Familial aggregation of early-onset cancers in early-onset breast cancer families.

The risk of early-onset (EO) breast cancer is known to be increased in relatives of EO breast cancer patients, but less is known about the familial risk of other EO cancers. We assessed familial risks of EO cancers (aged ≤40 years) other than breast cancer in 54 753 relatives of 5562 women with EO breast cancer (probands) by using a population-based cohort from Finland. Standardized incidence ratios (SIRs) and 95% confidence intervals (CI) were estimated by using gender-, age- and period-specific cancer incidences of the general population as reference. The risk of any cancer excluding breast cancer in first-degree relatives was comparable to population cancer risk (SIR 0.99, 95% CI: 0.84-1.16). Siblings' children of women with EO breast cancer were at an elevated risk of EO testicular and ovarian cancer (SIR = 1.74, 95% CI: 1.07-2.69 and 2.69, 95% CI: 1.08-5.53, respectively). The risk of EO pancreatic cancer was elevated in siblings of the probands (7.61, 95% CI: 1.57-22.23) and an increased risk of any other cancer than breast cancer was observed in children of the probands (1.27, 95% CI: 1.03-1.55). In conclusion, relatives of women with EO breast cancer are at higher familial risk of certain discordant EO cancers, with the risk extending beyond first-degree relatives.

Familial aggregation of early-onset cancers.

This registry-linkage study evaluates familial aggregation of cancer among relatives of a population-based series of early-onset (≤40 years) cancer patients in Finland. A cohort of 376,762 relatives of early-onset cancer patients diagnosed between 1970 and 2012 in 40,538 families was identified. Familial aggregation of early-onset breast, colorectal, brain and other central nervous system (CNS) cancer and melanoma was explored by standardized incidence ratios (SIR), stratified by relatedness. Gender-, age- and period-specific population cancer incidences were used as reference. Cumulative risks for siblings and offspring of the proband up to age ≤40 years were also estimated. Almost all early-onset cancers were sporadic (98% or more). Among first-degree relatives, SIR was largest in colorectal cancer (14, 95% confidence interval 9.72-18), and lowest in melanoma (1.93, 1.05-3.23). Highest relative-specific SIRs were observed for siblings in families, where also parent had concordant cancer, 90 (43-165) for colorectal cancer and 29 (11-64) for CNS cancer. In spouses, all SIRs were at population level. Cumulative risk of colorectal cancer by age 41 was 0.98% in siblings and 0.10% in population, while in breast cancer the corresponding risks were 2.05% and 0.56%. In conclusion, early-onset cancers are mainly sporadic. Findings support high familial aggregation in early-onset colorectal and CNS cancers. Familial aggregation in multiplex families with CNS cancers was mainly attributed to neurofibromatosis and in colorectal cancer to FAP- and HNPCC-syndromes. The pattern of familial aggregation of early-onset breast cancer could be seen to support very early exposure to environmental factors and/or rare genetic factors.

Evaluation of the effects of a probiotic supplementation with respect to placebo on intestinal microflora and secretory IgA production, during antibiotic therapy, in children affected by recurrent airway infections and skin symptoms.

Antibiotic therapy, especially in pediatric patients, is often associated with significant modifications of the gut microflora, which can lead to intestinal dysbiosis and influence intestinal physiology and immune system functionality. Herein we report the results from a double blind controlled clinical trial in 77 pediatric patients affected by recurrent airway infections, receiving antibiotic therapy with amoxicillin and clavulanic acid. A group was treated with an oral probiotic preparation composed of Lactobacillus paracasei ssp.paracasei CRL-431, Bifidobacterium BB-12, Streptococcus thermophilus TH-4 and a fructooligosaccharide (FOS) during and after antibiotic therapy for seven days, while the other group received placebo. The study revealed a reduction in the Clostridia population, with a contemporary increase in Bifidobacteria and Lactobacilli in fecal samples in the probiotic group and an increase in the Enterobacteria population in the placebo group. Moreover, there was a decreasing trend in secretory IgA production in the probiotic group. Some relevant, but not statistically significant probiotic supplementation effects were identified.

Pearls & Oy-sters: Thyroid-Associated Ophthalmopathy and Transient Neuromuscular Junction Disorder Due to Graves Disease.

The concomitant presentation of thyroid-associated ophthalmopathy (TAO) and ocular myasthenia gravis is well documented. In the course of Graves disease (GD), symptomatic transient neuromuscular junction disorder may occur due to the effect of thyroid hormones at the neuromuscular synapse. Diagnostic clues are the clinical and electrophysiologic remission synchronous with restoration of euthyroidism. Furthermore, the occurrence of thymic hyperplasia in GD poses further diagnostic and therapeutic considerations. These points are discussed in the case report of a 43-year-old male patient suffering from TAO and transient neuromuscular junction disorder due to GD.

Transcriptional repression and enhancer decommissioning silence cell cycle genes in postmitotic tissues.

The mechanisms that maintain a non-cycling status in postmitotic tissues are not well understood. Many cell cycle genes have promoters and enhancers that remain accessible even when cells are terminally differentiated and in a non-cycling state, suggesting their repression must be maintained long term. In contrast, enhancer decommissioning has been observed for rate-limiting cell cycle genes in the Drosophila wing, a tissue where the cells die soon after eclosion, but it has been unclear if this also occurs in other contexts of terminal differentiation. In this study, we show that enhancer decommissioning also occurs at specific, rate-limiting cell cycle genes in the long-lived tissues of the Drosophila eye and brain, and we propose this loss of chromatin accessibility may help maintain a robust postmitotic state. We examined the decommissioned enhancers at specific rate-limiting cell cycle genes and show that they encode dynamic temporal and spatial expression patterns that include shared, as well as tissue-specific elements, resulting in broad gene expression with developmentally controlled temporal regulation. We extend our analysis to cell cycle gene expression and chromatin accessibility in the mammalian retina using a published dataset, and find that the principles of cell cycle gene regulation identified in terminally differentiating Drosophila tissues are conserved in the differentiating mammalian retina. We propose a robust, non-cycling status is maintained in long-lived postmitotic tissues through a combination of stable repression at most cell cycle gens, alongside enhancer decommissioning at specific rate-limiting cell cycle genes.

Pregnancy in a methylmalonic acidemia patient with kidney transplantation: a case report.

Presently pregnancy is no more exceptional in women with metabolic diseases. However, it still poses significant medical problems both before and after childbirth. The challenge is even greater if the mother has undergone organ transplantation, because of her metabolic disease. We report on a case of pregnancy in a patient 29-year-old with methylmalonic acidemia cblA type (OMIM 251100) who received a renal transplantation at the age of 17 for end-stage renal disease (ESRD) caused by her primary disease. During pregnancy neither metabolic crises nor renal function changes were observed in the mother, with the only exception of a mild increase of her systemic blood pressure. To the fetus pregnancy was uneventful and during the first 30 months after birth the baby's neuropsychomotor development was normal and there were no episodes of metabolic derangement. This is evidence that methylmalonicacidemia cblA, even when treated with renal transplantation for inherent ESRD, is no contraindication to pregnancy. It is even possible that a functioning transplanted kidney contributes to improve metabolic parameters.

The role of gut microbiota in human obesity: recent findings and future perspectives.

AIMS: In recent years, gut microbiota have gained a growing interest as an environmental factor that may affect the predisposition toward adiposity. In this review, we describe and discuss the research that has focused on the involvement of gut microbiota in human obesity. We also summarize the current knowledge concerning the health effects of the composition of gut microbiota, acquired using the most recent methodological approaches, and the potential influence of gut microbiota on adiposity, as revealed by animal studies. DATA SYNTHESIS: Original research studies that were published in English or French until December 2011 were selected through a computer-assisted literature search. The studies conducted to date show that there are differences in the gut microbiota between obese and normal-weight experimental animals. There is also evidence that a high-fat diet may induce changes in gut microbiota in animal models regardless of the presence of obesity. In humans, obesity has been associated with reduced bacterial diversity and an altered representation of bacterial species, but the identified differences are not homogeneous among the studies. CONCLUSIONS: The question remains as to whether changes in the intestinal microbial community are one of the environmental causes of overweight and obesity or if they are a consequence of obesity, specifically of the unbalanced diet that often accompanies the development of excess weight gain. In the future, larger studies on the potential role of intestinal microbiota in human obesity should be conducted at the species level using standardized analytical techniques and taking all of the possible confounding variables into account.

A tissue dissociation method for ATAC-seq and CUT&RUN in Drosophila pupal tissues.

Chromatin accessibility, histone modifications, and transcription factor binding are highly dynamic during Drosophila metamorphosis and drive global changes in gene expression as larval tissues differentiate into adult structures. Unfortunately, the presence of pupa cuticle on many Drosophila tissues during metamorphosis prevents enzyme access to cells and has limited the use of enzymatic in situ methods for assessing chromatin accessibility and histone modifications. Here, we present a dissociation method for cuticle-bound pupal tissues that is compatible for use with ATAC-Seq and CUT&RUN to interrogate chromatin accessibility and histone modifications. We show this method provides comparable chromatin accessibility data to the non-enzymatic approach FAIRE-seq, with only a fraction of the amount of input tissue required. This approach is also compatible with CUT&RUN, which allows genome-wide mapping of histone modifications with less than 1/10th of the tissue input required for more conventional approaches such as Chromatin Immunoprecipitation Sequencing (ChIP-seq). Our protocol makes it possible to use newer, more sensitive enzymatic in situ approaches to interrogate gene regulatory networks during Drosophila metamorphosis.

Serum osteopontin and CD44 levels in lymphoreticular malignancies in children.

OBJECTIVE: Osteopontin (OPN) is an adhesive glycoprotein that interacts with a variety of cell surface receptors, including several integrins and CD44. OPN is expressed and secreted by numerous human malignancies. CD44 play an important role in tumor growth and metastasis. We aimed to evaluate serum levels of osteopontin and CD44 in patients with lymphorethicular malignancies in childhood. METHODS: We studied serum levels of CD44 and OPN levels of 54 patients (26, 18 and 10 patients with non-Hodgkin's lymphoma (NHL), Hodgkin's lymphoma (HL) and acute lymphoblastic leukemia (ALL), respectively) at the diagnosis. RESULTS: The mean levels of OPN were significantly higher in patients (5.42±8.24 ng/ml) than in controls (3.89 ±1.96 ng/ml). The mean levels of CD44 levels were also significantly higher in patients (3.82±2.31 ng/ml) than in controls (1.96±0.62 ng/ml), and significantly higher in the advanced stages than in early stages. The mean levels of the CD44 in NHL, HL and ALL were 3.49±2.00, 3.56±1.74, and 5.15±3.50 respectively. OPN and CD44 levels were found to be increased in parallel (p=0.003). A more advanced disease and/or poor prognostic factors were seen in 9 patients who had both serum CD44 and OPN levels higher than 2SD of the control. CONCLUSION: Elevated levels of both CD44 and OPN at the diagnosis may predict an unfavorable outcome in childhood leukemias and lymphomas (Tab. 2, Fig. 3, Ref. 44).

Genetic and epigenetic alterations in the GNAS locus and clinical consequences in Pseudohypoparathyroidism: Italian common healthcare pathways adoption.

BACKGROUND: Genetic and epigenetic alterations in the GNAS locus are responsible for the Gsα protein dysfunctions causing Pseudohypoparathyroidism (PHP) type Ia/c and Ib, respectively. For these heterogeneous diseases characterized by multiple hormone resistances and Albright's Hereditary Osteodystrophy (AHO) the current classification results inadequate because of the clinical overlap between molecular subtypes and a standard clinical approach is still missing. In the present paper several members of the Study Group Endocrine diseases due to altered function of Gsα protein of the Italian Society of Pediatric Endocrinology and Diabetology (ISPED) have reviewed and updated the clinical-molecular data of the largest case series of (epi)/genetically characterized AHO/PHP patients; they then produced a common healthcare pathway for patients with these disorders. METHODS: The molecular analysis of the GNAS gene and locus identified the causal alteration in 74 subjects (46 genetic and 28 epigenetic mutations). The clinical data at the diagnosis and their evolution during up to 15 years follow-up were collected using two different cards. RESULTS: In patients with genetic mutations the growth impairment worsen during the time, while obesity prevalence decreases; subcutaneous ossifications seem specific for this group. Brachydactyly has been detected in half of the subjects with epigenetic alterations, in which the disease overts later in life, often with symptomatic hypocalcaemia, and also early TSH and GHRH resistances have been recorded. CONCLUSIONS: A dedicated healthcare pathway addressing all these aspects in a systematic way would improve the clinical management, allowing an earlier recognition of some PHP features, the optimization of their medical treatment and a better clinical-oriented molecular analysis. Furthermore, standardized follow-up data would provide new insight into less known aspects.

Assessment of susceptibility of mould isolates towards biocides.

OBJECTIVES: The aim of this study was to determine the susceptibility of 77 mould strains: Aspergillus fumigatus (20), Aspergillus flavus (8), Aspergillus niger (4), Aspergillus ochraceus (2), Penicillium citrinum (15), Penicillium crysogenum (14), Penicillium aurantiogriseum (1), Penicillium roquefortii (4), Penicillium paneum (2), Rhizopus spp. (3), Tricoderma spp. (1) and Mucor spp. (3) to biocides. METHODS: MIC determination was determined based on CLSI methodology. RESULTS: For hospital acquired strains, MIC50 was 0.5mg/L, MIC90 was 1mg/L for chlorhexidine (CHX); MIC50 was 0.5mg/L, and MIC90 was 1mg/L for benzalkonium chloride (BZC); MIC50 was 1mg/L, and MIC90 was 2mg/L for triclosan (TRC); MIC50 was 1024mg/L, and MIC90 was 2048mg/L for sodium hypochloride (SHC). For feed and food isolates MIC50 was 2mg/L, MIC90 was 8mg/L for CHX, MIC50 was 2mg/L, and MIC90 was 4mg/L for BZC, MIC50 was 2mg/L, and MIC90 was 4mg/L for TRC, MIC50 was 256mg/L, and MIC90 was 512mg/L for SHC. CONCLUSION: We can conclude that food isolates presented slightly higher MIC50 and MIC90 values for CHX, BNZ and TRC, but not for SHC.

Genetic analysis of the replication region of the Lactobacillus plasmid vector pPSC22.

The sequence and genetic organization of the 1,600-bp replication region of the Lactobacillus vector pPSC22, a plasmid derived from a 7-kb cryptic plasmid of L. plantarum used for the cloning of heterologous genes in several lactobacilli, were determined. Sequence analysis revealed the presence of a plus origin of replication containing the two functional elements nic and bind, required for initiation of the leading strands typical of the rolling circle (RC)-replicating plasmids belonging to the pLS1 family. Two open reading frames (copA and repA) were located within the Lactobacillus portion of pPSC22. The repA gene product, a 234-amino acid protein, showed homologies with the Rep protein of the streptococcal plasmid pLS1 and contained the three conserved domains detected in most Rep proteins of RC-replicating plasmids and ss-coliphages. The genetic organization of the replication region of pPSC22 shared relevant homologies with the lactococcal plasmids pWVO1 and pFX2.

Renal function in patients undergoing cardiopulmonary bypass operations.

We evaluate short- and long-term effects on renal functional reserve of cardiopulmonary bypass in 11 patients. A selected group (persistence of renal functional reserve before operation) of 11 adult patients undergoing cardiopulmonary bypass for aorta-coronary bypass were studied. Renal functional reserve tests were performed in all patients before the operation, at postoperative day 9, and at 6 months after operation. Basal glomerular filtration rate did not show significant changes at 9 days and at 6 months after operation. On the contrary, renal functional reserve was absent at 9 days, but it was restored to preoperative levels at 6 months after operation. In conclusion, our data indicate that cardiopulmonary bypass probably causes renal damage that is not sufficient to influence routine renal function parameters.

Autologous protein reinfusion in severe ovary hyperstimulation syndrome.

BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) is one of the most serious complications of ovulation induction by exogenous gonadotropins. The pathophysiologic factors of this syndrome are not well known. Increased capillary permeability causes third space fluid shift, which is responsible for ascites, pleural fluid, and edemas. Severe OHSS may result in renal failure, hypovolemic shock, thromboembolic disease, respiratory distress, and may cause death. It has been observed that paracentesis is efficacious, provided that care is taken to reinfuse protein lost in the peritoneal exudate. For this reason, in three patients with severe OHSS we have used a dialytic technique of reinfusion of concentrated ascitic fluid. STUDY DESIGN: We treated three patients with severe OHSS (grade 6). Through sonography-guided paracentesis, the ascitic fluid was concentrated by ultrafiltration and reinfused. This treatment was instituted and performed once only. Ultrafiltration was obtained with a common high-flow dialyzer (polyacrylonitryle membrane). The concentrated fluid was returned to the patient in a peripheral vein. We have limited further treatment to restoration of fluid and electrolyte balance, avoiding in particular potentially teratogenic drugs. RESULTS: In all three patients, a progressive increase of diuresis was evident during treatment and subjective improvement was almost immediate. Fifteen days after treatment, hematologic and biochemical parameters had returned within normal limits. CONCLUSIONS: In treating severe OHSS, we have used the technique of reinfusion of concentrated ascitic fluid to avoid protein depletion induced by paracentesis. We have been able to successfully restore to normal the hematologic and biochemical imbalance with one treatment. Use of the technique described herein should be limited to carefully selected instances and treatment should be performed in an intensive care unit.

Description of new "bowel-sparing" techniques for long strictures of Crohn's disease.

In the period of January 1993 to December 1995 we operated on 55 patients with various complications of Crohn's disease. In properly selected cases, obstructive complications of Crohn's disease can be treated effectively by strictureplasty. Long strictures, even if a narrow lumen is still present, are commonly managed by resection, as classic strictureplasties cannot be done; also Finney strictureplasty seems inadequate, as it creates a blind loop that favors bacterial overgrowth and fecal stasis. Three original "sparing bowel" surgical approaches are proposed as possible alternative in the treatment of long stricture in Crohn's disease. We perform side-to-side ileoileal plasty whenever we are faced with severe narrowing of a long segment of small bowel (>10 cm); side-to-side ileocolic plasty whenever very severe disease with narrowing of ileocaecal valve is present; and ileocaecal plasty when terminal ileitis involves the very distal end of the small bowel, but sparing or only minimally affecting the ileocaecal valve. The above-mentioned procedures are described in detail and the clinical outcomes related to the first 8-patient series of our institution are presented.

Red blood cell lipid peroxidation in predialysis chronic renal failure.

In hemodialysis patients the pentose-phosphate shunt activity is deficient. As a consequence, the lipid peroxidation of the erythrocyte membranes is increased as shown by the increase in malonyldialdehyde concentrations and is accompanied by a decrease of the level of vitamin E in RBC. In the present study we have found that increased lipid peroxidation of the erythrocyte membranes is present also in chronic renal failure patients in the predialysis state, provided that the serum creatinine levels are higher than 5 mg/dl.

Effects of alpha-tocopherol administration on red blood cell membrane lipid peroxidation in hemodialysis patients.

The result of vitamin E treatment in 19 uremic patients in chronic hemodialysis is evaluated. In particular, the levels of erythrocyte malonyldialdehyde (MDA) and vitamin E were determined, and the fatty acid composition of red blood cell (RBC) membrane before and after treatment with parenterally administered vitamin E. A decrease of RBC MDA levels, an increase of RBC vitamin E concentrations, and a decreased saturated fatty acid to unsaturated fatty acid ratio were found after treatment with vitamin E. There was a statistically significant increase of the packed RBC volume.

Stability of cyclopropane and conjugated linoleic acids during fatty acid quantification in lactic acid bacteria.

Seven methods commonly used for fatty acid analysis of microorganisms and foods were compared to establish the best for the analysis of lyophilized lactic acid bacteria. One of these methods involves fat extraction followed by methylation of fatty acids, while the other methods use a direct methylation of the samples, under different operating conditions (e.g., reaction temperature and time, reagents, and pH). Fatty acid methyl esters were identified by gas chromatography-mass spectrometry and quantified by on-column capillary gas chromatography. Two reliable methods for the analysis of fatty acids in bacteria were selected and further improved. They guarantee high recovery of classes of fragile fatty acids, such as cyclopropane and conjugated acids, and a high degree of methylation for all types of fatty acid esters. These two direct methylation methods have already been successfully applied to the analysis of fatty acids in foods. They represent a rapid and highly reliable alternative to classical time- and solvent-consuming methods and they give the fatty acid profile and the amount of each fatty acid. Using these methods, conjugated linoleic acids were identified and quantified in lactic acid bacteria.

Carcinoma of the head of the pancreas: review of 67 cases and comparison with 27 additional periampullary carcinomas.

BACKGROUND/AIMS: Carcinoma of the head of the pancreas is reported with increasing incidence. The classical Whipple procedure, modified by Traverso and Longmire, is a major surgical undertaking and there is question on its indication in the case of lesions which, although technically resectable, are not susceptible to cure. PATIENTS AND METHODS: We review 67 cases of carcinoma of the head of the pancreas comparing the results of radical vs. palliative surgery. The results were then compared with 27 additional cases of periampullary carcinomas. RESULTS: Long term survival was obviously higher after resection, due to the more advanced tumor stage in the palliation group. However, perioperative mortality, supposedly high in extensive resective surgery, was 8.7% (vs 11.9% in the palliation group). CONCLUSIONS: At present, extensive resective surgery carries unacceptable incidence of perioperative mortality. Therefore we suggest that indication for resection should be widened, as it may offer better chances of cure in resectable periampullary carcinomas and, even in non curable cases, it offers better quality of life and the advantages of tumor debulking.

Third segment cholangio-jejunostomy in the treatment of unresectable klatskin tumors.

BACKGROUND/AIMS: Cholangiocarcinoma of the hepatic hilum is a seldom curable lesion when detected and several methods of palliation have been suggested. Bismuth has proposed an intrahepatic cholangiojejunostomy on the third segment of the liver, which in his experience obtains effective biliary drainage with better quality of life compared with other forms of palliation. PATIENTS AND MATERIALS: We have used this technique in nine cases. We evaluate results, mortality and morbidity, in comparison with other authors' reported series. RESULTS: In the early postoperative period one patient died, two patients suffered from immediate postoperative complications, and jaundice resolved completely in six patients. Long term survival was influenced by the underlying disease. CONCLUSION: Our findings support Bismuth preference for this technique of surgical palliation for non resectable tumors of the biliary tract.