Role of Endoscopic Biopsies and Morphologic Features in Predicting Microsatellite Instability Status in Gastric Cancer: A Multicenter Comparative Study of Endoscopic Biopsies and Surgical Specimens.

Evaluation of mismatch repair (MMR) protein and microsatellite instability (MSI) status plays a pivotal role in the management of gastric cancer (GC) patients. In this study, we aimed to evaluate the accuracy of gastric endoscopic biopsies (EBs) in predicting MMR/MSI status and to uncover histopathologic features associated with MSI. A multicentric series of 140 GCs was collected retrospectively, in which EB and matched surgical specimens (SSs) were available. Laurén and WHO classifications were applied and detailed morphologic characterization was performed. EB/SS were analyzed by immunohistochemistry (IHC) for MMR status and by multiplex polymerase chain reaction (mPCR) for MSI status. IHC allowed accurate evaluation of MMR status in EB (sensitivity: 97.3%; specificity: 98.0%) and high concordance rates between EB and SS (Cohen κ=94.5%). By contrast, mPCR (Idylla MSI Test) showed lower sensitivity in evaluating MSI status (91.3% vs. 97.3%), while maintaining maximal specificity (100.0%). These results suggest a role of IHC as a screening method for MMR status in EB and the use of mPCR as a confirmatory test. Although Laurén/WHO classifications were not able to discriminate GC cases with MSI, we identified specific histopathologic features that are significantly associated with MMR/MSI status in GC, despite the morphologic heterogeneity of GC cases harboring this molecular phenotype. In SS, these features included the presence of mucinous and/or solid components ( P =0.034 and <0.001) and the presence of neutrophil-rich stroma, distant from tumor ulceration/perforation ( P <0.001). In EB, both solid areas and extracellular mucin lakes were also discriminating features for the identification of MSI-high cases ( P =0.002 and 0.045).

Performance of Immunohistochemical and Molecular Methods in Detecting Microsatellite Instability in Gastric Cancer: A Multicenter Study.

INTRODUCTION: Microsatellite instability (MSI) is an important prognostic molecular biomarker for gastric cancer (GC). MSI status may be detected by immunohistochemistry (IHC) for mismatch repair (MMR) proteins and polymerase chain reaction (PCR). Idylla™ MSI assay has not been validated for GC but may prove to be a valid alternative. METHODS: In a series of 140 GC cases, MSI status was evaluated by IHC for MLH1, PMS2, MSH2, and MSH6; gold-standard pentaplex PCR panel (PPP) (BAT-25, BAT-26, NR-21, NR-24, and NR-27); and Idylla. Statistical analysis was performed using SPSS 27.0. RESULTS: PPP identified 102 microsatellite stable (MSS) cases and 38 MSI-high cases. Only 3 cases showed discordant results. Compared with PPP, the sensitivity was 100% for IHC and 94.7% for Idylla. Specificity was 99% for IHC and 100% for Idylla. MLH1 IHC alone showed sensitivity and specificity of 97.4% and 98.0%, respectively. IHC identified three indeterminate cases; all were MSS according to PPP and Idylla. CONCLUSION: IHC for MMR proteins represents an optimal screening tool for MSI status in GC. If resources are limited, isolated MLH1 evaluation may constitute a valuable option for preliminary screening. Idylla may help detect rare MSS cases with MMR-loss and define MSI status in indeterminate cases.

EBV and MSI Status in Gastric Cancer: Does It Matter?

We investigated the impactof microsatellite instability (MSI) and Epstein-Barr virus (EBV) status in gastric cancer (GC), regarding response to perioperative chemotherapy (POPChT), overall survival (OS), and progression-free survival (PFS). We included 137 cases of operated GC, 51 of which were submitted to POPChT. MSI status was determined by multiplex PCR and EBV status by EBV-encoded RNA in situ hybridization. Thirty-seven (27%) cases presented as MSI-high, and seven (5.1%) were EBV+. Concerning tumor regression after POPChT, no differences were observed between the molecular subtypes, but females were more likely to respond (p = 0.062). No significant differences were found in OS or PFS between different subtypes. In multivariate analysis, age (HR 1.02, IC 95% 1.002-1.056, p = 0.033) and positive lymph nodes (HR 1.82, IC 95% 1.034-3.211, p = 0.038) were the only prognostic factors for OS. However, females with MSI-high tumors treated with POPChT demonstrated a significantly increased OS compared to females with MSS tumors (p = 0.031). In conclusion, we found a high proportion of MSI-high cases. MSI and EBV status did not influence OS or PFS either in patients submitted to POPChT or surgery alone. However, superior survival of females with MSI-high tumors suggests that sex disparities and molecular classification may influence treatment options in GC.

Expression of Thomsen-Friedenreich Antigen in Colorectal Cancer and Association with Microsatellite Instability.

Microsatellite instability (MSI) is a molecular phenotype due to a deficient DNA mismatch repair (dMMR). In colorectal cancer (CRC), dMMR/MSI is associated with several clinical and histopathological features, influences prognosis, and is a predictive factor of response to therapy. In daily practice, dMMR/MSI profiles are identified by immunohistochemistry and/or multiplex PCR. The Thomsen-Friedenreich (TF) antigen was previously found to be a potential single marker to identify MSI-high gastric cancers. Therefore, in this study, we aimed to disclose a possible association between TF expression and MSI status in CRC. Furthermore, we evaluated the relationship between TF expression and other clinicopathological features, including patient survival. We evaluated the expression of the TF antigen in a cohort of 25 MSI-high and 71 microsatellite stable (MSS) CRCs. No association was observed between the expression of the TF antigen and MSI-high status in CRC. The survival analysis revealed that patients with MSI-high CRC showed improved survival when the TF antigen was expressed. This finding holds promise as it indicates the potential use of the TF antigen as a biomarker of better prognosis in MSI-high CRCs that should be validated in an independent and larger CRC cohort.

Sentinel node total tumour load as a predictive factor for non-sentinel node status in early breast cancer patients - The porttle study.

OSNA is a molecular assay for the detection of sentinel node metastasis. TTL emerged as a concept that seems to accurately predict the status of the NSN. Authors tried to confirm this motion. This is a retrospective and multicentric study that analyzed 2164 patients, 579 of whom had positive SN and completion AD. Logistic regression models were performed in order to identify a suitable cutoff to identify patients who benefit from AD. Univariate and multivariate regression analysis showed a relationship between TTL>30000 and the presence of NSN metastasis (OR 2.84, CI 1.99-4.08, p < 0.001). Logistic regression indicated that the cutoff of 30000 copies/µL better discriminates patients with NSN positivity and allows wide use of these criteria. This cutoff value may safely assist clinicians and patients to decide to proceed or not with an AD.

Survey on pathogenic Escherichia coli and Salmonella spp. in captive cockatiels (Nymphicus hollandicus).

We surveyed healthy captive cockatiels (Nymphicus hollandicus) for Escherichia coli and Salmonella spp. Cloacal swabs were collected from 94 cockatiels kept in commercial breeders, private residencies and pet shops in the cities of São Paulo/SP and Niterói/RJ (Brazil). Three strains of E. coli from each individual were tested for the presence of ExPEC-, APEC- and DEC-related genes. We evaluated the blaTEM, blaSHV, blaOXA, blaCMY, blaCTX-M, tetA, tetB, aadA, aphA, strAB, sul1, sul2, sul3, qnrA, qnrD, qnrB, qnrS, oqxAB, aac (6)'-Ib-cr, qepA resistance genes and markers for plasmid incompatibility groups. Salmonella spp. was not detected. E. coli was isolated in 10% of the animals (9/94). Four APEC genes (ironN, ompT, iss and hlyF) were detected in two strains (2/27-7%), and iss (1/27-4%) in one isolate. The highest resistance rates were observed with amoxicillin (22/27-82%), ampicillin (21/27-79%), streptomycin (18/27-67%), tetracycline (11/27-41%). Multiresistance was verified in 59% (16/27) of the isolates. We detected strAB, blaTEM, tetA, tetB, aadA, aphaA, sul1, sul2, sul3 resistance genes and plasmid Inc groups in 20 (74%) of the strains. E. coli isolated from these cockatiels are of epidemiological importance, since these pets could transmit pathogenic and multiresistant microorganisms to humans and other animals.

Survey on pathogenic Escherichia coli and Salmonella spp. in captive cockatiels (Nymphicus hollandicus)

Abstract We surveyed healthy captive cockatiels (Nymphicus hollandicus) for Escherichia coli and Salmonella spp. Cloacal swabs were collected from 94 cockatiels kept in commercial breeders, private residencies and pet shops in the cities of São Paulo/SP and Niterói/RJ (Brazil). Three strains of E. coli from each individual were tested for the presence of ExPEC-, APEC- and DEC-related genes. We evaluated the blaTEM, blaSHV, blaOXA, blaCMY, blaCTX-M, tetA, tetB, aadA, aphA, strAB, sul1, sul2, sul3, qnrA, qnrD, qnrB, qnrS, oqxAB, aac (6)′-Ib-cr, qepA resistance genes and markers for plasmid incompatibility groups. Salmonella spp. was not detected. E. coli was isolated in 10% of the animals (9/94). Four APEC genes (ironN, ompT, iss and hlyF) were detected in two strains (2/27-7%), and iss (1/27-4%) in one isolate. The highest resistance rates were observed with amoxicillin (22/27-82%), ampicillin (21/27-79%), streptomycin (18/27-67%), tetracycline (11/27-41%). Multiresistance was verified in 59% (16/27) of the isolates. We detected strAB, bla TEM, tetA, tetB, aadA, aphaA, sul1, sul2, sul3 resistance genes and plasmid Inc groups in 20 (74%) of the strains. E. coli isolated from these cockatiels are of epidemiological importance, since these pets could transmit pathogenic and multiresistant microorganisms to humans and other animals.

Survey on pathogenic Escherichia coli and Salmonella spp. in captive cockatiels (Nymphicus hollandicus)

Abstract We surveyed healthy captive cockatiels (Nymphicus hollandicus) for Escherichia coli and Salmonella spp. Cloacal swabs were collected from 94 cockatiels kept in commercial breeders, private residencies and pet shops in the cities of São Paulo/SP and Niterói/RJ (Brazil). Three strains of E. coli from each individual were tested for the presence of ExPEC-, APEC- and DEC-related genes. We evaluated the blaTEM, blaSHV, blaOXA, blaCMY, blaCTX-M, tetA, tetB, aadA, aphA, strAB, sul1, sul2, sul3, qnrA, qnrD, qnrB, qnrS, oqxAB, aac (6)-Ib-cr, qepA resistance genes and markers for plasmid incompatibility groups. Salmonella spp. was not detected. E. coli was isolated in 10% of the animals (9/94). Four APEC genes (ironN, ompT, iss and hlyF) were detected in two strains (2/277%), and iss (1/274%) in one isolate. The highest resistance rates were observed with amoxicillin (22/2782%), ampicillin (21/2779%), streptomycin (18/2767%), tetracycline (11/2741%). Multiresistance was verified in 59% (16/27) of the isolates. We detected strAB, bla TEM, tetA, tetB, aadA, aphaA, sul1, sul2, sul3 resistance genes and plasmid Inc groups in 20 (74%) of the strains. E. coli isolated from these cockatiels are of epidemiological importance, since these pets could transmit pathogenic and multiresistant microorganisms to humans and other animals.

Anal HR-HPV prevalence assessed with the cobas® HPV Test in an HIV-infected population of men who have sex with men.

BACKGROUND: Anal squamous cell cancer is an HPV associated disease, highly prevalent in HIV-infected men who have sex with men (MSM). We evaluate the prevalence of HPV infection in an HIV-positive MSM population attending our hospital. Cytological results are correlated with an HR-HPV genotype. METHODS: From February 2013 to July 2014, 95 anal Thin Prep cytologies and 17 histological specimens were evaluated in 89 HIV-infected MSM. HPV detection (cobas® HPV Test) was done in 78 patients. Patients with a positive cytological result were biopsied according to the recommendations of the European Society for Medical Oncology Clinical Practical Guidelines. The STATVIEW statistical computer program was used. RESULTS: The mean age was 37 years. Cytological results: 40% - NILM, 17.9% - ASCUS, 24.2% - LSIL, 4.2% - HSIL and 1.1% - AGC. The overall prevalence of HR-HPV was of 80.8%. In 76.9% of the cases, infection occurred with multiple HR-HPV genotypes. The "other" HR-HPV pattern was the most prevalent HR-HPV genotype. Considering age, HPV-16 prevalence was 50.0% and 28.6% in patients under and over 30 years, respectively. A histological specimen was obtained from 17 patients and 82.4% of them had AIN. CONCLUSION: 1 - HIV-infected MSM is a highly HR-HPV genotype infected population; 2 - The multiple infection pattern was the most prevalent; 3 - "Other", an HPV subtype was the most prevalent pattern; 4 - The prevalence of HPV infection and multiplicity of infection was directly related to the severity of the cytological abnormalities and age; 5 - HPV-16 prevalence was more prevalent in patients under 30 years.

Reed-Sternberg cells form by abscission failure in the presence of functional Aurora B kinase.

Large multinucleated Reed-Sternberg cells (RS) and large mononucleated Hodgkin cells (H) are traditionally considered to be the neoplastic population in classical Hodgkin lymphoma, (cHL) and postulated to promote the disease. However, the contribution of these larger cells to the progression of cHL remains debatable. We used established cHL cell lines and cHL cellular fractions composed of small mononucleated cells only or enriched in large RS/H cells to investigate RS/H cell origin and to characterize the cells which they derive from. We confirm that the small mononucleated cells give rise to RS/H cells, and we show that the latter proliferate significantly more slowly than the small cells. By using live-cell imaging, we demonstrate that binucleated RS cells are generated by failure of abscission when a few small cells attempt to divide. Finally, our results reveal that the small mononucleated cells are chromosomally unstable, but this is unlikely to be related to a malfunctioning chromosomal passenger protein complex. We propose that the small mononucleated cells, rather than the RS/H cells, are the main drivers of cHL.

Genotypes and prevalence of HPV single and multiple concurrent infections in women with HSIL.

The contribution of human papillomavirus (HPV) types to the carcinogenesis of cervical cancer has been established for a long time. However, the role of phylogenetically related and rare variants remains uncertain, as well as the influence of concurrent multiple HPV genotypes infection. We aimed at studying the prevalence of several HPV genotypes infecting women with single versus concurrent multiple HPV genotypes infection with a HSIL diagnosis in a cervical cytology. We conducted a cross-sectional study using Thin-Prep(®) liquid-based cervical cytology specimens with the diagnosis of high-grade squamous intraepithelial lesion (HSIL), in which HPV genotype was sequentially tested. Genotypes were determined with a PapilloCheck(®) system, a DNA-Chip for the type-specific identification of 18 high-risk and six low-risk types of HPV. Of the total study population, 176 cases had a diagnosis of HSIL and positive HPV genotyping result, being HPV16 the most prevalent genotype (48.86%; 95%CI: 41.58-56.19) followed by HPV31 (14.20%; 95%CI: 9.75-20.18). Concurrent multiple HPV genotypes were detected in 36.93% (95%CI: 30.15-44.27) of the patients. The prevalence of the 10 most common HPV genotypes detected varied significantly according to the presence of single vs. concurrent multiple HPV genotypes (P = 0.022). Moreover, women with concurrent multiple HPV genotypes were on average 3.53 (95%CI: 0.43-6.64) years younger than women with single genotype infection. Our results suggest that women with multiple genotype HPV infection differ in terms of age and distribution of the most prevalent HPV genotypes. Additionally, we provide further evidence of the predominance of HPV16 in HSIL lesions of the uterine cervix.

Advanced technologies for oral controlled release: cyclodextrins for oral controlled release.

Cyclodextrins (CDs) are used in oral pharmaceutical formulations, by means of inclusion complexes formation, with the following advantages for the drugs: (1) solubility, dissolution rate, stability, and bioavailability enhancement; (2) to modify the drug release site and/or time profile; and (3) to reduce or prevent gastrointestinal side effects and unpleasant smell or taste, to prevent drug-drug or drug-additive interactions, or even to convert oil and liquid drugs into microcrystalline or amorphous powders. A more recent trend focuses on the use of CDs as nanocarriers, a strategy that aims to design versatile delivery systems that can encapsulate drugs with better physicochemical properties for oral delivery. Thus, the aim of this work was to review the applications of the CDs and their hydrophilic derivatives on the solubility enhancement of poorly water-soluble drugs in order to increase their dissolution rate and get immediate release, as well as their ability to control (to prolong or to delay) the release of drugs from solid dosage forms, either as complexes with the hydrophilic (e.g., as osmotic pumps) and/or hydrophobic CDs. New controlled delivery systems based on nanotechnology carriers (nanoparticles and conjugates) have also been reviewed.

Characterization and removal of specific organic constituents in an UASB-trickling-filter system treating domestic wastewater.

This paper presents the characterization of specific organic constituents (carbohydrates, proteins and lipids) in raw sewage and in the anaerobic and aerobic effluents of a demo-scale (500 inhabitants) UASB- trickling-filter system. The evaluation of such parameters was carried out for two operating conditions, either without sludge recirculation (experiment I) from the trickling filter to the UASB reactor or with sludge recirculation (experiment II), for sludge thickening and stabilization, in the anaerobic reactor. The results showed that the contribution of acetic acid, carbohydrates, proteins and lipids amounted for approximately 70% of the total COD fed to the UASB during experiment I, whereas during experiment II these constituents amounted for only around 40% of the total COD. Although very high BOD and COD overall removal efficiencies were observed for the treatment system (around 90% and 80%, respectively), it was possible to infer that these efficiencies were mainly related to the removal of carbohydrates and lipids (around 80% removal), and of other non-identified constituents. The removal of proteins was much lower (around 50% removal), and the relative contribution of proteins to the total COD increased along the treatment course, being responsible for most of the residual COD of the treatment units. In the present system configuration, the UASB reactor played a major role in the removal of carbohydrates, whereas the trickling filter was very effective in the removal of lipids. The return of aerobic sludge for thickening and stabilization in the UASB reactor did not affect its performance.

Efeito do retorno de lodo aeróbio sobre as características da biomassa presente em reatores UASB tratando esgoto sanitário / Effect of aerobic sludge return on the characteristics of anaerobic biomass present in UASB reactors treating domestic sewage

O presente trabalho buscou avaliar a influência do retorno do lodo produzido em filtros biológicos percoladores (FBP) sobre as características da biomassa em reatores UASB utilizados para o tratamento de esgoto sanitário. Os experimentos foram conduzidos em reatores em escala piloto e em escala de demonstração, sendo que os reatores UASB foram operados com e sem retorno do lodo produzido nos FBP. Os resultados obtidos indicaram uma diminuição no diâmetro das partículas de lodo, durante as fases com retorno de lodo, mas apenas para as amostras de lodo tomadas nos pontos mais altos dos reatores anaeróbios. A estabilidade e a atividade metanogênica específica do lodo anaeróbio praticamente não se modificaram com o retorno de lodo. Conforme esperado, a produção específica de lodo anaeróbio aumentou com o retorno de lodo.

This work aimed at evaluating the influence of the return of excess aerobic sludge produced in trickling filters (TF) upon the biomass characteristics in UASB reactors used for the treatment of domestic sewage. The experiments were carried out in pilot and demonstration-scale reactors, with the UASB reactors being operated with and without the return of sludge produced in the TF. Smaller sludge particle sizes were observed when the aerobic sludge was returned to the UASB reactor, but only for sludge samples collected in the upper parts of the anaerobic reactors. Sludge stability and specific methanogenic activity were not affected by the return of sludge. As expected, the specific anaerobic sludge production increased with the return of sludge.

Triagem de desenvolvimento neuropsicomotor em instituições de educação infantil segundo o teste de Denver II / Neuropsychological development screening in child's educational institutions according do Denver II Test

Desenvolvimento é um direito fundamental de qualquer criança e depende da interação entre herança genética e condições ambientais. A partir desta premissa, triou-se o desenvolvimento de 66 crianças (37 meninos e 29 meninas) de 2 a 3 anos que frequentavam 4 instituições públicas de educação infantil (IEIs) na cidade de São Paulo. Usou-se o teste de Triagem de Desenvolvimento de Denver II. A maior parte da amostra (62, 74 por cento) ganha até 0,5 SM per capita por mês. As adequações de cada área do desenvolvimento foram: pessoal-social (N=54; 81, 8 por cento), motor fino (N=53; 80, 3 por cento), motor grosso (N=53; 80, 3 por cento) e linguagem (N=49; 74, 2 por cento). Nas áreas de linguagem e pessoal-social, as crianças obtiveram os piores resultados e na motora, os melhores...

A utilização do biofeedback no tratamento fisioterápico da paralisia facial periférica / Physical therapy for facial paralysis using the biofeedback

A paralisia facial periférica (PFP) é caracterizada como uma lesão do nervo facial que altera as funções da musculatura da face podendo ser causada por fatores traumáticos, infecciosos, idiopáticos e outros. Naqueles casos que evoluem para a forma crônica, as complicações mais comuns são o ressecamento corneano e as sincinesias. Além disso, essa doença gera incapacidades físicas, psicológicas e sociais. Várias técnicas fisioterápicas foram propostas para o tratamento da PFP, porém, muitas não consideram a especificidade do sistema neuromotor facial e nenhuma se destaca como sendo a mais efetiva. Esse artigo faz uma revisão bibliográfica sobre o uso do biofeedback na PFP, ressaltando técnicas, características e efeitos do tratamento. O biofeedback é uma técnica que utiliza referências visuais ou auditivas por meio da eletromiografia, dos espelho ou de outros recursos, para fornecer ao indivíduo informações sobre sua performance motora. Essa técnica associada exercícios específicos tem sido apontado como benéfico no tratamento da PFP. Os principais efeitos alcançados são a melhora do controle e da coordenação do movimento e a redução da assimetria e sincinesia, por um processo de aprendizado motor. Fatores como início precoce, maior duração do tratamento, acompanhamento posterior e adesão dos sujeitos parecem influenciar a obtenção de resultados positivos. Apesar das inúmeras vantagens apontadas na literatura para a utilização do biofeedback na PFP, os estudos revisados não consideram a heterogeneidade dos sujeitos submetidos ao tratamento e não determinam quais características realmente alteram a recuperação dos indivíduos em tratamento com biofeedback. Além disso, os efeitos do biofeedback a longo prazo não estão estabelecidos e a manutenção de um programa domiciliar orientado parece ser necessário.

Avaliaçäo do grau de insatisfação pós laqueadura tubária no Centro de Atençäo Integral à Saúde da Mulher - Faculdade de Medicina do ABC / Evaluation of the regret rates after tubal ligation at Centro de Atençäo Integral à Saúde da mulher - Faculdade de Medicina do ABC

O trabalho tem por objetivo avaliar o índice de insatisfaçäo pós-laqueadura tubária e propor meios para diminuir essa incidência. 310 pacientes do CAISM-ABC submetidas à laqueadura tubária entre os anos de 1971 e 1995, que foram entrevistadas e divididas em dois grupos: 1- Pacientes que fizeram parte do programa e realizaram a cirurgia em outros serviços. O índice de insatisfaçäo foi de 19,48 por cento tendo como causa principal o desejo de ter mais filhos. Os autores concluem que uma orientaçäo adequada a respeito do método é essencial para diminuir o índice de insatisfaçäo e propöem um programa de orientaçäo multidisciplinar.