Pregnancy outcomes following in vitro fertilization frozen embryo transfer (IVF-FET) with or without preimplantation genetic testing for aneuploidy (PGT-A) in women with recurrent pregnancy loss (RPL): a SART-CORS study.

STUDY QUESTION: Can preimplantation genetic testing for aneuploidy (PGT-A) improve the live birth rate in patients with recurrent pregnancy loss (RPL)? SUMMARY ANSWER: PGT-A use was associated with improved live birth rates in couples with recurrent pregnancy loss undergoing frozen embryo transfer (IVF-FET). WHAT IS KNOWN ALREADY: Euploid embryo transfer is thought to optimize outcomes in some couples with infertility. There is insufficient evidence, however, supporting this approach to management of recurrent pregnancy loss. STUDY DESIGN, SIZE, DURATION: This study included data collected by the Society of Assisted Reproductive Technologies Clinical Outcomes Reporting System (SART-CORS) for IVF-FET cycles between years 2010 through 2016. A total of 12 631 FET cycles in 10 060 couples were included in this analysis designed to assess the utility of PGT-A in couples with RPL undergoing FET, including 4287 cycles in couples with tubal disease who formed a control group. PARTICIPANTS/MATERIALS, SETTING, METHODS: The experimental group included couples with RPL (strictly defined as a history of 3 or more pregnancy losses) undergoing FET with or without PGT-A. The primary outcome was live birth rate. Secondary outcomes included rates of clinical pregnancy, spontaneous abortion, and biochemical pregnancy loss. Differences were analyzed using generalized estimating equations logistic regression models to account for multiple cycles per patient. Covariates included in the model were age, gravidity, geographic region, race/ethnicity, smoking history, and indication for assisted reproductive technologies. Analyses were stratified for age groups as defined by SART: <35 years, 35-37 years, 38-40 years, 41-42 years, and >42 years. MAIN RESULTS AND THE ROLE OF CHANCE: In women with a diagnosis of RPL, the adjusted odds ratio (OR) comparing IVF-FET with PGT-A versus without PGT-A for live birth outcome was 1.31 (95% CI: 1.12, 1.52) for age <35 years, 1.45 (95% CI: 1.21, 1.75) for ages 35-37 years, 1.89 (95% CI: 1.56, 2.29) for ages 38-40, 2.62 (95% CI: 1.94-3.53) for ages 41-42, and 3.80 (95% CI: 2.52, 5.72) for ages >42 years. For clinical pregnancy, the OR was 1.26 (95% CI: 1.08, 1.48) for age <35 years, 1.37 (95% CI: 1.14, 1.64) for ages 35-37 years, 1.68 (95% CI: 1.40, 2.03) for ages 38-40 years, 2.19 (95% CI: 1.65, 2.90) for ages 41-42, and 2.31 (95% CI: 1.60, 3.32) for ages >42 years. Finally, for spontaneous abortion, the OR was 0.95 (95% CI: 0.74, 1.21) for age <35 years, 0.85 (95% CI: 0.65, 1.11) for ages 35-37 years, 0.81 (95% CI: 0.60, 1.08) for ages 38-40, 0.86 (95% CI: 0.58, 1.27) for ages 41-42, and 0.58 (95% CI: 0.32, 1.07) for ages >42 years. LIMITATIONS, REASONS FOR CAUTION: The retrospective collection of data including only women with recurrent pregnancy loss undergoing FET presents a limitation of this study, and results may not be generalizable to all couples with recurrent pregnancy loss. Also, data regarding evaluation and treatment for RPL for the included women is unavailable. WIDER IMPLICATIONS OF THE FINDINGS: This is the largest study to date assessing the utility of PGT-A in women with RPL. PGT-A was associated with improvement in live birth and clinical pregnancy in women with RPL, with the largest difference noted in the group of women with age greater than 42 years. Couples with RPL warrant counseling on all management options to reduce subsequent miscarriage, which may include IVF with PGT-A for euploid embryo selection. STUDY FUNDING/COMPETING INTEREST(S): There are no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Real-world efficacy and safety of lenvatinib: data from a compassionate use in the treatment of radioactive iodine-refractory differentiated thyroid cancer patients in Italy.

BACKGROUND: Lenvatinib is a multi-kinase inhibitor approved for patients with radioactive iodine (RAI)-resistant differentiated thyroid cancer (DTC). Before the drug approval from the Italian National Regulatory Agency, a compassionate use programme has been run in Italy. This retrospective study aimed to analyse data from the first series of patients treated with lenvatinib in Italy. METHODS: The primary aim was to assess the response rate (RR) and progression-free survival (PFS). Secondary end-points include overall survival (OS) and toxicity data. RESULTS: From November 2014 to September 2016, 94 patients were treated in 16 Italian sites. Seventeen percent of patients had one or more comorbidities, hypertension being the most common (60%). Ninety-eight percent of patients were treated by surgery, followed by RAI in 98% of cases. Sixty-four percent of patients received a previous systemic treatment. Lenvatinib was started at 24 mg in 64 subjects. Partial response and stable disease were observed in 36% and in 41% of subjects, respectively; progression was recorded in 14% of patients. Drug-related side-effects were common; the most common were fatigue (13.6%) and hypertension (11.6%). Overall, median PFS and OS were 10.8 months (95% confidence interval [CI], 7.7-12.6) and 23.8 months (95% CI, 19.7-25.0) respectively. CONCLUSION: Lenvatinib is active and safe in unselected, RAI-refractory, progressive DTC patients in real-life setting. RR and PFS seem to be less favourable than those observed in the SELECT trial, likely due to a negative selection that included heavily pretreated patients or with poor performance status.

Long-term efficacy and safety of fourth-line multikinase inhibitor treatment with lenvatinib in a young papillary thyroid carcinoma patient.

Lenvatinib, a multikinase inhibitor, is approved for the treatment of patients with radioiodine-refractory metastatic thyroid cancer on the basis of a Phase III, prospective, double-blind, randomized, placebo-controlled trial that showed longer progression-free survival in the drug-treated arm. Here, we report the case of a young papillary thyroid cancer patient, pretreated with three other kinase inhibitors, who achieved a long-term clinical benefit from lenvatinib in the fourth-line setting.

Human exposure to lead, cadmium and mercury through fish and seafood product consumption in Italy: a pilot evaluation.

The presence of selected toxic heavy metals, such as cadmium (Cd), lead (Pb) and mercury (Hg), was investigated in fish and seafood products, namely, blue mussel, carpet shell clam, European squid, veined squid, deep-water rose shrimp, red mullet, European seabass, gilthead seabream, Atlantic cod, European hake, Atlantic bluefin tuna and swordfish so as to assess their human exposure through diet. Metals were detected by quadrupole inductively coupled plasma mass spectrometry (Q-ICP-MS) and hydride generation atomic absorption spectrometry (Hg-AAS). Measurements of Cd, Pb and Hg were performed by means of analytical methods validated in compliance with UNI CEI EN ISO/IEC 17025 [2005. General requirements for the competence of testing and calibration laboratories. Milano (Italy): UNI Ente Nazionale Italiano di Unificazione]. The exposure assessment was undertaken matching the levels of Cd, Pb and total Hg with consumption data related to fish and seafood products selected for this purpose. In order to establish human health implications, the estimated weekly intakes (EWIs) for Cd, Pb and Hg were compared with the standard tolerable weekly intakes (TWI) for Cd and provisional tolerable weekly intakes (PTWIs) for Pb and Hg stipulated by the European Food Safety Authority (EFSA) and the Food and Agriculture Organization/World Health Organization (FAO/WHO) Joint Expert Committee on Food Additives (JECFA). The found metal concentrations were largely below the maximum levels (MLs) established at the European Union level with the exception of Cd. This metal exceeded the MLs in squid, red mullet, European hake and Atlantic cod. Squid and blue mussel showed the highest Pb concentrations which accounted for 60% and 10% of the MLs, respectively. Highest Hg levels were found in predatory fish. The concentrations of Hg in swordfish, Atlantic bluefin tuna and red mullet accounted for 50%, 30% and 30% of the MLs, respectively. The EWIs for Cd, Pb and Hg related to the consumption of fish and seafood products by the median of the Italian total population accounted for 20%, 1.5% and 10% of the standard TWI for Cd as well as PTWIs for Pb and Hg, respectively. Furthermore, the EWIs estimated using consumption data concerning Italian consumers did not exceed the standard TWI and PTWIs, except for Cd at 95th percentile.

Validação do procedimento operacional padrão (POP) de colheita de alimentos importados / Validation of the operational procedure standard (SOP) of imported food

O objetivo desse trabalho foi avaliar o Procedimento Operacional Padrão (POP) de colheita de alimentos importados para verificar a sua legitimidade como instrumento nas ações de vigilância sanitária. O POP visa o controle e a certificação da qualidade dos alimentos importados e foi elaborado por técnicos da Coordenação de Vigilância Sanitária de Portos, Aeroportos e Fronteiras do Estado de São Paulo (CVSPAF/SP) , sendo revisado, juntamente, com a equipe técnica da Divisão de Bromatologia e Química, do Instituto Adolfo Lutz, de São Paulo (IAL/SP). A validação do POP foi realizada na avaliação das condições higiênico-sanitárias de 17 ingredientes e 10 aditivos, produtos alimentícios importados, armazenados numa Estação Aduaneira Interior – EADI, do Estado de São Paulo, que sofreu intempérie decorrente de forte tempestade, com possibilidade de comprometimento da integridade desses produtos.(...) A elaboração desse POP estabeleceu uma adoção de medidas rigorosas, com base legal que garantem a correta ação da vigilância sanitária para a certificação da qualidade dos produtos alimentícios importados.

Evolutionary cytogenetics of the Hoplias lacerdae, Miranda Ribeiro, 1908 group: a particular pathway concerning the other Erythrinidae fish

The taxonomy/systematics of the Erythrinidae fish is still imprecise, with several doubts on their relationships. Karyotypes and chromosomal characteristics of some species of the Hoplias lacerdae group (Erythrinidae), from different Brazilian hydrographic basins and pisciculture stations, were analyzed in the present study, using conventional Giemsa staining, C-banding, silver staining, Mithramycin and Distamycin/DAPI fluorochromes, and fluorescent in situ hybridization (FISH). A diploid chromosome number of 2n = 50 and karyotypes composed of meta- and submetacentric chromosomes without sex-related differences were found. Only one active NOR (Nucleolar Organizer Region) site was found, which was identified by silver staining (Ag-NOR) and FISH, located on the chromosome pair 11, although additional 45S rDNA sites were also mapped on other chromosome pairs only by FISH. The Ag-NOR of the chromosome pair 11 was found to be GC-rich, appearing positive after Mithramycin staining. Mithramycin-positive/DAPI-negative sites were also observed in the centromeric/pericentomeric regions of the chromosome pairs 4, 6, 15, and 19, which have also affinity to silver nitrate. However, these four sites were not detected by FISH with the rDNA probe, indicating to be only argentophilic GC-rich heterochromatic regions. Chromosome data show that the karyotype evolution in Hoplias lacerdae group is relatively conserved and follows a particular pathway concerning the other Erythrinidae fishes, such as Hoplias malabaricus, Hoplerythrinus unitaeniatus, and Erythrinus erythrinus, in which polytypic karyotypes are found. Thus, the H. lacerdae group shows chromosome features that are not closely related to those of the congeneric H. malabaricus group. These finds, together with genetic and morphologic data, are important tools to be considered in a major revision of the Erythrinidae family, as well as for conservation programs.

A taxonomia dos Erythrinidae encontra-se ainda por ser totalmente esclarecida, existindo várias dúvidas sobre suas relações filogenéticas. No presente estudo, foram analisados os cariótipos e características cromossômicas de algumas espécies do grupo Hoplias lacerdae (Erythrinidae), de diferentes bacias hidrográficas do Brasil e estações de piscicultura, empregando-se a coloração Giemsa convencional, bandamento C, coloração com nitrato de prata, fluorocromos Mitramicina e Distamicina/DAPI, e hibridação fluorescente in situ (FISH). O número diplóide de 2n = 50 cromossomos foi constante, sendo os cariótipos formados por cromossomos de dois braços, meta- e submetacêntricos, sem diferenciação relacionada ao sexo. Foi identificado apenas um sítio ativo de região organizadora de nucléolo (NOR), presente no par cromossômico no. 11, identificado tanto pela coloração com nitrato de prata (Ag-NOR) como por FISH com sonda de DNAr 45S, embora alguns sítios adicionais de DNAr tenham sido identificados em outros pares de cromossomos, apenas por FISH. O sítio de Ag-NOR no cromossomo 11 mostrou-se rico em pares de bases GC, sendo Mitramicina-positivo. Sítios Mitramicina-positivos/DAPI-negativos foram igualmente observados nas regiões centroméricas/pericentroméricas dos pares cromossômicos 4, 6, 15 e 19, os quais mostraram também afinidade pelo nitrato de prata. Contudo, estes quatro sítios não foram evidenciados por FISH com sonda de DNAr 45S, indicando serem somente regiões de heterocromatina GC-rica, com afinidade pela prata. Os dados cromossômicos mostram que a evolução cariotípica no grupo Hoplias lacerdae é relativamente conservada e segue um comportamento particular em relação aos outros eritrinídeos, como Hoplias malabaricus, Hoplerythrinus unitaeniatus e Erythrinus erythrinus, nos quais foram encontrados cariótipos politípicos. Assim sendo, o grupo H. lacerdae mostra características cromossômicas discordantes daquelas evidenciadas pelo grupo...

Experience at Italian National Institute of Health in the assessment of the quality in telemedicine: submission to a quality control process.

For applying a quality control of the telemedicine (TM) applications, in order to prevent device and/or service failure and to standardize some features of TM applications, a telemedicine quality control (TM-QC) process was proposed by authors. This TM-QC process was submitted to some TM manufacturers in order to assess their products/services and applied in two following phases, during which a set of procedures and tools were delivered to the TM suppliers. In the 1 degrees phase an Informative Questionnaire (InQu) and the Classification Form (ClFo) were delivered: the InQu gathered all the relevant information about the TM application in exam, the ClFo classified the TM product as belonging to one category (or application area) of TM. In the 2nd phase a Technical File (TF) and a Quality Assessment CheckList (QACL) were delivered: the TF was intended as a technical dossier of product; the QACL was a checklist of requirements that TM products must be agreed. The number of met requirements was computed and a Quality Level (QL), ranging from Basic Quality Level (QL1-B) to the Excellent Quality Level (QL7-E), was assigned to the examined TM products. The tested TM products presented in this paper, reached at least the minimum quality level (QL1-B). The TM-QC process allowed the definition of standardized quality levels for the TM products/services and the proposed procedures should be useful for promoting the using of standardized TM applications, so to speed up the integration process into the National Health Service.

Evolutionary cytogenetics of the Hoplias lacerdae, Miranda Ribeiro, 1908 group: a particular pathway concerning the other Erythrinidae fish

The taxonomy/systematics of the Erythrinidae fish is still imprecise, with several doubts on their relationships. Karyotypes and chromosomal characteristics of some species of the Hoplias lacerdae group (Erythrinidae), from different Brazilian hydrographic basins and pisciculture stations, were analyzed in the present study, using conventional Giemsa staining, C-banding, silver staining, Mithramycin and Distamycin/DAPI fluorochromes, and fluorescent in situ hybridization (FISH). A diploid chromosome number of 2n = 50 and karyotypes composed of meta- and submetacentric chromosomes without sex-related differences were found. Only one active NOR (Nucleolar Organizer Region) site was found, which was identified by silver staining (Ag-NOR) and FISH, located on the chromosome pair 11, although additional 45S rDNA sites were also mapped on other chromosome pairs only by FISH. The Ag-NOR of the chromosome pair 11 was found to be GC-rich, appearing positive after Mithramycin staining. Mithramycin-positive/DAPI-negative sites were also observed in the centromeric/pericentomeric regions of the chromosome pairs 4, 6, 15, and 19, which have also affinity to silver nitrate. However, these four sites were not detected by FISH with the rDNA probe, indicating to be only argentophilic GC-rich heterochromatic regions. Chromosome data show that the karyotype evolution in Hoplias lacerdae group is relatively conserved and follows a particular pathway concerning the other Erythrinidae fishes, such as Hoplias malabaricus, Hoplerythrinus unitaeniatus, and Erythrinus erythrinus, in which polytypic karyotypes are found. Thus, the H. lacerdae group shows chromosome features that are not closely related to those of the congeneric H. malabaricus group. These finds, together with genetic and morphologic data, are important tools to be considered in a major revision of the Erythrinidae family, as well as for conservation programs.

A taxonomia dos Erythrinidae encontra-se ainda por ser totalmente esclarecida, existindo várias dúvidas sobre suas relações filogenéticas. No presente estudo, foram analisados os cariótipos e características cromossômicas de algumas espécies do grupo Hoplias lacerdae (Erythrinidae), de diferentes bacias hidrográficas do Brasil e estações de piscicultura, empregando-se a coloração Giemsa convencional, bandamento C, coloração com nitrato de prata, fluorocromos Mitramicina e Distamicina/DAPI, e hibridação fluorescente in situ (FISH). O número diplóide de 2n = 50 cromossomos foi constante, sendo os cariótipos formados por cromossomos de dois braços, meta- e submetacêntricos, sem diferenciação relacionada ao sexo. Foi identificado apenas um sítio ativo de região organizadora de nucléolo (NOR), presente no par cromossômico no. 11, identificado tanto pela coloração com nitrato de prata (Ag-NOR) como por FISH com sonda de DNAr 45S, embora alguns sítios adicionais de DNAr tenham sido identificados em outros pares de cromossomos, apenas por FISH. O sítio de Ag-NOR no cromossomo 11 mostrou-se rico em pares de bases GC, sendo Mitramicina-positivo. Sítios Mitramicina-positivos/DAPI-negativos foram igualmente observados nas regiões centroméricas/pericentroméricas dos pares cromossômicos 4, 6, 15 e 19, os quais mostraram também afinidade pelo nitrato de prata. Contudo, estes quatro sítios não foram evidenciados por FISH com sonda de DNAr 45S, indicando serem somente regiões de heterocromatina GC-rica, com afinidade pela prata. Os dados cromossômicos mostram que a evolução cariotípica no grupo Hoplias lacerdae é relativamente conservada e segue um comportamento particular em relação aos outros eritrinídeos, como Hoplias malabaricus, Hoplerythrinus unitaeniatus e Erythrinus erythrinus, nos quais foram encontrados cariótipos politípicos. Assim sendo, o grupo H. lacerdae mostra características cromossômicas discordantes daquelas evidenciadas pelo grupo congenérico H. malabaricus. Estes dados, juntamente com os outros dados genéticos e morfológicos, constituem ferramentas importantes a serem consideradas em uma revisão ampla da família Erythrinidae, assim como para programas de conservação.

Quality assessment and cataloguing of telemedicine applications.

We created a Web catalogue of approved telemedicine systems that authoritative Italian research bodies had made available for more general use. The evaluation process was divided into two stages: (1) classification of the telemedicine systems and rough preliminary evaluation; (2) assessment of the telemedicine products and services. The scoring method was applied to four well-known telemedicine systems that had been tested in health-care settings: an echocardiology teleconsulting and analysis system; a ward nursing management system; a virtual cooperative system for the management of oncology patients and a telepathology system based on remotely controlled microscopy. After technical revision during the standardization/qualification process, the applications were transferred successfully to eight new health-care facilities. The methodology achieved the main goal of providing effective tools, such as a set of quality control procedures for telemedicine and telehealth projects and a Web catalogue of telemedicine applications with a standardized level of quality, available to all interested parties.

Chromosomal analysis of Astyanax fasciatus (PISCES, CHARACIDAE) from the Araguari river, Uberlândia, MG, Brazil

O gênero Astyanax é um dos mais numerosos da família Characidae, englobando um grande número de espécies muito semelhantes na forma, mas que apresentam inúmeras variações cariotípicas no número e/ou estrutura cromossômica. Encontram-se descritas na literatura populações de A. fasciatus com o número diplóide variando de 2n = 45 a 2n = 48 cromossomos. No presente trabalho, espécimes de Astyanax fasciatus da população da calha do rio Araguari (bacia do Alto Paraná), foram caracterizados citogeneticamente. O número diplóide observado foi 2n = 46 cromossomos. As regiões organizadoras de nucléolo (NORs), detectadas com nitrato de prata, apresentaram um sistema múltiplo, com dois pares de cromossomos marcados. Esses dados estão de acordo com os descritos para outras populações da mesma espécie.

The immune response of women with prolonged exposure to electromagnetic fields produced by radiotelevision broadcasting stations.

Twelve women, five of them housewives, exposed in their residences to electromagnetic fields (EMFs)emitted by radio-television broadcasting stations for a mean period of 13 years, were investigated. The EMFs in the balconies of the homes were (mean + S.D.) 4.3 + 1.4 V/m in the year 2000 and 3.7 + 1.3 V/m in 2005, while the exposure in the nearby area was <2.0 V/m. The EMF exposed women showed in 2000 reduced blood NK lymphocytes as well as PHA stimulated PBMC proliferation and IL-2 and IFN-gamma release. In the year 2005, the EMF exposed women and 48 control women with similar ages(mean 43 years), smoking habits, atopy and social level were investigated. State (temporary) and trait(tendency of the personality) anxiety were determined by STAI I and II, respectively. Blood cytotoxic activity and lymphocyte subsets were also determined. The ratio STAI I/STAI II of the EMF exposed group was lower than that of the control group. The blood cytotoxic activity of the exposed women was lower (p<0.01), percent of B CD45+-CD19+ lymphocytes higher and percent of CD45+-CD3+-CD8+ cells lower (p<0.05). Moreover, cytotoxic activity/CD45+-CD16+-56+ NK lymphocytes of the controls was negatively correlated with STAI I and STAI II (p<0.001). In conclusion, this study demonstrates reduced blood cytotoxic activity and increased trait anxiety in relation to state anxiety in EMF exposed women. An effect of EMFs on immune functions, in part mediated by nervous mechanisms, may be hypothesized. However, the influence of lifestyle may not be excluded.

Chromosomal analysis of Astyanax fasciatus (PISCES, CHARACIDAE) from the Araguari river, Uberlândia, MG, Brazil

The genus Astyanax is one of the most numerous of the family Characidae, comprising a large number of similar-shaped species, but displaying innumerable karyotypic variations in its chromosome number and/or structure. The literature describes A. fasciatus populations with diploid chromosome numbers varying from 2n = 45 to 2n = 48. In this study, A. fasciatus specimens captured in the Araguari River (Alto Paraná basin) were cytogenetically characterized, revealing a diploid chromosome number of 2n = 46. The nucleolar organizing regions (NORs), detected with silver nitrate staining, showed a multiple system with two pairs of marked chromosomes. These findings are congruent with those of other studies involving populations of the same species.

O gênero Astyanax é um dos mais numerosos da família Characidae, englobando um grande número de espécies muito semelhantes na forma, mas que apresentam inúmeras variações cariotípicas no número e/ou estrutura cromossômica. Encontram-se descritas na literatura populações de A. fasciatus com o número diplóide variando de 2n = 45 a 2n = 48 cromossomos. No presente trabalho, espécimes de Astyanax fasciatus da população da calha do rio Araguari (bacia do Alto Paraná), foram caracterizados citogeneticamente. O número diplóide observado foi 2n = 46 cromossomos. As regiões organizadoras de nucléolo (NORs), detectadas com nitrato de prata, apresentaram um sistema múltiplo, com dois pares de cromossomos marcados. Esses dados estão de acordo com os descritos para outras populações da mesma espécie.

Infective endocarditis from Enterococcus faecalis complicating colonoscopy in Heyde's syndrome.

A case of infective endocarditis from Enterococcus faecalis after colonoscopy in a patient with aortic stenoinsufficiency and bleeding intestinal angiodysplasia (Heyde's syndrome) is reported.A 77 year old man with aortic stenoinsufficiency presented with enterorrhagia and underwent a colonoscopy, which showed normal findings. Fifteen days later he developed a moderate degree of fever. Blood cultures were positive for E faecalis. An echocardiogram showed aortic valve vegetations, and infective endocarditis was diagnosed and successfully treated by antibiotics. Some months later, intestinal bleeding recurred and intestinal resection was performed. Histopathology showed angiodysplasia. In patients with Heyde's syndrome antibiotic prophylaxis should be considered before colonoscopy.

Iron, hepatic stellate cells and fibrosis in chronic hepatitis C.

BACKGROUND/AIMS: In patients with chronic hepatitis C, hepatic iron concentration correlates with liver fibrosis. However, it is not clear whether this correlation merely reflects the presence of more active disease, or iron exacerbates chronic hepatitis C virus (HCV)-induced damage through activation of hepatic stellate cells and regeneration of hepatocytes. MATERIALS AND METHODS: We studied 72 HCV-positive patients, staged according to the Ishak's score system. We measured hepatic iron concentration with spectrophotometry and evaluated the number of hepatic stellate cells (using monoclonal antibody against alpha smooth muscle actin) and proliferating hepatocytes (using monoclonal antibody against Ki67). Iron and ferritin serum levels were also determined. RESULTS: Hepatic iron concentration correlated statistically with ferritin serum level (r = 0.59, P < 0.001), with grading (r = 0.47, P < 0.001) and staging (r = 0.51, P < 0.001) scores for chronic hepatitis in the whole group of patients. Hepatic iron concentration correlated positively with stellate cell number (r = 0.55, P = 0.004) and Ki67-positive hepatocyte number (r = 0.36, P = 0.08) in patients with chronic hepatitis C and low grading score (< 3). CONCLUSIONS: In patients with chronic hepatitis C and low grading score, hepatic iron could play a role in the activation of hepatic stellate cells and in the progression of fibrosis.

Parathyroid hormone activation of map kinase in rat duodenal cells is mediated by 3',5'-cyclic AMP and Ca(2+).

In a previous study, we demonstrated that parathyroid hormone (PTH) stimulates in rat duodenal cells (enterocytes) the phosphorylation and activity of extracellular signal-regulated mitogen-activated protein kinase (MAPK) isoforms ERK1 and ERK2. As PTH activates adenylyl cyclase (AC) and phospholipase C and increases intracellular Ca(2+) in these cells, in the present study we evaluated the involvement of cAMP, Ca(2+) and protein kinase C (PKC) on PTH-induced MAPK activation. We found that MAPK phosphorylation by the hormone did not depend on PKC activation. PTH response could, however, be mimicked by addition of forskolin (5-15 microM), an AC activator, or Sp-cAMP (50-100 microM), a cAMP agonist, and suppressed to a great extent by the AC inhibitor, compound Sq-22536 (0.2-0.4 mM) and the cAMP antagonist Rp-cAMP (0.2 mM). Removal of external Ca(2+) (EGTA 0.5 mM), chelation of intracellular Ca(2+) with BAPTA (5 microM), or blockade of L-type Ca(2+)-channels with verapamil (10 microM) significantly decreased PTH-activation of MAPK. Furthermore, a similar degree of phosphorylation of MAPK was elicited by the Ca(2+) mobilizing agent thapsigargin, the Ca(2+) ionophore A23187, ionomycin and membrane depolarization with high K(+). Inclusion of the calmodulin inhibitor fluphenazine (50 microM) did not prevent hormone effects on MAPK. Taken together, these results indicate that cAMP and Ca(2+) play a role upstream in the signaling mechanism leading to MAPK activation by PTH in rat enterocytes. As Ca(2+) and cAMP antagonists did not block totally PTH-induced MAPK phosphorylation, it is possible that linking of the hormone signal to the MAPK pathway may additionally involve Src, which has been previously shown to be rapidly activated by PTH. Of physiological significance, in agreement with the mitogenic role of the MAPK cascade, PTH increased enterocyte DNA synthesis, and this effect was blocked by the specific inhibitor of MAPK kinase (MEK) PD098059, indicating that hormone modulation of MAPK through these messenger systems stimulates duodenal cell proliferation.

Clinical analysis of families with heart, midline, and laterality defects.

Disturbances of the normal asymmetric placement of organs, such as polysplenia or situs inversus, have been defined traditionally as laterality defects. However, there is compelling evidence from vertebrate models and human birth defects to hypothesize that defects of the midline, isolated congenital heart defects, and laterality defects are etiologically related. We present the clinical characteristics of three families that exhibit a variety of midline defects and isolated heart defects in addition to laterality defects. These observations suggest that the phenotypic consequences of mutations causing laterality defects include defects of the midline as well as isolated heart defects. To further explore the relationship between midline, heart, and laterality defects, it is imperative that detailed phenotyping of individuals and families with laterality defects be done and a classification system created to facilitate identification of genes causing human laterality disorders.

Cardiac involvement in patients with Wegener's granulomatosis.

Wegener's granulomatosis (WG) is a systemic necrotizing vasculitis, which could potentially affect any organ system. However, there have only been a few reports on cardiac involvement. We described the echocardiographic findings in nine patients affected by WG. A complete M-mode, two-dimensional, Doppler and color-Doppler transthoracic echocardiogram was performed in nine patients (seven females and two males) affected by WG. In each patient, cardiac abnormality, for example, valvular damage, left ventricular global systolic dysfunction, or pericardial effusion, was detected. In particular, heart valve disease was found in eight patients, and in three cases, aortic valve insufficiency, which was severe enough to require surgical valve replacement, was observed. Cardiac involvement in patients with WG is common. In particular, there is a high frequency of aortic valve abnormalities. Thus, an echocardiographic study should be routinely performed.

Involvement of tyrosine kinase activity in 1alpha,25(OH)2-vitamin D3 signal transduction in skeletal muscle cells.

In cultured chick skeletal muscle cells loaded with Fura-2, the tyrosine kinase inhibitors herbimycin A and genistein abolished both the fast inositol 1,4,5-trisphosphatedependent Ca(2+) release from internal stores and extracellular Ca(2+) influx induced by 1alpha, 25(OH)(2)-vitamin D(3) (1alpha,25(OH)(2)D(3)). Daidzein, an inactive analog of genistein, was without effects. Tyrosine phosphatase inhibition by orthovanadate increased cytosolic Ca(2+). Anti-phosphotyrosine immunoblot analysis revealed that 1alpha, 25(OH)(2)D(3) rapidly (0.5-10 min) stimulates in a concentrationdependent fashion (0.1-10 nm) tyrosine phosphorylation of several myoblast proteins, among which the major targets of the hormone could be immunochemically identified as phospholipase Cgamma (127 kDa), which mediates intracellular store Ca(2+) mobilization and external Ca(2+) influx, and the growth-related proteins mitogen-activated protein (MAP) kinase (42/44 kDa) and c-myc (65 kDa). Genistein suppressed the increase in phosphorylation and concomitant elevation of MAPK activity elicited by the sterol. Both genistein and the MAPK kinase (MEK) inhibitor PD98059 abolished stimulation of DNA synthesis by 1alpha,25(OH)(2)D(3). The sterol-induced increase in tyrosine phosphorylation of c-myc, a finding not reported before for cell growth regulators, was totally suppressed by the specific Src inhibitor PP1. These results demonstrate that tyrosine phosphorylation is a previously unrecognized mechanism involved in 1alpha,25(OH)(2)D(3) regulation of Ca(2+) homeostasis in hormone target cells. In addition, the data involve tyrosine kinase cascades in the mitogenic effects of 1alpha, 25(OH)(2)D(3) on skeletal muscle cells.

Flutamide-induced acute hepatitis: investigation on the role of immunoallergic mechanisms.

Flutamide is a nonsteroidal antiandrogen drug used in the treatment of prostatic cancer. Hepatotoxic reactions due to flutamide have been reported with an incidence ranging from 1% to 5%. These reactions are usually reversible upon withdrawal of the drug but can occasionally be life-threatening. The mechanism of flutamide-associated hepatotoxicity is not well established. We report a case of a 69-year-old man with prostatic carcinoma in whom flutamide induced an acute hepatitis which resolved completely soon after drug withdrawal. In this patient, we have studied the possible involvement of an immunological mechanism in causing flutamide hepatitis by investigating the presence of circulating antibodies directed against reactive metabolites of flutamide bound to liver proteins with enzyme-linked immunosorbent assay technique. Although, in the present case, we have failed to detect IgG reacting with rat liver microsomes incubated in vitro with flutamide, this does not completely rule out the possibility of an immunological involvement in flutamide hepatotoxicity. The possibility of severe flutamide-related injury, independently of the underlying pathogenic mechanism, strongly suggests the need for careful monitoring of liver enzymes in patients taking this drug.

Matérias estranhas em doces de amendoim e de leite vendidos por ambulantes na Cidade de Säo Paulo / Light filth in peanuts and \"milk sweet\" bars, sold by street merchants in the city of Säo Paulo

Foram analisadas 351 amostras de doces de amendoim e 157 de doces de leite, no período de janeiro/93 a janeiro/94, adquiridas mensalmente em dez barracas de ambulantes de doces, distribuídas em quatro regiöes da Cidade de Säo Paulo/SP. O objetivo do trabalho foi a adequaçäo de métodos para pesquisa de matérias estranhas e avaliaçäo das condiçöes sanitárias desses produtos, durante as quatro estaçöes do ano, utilizando-se métodos da "Association of Official Analytical Chemists", 15 ed., 1990. Os métodos permitiram obter material adequado para leitura e identificaçäo das matérias estranhas, sem resíduos interferentes que dificultassem o diagnóstico da análise. Do total de amostras de doces de amendoim e de leite, 32,8 (por cento) e 28,7 (por cento), respectivamente, estavam aprovadas, segundo as normas legais vigentes brasileiras. Das 351 amostras de doces de amendoim, 60,7 (por cento) continham fragmentos de insetos, 11,7 (por cento) ácaros e 6,8 (por cento) estavam contaminadas com pêlos de roedor. Das 157 amostras de doce de leite, 58,6 (por cento) estavam contaminadas por fragmentos de insetos, 26,8 (por cento) por ácaros e 5,7 (por cento) por pêlos de roedor. Os números médios de fragmentos de insetos, ácaros e pêlos de roedor, em relaçäo às amostras condenadas, foram 8,6 (por cento); 11,3 e 1,4 para os doces de leite e 6,5; e 1,1 para os doces de amendoim, respectivamente. As estaçöes do ano que apresentaram o maior percentual de anostras positivas para fragmentos de insetos e pêlos de roedor foram o outono e o inverno, para os doces de amendoim, e o outono para os doces de leite; para ácaros, o maior percentual de amostras positivas ocorreu no veräo