Proteomic analysis of infiltrating ductal carcinoma tissues by coupled 2-D DIGE/MS/MS analysis.

There is a growing interest in protein expression profiling aiming to identify novel diagnostic markers in breast cancer. Proteomic approaches such as two-dimensional differential gel electrophoresis coupled with tandem mass spectrometry analysis (2-D DIGE/MS/MS) have been used successfully for the identification of candidate biomarkers for screening, diagnosis, prognosis and monitoring of treatment response in various types of cancer. Identifying previously unknown proteins of potential clinical relevance will ultimately help in reaching effective ways to manage the disease. We analyzed breast cancer tissues from five tumor and five normal tissue samples from ten breast cancer subjects with infiltrating ductal carcinoma (IDC) by 2-D DIGE using two types of immobilized pH gradient (IPG) strips: pH 3-10 and pH 4-7. From all the spots detected, differentially expressed (p < 0.05 and ratio > 2) were 50 spots. Of these, 39 proteins were successfully identified by MS, representing 29 different proteins. Ten proteins were overexpressed in the tumor samples. The 2-D DIGE/MS/MS analysis revealed an increase in the expression levels in tumor samples of several proteins not previously associated with breast cancer, such as: macrophage-capping protein (CAPG), phosphomannomutase 2 (PMM2), ATPase ASN1, methylthioribose-1-phosphate isomerase (MRI1), peptidyl-prolyl cis-trans isomerase FKBP4, cellular retinoic acid-binding protein 2 (CRABP2), lamin B1 and keratin, type II cytoskeletal 8 (KRT8). Ingenuity Pathway Analysis (IPA) revealed highly significant (p = 10(-26)) interactions between the identified proteins and their association with cancer. These proteins are involved in many diverse pathways and have established roles in cellular metabolism. It remains the goal of future work to test the suitability of the identified proteins in samples of larger and independent patient groups.

Proteomic analysis of seminal plasma in men with different spermatogenic impairment.

Seminal plasma is a potential source of biomarkers for many disorders of the male reproductive system including male infertility. The identification and characterisation of differentially expressed proteins in seminal plasma of man with normal and impaired spermatogenesis can help in the elucidation of the molecular basis of male infertility. We compared the protein expression profiles of seminal plasma from four different groups of men as follows: normozoospermic, asthenozoospermic, oligozoospermic and azoospermic groups, using two-dimensional differential gel electrophoresis (2-D DIGE). We found eight proteins with statistically significant increased expression in azoospermia compared with at least one of the other studied groups. The differentially expressed spots were fibronectin, prostatic acid phosphatase (PAP), proteasome subunit alpha type-3, beta-2-microglobulin, galectin-3-binding protein, prolactin-inducible protein and cytosolic nonspecific dipeptidase. Notably, PAP was increased in patients with azoospermia compared with that of all other groups. We have observed no statistically significant differences in protein expression between three of the groups: normozoospermic, oligozoospermic and asthenozoospermic. We suggest that the identified panel of proteins in our study especially PAP have a strong potential to be used as azoospermia markers. However, further investigations will be necessary to validate these markers in samples of larger and independent patient cohorts and to clarify their role in the pathogenesis of male infertility.

Evaluation of probiotic administration on the immune response of coccidiosis-vaccinated broilers.

Probiotics are nonpathogenic bacteria that can promote bird health by reducing pathogen colonization. Researchers have previously demonstrated that the avian immune response can be modulated with probiotics, which may provide a mechanism for the reported reductions in pathogens. We examined phagocyte oxidative burst and cell proliferation of vaccinated broilers administered probiotics. We hypothesized that the combination of probiotic bacteria and a vaccine would affect immune function. Two studies were conducted to evaluate this interaction in broilers. Treatments consisted of a negative control, probiotic, vaccine, or a probiotic + vaccine. Peripheral blood was collected on d 7, 14, and 21 of age. Heterophils and monocytes were evaluated for oxidative burst and lymphocytes were assayed for proliferation. In study 1, heterophil oxidative burst was higher (P ≤ 0.05) in each treatment that received probiotic on d 14 when compared with the negative control. On d 21, an enhanced (P ≤ 0.05) heterophil oxidative burst was observed in the probiotic treatment when compared with the other treatments. On d 14, monocyte oxidative burst was greater (P ≤ 0.05) in the probiotic + vaccine treatment when compared with all other treatments. An increase (P ≤ 0.05) in lymphocyte proliferation was observed among all treatments on d 7 when compared with the negative control. Both vaccine treatments had significant lymphocyte proliferation on d 14 when compared with the negative control. In study 2, the probiotic treatment was associated with greater levels in heterophil oxidative burst on d 7 when compared with all other treatments. On d 21, an increase (P ≤ 0.05) in heterophil oxidative burst was seen in the vaccine treatment when compared with the negative control. On d 7, increased (P ≤ 0.05) monocyte oxidative burst was observed in the vaccine treatment when compared with the negative control. No significant differences were observed in lymphocyte proliferation in any of the treatment groups. These data suggest that probiotics can modulate the immune response and may play a role in vaccination.

Neurocognitive predictors of functional outcome two to 13 years after identification as ultra-high risk for psychosis.

BACKGROUND AND AIM: Little is known about the relationship between neurocognitive performance and functional outcome before the onset of frank psychosis. This longitudinal study aimed to investigate neurocognitive predictors of poor functional outcome in a group identified as ultra-high risk (UHR) for psychosis between two and 13 years prior. METHOD: Individuals (N=230) identified as UHR for psychosis at the PACE Clinic in Melbourne completed assessment of psychopathology, functioning and neurocognition at baseline and follow-up. The mean length of follow-up was 7.26 years (SD 3.05). RESULTS: Forty-one individuals with the poorest functional outcome were identified. Only 48.8% of this group had transitioned to psychosis. Poor functional outcome was associated with reduced performance at baseline in the specific neurocognitive domains of verbal learning and memory, processing speed and attention, and verbal fluency, but not global cognitive impairment. Reduced performance on a verbal story recall task, in combination with higher negative symptoms at baseline, was the best predictor of later poor outcome. Baseline positive psychotic symptoms and GAF scores were not associated with later poor outcome. DISCUSSION: To date, this is the longest follow-up study of an UHR sample. Poor functional outcome was associated with specific neurocognitive decrements, regardless of transition to psychosis. The detection of individuals with poor functioning at follow-up, against a background of previously identified risk factors for psychotic disorder, may yield a valid group in which to study biomarkers and treatment of schizophrenia.

Assessing the health perspectives of unique populations of adolescents: a focus group study.

Few articles report on youth's perceptions about health-related issues and needs and even fewer have given voice to youth with disabilities, chronic health conditions, or youth in therapeutic foster care. The purpose of this study was to explore perceptions of health issues from the perspectives of youth placed in therapeutic foster care, youth with chronic medical conditions, and youth with disabilities. Twelve focus groups with 67 youth aged 13-20 were conducted in rural, suburban, and urban communities in the Commonwealth of Virginia. Data analysis revealed both within unique group themes and themes that cut across unique groups. Youth in therapeutic foster care with chronic health conditions or with disabilities perceive themselves as being different from their peers. Participants in each group expressed concerns about the unique issues that they confront daily and how these challenges differentiate them from youth who do not share their life experiences. They expressed the desire for other youth to understand them better and to not treat them differently. Discussion and implications focus on the need to create programs to support the unique needs of these youth.

Antifibroblast antibodies from systemic sclerosis patients bind to {alpha}-enolase and are associated with interstitial lung disease.

OBJECTIVE: To identify target antigens of antifibroblast antibodies (AFA) in systemic sclerosis (SSc) patients. PATIENTS AND METHODS: In the first part, sera from 24 SSc patients (12 with pulmonary arterial hypertension (PAH) and 12 without) and 36 idiopathic PAH patients, tested in pooled sera for groups of three, were compared with a sera pool from 14 healthy controls (HC). Serum IgG reactivity was analysed by the use of a two-dimensional electrophoresis and immunoblotting technique with normal human fibroblasts antigens. In the second part, serum IgG reactivity for two groups: 158 SSc, 67 idiopathic PAH and 100 HC; and 35 SSc and 50 HC was tested against alpha-enolase from Saccharomyces cerevisiae and recombinant human (rHu) alpha-enolase, respectively, on ELISA. RESULTS: In the first part, alpha-enolase was identified as a main target antigen of AFA from SSc patients. In the second part, 37/158 (23%) SSc patients, 6/67 (9%) idiopathic PAH patients and 4/100 (4%) HC (p<0.001) had anti-S cerevisiae alpha-enolase antibodies; 12/35 (34%) SSc patients and 3/50 (6%) HC had anti-rHu alpha-enolase antibodies (p = 0.001). In SSc, the presence of anti-S cerevisiae alpha-enolase antibodies was associated with interstitial lung disease (ILD), decreased total lung capacity (73.2% vs 89.7%; p<0.001) and diffusion capacity for carbon monoxide (47.4% vs 62.3%; p<0.001), and antitopoisomerase 1 antibodies (46% vs 21%; p = 0.005) but not anticentromere antibodies (11% vs 34%; p = 0.006). Results were similar with rHu alpha-enolase testing. CONCLUSION: In SSc, AFA recognise alpha-enolase and are associated with ILD and antitopoisomerase antibodies.

Increased level of Eimeria sensitivity to diclazuril after using a live coccidial vaccine.

Anticoccidial vaccine and an anticoccidial drug rotation program were compared to determine which program was more effective in producing coccidia populations sensitive of 1 ppm diclazuril. The study used an anticoccidial drug-sensitivity battery test (AST) to determine the baseline level of diclazuril sensitivity to field isolates of Eimeria spp. from seven broiler complexes that had used diclazuril. Based on percentage reduction in weight gain and lesion scores, 25% or fewer of the isolates were effectively controlled by diclazuril. Following the baseline sampling, four of the complexes switched to a nondiclazuril in-feed anticoccidial drug program and three of the complexes switched to a vaccination program for two broiler grow-out cycles as the sole coccidiosis-control program. This study demonstrated that the vaccine used (Coccivac-B) contained anticoccidial drug-sensitive strains. Eimeria isolates were subsequently collected from the identical houses and diclazuril AST results were compared with the baseline AST results. Following the two grow-out cycles, sensitivity of the isolates to diclazuril from the four complexes that continued to use in-feed anticoccidial drugs remained essentially unchanged. The isolates from the three complexes that switched to the vaccination program demonstrated a marked increase in diclazuril sensitivity, with 60%-100% of the isolates from each complex effectively controlled by diclazuril. Vaccination with the anticoccidial drug-sensitive strains produced a measurable increase in the level of sensitivity to diclazuril.

Men's adjustment to their partners' breast cancer: a dyadic coping perspective.

The continuing increase in cancer rates among women in the United States is forcing more men to experience the impact of breast cancer on their relationships. Using 71 male partners of newly diagnosed breast cancer patients, this study assessed how dyadic coping strategies affected men's adjustment to their partners' illness. While their partners were undergoing treatment, participants completed standardized instruments that measured emotional well-being, illness intrusiveness, and dyadic coping styles. Regression analysis revealed significant associations between coping styles and illness intrusiveness. In addition, depression predisposed men to poorer adjustment and affected their coping patterns. The findings emphasize that social workers must work with patients and partners to develop positive couple coping strategies. Practice implications for social workers are addressed.

The Influence of relational factors on men's adjustment to their partners' newly-diagnosed breast cancer.

Steadily rising breast cancer rates in America's women are forcing more men to confront challenges of living with a partner afflicted with this disease. This study assesses teh impact of mutual emotional support as perceived by male partners on their adjustment to the diagnosis and the illness and on interactions between their perceived emotional support nd their coping methods. Seventy-one male partners of newly-diagnosed breast cancer patients completed standardized instruments that measured emotional wellbeing, illness intrusiveness, emotional support, dyadic coping styles and demographic factors. Regression analysis revealed significant associations between perceived emotional support and men's coping strategies, and between coping styles and illness intrusiveness. Also, a history of depression predisposes men to poorer adjustment and affects their coping patterns. Findings suggest that as the health care system continues to relocate burden of care to partners and families, social workers must increase their understanding of how to effectively assist patients' partners. This study emphasizes the need to work with patients and partners to develop positive coping strategies as a couple.

Mushroom poisoning--from diarrhea to liver transplantation.

Mushroom poisoning from the genus Amanita is a medical emergency, with Amanita phalloides being the most common species. The typical symptoms of nausea, vomiting, abdominal pain, and diarrhea are nonspecific and can be mistaken for gastroenteritis. If not adequately treated, hepatic and renal failure may ensue within several days of ingestion. In this case series, patients poisoned with Amanita virosa are described with a spectrum of clinical presentations and outcomes ranging from complete recovery to fulminant hepatic failure. Although there are no controlled clinical trials, a few anecdotal studies provide the basis for regimens recommended to treat Amanita poisoning. Use of i.v. penicillin G is supported by most reports. Silibinin, although preferred over penicillin, is not easily available in the United States. In those with acute liver failure, liver transplantation can be life saving.

Tec kinases: modulators of lymphocyte signaling and development.

The Tec kinases are implicated as important components of the antigen receptor signaling required for proper lymphocyte activation and development. Recent data suggest that these kinases contribute to multiprotein complexes containing LAT and SLP-76 in T cells, and BLNK/SLP-65 in B cells, which are required for activation of PLC-gamma and downstream pathways.

Tec family kinases modulate thresholds for thymocyte development and selection.

Tec family kinases are implicated in T cell receptor (TCR) signaling, and combined mutation of inducible T cell kinase (Itk) and resting lymphocyte kinase (Rlk)/Txk in mice dramatically impairs mature T cell function. Nonetheless, mutation of these kinases still permits T cell development. While itk(-)(/)- mice exhibit mild reductions in T cells with decreased CD4/CD8 cell ratios, rlk(-)(/)-itk(-)(/)- mice have improved total T cell numbers yet maintain decreased CD4/CD8 ratios. Using TCR transgenics and an in vitro thymocyte deletion model, we demonstrate that mutation of Tec kinases causes graded defects in thymocyte selection, leading to a switch from negative to positive selection in rlk(-)(/)-itk(-)(/)- animals. The reduction in both positive and negative selection and decreased CD4/CD8 ratios correlates with decreased biochemical parameters of TCR signaling, specifically defects in capacitive Ca(2+) influx and activation of the mitogen-activated kinases extracellular signal-regulated kinase 1 and 2. Thus, Tec kinases influence cell fate determination by modulating TCR signaling, leading to altered thresholds for thymocyte selection. These results provide support for a quantitative model for thymic development and provide evidence that defects in negative selection can substantially alter thymic cellularity.

Conditional deletion of the Bcl-x gene from erythroid cells results in hemolytic anemia and profound splenomegaly.

Bcl-x is a member of the Bcl2 family and has been suggested to be important for the survival and maturation of various cell types including the erythroid lineage. To define the consequences of Bcl-x loss in erythroid cells and other adult tissues, we have generated mice conditionally deficient in the Bcl-x gene using the Cre-loxP recombination system. The temporal and spatial excision of the floxed Bcl-x locus was achieved by expressing the Cre recombinase gene under control of the MMTV-LTR. By the age of five weeks, Bcl-x conditional mutant mice exhibited hyperproliferation of megakaryocytes and a decline in the number of circulating platelets. Three-month-old animals suffered from severe hemolytic anemia, hyperplasia of immature erythroid cells and profound enlargement of the spleen. We demonstrate that Bcl-x is only required for the survival of erythroid cells at the end of maturation, which includes enucleated reticulocytes in circulation. The extensive proliferation of immature erythroid cells in the spleen and bone marrow might be the result of a fast turnover of late red blood cell precursors and accelerated erythropoiesis in response to tissue hypoxia. The increase in cell death of late erythroid cells is independent from the proapoptotic factor Bax, as demonstrated in conditional double mutant mice for Bcl-x and Bax. Mice conditionally deficient in Bcl-x permitted us for the first time to study the effects of Bcl-x deficiency on cell proliferation, maturation and survival under physiological conditions in an adult animal.

A report of congenital indifference to pain as diagnosed in infancy.

As physicians and nurses, we have a fundamental obligation to consider patient welfare above all else. The case presented demonstrates the depth of this obligation. Our patient, who was diagnosed during infancy with congenital indifference to pain, can neither feel nor perceive the meaning of pain. Congenital indifference to pain is a rare disorder, which is characterized by normal intelligence, development, and sensation despite an absence of pain perception. There is no cure and treatment difficulties are plentiful. Educating and maintaining an open, consistent relationship with the family and caregivers is essential to reduce the morbidity of this condition.

Treating gastro-oesophageal reflux disease during pregnancy and lactation: what are the safest therapy options?

Gastro-oesophageal reflux and heartburn are reported by 45 to 85% of women during pregnancy. Typically, the heartburn of pregnancy is new onset and is precipitated by the hormonal effects of estrogen and progesterone on lower oesophageal sphincter function. In mild cases, the patient should be reassured that reflux is commonly encountered during a normal pregnancy: lifestyle and dietary modifications may be all that are required. In a pregnant woman with moderate to severe reflux symptoms, the physician must discuss with the patient the benefits versus the risks of using drug therapy. Medications used for treating gastro-oesophageal reflux are not routinely or vigorously tested in randomised, controlled trials in women who are pregnant because of ethical and medico-legal concerns. Safety data are based on animal studies, human case reports and cohort studies as offered by physicians, pharmaceutical companies and regulatory authorities. If drug therapy is required, first-line therapy should consist of nonsystemically absorbed medications, including antacids or sucralfate, which offer little, if any, risk to the fetus. Systemic therapy with histamine H2 receptor antagonists (avoiding nizatidine) or prokinetic drugs (metoclopramide, cisapride) should be reserved for patients with more severe symptoms. Proton pump inhibitors are not recommended during pregnancy except for severe intractable cases of gastrooesophageal reflux or possibly prior to anaesthesia during labour and delivery. In these rare situations, animal teratogenicity studies suggests that lansoprazole may be the best choice. Use of the least possible amount of systemic drug needed to ameliorate the patient's symptoms is clearly the best for therapy. If reflux symptoms are intractable or atypical, endoscopy can safely be performed with conscious sedation and careful monitoring the mother and fetus.

Measuring the spin polarization of a metal with a superconducting point contact

A superconducting point contact is used to determine the spin polarization at the Fermi energy of several metals. Because the process of supercurrent conversion at a superconductor-metal interface (Andreev reflection) is limited by the minority spin population near the Fermi surface, the differential conductance of the point contact can reveal the spin polarization of the metal. This technique has been applied to a variety of metals where the spin polarization ranges from 35 to 90 percent: Ni0.8Fe0.2, Ni, Co, Fe, NiMnSb, La0.7Sr0.3MnO3, and CrO2.

Portal decompression by transjugular intrahepatic portosystemic shunt and changes in serum-ascites albumin gradient.

The serum ascites albumin gradient (SAAG) is widely used to help determine the cause of ascites formation. A serum ascites albumin gradient of > or = 1.1 g/dL reliably distinguishes portal hypertension-related ascites from other causes. To date, there are no published data on the impact of portal decompression on this gradient. The recent development of transjugular intrahepatic portosystemic shunt (TIPS) allows for nonsurgical decompression of portal hypertension by radiologically creating a portosystemic shunt. This study examines the short-term impact of portal decompression on the serum ascites albumin gradient (SAAG) in patients with portal hypertension-related ascites undergoing transjugular intrahepatic portosystemic shunt. Portal pressure measurements were obtained before and after TIPS placement. Serum ascites albumin gradient was determined before and at 6 and 24 hours post-TIPS placement. Fifteen patients were enrolled in the study. The mean portosystemic gradient (PSG) before TIPS was 21.0 +/- 9.2 mmHg, whereas the post-TIPS mean PSG was reduced to 11.0 +/- 6.3 mmHg, consistent with portal decompression (p = 0.005). The mean pre-TIPS serum ascites albumin gradient was 1.9 +/- 0.5 g/dL and was reduced to 1.7 +/- 0.5 g/dL at 6 hours (p = 0.003) and 1.4 +/- 0.4 g/dL at 24 hours (p = 0.002) after TIPS placement. These findings further solidify the association between the SAAG and portal hypertension.

Nausea and vomiting of pregnancy.

NVP is a spectrum of disorders ranging from the physiologically typical mild to moderate nausea and vomiting that is usually self-limited, to the pathologic, intractable symptoms of hyperemesis gravidarum that are associated with metabolic and electrolyte disturbances and weight loss. Up to 90% of pregnant women experience NVP. The pathogenesis remains poorly understood with multifactorial theories proposed combining both biologic and psychological factors. Diagnosing this syndrome is straightforward, but other organic sources should be excluded when symptoms are severe or prolonged. The overall prognosis is excellent for typical NVP, but whether hyperemesis gravidarum increases the risk to the fetus is controversial. Initial management should be conservative, including reassurance of the transient nature of the symptoms and the good prognosis, in addition to dietary modifications. Pharmacologic therapy is reserved for patients with persistent symptoms and is appropriate after discussion of the risks and benefits with informed consent. Alternative treatments, including psychotherapy and other nonpharmacologic modalities, are less proven but potentially safe and effective, thus providing additional therapeutic options. In refractory cases, nutritional supplementation becomes life-saving for both the mother and the fetus. Therapeutic abortion is a rare and last resort, to be used only when maternal life is threatened.

A preliminary analysis of interactive effects between common classroom contingencies and methylphenidate.

To assess the drug-behavior interaction effects with an 8-year-old boy wih attention deficit hyperactivity disorder, common classroom antecedent (e.g., seating arrangement) and consequent (e.g., peer prompts) stimuli were alternated within a school day while drug conditions (methylphenidate vs. placebo) were alternated across days. The results suggested that peer attention maintained disruptive behavior when methylphenidate was absent but not when it was present.

A comparison of reinforcer assessment methods: the utility of verbal and pictorial choice procedures.

We compared three methods of stimulus preference assessment for verbal children and specifically evaluated the utility of a verbal choice procedure for assessing relative reinforcer value. Using a token system, relative preference for five categories of reinforcers, representing 15 different stimuli, was assessed by three methods: a reinforcer survey, a verbal stimulus-choice questionnaire, and a pictorial stimulus-choice procedure. Results showed that the verbal and pictorial stimulus-choice assessments accurately identified high- and low-preference categories for 3 of 4 participants. Survey results alone often rated multiple categories as high preference, were less likely to identify low-preference categories, and were less likely to correspond with the results of a reinforcer assessment.