Untargeted NMR-based metabolomics analysis of kidney allograft perfusates identifies a signature of delayed graft function.

INTRODUCTION: Kidney transplantation (KTx) necessarily conveys an ischemia/reperfusion (I/R) process, which impacts on allograft outcomes. Delayed graft function (DGF) is defined as a non-decrease of serum creatinine by at least 10% daily on 3 consecutive days during the first 7 days post-KTx. DGF significantly conditions both short- and long-term graft outcomes. Still there is a lack of DGF predictive biomarkers. OBJECTIVES: This study aimed to explore the potential of kidney graft perfusate metabolomics to predict DGF occurrence. METHODS: 49 human perfusates from grafts categorized upon donor type [donation after brain death (DBD)/donation after circulatory death (DCD)] and DGF occurrence and 19 perfusates from a murine model classified upon death type (DBD/DCD) were collected and analyzed by NMR-based metabolomics. RESULTS: The multivariate analysis of the murine data highlighted significant differences between perfusate metabolomes of DBD versus DCD. These differences were similarly observed in the human perfusates. After correcting for the type of donor, multivariate analysis of human data demonstrated a metabolomics signature that could be correlated with DGF occurrence. CONCLUSIONS: The metabolome of kidney grafts is influenced by the donor's type in both human and pre-clinical studies and could be correlated with DGF in the human DBD cohort. Thus, metabolomic analysis of perfusate applied prior to KTx may represent a new predictive tool for clinicians in a more personalized management of DGF. Moreover, our data paves the way to better understand the impact of donor's types on the biochemical events occurring between death and the hypothermic storage.

Comparison of X-chromosome inactivation in Duchenne muscle/myocardium-manifesting carriers, non-manifesting carriers and related daughters.

Female carriers of Duchenne muscular dystrophy (DMD) are usually asymptomatic. However, 2.5-7.8% of them may present muscle symptoms and cardiomyopathy, attributed to a reduced production of dystrophin, probably because of skewed patterns of X-chromosome inactivation (XCI). To evaluate the role of XCI in symptomatic (at muscle or heart level) and asymptomatic DMD carriers, 44 subjects were selected from our database (12 manifesting, 21 non-manifesting, 11 healthy females), and XCI pattern determined in the lymphocytes by the androgen receptor methylation-based assay. The results showed that DMD-manifesting carriers had a preferential inactivation of the X-chromosome carrying the normal allele, while non-manifesting carriers and healthy females showed a random XCI pattern. Moreover, when comparing muscle with heart manifesting carriers, the former group showed a higher degree of skewing. No concordance in XCI was found between mothers and daughters, when symptomatic/asymptomatic mother-daughter pairs were analyzed. The results confirm that DMD clinical manifestations in carriers are associated with non-random patterns of X inactivation.

Frenulectomy: proposal of a new surgical approach and case report.

The frenula of the oral cavity represent the insertion of perioral muscles in jaws, consisting of a fold of the triangular fibro-connective tissue covered with mucous membrane and usually inserted at the mucogingival line. The purpose of this work, after a review of literature, was to provide the clinical signs above the median frenum removal order for the orthodontic closure of a interincisor diastema and to present a new surgical technique.

A preamplification approach to GMO detection in processed foods.

DNA is widely used as a target for GMO analysis because of its stability and high detectability. Real-time PCR is the method routinely used in most analytical laboratories due to its quantitative performance and great sensitivity. Accurate DNA detection and quantification is dependent on the specificity and sensitivity of the amplification protocol as well as on the quality and quantity of the DNA used in the PCR reaction. In order to enhance the sensitivity of real-time PCR and consequently expand the number of analyzable target genes, we applied a preamplification technique to processed foods where DNA can be present in low amounts and/or in degraded forms thereby affecting the reliability of qualitative and quantitative results. The preamplification procedure utilizes a pool of primers targeting genes of interest and is followed by real-time PCR reactions specific for each gene. An improvement of Ct values was found comparing preamplified vs. non-preamplified DNA. The strategy reported in the present study will be also applicable to other fields requiring quantitative DNA testing by real-time PCR.

Efficient immunization of rhesus macaques with an HCV candidate vaccine by heterologous priming-boosting with novel adenoviral vectors based on different serotypes.

Efficient vaccination against viral agents requires a strong T-cell-mediated immune response to clear viral-infected cells. Optimal vaccination can be achieved by administration of recombinant viral vectors encoding phatogen antigens. Adenoviral vectors have attracted considerable attention as potential viral vectors for genetic vaccination owing to their favorable safety profile and potent transduction efficiency following intramuscular injection. However, the neutralizing antibody response against adenoviral capsid proteins following adenoviral vectors injection limits the success of vaccination protocols based on multiple administrations of the same adenoviral serotype. In this work, we describe efficient immunization of rhesus macaques, the preferred model for preclinical assessment, with an HCV candidate vaccine by heterologous priming-boosting with adenoviral vectors based on different serotypes. The induced responses are broad and show significant cross-strain reactivity. Boosting can be delayed for over 2 years after priming, indicating that there is long-term maintenance of resting memory cells.

Neural retina of chick embryo in organ culture: effects of blockade of growth factors by suramin.

The neural retina is a highly organized organ whose final histoarchitecture depends on the presence of diverse growth factors and on their interactions with extracellular matrix components. However, the role of growth factors on retinal development is not fully understood. Suramin has been shown to produce diverse cellular effects via the simultaneous block of the action of several growth factors. We have therefore studied the effects of suramin on organotypic culture of chick embryo neural retina in order to gain further insights into the participation of growth factors in neural retinal development. Neural retina was incubated for 24 h with suramin at 50-200 microM and then processed to determine cell proliferation, nuclear morphology, and actin distribution. Suramin provoked extensive morphological changes revealed by a decrease in BrdU incorporation, alterations in cellular organization, and disruption of the outer limiting membrane, with the emergence of cellular elements through it. All of these effects were dose-dependent and markedly attenuated by the simultaneous presence of suramin and fibroblast growth factor 2 (FGF-2) in the culture medium. These findings indicate that suramin induces pleiotropic effects on the histoarchitecture of the chicken neural retina in organ culture and suggest that FGF-2 is one of the biological modulators involved in the maintenance of the structural organization of the chicken neural retina.

Tamoxifen as primary treatment of breast cancer in elderly patients.

The authors report on a series of 120 elderly (age over 69) women with primary breast treated with tamoxifen alone. Treatment schedule was 160 mg on day 1 followed by a daily maintenance dose of 20 mg. Compliance to treatment was optimal and side effects were minimal. The best results achieved after at least six months of treatments were complete response in 12, a partial response in 46 and minor response in 10 patients, whereas stable disease or progression was observed in 43 or 9 patients, respectively. Response duration was limited and progression was increasingly observed over time. After 6, 12, 24, 36, 48 and 60 months the proportion of subjects still showing response to treatment was 43%, 57%, 56%, 46%, 32% and 31%, whereas progression rate was 7%, 12%, 25%, 39%, 55% and 60%, respectively, the difference being accounted for by patients with stable disease. As determined in a subset of 27 subjects, treatment response was strongly associated with immunocytochemically assessed tumor estrogen receptor content, progression being 100%, 43% or 6% in subjects with 0%, 30-60% or > 60% immunostained cells, respectively. These results do not support primary hormone therapy as a current alternative to surgery, which should be the standard treatment in otherwise healthy elderly patients with operable breast cancer. When surgery is specifically contraindicated, hormone treatment should be proposed as an alternative only in subjects with high tumor estrogen receptor content.

[Prevalence of group B beta-hemolytic Streptococcus colonization in a sample of 23,312 pregnant women and newborn infants]. / La prevalenza della colonizzazione da streptococco beta-emolitico di gruppo B in un campione di 23.312 donne gravide e neonati.

We report the prevalence of colonization of Group B Streptococci in a given population referred to a limited area in the north-west of Italy. 23.312 pregnant women were tested. Group B Streptococci have been isolated from genital cultures in 0.18-13.2% (mean 8.18). The prevalence of Group B streptococcal colonization from ear, throat and ocular cultures of newborn infants from colonized mothers was 11.55%. Incidence of infection in neonates has varied from 0 to 2.33% (1.5 per 1000 live births).

Response of primary breast cancer to tamoxifen alone in elderly women.

The authors report on a consecutive series of 62 evaluable, elderly (age greater than 69 years) women with primary operable breast cancer treated only with tamoxifen. The treatment schedule was 160 mg on day 1 followed by a daily maintenance dose of 20 mg. Compliance to treatment was excellent, and subjective side effects were minimal. The best response achieved after at least 6 months of treatment was complete in 7, partial in 22 and minor in 3, whereas stable disease or progression was observed in 28 and 2 patients, respectively. The response rate decreased and progressions increased with time. At 6, 12, 24, 36 and 48 months of treatment, the complete + partial response rates were 30.6%, 45%, 45.6%, 38.7% and 25% and the progressive disease rates 4.8%, 19.6%, 34.8%, 45.2% and 66.6%, respectively. These results do not confirm some previous reports of a high response to tamoxifen. This difference is only partially explained by the use of mammography, which is more sensitive than palpation, to assess tumor size and treatment response. The present study does not support primary hormone therapy as a current alternative to surgery, which should be the standard treatment in otherwise healthy elderly patients with operable breast cancer.

The risk of breast cancer subsequent to histologic diagnosis of benign intraductal papilloma follow-up study of 339 cases.

The risk of developing breast cancer was investigated in 339 patients in whom histologically confirmed intraductal papilloma had been surgically removed. Follow-up ranged from 2 to 14 years, (average, 6.62 years). Ten breast cancers were observed, whereas 3 were expected on the basis of age-specific incidence rates drawn from local cancer registries (relative risk = 3.33, 95% confidence interval = 1.60-6.13). No significant difference in the relative risk was observed as far as papilloma type (single or multiple) was concerned. All breast cancers observed occurred in the same breast as the papilloma. Women in whom a benign intraductal papilloma has been excised should be carefully followed since they at are higher risk for ipsilateral breast cancer.

Aderências em microcirurgia tubária / Adhesions in tubal microsurgery

No Setor de Reproduçäo Humana da Disciplina de Ginecologia da Escola Paulista de Medicina os AA. avaliam os resultados de microcirurgia tubária em 202 pacientes, com e sem uso de Dextran 70%. Nos 202 casos referem permeabilizaçäo tubária em 90% dos casos. As taxas de gravidez pós-cirúrgicas foram de 27% no grupo sem o uso de Dextran e de 33,9% naqueles em que essa soluçäo glicosada foi utilizada. Concluem afirmando ser o uso do Dextran 70 útil para reduzir as aderências tubárias pós-operatórias

Localization of basic fibroblast growth factor binding sites in the chick embryonic neural retina.

We have investigated the localization of basic fibroblast growth factor (bFGF) binding sites during the development of the neural retina in the chick embryo. The specificity of the affinity of bFGF for its receptors was assessed by competition experiments with unlabelled growth factor or with heparin, as well as by heparitinase treatment of the samples. Two different types of binding sites were observed in the neural retina by light-microscopic autoradiography. The first type, localized mainly to basement membranes, was highly sensitive to heparitinase digestion and to competition with heparin. It was not developmentally regulated. The second type of binding site, resistant to heparin competition, appeared to be associated with retinal cells from the earliest stages studied (3-day-old embryo, stages 21-22 of Hamburger and Hamilton). Its distribution was found to vary during embryonic development, paralleling layering of the neural retina. Binding of bFGF to the latter sites was observed throughout the retinal neuroepithelium at early stages but displayed a distinct pattern at the time when the inner and outer plexiform layers were formed. During the development of the inner plexiform layer, a banded pattern of bFGF binding was observed. These bands, lying parallel to the vitreal surface, seemed to codistribute with the synaptic bands existing in the inner plexiform layer. The presence of intra-retinal bFGF binding sites whose distribution varies with embryonic development suggests a regulatory mechanism involving differential actions of bFGF on neural retinal cells.

Mammographic parenchymal patterns and breast cancer risk. A case control study of a population-based screening experience.

The authors investigated the association between mammographic parenchymal patterns and breast cancer risk by a case control study. Three-hundred and thirty-four cancers, either screen or interval detected in the course of a screening program, were assumed as cases and compared to 668 non-cancer controls matched by age, residence and date of screening examination. Parenchymal patterns were classified by retrospective blind review of randomly mixed films by two independent readers. A high agreement was achieved on a two-group scale (N1-P1 vs P2-DY) inter and intraobserver consistency being 0.94 and 0.95, respectively. The relative risk of breast cancer exposure (RR) with respect to N1 pattern was 1.99 for P1, 3.59 for P2 and 4.18 for DY patterns. The RR of P2-DY with respect to N1-P1 patterns was 2.62. The results did not change after stratification by age group. Parenchymal patterns do not seem to be a reliable criterion for selecting high risk women for screening as in this series only 57% of all cancers occurred in high risk (P2-DY) subjects.

Microcirurgia tubária na clínica ginecológica da Escola Paulista de Medicina / Tubal microsurgery at the gynecologic clinic of the Escola Paulista de Medicina

Os AA avaliaram o resultado da microcirurgia tubária em 196 casos operados na Clínica Ginecológica da Escola Paulista de Medicina, no período de 10 anos; de 1978 a 1987. Separam os casos de acordo com o tipo de cirurgia em Salpingostomia: 52 vezes; lise de aderências (salpingolise): 48; anastomose ístmico ístmica: 62 e reimplante tubo-uterino: 34. Descrevem as técnicas e concluem expondo seus resultados em termos de permeabilizaçäo e de gravidez. Afirmam que cerca de 25% dos casos resultam em gestaçäo, na média de todos os casos e especificam as porcentagens de cada tipo de cirurgia realizada