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Background: Since failure in recognition of abnormal cells by the immune system has an important role in bladder cancer development and progression, this study aimed to evaluate whether PD1 (c.627+252C>T) and PD1.5 (c.804C>T) single-nucleotide variants (SNVs) in PDCD1 gene, enrolled in modulation of T lymphocyte activity, influence risk, clinicopathological aspects, and outcome of non-muscle-invasive bladder cancer (NMIBC) patients. Material and methods: DNA genotyping by real-time polymerase chain reaction was offered to 160 non muscle invasive bladder cancer (NMIBC) patients and 250 controls. One hundred and twenty-seven patients treated with bladder transurethral resection and intravesical bacillus Calmette-Guérin were enrolled in survival analyses. Results: Individuals with PD1.5 CC genotype had 2.3-fold increased risk of developing NMIBC. Similar genotype and haplotype frequencies were seen in patients stratified by clinicopathological aspects. Patients with T allele, CT or TT plus CT or TT genotype and TT haplotype of PD1 and PD1.5 SNVs had up to 4.0-times greater chances of presenting NMIBC relapse and death by any cause than the remaining patients, but analysis of NMIBC specific survival was not possible in study due to the small number of patients evolving to death during follow up. Conclusions: Our data presented for the first time, preliminary evidence that inherited abnormality in regulation of T lymphocyte activity alters NMIBC risk.

Ultraviolet light exposure and cutaneous pigmentation are important host risk factors for cutaneous melanoma (CM), and it is well known that inherited ability to produce melanin varies in humans. The study aimed to identify single-nucleotide variants (SNVs) on pigmentation-related genes with importance in risk and clinicopathological aspects of CM. The study was conducted in two stages. In stage 1, 103 CM patients and 103 controls were analyzed using Genome-Wide Human SNV Arrays in order to identify SNVs in pigmentation-related genes, and the most important SNVs were selected for data validation in stage 2 by real-time polymerase-chain reaction in 247 CM patients and 280 controls. ADCY3 c.675+9196T>G, CREB1 c.303+373G>A, and MITF c.938-325G>A were selected for data validation among 74 SNVs. Individuals with CREB1 GA or AA genotype and allele “A” were under 1.79 and 1.47-fold increased risks of CM than others, respectively. Excesses of CREB1 AA and MITF AA genotype were seen in patients with tumors at Clark levels III to V (27.8% versus 13.7%) and at III or IV stages (46.1% versus 24.9%) compared to others, respectively. When compared to others, patients with ADCY3 TT had 1.89 more chances of presenting CM progression, and those with MITF GA or AA had 2.20 more chances of evolving to death by CM. Our data provide, for the first time, preliminary evidence that inherited abnormalities in ADCY3, CREB1, and MITF pigmentation-related genes, not only can increase the risk to CM, but also influence CM patients’ clinicopathological features.

This study aimed to evaluate whether PD1.1 (c.-606G>A), PD1 (c.627 + 252C>T), PD1.5 (c.804C>T), and PD1.9 (c.644C>T) single nucleotide polymorphisms of PDCD1 gene influence the risk, clinicopathological aspects, and survival of cutaneous melanoma (CM). Individuals with phototype I or II and PD1 CC genotype were under 5.89-fold increased risk of developing CM. PD1.5 TT genotype increased PDCD1 expression (2.49 versus 1.28 arbitrary units, p = .03) and PD1.5 CT or TT genotype and allele T increased PD1 expression in TCD4+ lymphocytes (16.6 versus 12.5%, p = .01; 17.0 versus 13.1%, p = .006). At 60 months of follow-up, short recurrence-free survival was seen in patients with PD1.1 AA genotype (33.3 versus 71.8%, p = .03). Patients with PD1.1 AA and PD1.5 CC genotype had 4.21 and 2.62 more chances of presenting relapse and evolving death by disease in Cox analyses, respectively. Our data provide preliminary evidence that abnormalities in regulation of T lymphocyte alter CM risk, clinical aspects, and prognosis.

Genetic susceptibility observed in Hodgkin lymphoma (HL) – one common cancer in young adults – relates to dysfunction of immune system [1].The receptor programed death-1 (PD-1) in T lymphocytes has...

We aimed to evaluate whether variants in repair (XPD Asp312Asn, XPD Lys751Gln) and detoxification (GSTM1, GSTT1) genes alter risk, clinicopathological aspects and survival of cutaneous melanoma (CM). Genotyping was performed in 229 CM patients and 258 controls. Individuals with XPD 312Asp/Asn or Asn/Asn plus GSTT1 null genotype were under 2.00 (95% CI: 1.06-3.79), and XPD 312Asn/Gln haplotype was under 1.44-fold (95% CI: 0.99-2.08) increased risks to CM than others. Individuals with GSTM1 plus GSTT1 null genotype had 9.61-fold (95% CI: 2.28-40.38) increased risk of metastatic CM. At 60 months of follow-up, patients with XPD 751Gln/Gln plus GSTT1 null and GSTM1 null plus GSTT1 null genotype presented 7.36 and 3.05 more chances of evolving to death in multivariate Cox analysis, respectively. In conclusion, our data indicate, for the first time, that specific variant combinations of XPD, GSTM1 and GSTT1 may increase susceptibility to CM and influence patients' clinicopathological features and survival.

Background The JAK/STAT signaling pathway supports the development, progression and metastatic potential of cutaneous melanoma (CM). In normal cells the activation of STAT3 is rapid and transient, but in abnormal melanocytes the activation of the protein occurs in a constitutive manner favoring tumor expansion. STAT3 is coded by the polymorphic gene in humans and thus it is possible that normal individuals show inherited differences in pathway functionality. The aim of the study was to evaluate whether STAT3 c.*1671T>C and STAT3 c.-1937C>G variants influence the risk of CM patients and its functional consequences. Methods We evaluated 248 MC patients and 274 controls. DNA was analyzed by real-time polymerase chain reaction (PCR) for genotyping. Luciferase assay and gene expression in a genetically modified SK-MEL-28 melanoma cell line to present the STAT3 c.-1937C>G ancestral and variant genotypes were realized in study. Differences between groups were assessed by the Fisher or chi-square test. Comparisons of luciferase and gene expression were performed using t-tests and ANOVA or Mann-Whitney and Kruskal-Wallis tests. Results Individuals with STAT3 c.*1671TT and T allele, and STAT3 c.-1937CC genotype had 1.76 (95% CI: 1.08-2.85, P = 0.02), 1.42 (95% CI: 1.03-1.96, P = 0.02) and 1.67 (95% CI: 1.06-2.63, P = 0.02)-fold increased risks of CM than individuals with the other genotypes and allele, respectively. Individuals with TC haplotype of the STAT3 variants were at 1.70 (95% CI: 1.22-2.36, P = 0.001)-fold increased risk of CM than those with other haplotypes. In a genetically modified melanoma cell line to present the distinct genotypes of STAT3 c.-1937C> G, we observed a 30% increase in the promoter activity of the gene and increase of mRNA in cells with the CC genotype compared to those with the GG genotype. Conclusions The data present, for the first time, preliminary evidence that inherited abnormalities in the JAK/STAT pathway, related to STAT3 gene, alter the CM risk. These results may contribute to identify individuals at high risk for CM, who deserve special measures for prevention or early diagnosis. Legal entity responsible for the study The authors. Funding FAPESP and CAPES. Disclosure All authors have declared no conflicts of interest.

Background Cutaneous melanoma (CM) deserves prominence among tumors due to its high mortality. Single nucleotide variants (SNVs) in genes of immune system pathways in T lymphocytes and abnormal melanocytes may favor tumor proliferation and progression. The aim of the present study was to verify whether SNVs JAK1 c.1648 + 1272G>A and c.991-27C>T and JAK2 c.-1132G>T and c.-139G>A alter the risk of CM and prognosis of CM patients. Methods We analysed 248 patients with CM seen at diagnosis at the General Hospital and 274 blood donors (controls) from the Haematology and Haematology Centre of the University of Campinas (UNICAMP). Genotypes of SNVs were identified in DNA of blood samples by real time polimerase chain reaction (RT-PCR), and gene expressions were evaluated by quantitative PCR (qPCR). The statistical meaning of diferences between groups was calculated by t test and Mann-Whitney test. The logistic regression was used to obtain odds ratio (ORs) adjusted for age, cutaneous phototype, nevi and sun exposure, considering the 95% confidence interval (CI). The progression free survival and overall survival were obtained using the Kaplan-Meier estimates and differences between curves were analyzed by the log-rank test. The prognostic value of each variable in patients survival was verified through the univariate and multivariate Cox analyses. Results Patients with the JAK1 c.991-27CC genotype (1,22 vs. 0,75 UA, P = 0,01) and allele C (1,11 vs. 0,75 UA, P = 0,02) showed higher mRNA than others. The median follow-up of CM patients was 93 months (variation: 5-221 months). In univariate Cox analysis, patients with the JAK1 c.1648 + 1272GG and JAK1 c.991-27CC isolated genotypes and JAK1 c.1648 + 1272GG plus JAK1 c.991-27CC combined genotype had 1,78, 1,71 and 1,82 more chances of presenting progression of disease, respectively. Patients with JAK1 c.1648 + 1272GG plus JAK1 c.991-27CC plus JAK2 c.-1132GG plus JAK2 c.-139GG combined genotype had 2,11 more chance of presenting disease progression than others. Conclusions Our data suggest, for the first time, that JAK1 c.1648 + 1272G>A and c.991-27C>T and JAK2 c.-1132G>T and c.-139G>A SNVs of the JAK/STAT intracellular signaling pathway can alter JAK1 expression and prognosis of CM patients. Legal entity responsible for the study The authors. Funding CAPES, FAPESP. Disclosure All authors have declared no conflicts of interest.

Orientador: Carmen Silvia Passos Lima Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas Resumo: Melanócitos anormais podem ser eliminados do organismo por atividade de linfócitos T. O gene PDCD1 codifica o receptor PD1 em linfócitos T, relacionado à regulação do sistema imune, e é polimórfico em humanos. Assim, é possível que indivíduos saudáveis herdem habilidades distintas para eliminar melanócitos anormais e desenvolver melanoma cutâneo (MC). Os objetivos do estudo foram os de verificar se os polimorfismos de nucleotídeo único (SNPs) PD1.1 c.-606G>A, PD1 c.627+252C>T, PD1.5 c.804C>T e PD1.9 c.644C>T do gene PDCD1 influenciam no risco de MC, nas manifestações clínicas e biológicas do tumor, na expressão do gene PDCD1 e na sobrevida dos pacientes com MC. Foram avaliados 250 pacientes com MC e 250 controles (doadores de sangue). Os genótipos dos SNPs foram identificados em DNA em leucócitos do sangue periférico, por meio da reação em cadeia da polimerase (PCR) em tempo real. A expressão do gene PDCD1 foi avaliada por PCR quantitativo. As diferenças entre grupos foram avaliadas por meio do teste de Fisher ou qui-quadrado. Os riscos de ocorrência de MC a que os indivíduos foram submetidos, foram obtidas por meio das razões das chances (ORs). As comparações da expressão gênica de indivíduos com os genótipos distintos de cada SNPs foram realizadas por meio do teste de Mann-Whitney. A sobrevida livre de recidiva (SLR) e a global (SG) dos pacientes foram avaliadas pelo método de Kaplan-Meier/teste log-rank e por análises univariada e multivariada de Cox. Indivíduos com o genótipo PD1 CC isolado e associado ao genótipo PD1.5 CC estiveram sob riscos 2,20 e 2,51 vezes maiores de desenvolvimento do MC do que os demais, respectivamente. Indivíduos com o fototipo I ou II e com o genótipo CC do SNP PD1 e CC + CC dos SNPs PD1 e PD1.5 estiveram sob riscos 5,89 vezes e 6,71 vezes maiores de MC do que os demais. O genótipo PD1.5 TT foi associado com maior expressão do gene PDCD1 do que os genótipos CT ou CC (1,28 versus 2,49 UA; p= 0.03). Os pacientes com o genótipo AA do PD1.1 apresentaram menor SLR do que os com os genótipos GA ou GG. Acreditamos que os nossos resultados possam contribuir para identificar indivíduos com alto risco para a ocorrência do MC e de portadores do tumor com prognóstico desfavorável, que mereçam receber atenção especial na prevenção, diagnóstico precoce e terapêutica diferenciada Abstract: Abnormal melanocytes can be eliminated from the body by T lymphocytes. The PDCD1 gene encoding the receptor PD1 on T cells is related to the immune system regulation, and it is polymorphic in humans. Thus, it is possible for healthy individuals to inherit different abilities for eliminating abnormal melanocytes and developing cutaneous melanoma (CM). This study aimed to assess whether the single nucleotide polymorphisms (SNPs) PD1.1 c.-606G>A, PD1 c.627+252C>T, PD1.5 c.804C>T and PD1.9 c.644C>T of PDCD1 gene influence the risk of CM, clinical and biological manifestations of the tumor, expression of PDCD1 gene, and survival of patients with CM. We evaluated 250 patients with CM and 250 controls (blood donors). Genotypes of SNPs were identified in DNA from peripheral blood leukocytes, by real-time polymerase chain reaction (PCR). The PDCD1 gene expression was assessed by quantitative PCR. The statistical significance of differences between groups was calculated using the Fisher's exact or chi-square test. The occurrence of CM risks in patients and controls, was accessed by the odds ratios (ORs). Comparisons of gene expression in individuals with different genotypes for each SNP were performed using the Mann-Whitney test. The recurrence-free survival (RFS) and overall (OS) of patients were evaluated by the Kaplan-Meier method/log-rank test, univariate and multivariate analyses of Cox. Individuals with CC genotype PD1 isolated and associated with PD1.5 CC genotype were under 2.20 and 2.51 times greater risk of developing CM than others, respectively. Indivíduas with I or II fototype and the PD1 CC genotype and CC + CC of PD1 e PD1.5 SNPs were under 5.89 times and 6.71 times greater risks of developing CM than others. PD1.5 TT genotype was associated with increased expression of PDCD1 gene than CT or CC genotype (1.28 versus 2.49 UA; p= 0.03). Patients with PD1.1 AA genotype had lower RFS than those with GA or GG genotypes. We believe that our results may help to identify individuals at high risk of developing CM and tumor patients with poor prognosis, deserving special attention in prevention, early diagnosis and differentiated treatment Mestrado Fisiopatologia Médica Mestra em Ciências FAPESP 2014/10042-5

This study aimed to evaluate whether XPC A2920C, XPF T30028C, TP53 Arg72Pro, and GSTP1 Ile105Val polymorphisms alter outcomes of cutaneous melanoma (CM) patients. DNA from 237 CM patients seen at the University of Campinas Teaching Hospital from April 2000 to February 2014 was analyzed by polymerase chain reaction and restriction fragment length polymorphism assays. The prognostic impact of genotypes of polymorphisms on progression-free survival (PFS) and overall survival (OS) of CM patients were examined using the Kaplan-Meier probability estimates and univariate and multivariate Cox regression analyses. At 60 months of follow-up, shorter PFS and OS were seen in patients with XPF CC genotype (48.9 vs. 66.7 %, P = 0.002; 77.9 vs. 83.5 %, P = 0.006, respectively) and XPF CC + TP53 ArgArg (43.6 vs. 65.9 %, P = 0.007; 71.6 vs. 84.8 %, P = 0.006, respectively) compared with those with remaining genotypes (Kaplan-Meier estimates). Patients with XPF CC (hazard ratio (HR) 2.45, P = 0.002; HR 3.77, P = 0.005) and XPF CC + TP53 ArgArg (HR 2.67, P = 0.009; HR 4.04, P = 0.03) genotypes had more chance to present tumor progression in univariate and multivariate analyses, whereas patients with XPF CC (HR 2.78, P = 0.009) and XPF CC + TP53 ArgArg (HR 3.84, P = 0.01) genotypes were under greater risk of progressing to death in univariate analysis, compared with those with the remaining genotypes. The data suggest, for the first time, that inherited abnormalities in DNA repair pathway related to XPF 30028C and TP53 Arg72Pro polymorphisms act as prognostic factors for PFS and OS of CM patients.

Our aim was to evaluate complications and long-term functional outcome in patients who had sphincter reconstruction using the gluteus maximus muscle as the neosphincter after abdominoperineal resection for rectal cancer treatment.Seven patients underwent reconstruction from 2000 to 2010. First, the sigmoid colon was brought down to the perineum as a perineal colostomy, with the procedure protected by a loop ileostomy. Reconstruction of the sphincter mechanism using the gluteus maximus took place 3 months later, and after another 8-12 weeks, the loop ileostomy was closed. We studied the functional outcome of these interventions with follow-up interviews of patients and objectively assessed anorectal function using manometry and the Cleveland Clinic Florida (Jorge-Wexner) fecal incontinence score.The mean follow-up was 56 months (median 47; range 10-123 months). One patient had a perianal wound infection and another had fibrotic stricture in the colocutaneous anastomosis that required several digital dilatations. Anorectal manometry at 3-month follow-up showed resting pressures from 10 to 18 mm Hg and voluntary contraction pressures from 68 to 187 mm Hg. Four patients had excellent sphincter function (Jorge-Wexner scores ≤5).Our preliminary results show that sphincter reconstruction by means of gluteus maximus transposition can be effective in restoring gastrointestinal continuity and recovering fecal continence in patients who have undergone APR with permanent colostomy for rectal cancer. Furthermore, the reconstruction procedure can be performed 2-4 years after the APR.

9038 Background: XPC, XPF, TP53 and GSTP1 genes act on repair and detoxification of ultraviolet (UV) radiation-induced DNA damage, which are related to cutaneous melanoma (CM) development and progr...

Aim: XPC, XPF and P53 genes act on nucleotide excision repair of ultraviolet radiation-induced DNA damage and are involved in cutaneous melanoma (CM) development. The DNA repair ability is variable in humans.Variant C and wild Arg alleles of XPC A2920C and P53 Arg72Pro polymorphims encode proteins with lower activities in DNA repair than those encoded by other alleles, respectively. Variant C allele of XPF T30028C influences mRNA stability and affects levels of protein expression. This study aimed to evaluate whether these polymorphisms are associated with relapse-free survival (RFS) and overall survival (OS) of CM patients. Methods: We analyzed 232 CM patients diagnosed from 1989 to 2013 at a University Hospital. Genomic DNA of patients was analyzed by polymerase chain reaction, and enzymatic digestion, for discrimination of pertinent genotypes. RFS and OS were calculated using the univariate Kaplan-Meier estimate probabilities, and differences between survival curves were analyzed by the log-rank test. OS was calculated from the date of diagnosis until the date of death or last follow-up. Results: The median period of observation of patients in the study was 57 months (range: 1-285). At 60 months of follow-up, we observed that RFS of patients with XPF TT + TC genotypes was higher than that found in patients with CC genotype (73% vs 30%, P = 0.007). At this time, RFS was also higher in patients with XPF TT + TC plus P53 ArgPro + ProPro genotypes than in those with XPF CC plus P53 ArgArg genotypes (74% vs 19%, P = 0.004). At 120 months of follow-up, we observed that OS of patients with XPF TT + TC genotypes was higher than that observed in patients with CC genotype (86% vs 63%, P = 0.05). OS of patients with XPF TT + TC plus P53 ArgPro + ProPro genotypes was also higher than that found in patients with XPF CC plus P53 ArgArg genotypes (89% vs 68%, P = 0.07) at this time. In contrast, RFS and OS were worse in patients with Breslow thickness ≥1.5 mm (87% vs 52%, P = 0.0001; 88% vs 79%, P = 0.008), Clark level IV + V (79% vs 60%, P = 0.004; 86% vs 78%, P = 0.02) and advantage stage III + IV (77% vs 48% P = 0.0001; 88% vs 66% P = 0.0001), respectively. Conclusions: The data found herein provide evidence, for the first time, that inherited abnormality in DNA repair pathway related to XPF and P53 polymorphisms influence RFS and OS in CM patients. Disclosure: All authors have declared no conflicts of interest.

A total of 124 women who re-inserted a TCu-200B intrauterine device (IUD) following an expulsion were followed up for 1 year after the IUD re-insertion. The cumulative expulsion rate at 6 months was 21.7 and 31.4 per 100 women at the end of the first year. Women who expelled the first IUD within the first 3 months of use had a significantly higher expulsion rate with the second IUD. Our conclusion is that women who expelled a copper IUD are at a significantly higher risk of expelling the re-inserted IUD than the first IUD. Health workers must also be informed about the high probability of another expulsion when re-inserting an IUD after an expulsion.Researchers analyzed retrospective data on 124 women aged 16-35 who had experienced spontaneous expulsion of the copper T-200 B IUD and then re-insertion of the same type of IUD at the family planning clinic of the State University of Campinas in Brazil during August 1986-December 1992. They were followed for 12 months after IUD re-insertion. Two senior nurses/midwives and two senior physicians performed 86 and 38 of the IUD re-insertions, respectively. Even though the re-expulsion rate was higher when performed by physicians than nurses (34.2% vs. 26.7%), the difference was not statistically significant. The net cumulative re-expulsion rates at 6 and 12 months were high (21.7% and 31.4%, respectively). There were few removals for medical and personal reasons. The 12-month continuation rate stood at 64.3%. Women who had experienced the first expulsion within 3 months after insertion had a significantly higher re-expulsion rate than those whose first expulsion occurred 3 months after insertion (41% vs. 18%; p 0.001). Both these re-expulsion rates were higher than the expulsion rate during the first period of IUD use in the same family planning clinic (3.1-3.9%). These findings suggest that women who have already experienced expulsion of a copper IUD face a much higher risk of expelling the re-inserted IUD than the first IUD. They also indicate that an expulsion after IUD re-insertion due to an earlier expulsion is more likely to happen than after a re-insertion due to life-span expiration. Providers should perform careful follow-up and adequate counseling to all IUD women who have had an IUD re-inserted after expulsion during the first year. The providers should also be informed about the high probability of another expulsion when re-inserting an IUD after an expulsion.