DAHs can show
that such actions are not necessary to prevent WFD before the airplane
reaches the LOV. Many LOVs, however, do depend on accomplishment of
future maintenance actions. As stated in the WFD rule, any maintenance
actions necessary to reach the LOV will be mandated by airworthiness
directives through separate rulemaking actions.
    In the context of WFD, this action is necessary to enable DAHs to
propose LOVs that allow operators the longest operational lives for
their airplanes, and still ensure that WFD will not occur. This
approach allows for an implementation strategy that provides
flexibility to DAHs in determining the timing of service information
development (with FAA approval), while providing operators with
certainty regarding the LOV applicable to their airplanes.
    On June 24, 2005, we issued AD 2005-13-30, Amendment 39-14167 (70
FR 36829, June 27, 2005) (``AD 2005-13-30''), for all Boeing Model 737-
100, -200, and -200C series airplanes. AD 2005-13-30 requires
repetitive inspections to detect discrepancies of certain fuselage skin
panels located just aft of the wheel well, and repair if necessary. AD
2005-13-30 resulted from reports of fatigue cracking of the skins and
doublers located aft of the wing, between body station (BS) 727 and BS
1016, and between body stringers S-14 and S-25 on numerous Boeing Model
737-100, -200, and -200C series airplanes. On some airplanes,
reinforcing angles had been installed on the skin doublers; however,
cracking was detected on both modified and unmodified airplanes. The
cracking has been attributed to fatigue from a combination of shear
stresses due to repeated wrinkling of the skin, and the skin chem-
milled pockets configuration. We issued AD 2005-13-30 to detect and
correct fatigue cracking of the fuselage skin panels, which could cause
rapid decompression of the airplane.

Actions Since AD 2005-13-30 Was Issued

    Since we issued AD 2005-13-30, an evaluation by the DAH indicates
that the fuselage skin is subject to WFD, and we have received reports
of cracks at the chem-milled steps in the fuselage skin.

Related Service Information Under 1 CFR Part 51

    We reviewed Boeing Special Attention Service Bulletin 737-53-1065,
Revision 3, dated June 30, 2015. The service information describes
procedures for inspection and repair of the fuselage skin panels
between BS 727 and BS 1016, and between stringers S-14 and S-25; and
also describes procedures for skin panel replacement. This service
information is reasonably available because the interested parties have
access to it through their normal course of business or by the means
identified in the ADDRESSES section.

FAA's Determination

    We are proposing this AD because we evaluated all the relevant
information and determined the unsafe condition described previously is
likely to exist or develop in other products of the same type design.

Proposed AD Requirements

    Although this proposed AD does not explicitly restate the
requirements of AD 2005-13-30, this proposed AD would retain certain
requirements of AD 2005-13-30. Those requirements are referenced in the
service information identified previously, which, in turn, is
referenced in paragraph (h) of this proposed AD.
    This proposed AD would also require accomplishing the actions
specified in the service information described previously, except as
discussed under ``Differences Between this Proposed AD and the Service
Information.'' For airplanes having line numbers 1 through 291, this
proposed AD would require actions done in accordance with a method
approved by the Manager, Seattle ACO, FAA.
    The phrase ``related investigative actions'' is used in this
proposed AD. ``Related investigative actions'' are follow-on actions
that (1) are related to the primary actions, and (2) further
investigate the nature of any condition found. Related investigative
actions in an AD could include, for example, inspections.
    The phrase ``corrective actions'' is used in this proposed AD.
``Corrective actions'' are actions that correct or address any
condition found. Corrective actions in an AD could include, for
example, repairs.

Differences Between This Proposed AD and the Service Information

    Boeing Special Attention Service Bulletin 737-53-1065, Revision 3,
dated June 30, 2015, specifies to contact the manufacturer for
instructions on how to repair certain conditions and also to obtain
certain work instructions, but this proposed AD would require repairing
those conditions and also to obtain those work instructions in one of
the following ways:
     In accordance with a method that we approve; or
     Using data that meet the certification basis of the
airplane, and that have been approved by the Boeing Commercial
Airplanes Organization Designation Authorization (ODA) whom we have
authorized to make those findings.

Explanation of Compliance Time

    The compliance time for the replacement specified in this proposed
AD for addressing WFD was established to ensure that discrepant
structure is replaced before WFD develops in airplanes. Standard
inspection techniques cannot be relied on to detect WFD before it
becomes a hazard to

flight. We will not grant any extensions of the compliance time to
complete any AD-mandated service bulletin related to WFD without
extensive new data that would substantiate and clearly warrant such an
extension.

Costs of Compliance

    We estimate that this proposed AD affects 9 airplanes of U.S.
registry.
    We estimate the following costs to comply with this proposed AD:

                                                 Estimated Costs
----------------------------------------------------------------------------------------------------------------
                                                                                Cost on U.S.
             Action                   Labor cost          Parts cost       Cost per product        operators
----------------------------------------------------------------------------------------------------------------
Inspection [actions retained      Up to 88 work-      $0................  Up to $7,480 per    Up to $67,320 per
 from AD 2005-13-30].              hours x $85 per                         inspection cycle.   inspection cycle.
                                   hour = $7,480 per
                                   inspection cycle.
Inspection [new proposed action]  Up to 1,914 work-   $0................  Up to $162,690 per  Up to $1,464,210
                                   hours x $85 per                         inspection cycle.   per inspection
                                   hour = $162,690                                             cycle.
                                   per inspection
                                   cycle.
Skin panel replacement [new       688 work-hours x    $96,000...........  $154,480..........  $1,390,320.
 proposed action].                 $85 per hour =
                                   $58,480.
----------------------------------------------------------------------------------------------------------------

    We estimate the following costs to do any necessary repairs that
would be required based on the results of the proposed inspection. We
have no way of determining the number of aircraft that might need these
repairs:

                                               On-Condition Costs
----------------------------------------------------------------------------------------------------------------
                                                                                Cost per
                    Action                                 Labor cost               Parts cost        product
----------------------------------------------------------------------------------------------------------------
Time-limited repair...........................  24 work-hours x $85 per hour =               (1)      \1\ $2,040
                                                 $2,040.
Permanent repair..............................  43 work-hours x $85 per hour =               (1)      \1\ $3,655
                                                 $3,655.
Permanent repair inspection...................  7 work-hours x $85 per hour =                (1)        \1\ $595
                                                 $595.
----------------------------------------------------------------------------------------------------------------
\1\ We have received no definitive data that would enable us to provide parts cost estimates for the on-
  condition actions specified in this proposed AD.

    According to the manufacturer, some of the costs of this proposed
AD may be covered under warranty, thereby reducing the cost impact on
affected individuals. We do not control warranty coverage for affected
individuals. As a result, we have included all available costs in our
cost estimate.

Authority for This Rulemaking

    Title 49 of the United States Code specifies the FAA's authority to
issue rules on aviation safety. Subtitle I, Section 106, describes the
authority of the FAA Administrator. Subtitle VII, Aviation Programs,
describes in more detail the scope of the Agency's authority.
    We are issuing this rulemaking under the authority described in
Subtitle VII, Part A, Subpart III, Section 44701, ``General
requirements.'' Under that section, Congress charges the FAA with
promoting safe flight of civil aircraft in air commerce by prescribing
regulations for practices, methods, and procedures the Administrator
finds necessary for safety in air commerce. This regulation is within
the scope of that authority because it addresses an unsafe condition
that is likely to exist or develop on products identified in this
rulemaking action.

Regulatory Findings

    We have determined that this proposed AD would not have federalism
implications under Executive Order 13132. This proposed AD would not
have a substantial direct effect on the States, on the relationship
between the national Government and the States, or on the distribution
of power and responsibilities among the various levels of government.
    For the reasons discussed above, I certify that the proposed
regulation:
    (1) Is not a ``significant regulatory action'' under Executive
Order 12866,
    (2) Is not a ``significant rule'' under the DOT Regulatory Policies
and Procedures (44 FR 11034, February 26, 1979),
    (3) Will not affect intrastate aviation in Alaska, and
    (4) Will not have a significant economic impact, positive or
negative, on a substantial number of small entities under the criteria
of the Regulatory Flexibility Act.

List of Subjects in 14 CFR Part 39

    Air transportation, Aircraft, Aviation safety, Incorporation by
reference, Safety.

The Proposed Amendment

    Accordingly, under the authority delegated to me by the
Administrator, the FAA proposes to amend 14 CFR part 39 as follows:

PART 39--AIRWORTHINESS DIRECTIVES

0
1. The authority citation for part 39 continues to read as follows:

    Authority: 49 U.S.C. 106(g), 40113, 44701.

Sec.  39.13  [Amended]

0
2. The FAA amends Sec.  39.13 by removing Airworthiness Directive (AD)
2005-13-30, Amendment 39-14167 (70 FR 36829, June 27, 2005), and adding
the following new AD:

The Boeing Company: Docket No. FAA-2016-6430; Directorate Identifier
2015-NM-176-AD.

 (a) Comments Due Date

    The FAA must receive comments on this AD action by June 27,
2016.

 (b) Affected ADs

    This AD replaces AD 2005-13-30, Amendment 39-14167 (70 FR 36829,
June 27, 2005) (``AD 2005-13-30'').

 (c) Applicability

    This AD applies to all The Boeing Company Model 737-100, -200,
and -200C series airplanes, certificated in any category.

 (d) Subject

    Air Transport Association (ATA) of America Code 53, Fuselage.

 (e) Unsafe Condition

    This AD was prompted by an evaluation by the design approval
holder (DAH) indicating that the fuselage skin is subject to
widespread fatigue damage (WFD), and reports of cracks at the chem-
milled steps in the fuselage skin. We are issuing this AD to detect
and correct fatigue cracking of the fuselage skin panels, which
could cause rapid decompression of the airplane.

 (f) Compliance

    Comply with this AD within the compliance times specified,
unless already done.

 (g) Actions for Group 1 Airplanes

    For Group 1 airplanes identified in Boeing Special Attention
Service Bulletin 737-53-1065, Revision 3, dated June 30, 2015:
Within 120 days after the effective date of this AD, accomplish
actions to correct the unsafe condition (e.g., inspections, repairs,
modifications, and related investigative and corrective actions)
using a method approved in accordance with the procedures specified
in paragraph (o) of this AD.

 (h) Inspections, Related Investigative and Corrective Actions

    Except for Group 1 airplanes identified in Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015: At the applicable times specified in tables 1 and 2 of
paragraph 1.E., ``Compliance,'' of Boeing Special Attention Service
Bulletin 737-53-1065, Revision 3, dated June 30, 2015, except as
required by paragraphs (i)(1) and (i)(2) of this AD: Do the
applicable inspections to detect cracks in the fuselage skin panels;
and do all applicable related investigative and corrective actions;
in accordance with the Accomplishment Instructions of Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015, except as required by paragraphs (i)(3) and (i)(4) of this AD.
Do all applicable related investigative and corrective actions
before further flight. Repeat the applicable inspections thereafter
at the applicable intervals specified Boeing Special Attention
Service Bulletin 737-53-1065, Revision 3, dated June 30, 2015.
Accomplishment of a repair in accordance with ``Part 3: Repair'' of
the Accomplishment Instructions of Boeing Special Attention Service
Bulletin 737-53-1065, Revision 3, dated June 30, 2015, except as
required by paragraph (i)(3) of this AD, is terminating action for
the repetitive inspections required by this paragraph at the
repaired locations only.

 (i) Exceptions to Boeing Special Attention Service Bulletin 737-53-
1065, Revision 3, Dated June 30, 2015

    (1) Where Boeing Special Attention Service Bulletin 737-53-1065,
Revision 3, dated June 30, 2015, specifies compliance times ``after
the Revision 3 date of this service bulletin,'' this AD requires
compliance within the specified compliance times after the effective
date of this AD.
    (2) The Condition column of paragraph 1.E., ``Compliance,'' of
Boeing Special Attention Service Bulletin 737-53-1065, Revision 3,
dated June 30, 2015, refers to airplanes in certain configurations
as of the ``issue date of Revision 3 of this service bulletin.''
However, this AD applies to airplanes in the specified
configurations ``as of the effective date of this AD.''
    (3) Where Boeing Special Attention Service Bulletin 737-53-1065,
Revision 3, dated June 30, 2015, specifies contacting Boeing for
repair instructions or work instructions, before further flight,
repair or perform the work instructions using a method approved in
accordance with the procedures specified in paragraph (o) of this
AD, except as required by paragraph (i)(4) of this AD.
    (4) For airplanes on which an operator has a record that a skin
panel was replaced with a production skin panel at or before 59,000
total flight cycles: At the applicable time for the next inspection
as specified in tables 1 and 2 of paragraph 1.E., ``Compliance,''
Boeing Special Attention Service Bulletin 737-53-1065, Revision 3,
dated June 30, 2015, except as provided by paragraphs (i)(1) and
(i)(2) of this AD: Perform inspections and applicable corrective
actions using a method approved in accordance with the procedures
specified in paragraph (o) of this AD.

 (j) Actions for Airplanes With a Time Limited Repair Installed

    Except for Group 1 airplanes identified in Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015: Do the applicable actions required by paragraphs (j)(1) and
(j)(2) of this AD.
    (1) For airplanes with a time limited repair installed as
specified in Boeing Special Attention Service Bulletin 737-53-1065,
Revision 2, dated April 19, 2001: At the applicable times specified
in table 3 of paragraph 1.E., ``Compliance,'' of Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015, except as provided by paragraphs (i)(1) and (i)(2) of this AD:
Do the actions specified in paragraphs (j)(1)(i) and (j)(1)(ii) of
this AD.
    (i) Do the applicable inspections to detect missing or loose
fasteners and any disbonding or cracking of bonded doublers; and do
all applicable related investigative and corrective actions; in
accordance with the Accomplishment Instructions of Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015, except as required by paragraph (i)(3) of this AD. Do all
applicable related investigative and corrective actions before
further flight. Repeat the applicable inspections thereafter at the
applicable intervals specified Boeing Special Attention Service
Bulletin 737-53-1065, Revision 3, dated June 30, 2015.
    (ii) Make the time limited repair permanent; and do all
applicable related investigative and corrective actions; in
accordance the Accomplishment Instructions of Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015, except as required by paragraph (i)(3) of this AD. Do all
applicable related investigative and corrective actions before
further flight. Accomplishing the permanent repair required by this
paragraph terminates the inspections required by paragraph (j)(1)(i)
of this AD for the permanently repaired area only.
    (2) For airplanes with a time limited repair installed as
specified in Boeing Special Attention Service Bulletin 737-53-1065,
Revision 3, dated June 30, 2015: At the applicable times specified
in table 4 of paragraph 1.E., ``Compliance,'' of Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015: Do the actions specified in paragraphs (j)(2)(i) and
(j)(2)(ii) of this AD.
    (i) Do the applicable inspections to detect missing or loose
fasteners and any disbonding or cracking of bonded doublers; and do
all applicable related investigative and corrective actions; in
accordance with the Accomplishment Instructions of Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015, except as required by paragraph (i)(3) of this AD. Do all
applicable related investigative and corrective actions before
further flight. Repeat the applicable inspections thereafter at the
applicable intervals specified Boeing Special Attention Service
Bulletin 737-53-1065, Revision 3, dated June 30, 2015.
    (ii) Make the time limited repair permanent; and do all
applicable related investigative and corrective actions; in
accordance the Accomplishment Instructions of Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015, except as required by paragraph (i)(3) of this AD. Do all
applicable related investigative and corrective actions before
further flight. Accomplishing the permanent repair required by this
paragraph terminates the inspections required by paragraph (j)(2)(i)
of this AD for the permanently repaired area only.

 (k) Modification of Certain Permanent Repairs

    Except for Group 1 airplanes identified in Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015: For airplanes with an existing time limited repair that was
made permanent as specified in Boeing Special Attention Service
Bulletin 737-53-1065, Revision 2, dated April 19, 2001: At the
applicable times specified in table 5 of paragraph 1.E.,
``Compliance,'' of Boeing Special Attention Service Bulletin 737-53-
1065, Revision 3, dated June 30, 2015, except as required by
paragraph (i)(1) of this AD: Modify the existing permanent repair;
and do all applicable related investigative and corrective actions;
in accordance the Accomplishment Instructions of Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015, except as required by paragraph (i)(3) of this AD. Do all
applicable related investigative and corrective actions before
further flight.

 (l) Certain Post-Repair Inspections

    For airplanes with a permanent repair installed as specified in
Boeing Special Attention Service Bulletin 737-53-1065, Revision 3,
dated June 30, 2015: At the applicable time specified in table 6 of
paragraph 1.E., ``Compliance,'' of Boeing Special Attention Service
Bulletin 737-53-

1065, Revision 3, dated June 30, 2015: Do an external low frequency
eddy current (LFEC) inspection for cracking of the skin at the
critical fastener row of the repair doubler; and do all applicable
corrective actions; in accordance the Accomplishment Instructions of
Boeing Special Attention Service Bulletin 737-53-1065, Revision 3,
dated June 30, 2015, except as required by paragraph (i)(3) of this
AD. Do all applicable corrective actions before further flight.
Repeat the LFEC inspection thereafter at the applicable intervals
specified in Boeing Special Attention Service Bulletin 737-53-1065,
Revision 3, dated June 30, 2015.

 (m) Skin Panel Replacement

    Except for Group 1 airplanes identified in Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015: At the later of the times specified in paragraphs (m)(1) and
(m)(2) of this AD: Replace the applicable skin panels, and do all
applicable related investigative and corrective actions, in
accordance with the Accomplishment Instructions of Boeing Special
Attention Service Bulletin 737-53-1065, Revision 3, dated June 30,
2015. Do all applicable related investigative and corrective actions
before further flight. Doing the skin panel replacement required by
this paragraph terminates the inspection requirements of paragraph
(h) of this AD for that skin panel only, provided the skin panel was
replaced with a production skin panel after 59,000 total flight
cycles.
    (1) Before 60,000 total flight cycles, but not at or before
59,000 total flight cycles.
    (2) Within 6,000 flight cycles after the effective date of this
AD, but not at or before 59,000 total flight cycles.

 (n) Credit for Previous Actions

    This paragraph provides credit for the actions required by
paragraph (h) of this AD, if those actions were performed before the
effective date of this AD using Boeing Special Attention Service
Bulletin 737-53-1065, Revision 2, dated April 19, 2001, which was
incorporated by reference in AD 2005-13-30.

 (o) Alternative Methods of Compliance (AMOCs)

    (1) The Manager, Seattle Aircraft Certification Office (ACO),
FAA, has the authority to approve AMOCs for this AD, if requested
using the procedures found in 14 CFR 39.19. In accordance with 14
CFR 39.19, send your request to your principal inspector or local
Flight Standards District Office, as appropriate. If sending
information directly to the manager of the ACO, send it to the
attention of the person identified in paragraph (p)(1) of this AD.
Information may be emailed to<EMAIL_ADDRESS>    (2) Before using any approved AMOC, notify your appropriate
principal inspector, or lacking a principal inspector, the manager
of the local flight standards district office/certificate holding
district office.
    (3) An AMOC that provides an acceptable level of safety may be
used for any repair, modification, or alteration required by this AD
if it is approved by the Boeing Commercial Airplanes Organization
Designation Authorization (ODA) that has been authorized by the
Manager, Seattle ACO, to make those findings. To be approved, the
repair method, modification deviation, or alteration deviation must
meet the certification basis of the airplane, and the approval must
specifically refer to this AD.
    (4) AMOCs approved previously for AD 2005-13-30, are approved as
AMOCs for the corresponding provisions of paragraph (h) of this AD.

 (p) Related Information

    (1) For more information about this AD, contact Wade Sullivan,
Aerospace Engineer, Airframe Branch, ANM-120S, FAA, Seattle ACO,
1601 Lind Avenue SW., Renton, WA 98057-3356; phone:<PHONE_NUMBER>;
fax:<PHONE_NUMBER>; email<EMAIL_ADDRESS>    (2) For service information identified in this AD, contact
Boeing Commercial Airplanes, Attention: Data & Services Management,
P. O. Box 3707, MC 2H-65, Seattle, WA 98124-2207; telephone 206-544-
5000, extension 1; fax<PHONE_NUMBER>; Internet https://www.myboeingfleet.com. You may view this referenced service
information at the FAA, 1601 Lind Avenue SW., Renton, Washington.
For information on the availability of this material at the FAA,
call<PHONE_NUMBER>.

    Issued in Renton, Washington, on May 4, 2016.
Michael Kaszycki,
Acting Manager, Transport Airplane Directorate, Aircraft Certification
Service.
[FR Doc. 2016-11095 Filed 5-10-16; 8:45 am]
 BILLING CODE 4910-13-P