α-synuclein inhibits Snx3-retromer retrograde trafficking of the conserved membrane-bound proprotein convertase Kex2 in the secretory pathway of Saccharomyces cerevisiae.

We tested the ability of alpha-synuclein (α-syn) to inhibit Snx3-retromer-mediated retrograde trafficking of Kex2 and Ste13 between late endosomes and the trans-Golgi network (TGN) using a Saccharomyces cerevisiae model of Parkinson's disease. Kex2 and Ste13 are a conserved, membrane-bound proprotein convertase and dipeptidyl aminopeptidase, respectively, that process pro-α-factor and pro-killer toxin. Each of these proteins contains a cytosolic tail that binds to sorting nexin Snx3. Using a combination of techniques, including fluorescence microscopy, western blotting and a yeast mating assay, we found that α-syn disrupts Snx3-retromer trafficking of Kex2-GFP and GFP-Ste13 from the late endosome to the TGN, resulting in these two proteins transiting to the vacuole by default. Using three α-syn variants (A53T, A30P, and α-synΔC, which lacks residues 101-140), we further found that A53T and α-synΔC, but not A30P, reduce Snx3-retromer trafficking of Kex2-GFP, which is likely to be due to weaker binding of A30P to membranes. Degradation of Kex2 and Ste13 in the vacuole should result in the secretion of unprocessed, inactive forms of α-factor, which will reduce mating efficiency between MATa and MATα cells. We found that wild-type α-syn but not A30P significantly inhibited the secretion of α-factor. Collectively, our results support a model in which the membrane-binding ability of α-syn is necessary to disrupt Snx3-retromer retrograde recycling of these two conserved endopeptidases.

Loop Electrosurgical Excision Procedure or Cervical Conization to Exclude Cervical Cancer Before Simple Hysterectomy.

OBJECTIVE: The aim of the study was to determine which women require loop electrosurgical excision procedure (LEEP) or cervical conization (cone) to exclude cervical cancer after colposcopy for evaluation of abnormal cervical cancer screening tests yet before simple hysterectomy. MATERIALS AND METHODS: Review of electronic medical records from colposcopy clinics followed by chart review of women with cervical cancer was conducted. RESULTS: Of 18,537 cervical colposcopies for evaluation of abnormal cervical cancer screening tests, 0.6% (103/18,537) had cervical cancer; of 103 women with cervical cancer, 57 had neither cervical biopsy nor endocervical curettage (ECC) showing cancer or rule-out cancer (occult cancers) and were diagnosed by subsequent LEEP (n = 22), cone (n = 31), failed cone (n = 1), or hysterectomy (n = 3). The relative risk of occult cervical cancer at colposcopy for ECC of cervical intraepithelial neoplasia (CIN) 2 or CIN 3 versus ECC not CIN 2 or CIN 3 was 51.5 (5.0% vs 0.1%), for cervical biopsy of CIN 3 versus not CIN 3, was 34.5 (3.9% vs 0.1%), and for colposcopic impression of CIN 2, CIN 3, or cancer (CIN 2+) versus impression not CIN 2+, was 8.5 (1.9% vs 0.2%). If the 10.9% (2,018/18,537) of colposcopies with ECC of CIN 2 or CIN 3, cervical biopsy of CIN 3, or colposcopic impression of CIN 2+ had subsequent LEEP or cone, 96.5% (55/57) of occult cervical cancers would be detected before hysterectomy. CONCLUSIONS: After colposcopy, women with ECC of CIN 2 or CIN 3, cervical biopsy of CIN 3, or colposcopic impression of CIN 2+ require LEEP or cone before simple hysterectomy.

Are Clinicians Ready for Safe Use of Stratified Therapy in Primary Biliary Cholangitis (PBC)? A Study of Educational Awareness.

BACKGROUND: Primary Biliary Cholangitis (PBC, formerly cirrhosis), is a chronic cholestatic liver disease which until spring 2016 had a single licensed therapy, Ursodeoxycholic acid (UDCA). Approximately 30% of patients do not respond to UDCA, and are high-risk for progressing to end stage liver disease, transplantation or death. A new era of stratified medicine with second-line therapies to treat high-risk disease is emerging, with the first such second-line agent obeticholic acid recently receiving FDA and EMA approval and entering practice. Recent experience in the USA of inappropriate use and associated deaths has highlighted concerns as to whether clinicians have the knowledge to implement second-line therapies appropriately and safely. METHODS: Online survey of knowledge regarding optimal PBC management in Gastroenterologists and Hepatologists in the USA; the first 100 completed responses from each group used for analysis. RESULTS: 80% of Hepatologists felt they were highly competent in their understanding of the importance of early diagnosis and early UDCA therapy in PBC compared with 65% of gastroenterologists. However, only 36% of Hepatologists and 30% of gastroenterologists felt competent at assessing response to UDCA. Competence in knowledge (mode of action, efficacy, and side effects) of second-line therapies and enrollment into trials was low among both groups. CONCLUSION: Significant knowledge gaps in clinicians managing PBC presents a problem in optimizing care. It is perhaps not surprising that knowledge of emerging second-line therapies is low, however more concerning is sub-optimal use of UDCA in real-life practice and the lack of confidence at assessing treatment response which should be a routine part of clinical practice to assess risk of disease progression and will be key in delivering stratified medicine.

High-Grade Squamous Intraepithelial Lesion Cytology With Negative High-Risk Human Papillomavirus Tests Rarely Diagnoses Endometrial Cancer.

OBJECTIVES: We hypothesized that women with cervical cytologic results of high-grade squamous intraepithelial lesion (HSIL) and negative high-risk human papillomavirus (HR-HPV) test results would have a high risk of having endometrial cancer and would benefit from routine endometrial biopsy. MATERIALS AND METHODS: Reports of women with cytologic results of HSIL and negative HR-HPV test results were found in an electronic colposcopy database; their charts were reviewed. Rates of endometrial cancer for cytologic results of HSIL and negative HR-HPV test results were compared to a historical series for cytologic results of HSIL with positive HR-HPV and cytologic results of atypical glandular cells (AGCs) and negative HR-HPV test results. RESULTS: Between August 10, 1998, and April 20, 2013, 56 women were evaluated in our colposcopy clinics for cytologic results of HSIL and negative HR-HPV test results; of these 56 women, 1 (1.8%) was diagnosed with endometrial cancer. No endometrial cancer was diagnosed during the follow-up (median = 63 mo) after colposcopy. The risk for endometrial cancer with cytologic results of HSIL and negative HR-HPV test results (1.8%, 1/56) did not differ from that of a historical series from 2007 to 2009 from the same colposcopy clinic in 223 women with cytologic results of HSIL and positive HR-HPV test results (0.0%, 0/223; p = .2) and was lower than the risk for endometrial cancer from the historical series from 2007 to 2009 in women with cytologic results of AGC and negative HR-HPV test results (14.4%, 4/27; p = .04). CONCLUSIONS: Women with cytologic results of HSIL and negative HR-HPV test results are more like those with cytologic results of HSIL and positive HR-HPV test results than those with cytologic results of AGC and negative HR-HPV test results and would unlikely to benefit from routine endometrial biopsy at the time of colposcopy.

Factors That Virtually Exclude Cervical Cancer at Colposcopy.

OBJECTIVES: The objective of this work was to determine the risk of invasive cervical cancer at colposcopy based on the woman's age, associated cervical cytology, and colposcopic impression. METHODS: Review of electronic medical records from colposcopy clinics followed by chart review of women with cervical cancer. RESULTS: Between March 1, 1996, and April 23, 2013, 27,381 cervical colposcopies for evaluation of abnormal cervical cytology and/or positive high-risk human papillomavirus tests were performed. Median age at colposcopy was 32 years (range, 11-91 years). Biopsy at colposcopy or at subsequent loop electrocautery excision procedure, cervical conization, or hysterectomy diagnosed cervical cancer in 0.5% (132/27,381) of women. At colposcopy, the risk of cervical cancer for women younger than 30 years was 0.07% (8/12,131), for women with Cytology-Lo (low-grade squamous intraepithelial lesion, atypical squamous cells of uncertain significance, benign endometrial cells, or negative), it was 0.17% (43/25,779); and for women with normal colposcopic impressions, it was 0.17% (18/11,389). The risk of cervical cancer for women younger than 30 years with Cytology-Lo was 0.01% (1/11,663). CONCLUSIONS: At colposcopy, cervical cancer is not excluded by a normal colposcopic examination result, Cytology-Lo, or age younger than 30 years, but is virtually excluded by the combination of age younger than 30 years and Cytology-Lo.

Which Colposcopies Should Include Endocervical Curettage?

OBJECTIVES: Although endocervical curettage (ECC) is often performed at colposcopy, it remains unclear whether it should be done in all women, only women over a certain age, only women with unsatisfactory colposcopy, or only in women with normal colposcopic impressions. To clarify the indications for ECC, we determined the proportion of colposcopies with CIN 3, or cancer (CIN 3+) detected only by ECC showing CIN 2, CIN 3, or cancer (CIN 2+). METHODS: Review of electronic medical records from colposcopy clinics. RESULTS: Between March 1, 1996, and April 23, 2013, approximately 18,537 cervical colposcopies with no missing results evaluated women with abnormal cervical cytology and/or positive high-risk human papillomavirus tests. In 7.5% (1,398/18,537) of colposcopies, the final diagnosis, based on the worst biopsy from cervix; ECC; or subsequent loop electrocautery excision procedure, cervical conization, or hysterectomy, was CIN 3+. The proportion of colposcopies with CIN 3+ detected only by ECC showing CIN 2+ was 0.5% (101/18,537). Limiting the ECC to women with unsatisfactory colposcopy, colposcopic impressions suggestive of CIN 2+, or impressions of normal resulted in fewer ECC performed but also greatly decreased the proportion of CIN 3+ that could have been detected only by ECC showing CIN 2+. Limiting the ECC to colposcopies in women age 25 years and older resulted in 29.3% (5,433/18,537) fewer ECCs while detecting 96.0% (97/101) of the CIN 3+ that could have been detected only by ECC showing CIN 2 + . CONCLUSIONS: Endocervical curettage should be performed at colposcopy in women age 25 and older.

Yield and mode of diagnosis of cervical intraepithelial neoplasia 3 or cancer among women with negative cervical cytology and positive high-risk human papillomavirus test results.

OBJECTIVE: In women with negative cervical cytology and positive high-risk human papillomavirus (HR-HPV) test results, we compared the risk of cervical intraepithelial neoplasia 3 (CIN 3) or cancer (CIN 3+) in women with previous abnormal cervical cytology, CIN, or HR-HPV with that in women without this history, and we determined their cumulative risk of CIN 3+. MATERIALS AND METHODS: We reviewed colposcopies for negative cytology and positive HR-HPV test results from 2007 to 2009 (colposcopy was done for previous abnormal cytology, HR-HPV, or CIN or if negative cytology and positive HR-HPV test results for 20-35 months). Women with negative cytology and positive HR-HPV test results in 2007 were reviewed to determine their cumulative risk of CIN 3+. RESULTS: Of the 513 women with colposcopy for negative cytology and positive HR-HPV test results, 367 had previous abnormal cytology, CIN, or HR-HPV greater than 35 months ago and 146 had negative cytology and positive HR-HPV test results for 20 to 35 months. Risk of CIN 3+ for women with negative cytology and positive HR-HPV test results with previous abnormal cytology, HR-HPV, or CIN who did not have previous colposcopy was 11.9% (8/67); for similar women with previous colposcopy, it was 2.7% (8/300); and for women with positive HR-HPV test result for 20 to 35 months, it was 7.5% (11/146). In 2007 to 2009, risk of CIN 3+ for women with cytology of atypical squamous cells of undetermined significance (ASC-US) with positive HR-HPV test result was 3.9% (60/1,540). Of the 1,726 women with negative cytology and positive HR-HPV test result in 2007, 381 (22.1%) were lost to follow-up. Of the 1,345 women with median follow-up of 44 months, 602 (44.8%) had 1 or 2 subsequent negative cytology and negative HR-HPV test results and 180 had subsequent positive HR-HPV test result without biopsy. Of the 563 women with biopsy, there were 711 evaluations. Invasive cancer was found in 4 and CIN 3 in 54 (cumulative CIN 3+ of 4.3%, 58/1,345). Half (29/58) of the cumulative CIN 3+ was diagnosed after subsequent abnormal cytology and positive HR-HPV test result. CONCLUSIONS: If referral to colposcopy of women with cervical cytology of ASC-US and positive HR-HPV test result (CIN 3+, 3.9%) is justified, referral to colposcopy of women with negative cytology and positive HR-HPV test results is justified if they have previous abnormal cervical cytology, CIN, or HR-HPV greater than 35 months ago but have not had previous colposcopy (CIN 3+=11.9%) or are persistently HR-HPV positive for 20 to 35 months (CIN 3+, 7.5%). The risk of CIN 3+ in women with previous abnormal cytology, CIN, or HR-HPV who have previous colposcopy (2.7%) is lower because these women have incident rather than prevalent CIN 3+.

Utility of random cervical biopsy and endocervical curettage in a low-risk population.

OBJECTIVE: The study aimed to determine the increase in the yield of cervical intraepithelial neoplasia 3 (CIN 3) or cancer (CIN 3+) from random cervical biopsy in quadrants without visible lesions and endocervical curettage (ECC) in a low-prevalence setting. MATERIALS AND METHODS: Random biopsy and ECC (unless pregnant) have been obtained in the colposcopy clinic of the Southern California Permanente Medical Group (SCPMG)-Fontana since 2004. We reviewed the colposcopy experience of SCPMG-Fontana for January 1, 2007, to December 31, 2009, to determine the method of diagnosis of CIN 3+. RESULTS: Between January 1, 2007, and December 31, 2009, 4677 women with median age 32 years had 4932 colposcopies in the SCPMG-Fontana colposcopy clinics. Cervical intraepithelial neoplasia 3+ was diagnosed in 295 women. Cervical biopsy detected 64.4% of CIN 3+; ECC diagnosed 5.1%; loop electrocautery excision procedure (LEEP) or cervical conization for cervical biopsy and/or ECC of CIN 2 diagnosed 27.8%; LEEP for the cytology of high-grade squamous intraepithelial lesion with cervical biopsy result of negative or CIN 1 diagnosed 1.4%; and LEEP, cervical conization, or biopsy in follow-up of CIN 2 diagnosed 1.4%. Sixty-one of the 295 cases of CIN 3+ (20.7%) were diagnosed after evaluation of random cervical biopsy and/or ECC of CIN 2+. CONCLUSIONS: Random biopsy in cervical quadrants without visible lesions and ECC increased the yield of CIN 3+ in this low-risk colposcopy setting. Endocervical curettage can be omitted in women younger than 25 years.

Identification and characterization of rapidly accumulating sch9Δ suppressor mutations in Saccharomyces cerevisiae.

Nutrient sensing is important for cell growth, aging, and longevity. In Saccharomyces cerevisiae, Sch9, an AGC-family protein kinase, is a major nutrient sensing kinase homologous to mammalian Akt and S6 kinase. Sch9 integrates environmental cues with cell growth by functioning downstream of TORC1 and in parallel with the Ras/PKA pathway. Mutations in SCH9 lead to reduced cell growth in dextrose medium; however, reports on the ability of sch9Δ mutants to utilize non-fermentable carbon sources are inconsistent. Here, we show that sch9Δ mutant strains cannot grow on non-fermentable carbon sources and rapidly accumulate suppressor mutations, which reverse growth defects of sch9Δ mutants. sch9Δ induces gene expression of three transcription factors required for utilization of non-fermentable carbon sources, Cat8, Adr1, and Hap4, while sch9Δ suppressor mutations, termed sns1 and sns2, strongly decrease the gene expression of those transcription factors. Despite the genetic suppression interactions, both sch9Δ and sns1 (or sns2) homozygous mutants have severe defects in meiosis. By screening mutants defective in sporulation, we identified additional sch9Δ suppressor mutants with mutations in GPB1, GPB2, and MCK1. Using library complementation and genetic analysis, we identified SNS1 and SNS2 to be IRA2 and IRA1, respectively. Furthermore, we discovered that lifespan extension in sch9Δ mutants is dependent on IRA2 and that PKA inactivation greatly increases basal expression of CAT8, ADR1, and HAP4. Our results demonstrate that sch9Δ leads to complete loss of growth on non-fermentable carbon sources and mutations in MCK1 or genes encoding negative regulators of the Ras/PKA pathway reverse sch9Δ mutant phenotypes.

Engaging Communities to Reach Immigrant and Minority Populations: The Minnesota Immunization Networking Initiative (MINI), 2006-2017.

In Minneapolis-St Paul, Minnesota, factors such as cultural and linguistic diversity make it difficult for public health agencies to reach immigrant and racial/ethnic minority populations with health initiatives. Founded in 2006, the Minnesota Immunization Networking Initiative (MINI) is a community project that has provided more than 80 000 free influenza vaccinations to vulnerable populations, including immigrants and racial/ethnic minority groups. MINI administered 5910 vaccinations through 99 community-based vaccination clinics during the 2017-2018 influenza season and surveyed the clients in their own language about influenza vaccination knowledge and practices. Among those surveyed, 2545 (43.1%) were uninsured and 408 (6.9%) received a first-time influenza vaccination at the MINI clinic. A total of 2893 (49.0%) respondents heard about the clinic through their faith community. Lessons learned included the importance of building relationships with community leaders and involving them as full partners, holding clinics in community-based settings to bring vaccinations to clients, and reporting outcomes to partners.

Inclusão nos Estados Unidos: filosofia, implementação e capacitação de professores / Inclusion in the United States: philosophy; implementation, and capacitating teachers

A inclusão de alunos com deficiência em classes da educação geral é uma exigência importante das Leis para Indivíduos com Deficiência de 1975 e 1997 e da emenda da Lei para Indivíduos com Deficiência de 2004. Desde a promulgação dessas leis, as escolas públicas dos Estados Unidos têm sido obrigadas a incluir alunos com deficiência ao máximo possível em classes da educação geral onde eles possam estudar com pares não deficientes ao invés de passarem a maior parte de seu dia em classes segregadas com outros alunos com deficiência. Este trabalho discute a filosofia dos programas de inclusão e apresenta detalhes sobre programas de inclusão em um Estado específico dos Estados Unidos, o Estado do Arizona, Finalmente, descreve a necessidade de capacitação de professores para a área de educação geral e especial. Essa sessão demonstra porque os programas melhorados de capacitação de professores são necessários para aumentar o sucesso da inclusão nos Estados Unidos.

Inclusion of students with disabilities in general education classes is an important provision of the Individuals with Disabilities Education Act of 1975, 1997, and the Individuals with Disabilities Improvement Act of 2004. Since the inception of this law, public schools in the United States have been mandated to include students with disabilities to the maximum extent possible in general education classes where they can study with typical peers rather than spending most of their day in segregated classes with other students with disabilities. This paper will discuss the philosophy of these inclusion programs. In addition, the paper will present details about inclusion programs in a specific state within the United States, the state of Arizona. Finally, the teacher training need in the area of general education and special education collaboration will be described. This section will demonstrate why improved teacher preparation programs are needed in order to increase the success of inclusion in the United States.