Obese Ningxiang pig-derived microbiota rewires carnitine metabolism to promote muscle fatty acid deposition in lean DLY pigs.
The gut microbiota consistently shows strong correlations with lipid metabolism in humans and animals, and whether the gut microbiota contributes to muscle fatty acid (FA) deposition and meat traits in farm animals has not been fully resolved. In this study, we aimed to unveil the microbial mechanisms underlying muscle FA deposition in pigs. First, we systematically revealed the correlation between the gut microbiome and muscle FA levels in 43 obese Ningxiang pigs and 50 lean Duroc Landrace Yorkshire (DLY) pigs. Mutual fecal microbial transplantation showed that the obese Ningxiang pig-derived microbiota increased the muscle FA content and improved meat quality by reshaping the gut microbial composition in lean DLY pigs. Lactobacillus reuteri has been identified as a potential microbial biomarker in obese Ningxiang pig-derived microbiota-challenged DLY pigs. A gavage experiment using lean DLY pigs confirmed that L. reuteri XL0930 isolated from obese Ningxiang pigs was the causal species that increased the muscle FA content. Mechanistically, SLC22A5, a carnitine transporter, was downregulated in L. reuteri XL0930-fed DLY pigs, resulting in reduced muscle carnitine levels. Muscle and intestinal L-carnitine levels, which correlated with the muscle FA content, impeded fat synthesis and FA accumulation in in vitro and in vivo models. In conclusion, we uncovered an unexpected and important role of the obese Ningxiang pig-derived microbiota in regulating muscle FA metabolism via the SLC22A5-mediated carnitine system.


Does the 23-valent pneumococcal vaccine protect cochlear implant recipients?
The incidence of meningitis is increased in cochlear implant (CI) recipients. Besides malformations, immunological deficiencies are predisposing factors. Therefore, the immunological background of CI recipients and the immunogenicity of the recommended 23-valent pneumococcal polysaccharide vaccine (PPV-23) were investigated. Prospective study in a tertiary care referral center. One hundred twenty CI recipients who were at least 5 years of age were vaccinated with PPV-23. Levels of immunoglobulins G, A, and M (IgG, IgA, and IgM, respectively) and IgG subclasses IgG1-IgG4 before vaccination and serum concentrations of antibodies against seven pneumococcal serotypes before and 4 weeks after vaccination were determined. The cohort was subdivided by patient age into groups A1 (5-8 y), A2 (8-12 y), and A3 (>12 y). Geometric mean concentrations of pneumococcal antibodies before vaccination were remarkably low in all three groups, emphasizing the importance of vaccination in this risk group. All groups showed a statistically significant increase in geometric mean concentrations after immunization. For group A1 compared with groups A2 and A3, response was limited, especially for serotypes 6B (geometric mean concentration, 1.71 microg/mL; P = .0007), 23F (geometric mean concentration, 2.28 microg/mL; P = .04), and 14 (geometric mean concentration, 3.98 microg/mL; P = .0004). The percentages of patients reaching the presumed protective threshold of at least 1 microg/mL pneumococcal antibody concentration were at least 71.1% in group A1, 93.8% in group A2, and 90.5% in group A3. This raises the question of whether PPV-23 evokes satisfying seroprotection in CI recipients younger than 8 years of age. With regard to the increased risk for bacterial meningitis, the authors recommend priming CI recipients younger than 8 years of age with pneumococcal conjugate vaccine followed by a PPV-23 booster.


Endothelial nitric oxide synthase is not essential for the development of fibrosis and portal hypertension in bile duct ligated mice.
It is postulated that nitric oxide (NO) is responsible for the hyperdynamic circulation of portal hypertension. Therefore, we investigated induction of fibrosis and hyperdynamic circulation in endothelial NO synthase knock-out (KO) mice. Fibrosis was induced by bile duct ligation. Hemodynamic studies were performed after portal vein ligation. All studies were performed in wild-type (WT) and KO mice. Three to 4 weeks after bile duct ligation (BDL), both WT and KO groups had similar degrees of portal hypertension, 12 (9-14) and 11(8-15) mmHg, median (range), and liver function. Fibrosis increased from 0.0% in sham operated to 1.0 and 1.1% in WT and KO mice, respectively. Cardiac output was similar after portal vein ligation (20 and 17 ml/min in WT and KO mice, respectively). There was no difference in liver of mRNA for endothelin 1, inducible NO synthase (iNOS) and hem-oxygenase 1 (HO1); proteins of iNOS, HO1 and HO2; nor in endothelin A and B (EtA and EtB) receptor density between WT and KO mice after BDL. These results suggest that endothelial NO synthase is neither essential for the development of fibrosis and portal hypertension in bile duct ligated mice, nor for the hyperdynamic circulation associated with portal hypertension in the portal vein ligated mice.


Purification and characterization of antizyme inhibitor of ornithine decarboxylase from rat liver.
A protein inhibiting a protein inhibitor (antizyme) to ornithine decarboxylase (L-ornithine carboxy-lyase, EC 188.8.131.52) (ODC), antizyme inhibitor, was purified from the liver cytosol of thioacetamide-treated rats by procedures including antizyme affinity chromatography. Overall purification was roughly estimated to be about 17,000,000-fold and recovery was about 2.4%. The purified preparation showed one major protein band and a faint band corresponding in mobility to molecular weights of 51,000 and 53,500, respectively, on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Judging from the ornithine decarboxylase activity of the final preparation, the faint band may be ornithine decarboxylase. The apparent molecular weight of antizyme inhibitor estimated by gel filtration on Sephacryl S-200 was approx. 62,000, indicating that antizyme inhibitor may be composed of a single polypeptide chain. In order to examine the question of whether antizyme inhibitor is a protein derived from ornithine decarboxylase, an inactive ornithine decarboxylase, in an immunotitration study and analysis of the binding to antizyme were investigated. The results indicate that antizyme inhibitor may be a protein distinct from ornithine decarboxylase.


Ammonia fiber expansion-assisted deep eutectic solvent treatment for wheat straw fraction separation and bioconversion.
In this study, an ammonia fiber expansion (AFEX)-assisted deep eutectic solvent (DES) pretreatment method was developed for the rapid separation of wheat straw fractions, which reduced the pretreatment time for DES and improved the pretreatment efficiency. This study describes the feasibility of the AFEX-assisted DES pretreatment in terms of both progressive and parallel relationships and analyzes the subsequent enzymatic effect in generating glucose from cellulose. Ammonia fiber expansion-assisted DES one-pot pretreatment at 120 °C, for 1.5 h resulted in an enzymatic efficiency of 98.0 ± 3.1 %. Moreover, the enzyme efficiency remained greater than 85 % after three recovery cycle experiments. The comparison between regenerated-lignin (d-lignin) and alkaline-lignin showed that regenerated lignin has a lower molecular weight and belongs to para-hydroxy-phenyl-guaiacyl-syringyl (H-G-S) type lignin. This study developed is a green and efficient pretreatment process with great potential in the separation and utilization of biomass fractions.


Analysis of urinary kallikrein-related peptidase 13 for monitoring bladder cancer.
Bladder cancer (BC) is one of the 10 most common types of cancer worldwide, with approximately 550,000 new cases each year. Early detection and appropriate diagnosis are important factors in successful treatment of the disease. We used specific fluorogenic substrate for the quantitative determination of urine kallikrein 13 (KLK13) activity in healthy (n = 15) and BC (n = 54) patients. The proteolytic activity in individual urine samples was determined by fluorescence measurements. Then, immunoenzymatic analyses (ELISA, Western blot) were performed to confirm the presence of KLK13 in the tested samples. Urine samples from patients with G2 and G3 grade BC contained proteolytically active KLK13, as confirmed by kinetic analysis and immunochemical detection. KLK13 was not detected in the urine of patients with G1 grade BC. Previous clinical study reveals the KLK13 significance for BC prognosis as increased KLK13 expression was highlighted in bladder tumours compared to normal adjacent tissues. Our findings correlate to the report. KLK13 activity was confirmed in BC patients with G2 and G3 stage of disease development. Using specific chromogenic/fluorogenic peptides could be useful for the non-invasive disease monitoring of BC progress.


Functional characterization of fibrinolytic metalloproteinases (colombienases) isolated from Bothrops colombiensis venom.
Researchers trying to improve the safety and efficacy of fibrinolytic therapy have been isolating fibrinolytic enzymes from snake venoms. Two fibrinolytic enzymes, colombienases 1 and 2, with significant activity have been isolated from the venom of Bothrops colombiensis. The colombienases were characterized for various biological activities which include hemorrhagic, fibrinogenolytic, proteolytic, hemolytic, edematogenic and cytotoxic. Colombienases directly acted on fibrin by degrading fibrinogen Aα and Bβ chains without activating the fibrinolytic system (plasminogen/plasmin), additionally colombienase-2 degraded fibrinogen γ chains as well as the fibronectin molecule. Laminin and type IV collagen were colombienases resistant. Gelatin-zymogram activity was present in B. colombiensis venom (BcV) bands between 64 and 148 kDa. All activities were abolished by metalloproteinases inhibitors. Both enzymes lacked hemorrhagic, hemolytic, cytotoxic, plasminogen activator and coagulant activities. Both colombienases had direct fibrino(geno)lytic activity without other toxic side effects including in vivo hemorrhaging, which could be promising in terms of therapeutic potential as thrombolytic agents.


Direct Synthesis of Amides by Dehydrogenative Coupling of Amines with either Alcohols or Esters: Manganese Pincer Complex as Catalyst.
The first example of base-metal-catalysed synthesis of amides from the coupling of primary amines with either alcohols or esters is reported. The reactions are catalysed by a new manganese pincer complex and generate hydrogen gas as the sole byproduct, thus making the overall process atom-economical and sustainable.


Systematic analysis of solvents and other volatile substances by gas chromatography.
Four column packings for screening volatiles in biological material by gas chromatography are evaluated. Retention data are standardized by the calculation of retention indices, and packing materials are compared by discriminating power and identification power. A combination of 5% Carbowax 20M on Carbopack B and 0.3% Carbowax 20M on Carbopack C appears to be best suited for screening. Hydroxy-n-alkanes are used as reference substances for the calculation of retention indices.


Effect of posttranscriptional regulatory elements on transgene expression and virus production in the context of retrovirus vectors.
Ineffective transgene expression in a sufficient amount of target cells is still a limitation in retroviral vector mediated gene therapy. Thus, we systematically evaluated four genetic modulators, (i) the woodchuck posttranscriptional regulatory element (WPRE), (ii) the mouse RNA transport element (RTE), (iii) the constitutive transport element (CTE) of the simian retrovirus type 1 (SRV-1), and (iv) the 5' untranslated region of the human heat shock protein 70 (Hsp70 5'UTR), all of them involved in the posttranscriptional control of mRNA nucleo/cytoplasmatic transport, RNA stability, and translation efficiency, in an MLV-based retrovirus vector context. Insertion of the WPRE into the retrovirus vector resulted in enhancement of transgene expression (EGFP) both in transfected virus producing cells as well as in infected recipient cells irrespective of the location in the vector. The best effect was observed with two copies of the WPRE, 3' of the transgene and in the 3' untranslated region of the vector backbone. However, oligomerization of this element does not further increase transgene expression. Presence of the WPRE resulted also in an increase in virus production. Introduction of the CTE and/or RTE in the retroviral vector did not alter transgene expression and infectious particle production. Positive effects were observed only in vectors harboring the CTE and/or RTE in combination with the WPRE. The activity of the Hsp70 5'UTR as a translational enhancer was found to be negligible in the context of the retroviral vector. However, interference of the Hsp70 5'UTR strong secondary structure with the packaging sequence of the viral RNA was experimentally excluded as being the cause of this. These data suggest that only the WPRE is a suitable element for the improvement of transgene expression and oncoretroviral vector production.


Pulmonary fungal infections after bone marrow transplantation.
Of 319 pediatric patients treated with bone marrow transplantation (BMT) during a 10-year period, 27 developed pulmonary fungal infections (PFI). Only 2 patients (7%) survived. Twenty-three patients (85%) had been treated with systemic antifungal therapy immediately before or at the time of diagnosis. Nineteen patients (70%) were neutropenic, and 4 of the 8 patients who were not neutropenic were being treated with systemic steroids for graft vs. host disease (GVHD). Seven patients (26%) died within 7 days of diagnosis. The diagnosis was made ante-mortem in 9 patients (33%). Radiographic abnormalities were variable. At the onset of chest X-ray (CXR) change, the pulmonary infiltrates were unilateral in 14 patients (52%) and, at diagnosis, bilateral in 18 (66%). At diagnosis the infiltrates were interstitial in 3 patients (11%), alveolar in 20 (74%) and mixed in 4 (15%). Six patients (22%) developed cavitary lesions. The infecting agents were Aspergillus in 21 patients (78%), Candida in 7 (26%), Mucormycosis in 3 (11%), and Fusarium in 1 (4%). Five patients (19%) had mixed fungal infections and 7 (26%) had concurrent cytomegalovirus (CMV) pulmonary infections. Although the radiographic changes are often nonspecific in PFI, alveolar or nodular infiltrates in neutropenic patients or in those being treated for GVHD should strongly suggest a fungal etiology.


Development of a blocking ELISA for Caprine parainfluenza virus type 3.
Caprine parainfluenza virus type 3 (CPIV3) is a novel pathogen mainly causing respiratory diseases in goats. At present, there are no high throughput and rapid testing methods available for epidemiological investigation. In this study, we designed a modified method for selection of hybridomas that secrete monoclonal antibodies (mAb) specific for CPIV3. The monoclonal antibodies were obtained by combination of indirect enzyme-linked immunosorbent assay (iELISA) and blocking ELISA (bELISA). The technique was efficient to determine each mAb with specificity and sensitivity. One bELISA was validated for the serological diagnosis of CPIV3. After optimization conditions were established, a total of 205 reference goat sera were tested in parallel by bELISA and by virus neutralization (VN) for their relative performances. The cut-off point was ultimately defined as 33.6% by ROC curve analysis. The bELISA specificity and sensitivity were 99.2% and 98.7%, respectively, and agreement with the VN test was >99.0%. Furthermore, testing another 2919 goat sera by bELISA demonstrated 39.3% prevalence in the goat population, more sensitive than HI detection. This new bELISA would offer higher throughput, sensitivity, and specific detection for CPIV3, and will be of great value not only for surveillance, but also for monitoring the efficiency of vaccination programs in the future.


Preparation of natural indicator incorporated media and its logical use as a colorimetric biosensor for rapid and sensitive detection of Methicillin-resistant Staphylococcus aureus.
Herein, we report a rational design in preparation of natural indicator incorporated media used as a rapid, selective, sensitive and economic colorimetric biosensor for detection of Methicillin-resistant Staphylococcus aureus (MRSA). Anthocyanins obtained from red cabbage (Brassica oleracea) extract (RCE) are major components and acted as pH responsive indicators in the RCE media. The RCE media was tested for eight gram-positive, four gram-negative bacterial and one model fungal pathogens. We experimentally revealed how salt concentration and antibiotic (cefoxitin) in the RCE media influence the growth of pathogens and especially MRSA. Benefiting pH dependent color change function of anthocyanins is indispensable point of the RCE media in sensing of MRSA growth. The potential MRSA colorimetric sensing mechanism of anthocyanins relies on both protonation and decrease in electron density on structures of anthocyanins by acidic organic volatile compounds produced during growth of MRSA. The protonated anthocyanins with low electron density changed original purple color of the RCE media to pink color. We demonstrated that detection of MRSA was achieved by spectrophotometrically (one single cell) and visually with naked eyes (100 cells) in 2nd hrs and 6th hrs of incubation, respectively. We optimized concentrations of cefoxitin (from 1 μg/mL to 20 μg/mL) against MRSA and Methicillin-sensitive Staphylococcus aureus (MSSA) bacterial cell (3 McFarland) suspensions, then growth of MRSA was visually and clearly detected in the presence of 4 μg/mL cefoxitin between 90 min and 3 h. We persistently offer that the proposed RCE media can be a well alternative to currently used phenotypic methods in clinics owing to its much rapid, sensitive, selective and economic properties.


An Endoplasmic Reticulum Stress-MicroRNA-26a Feedback Circuit in NAFLD.
Endoplasmic reticulum (ER) stress is an adaptive response to excessive ER demand and contributes to the development of numerous diseases, including nonalcoholic fatty liver disease (NAFLD), which is hallmarked by the accumulation of lipid within hepatocytes. However, the underlying mechanisms remain elusive. MicroRNAs (miRNAs) play an indispensable role in various stress responses, but their implications in ER stress have not yet been systemically investigated. In this study, we identify a negative feedback loop consisting of hepatic ER stress and miR-26a in NAFLD pathogenesis. Combining miRNA dot blot array and quantitative PCR, we find that miR-26a is specifically induced by ER stress in liver cells. This induction of miR-26a is critical for cells to cope with ER stress. In human hepatoma cells and murine primary hepatocytes, overexpression of miR-26a markedly alleviates chemical-induced ER stress, as well as palmitate-triggered ER stress and lipid accumulation. Conversely, deficiency of miR-26a exhibits opposite effects. Mechanistically, miR-26a directly targets the eukaryotic initiation factor 2α, a core ER stress effector controlling cellular translation. Intriguingly, miR-26a is reduced in the livers of patients with NAFLD. Hepatocyte-specific restoration of miR-26a in mice significantly mitigates high-fat diet-induced ER stress and hepatic steatosis. In contrast, deficiency of miR-26a in mice exacerbates high-fat diet-induced ER stress, lipid accumulation, inflammation and hepatic steatosis. Our findings suggest ER stress-induced miR-26a up-regulation as a regulator for hepatic ER stress resolution, and highlight the ER stress/miR-26a/eukaryotic initiation factor 2α cascade as a promising therapeutic strategy for NAFLD.


Risk of Non-Hodgkin Lymphoma among Patients with Hepatitis B Virus and Hepatitis C Virus in Taiwan: A Nationwide Cohort Study.
Hepatitis B virus (HBV) and hepatitis C virus (HCV) are associated with an increased risk of developing non-Hodgkin lymphoma (NHL); however, adequate data corroborating these associations are lacking. Therefore, a study based on the national database was performed to investigate the correlation between HBV and HCV with NHL in Taiwan. This research was a retrospective cohort study using a nationally representative database established by the Health and Welfare Data Science Center of the Ministry of Health and Welfare, Taiwan. The participants were patients with HBV and HCV, analyzed using the propensity score matching method. The study results indicated that the incidence rate of NHL (0.13%) was significantly higher than that in patients from the general population. After controlling related variables, the hazard ratio (HR) of the incidence of NHL in patients with hepatitis was 2.37 (95% CI, 1.93-2.91). Furthermore, the incidence of NHL in patients with HBV was significantly higher than in patients from the general population (HR, 2.49; 95% CI, 1.94-3.19). The incidence of NHL in patients with HCV was significantly higher than in patients from the general population (HR, 2.36; 95% CI, 1.73-3.22). This study indicated that HBV and HCV significantly increase the risk of NHL.


Economic Evaluation of Implementing a Rapid Point-of-Care Screening Test for the Identification of Hepatitis C Virus under National Viral Hepatitis Control Programme in Tamil Nadu, South India.
Viral hepatitis is a crucial public health problem in India. Hepatitis C virus (HCV) elimination is a national priority and a key strategy has been adopted to strengthen the HCV diagnostics services to ensure early and accurate diagnosis. To conduct an economic evaluation of implementing a rapid point-of-care screening test for the identification of HCV among the selected key population under the National Viral Hepatitis Control Programme in Tamil Nadu, South India. Economic evaluation of a point-of-care screening test for HCV diagnosis among the key population attending the primary health care centers. A combination of decision tree and Markov model was developed to estimate cost-effectiveness of point-of-care screening test for HCV diagnosis at the primary health care centers. Total costs, quality-adjusted life years (QALYs) of the intervention and comparator, and incremental cost-effectiveness ratio (ICER) were calculated. The model parameter uncertainties which would influence the cost-effectiveness outcome has been evaluated by one-way sensitivity analysis and probabilistic sensitivity analysis. When compared to the tertiary level diagnostic strategy for HCV, the point-of-care screening for selected key population at primary health care level results in a gain of 57 undiscounted QALYs and 38 discounted QALYs, four undiscounted life years and two discounted life years. The negative ICER of the new strategy indicates that it is less expensive and more effective compared with the current HCV diagnosis strategy. The proposed strategy for HCV diagnosis in the selected key population in Tamil Nadu is dominant and cost-saving compared to the current strategy.


Chronic treatment with a benzodiazepine agonist in vivo increases the actions of the benzodiazepine partial inverse agonist, FG7142, on the hippocampal slice in vitro.
We have shown previously that chronic treatment of mice with a benzodiazepine agonist, flurazepam, increased the pharmacological actions of the partial inverse agonist, FG7142. We have investigated the neurophysiological basis for this using extracellular recordings of evoked field potentials in area CA1 of isolated hippocampal slices. The slices were prepared 48 h after the end of the chronic in vivo treatment, a time when no evidence of residual benzodiazepine agonist activity was found in the CNS. During perfusion with standard Ringer solution, no significant differences were seen between the field potentials in slices from flurazepam-treated mice and those from control animals. When FG7142 was added to the perfusion medium there was an increase in the secondary discharges that followed the initial population spikes, and an increase in paired pulse potentiation. These increases were significantly greater in slices from flurazepam-treated mice, compared with controls. The results show that the effects of the partial inverse agonist, FG7142, on an isolated neuronal preparation, were increased by chronic administration of a benzodiazepine agonist in vivo. This effect is suggested to be due to a decrease in GABAergic inhibition.


Creating a Care for the Caregiver Program in a Ten-Hospital Health System.
Critical care clinicians involved serious adverse events may experience a constellation of distressing emotions that may interfere with home and work life. Offering support after a serious adverse event may restore a clinician's ability to cope with the event, reestablish emotional balance and assist a clinician to function capably in the workplace and at home. A description of a care for the caregiver program implementation at a 10-hospital health system provides a roadmap to implement this program in other hospitals and health systems.


Increases of neuropeptide Y-like immunoreactivity in plasma during insulin-induced hypoglycemia in man.
Neuropeptide Y-like immunoreactivity (NPY-LI) in plasma during insulin-induced hypoglycemia was measured in 4 healthy male volunteers. Plasma NPY-LI increased from 167 +/- 11 pg/ml to 247 +/- 25 pg/ml 30 min after the administration of insulin (0.1 U/kg body weight IV), reached the maximum (296 +/- 6 pg/ml) 45 min after the insulin, and then decreased. These results suggest that NPY is released into the systemic circulation during insulin-induced hypoglycemia in man.


[Anorectal reconstruction by coloperineal anastomosis and dynamic double graciloplasty after abdomino-perineal resection].
The aim of this study was to assess the technical and functional results of total anorectal reconstruction with double dynamic graciloplasty after abdominoperineal resection (APR). From May 1995 to December 1996. 10 patients (6 males and 4 females), with a mean age of 54 years (range 39-74), underwent anorectal reconstruction for low rectal adenocarcinoma. All patients had preoperative radiotherapy and six had postoperative chemotherapy. The surgical procedure was performed in three stages: 1) APR, coloperineal anastomosis, double graciloplasty and ileostomy; 2) three months later, implantation of stimulator and leads; 3) after a two-months training period, the stoma was closed. There was no postoperative mortality. Early and late morbidity occurred in 5 patients: 2 colonic fistulas, 1 necrosis of colon, 1 ileostomy prolapse, 1 neosphincter stenosis, 1 sepsis of stimulator. No patient had recurrence of the disease (mean follow-up 16 months), but two patients died at 3 and 8 months, respectively from anorexia and pulmonary embolism. Seven patients were available for evaluation (2 fistula, 1 death). Before training, the resting pressure and the squeeze pressure were 30 and 175 cm H2O respectively. At the time of evaluation, the electrical stimulated pressure was 95 cm H2O. Six of these 7 patients were continent (5 with spontaneous defecation, 1 with enemas) and 1 was incontinent. Anorectal reconstruction with dynamic graciloplasty can be an alternative to permanent colostomy for selected patients after APR. However, there is a high morbidity and the quality of life of the patients must be evaluated.


The High-Risk Young People's Program. A summing up.
For over five years, 21 teaching hospitals and 54 community cosponsors provided health services to young people at risk for sociomedical problems, i.e., to young people living in communities characterized by high rates of teen pregnancy, sexually transmitted disease, drug abuse, alcohol abuse, accidents, homicide, suicide, and mental illness. With support from The Robert Wood Johnson Foundation, the 20 grantees of the High-Risk Young People's Program developed projects whose collective goal was to expand services to high-risk youth and improve their health. This goal was to be achieved by a direct provision of medical services, training health providers in the care of high-risk youth, consolidating categorical youth services into single, comprehensive care sites, and securing long-term support for these new services and training activities. During the grant period, 114 fellows, 974 residents, 453 medical students, and 126 graduate nurses trained at project sites. Patient visits, which totaled 47,203 the first year, reached 84,754 the second year, and were reported at 89,024 in the fourth year. Sixteen of the 20 projects secured 117 grants worth nearly $7 million. We conclude that the projects were successful in training health care providers and in securing additional support for Program purposes, but were less successful in expanding services beyond the initial pilot for clinical care.


A direct high-throughput protein quantification strategy facilitates discovery and characterization of a celastrol-derived BRD4 degrader.
We describe a generalizable time-resolved Förster resonance energy transfer (TR-FRET)-based platform to profile the cellular action of heterobifunctional degraders (or proteolysis-targeting chimeras [PROTACs]) that is capable of both accurately quantifying protein levels in whole-cell lysates in less than 1 h and measuring small-molecule target engagement to endogenous proteins, here specifically for human bromodomain-containing protein 4 (BRD4). The detection mix consists of a single primary antibody targeting the protein of interest, a luminescent donor-labeled anti-species nanobody, and a fluorescent acceptor ligand. Importantly, our strategy can readily be applied to other targets of interest and will greatly facilitate the cell-based profiling of small-molecule inhibitors and PROTACs in a high-throughput format with unmodified cell lines. We furthermore validate our platform in the characterization of celastrol, a p-quinone methide-containing pentacyclic triterpenoid, as a broad cysteine-targeting E3 ubiquitin ligase warhead for potent and efficient targeted protein degradation.


Quantification of the relative impairment in actions of insulin on hepatic glucose production and peripheral glucose uptake in non-insulin-dependent diabetes mellitus.
In non-insulin-dependent diabetes mellitus (NIDDM), both liver and peripheral tissues are resistant to insulin, but the relative severity and contribution of these abnormalities to fasting hyperglycemia are poorly understood. We, therefore, determined the dose-response characteristics for insulin-mediated suppression of hepatic glucose production (GP) and stimulation of peripheral glucose uptake (GU) in 14 NIDDM subjects and 14 age- and weight-matched nondiabetic volunteers (NV) using the glucose clamp sequential insulin infusion technique along with isotopic estimation of glucose flux. Postabsorptive rates of both GP (94 +/- 7 v 72 +/- 2 mg/M2/min in NV, P less than .01) and GU (88 +/- 5 v 72 +/- 2 in NV, P less than .01) were significantly increased in NIDDM subjects. The ED50 (half-maximally effective plasma insulin concentration) in NIDDM subjects for suppression of GP (64 +/- 14 microU/mL) and stimulation of GU (118 +/- 20 microU/mL were both increased more than twofold above normal (26 +/- 2 and 58 +/- 5 microU/mL, respectively, both P less than .01) and were significantly correlated with one another (r = .68, P less than .01). Although GP could be totally suppressed in the NIDDM subjects, their maximal GU was reduced 30% (287 +/- 20 v 372 +/- 15 mg/m2/min in NV, P less than .01). Nevertheless, at all physiologically relevant plasma insulin concentrations studied, there was comparable impairment in GP and GU responses. Moreover, fasting plasma glucose concentrations in NIDDM subjects were highly correlated with their increased basal rates of GP (r = .81, P less than .005) but not with their reduced GU.(ABSTRACT TRUNCATED AT 250 WORDS)


The distribution of degenerating axons after small lesions in the intact and isolated visual cortex of the cat.
The extent of the spread of axonal degeneration was investigated in the visual cortex of the cat after making small lesions restricted to the grey matter. Two series of experiments were undertaken. In the first, normal adult cats were used, and in the second, the cortex of the postlateral gyrus was isolated from its extrinsic afferents by surgical undercutting 3 months before making the lesions. The results were similar in the two series in most respects. 1. Horizontal fibres extended in considerable numbers for some 500 micrometer from the lesion, mainly in layers I, III/IV and V, a few reaching 2/3 mm. These fibres were better seen in the intact than in the isolated cortex. Their spread was usually asymmetrical, being greater posteromedially than anterolaterally. 2. Oblique axons ran downwards from the middle layers into layers V and VI, or upwards into layers I and II. 3. Axons arising from layers II to VI descended vertically into the white matter. Degeneration patterns after lesions in areas 17 and 18 were compared.


Preferential crystallization.
This contribution focuses primarily on chiral discrimination in the solid state, i.e. the formationof conglomerates, how to detect them and the various benefits that can be retrieved from chiral recognitionin crystal lattices.The core of the work is devoted to phase diagrams (mainly binary and ternary). The stable and metastableheterogeneous equilibria are depicted and used to understand different variants of preferential crystallizationand how the entrainment effect proceeds. Seeded and auto-seeded processes are analyzed minutely. The seedingprocedure has a significant impact on the control of stereoselective secondary nucleation and crystalgrowth.The review ends with a systematic analysis of the reasons that can put, sometimes, severe limitationson the entrainment effect.


A new relationship between cholesterolemia and cholesterol synthesis determined in rats fed an excess of cystine.
The present study deals with an attempt to describe how the plasma cholesterol level is related to input into the plasma of cholesterol synthesized in the liver and in the intestine. It has previously been shown in our laboratory that, for a given absorption of alimentary cholesterol, the rat plasma cholesterol level decreases when internal secretion of cholesterol (cholesterol synthesized in the organs and poured into the plasma) increases. This relationship was established using rats in which the major source of cholesterol synthesis was the intestine. We used rats fed a cystine-enriched diet (5%) which was previously shown to increase cholesterolemia and internal secretion of cholesterol. It was first demonstrated that a significant positive linear correlation exists between individual values of cholesterolemia and those of internal secretion of cholesterol. Secondly, using [14C]acetate as the cholesterol precursor it was shown that ingestion of the cystine-enriched diet increased hepatic but not intestinal cholesterogenesis. Individual values of cholesterolemia were linearly correlated to those of [14C]acetate incorporation into the hepatic sterols. Results obtained by this method were validated by determining the 13C-labeling pattern of cholesterol synthesized de novo by the liver and the intestine after [13C]acetate infusion. Indeed, this labelling indicated that the dilution of exogenous acetyl-CoA in the liver was not changed by cystine feeding, whereas that in the intestine was enhanced. It is concluded that the plasma cholesterol level varies with internal cholesterol secretion, depending on the organ which determines the variations of this secretion: it decreases when intestinal cholesterogenesis increases, whereas it increases when hepatic cholesterogenesis increases. Finally, the use of [14C]acetate coupled with lipoprotein analysis in rats fed the cystine-enriched diet, in control rats and in rats fed a cholesterol-enriched diet, allowed a new linear correlation to be demonstrated: between cholesterol concentration in LDL2 (lipoproteins of density 1.040-1.063 g/ml) and [14C]acetate incorporation into liver sterols. Our results suggest that LDL2 are produced by the liver in relation to cholesterogenesis in this organ.


Occult papillary thyroid microcarcinoma manifesting only as a symptomatic lateral cervical mass: report of a case.
We report a case of occult papillary thyroid carcinoma (PTC) manifesting as a solitary lateral cervical mass. Few such cases have been reported and, to our knowledge, this is the first report of cervical lymph node involvement from thyroid cancer being so massive that it is the cause of the local symptoms. The patient, a 64-year-old man, presented with vocal cord paralysis and voice alteration and was found to have a 4 cm lateral cervical mass infiltrating the ipsilateral internal jugular vein and recurrent laryngeal nerve. The diagnosis of PTC was made preoperatively following an open-biopsy of the lesion. The patient underwent total thyroidectomy with modified radical neck dissection, followed by radioactive iodine therapy. His postoperative course was uneventful and he remains well without any signs of recurrence 7 years after the operation.


Co-grafting of antiadhesive and antimicrobial agents onto UV-micropatterned copper surfaces.
Adhesion of proteins and bacteria was reduced by a factor close to one order of magnitude, and adhered bacteria were dramatically damaged on copper surfaces by grafting a PEG-modified PMMA-based (polymethyl methacrylate) copolymer together with an antimicrobial peptide. To obtain PEG and a peptide grafted together on the surface, a UV sensitive copolymer (containing PMMA, PEG and a UV sensitive reagent) was primary synthesized and deposited. After selective UV irradiation of this copolymer layer, an antimicrobial peptide, Magainin I, was grafted onto freed-polymer coated-copper surface via a spacer molecule (a mercapto carboxylic acid). The functionalization was characterized at each step by Polarization Modulation Reflection Absorption Infrared Spectroscopy (PM-RAIRS). The antiadhesive properties of the copolymer layer and antibacterial activity of the anchored Magainin I, were individually tested toward adsorption of bovine serum albumin (BSA) proteins, and against Gram positive bacteria, Listeria ivanovii, respectively. The results revealed that adhesion of both proteins and bacteria has been considerably reduced; moreover, the peptide still displays some antimicrobial activity after grafting. This work gives new ideas and perspectives to elaborate complex surface coatings where several agents are needed like for anti-biofilm or sensing applications.


Alopecia associated with unexpected leakage from electron cone.
Excessive irradiation due to unexpected leakage was found on a patient receiving electron beam therapy. The cause of this leakage was analyzed and the amount of leakage was measured for different electron beam energies. The highest leakage occurred with a 6 x 6 cm cone using a 12 MeV electron beam. The leakage dose measured along the side of the cone could be as great as 40%. Until the cones are modified or redesigned, it is advised that all patient setups be carefully reviewed to assure that no significant patient areas are in the side scatter region.


Bioinspired Micro Glue Threads Fabricated by Liquid Bridge-to-Solidification as an Effective Sensing Platform.
Spiders synthesize their web using a liquid bridge-to-solidification mechanism at the end of their glands. Inspired by this process, in this work, we fabricated micro-glue threads (μGTs, polymer microwires) by a simple "pinch and spread" process using just two fingertips. The μGTs exhibited excellent tensile strength (∼50 GPa), comparable to those of spider silk and biological fibers. The chemical, physical, and mechanical properties of the μGTs were investigated, and it was confirmed that the thickness of the μGTs could be controlled by ethanol treatment in varying concentrations. Moreover, electrically conductive μGTs were easily fabricated by simply mixing them with various nanomaterials such as gold nanoparticles, zinc oxide nanowires, and reduced graphene oxide (rGO). Interestingly, the conductive μGTs, fabricated using rGO, exhibited remarkable electrical conductivity (0.45 μS) compared to those fabricated using other materials. The conductive μGTs are applicable not only to NO2 gas sensing but also as electrical fuselike materials that melt when the humidity increases. Collectively, the results present μGTs as cost-effective, simple, and versatile materials, which enables their application in a variety of sensors.


Fetal growth and subsequent maternal risk of thyroid cancer.
Thyroid cancer has peak incidence among women of reproductive age, and growth factors, which have procarcinogenic properties, may play an important etiologic role. However, the association between fetal growth rate during a woman's pregnancy and her subsequent risk of thyroid cancer has not been previously examined. We conducted a national cohort study of 1,837,634 mothers who had a total of 3,588,497 live-births in Sweden in 1973-2008, followed up for thyroid cancer incidence through 2009. There were 2,202 mothers subsequently diagnosed with thyroid cancer in 36.8 million person-years of follow-up. After adjusting for maternal age, height, weight, smoking, and sociodemographic factors, high fetal growth (birth weight standardized for gestational age and sex) was associated with a subsequent increased risk of thyroid cancer in the mother (incidence rate ratio [IRR] per additional 1 standard deviation, 1.05; 95% CI, 1.01-1.09; p = 0.02). Each 1,000 g increase in the infant's birth weight was associated with a 13% increase in the mother's subsequent risk of thyroid cancer (IRR, 1.13; 95% CI, 1.05-1.22; p = 0.001). These findings appeared to involve both papillary and follicular subtypes, and did not vary significantly by the mother's height, weight or smoking status. In this large national cohort study, high fetal growth during a woman's pregnancy was independently associated with a subsequent increased risk of her developing thyroid cancer. If confirmed, these findings suggest an important role of maternal growth factors in the development of thyroid cancer, and potentially may help facilitate the identification of high-risk subgroups of women.


[Schwannoma of the vagus nerve: diagnostic strategy and therapeutic approach].
Scwhannomas are uncommon neurogenic tumors of the mediastinum. Most of them are located in the posterior mediastinum usually in the paralateral gutters. We report the case of an uncommon localisation of such a tumour appended to the right vagus nerve in the middle mediastinum. The patient was 50 years old and complained of thoracic pain, shortness of breath and dysphagia. CT scanning of thorax and abdomen revealed a heterogeneous mass in the middle mediastinum, which was not visible on plain radiography. Further investigation included transoesophageal ultrasound and PET scan. The diagnosis was confirmed by histopathology after mini video-assisted thoracotomy. The schwannoma was entirely removed without any post-surgical complications. We report a novel case of schwannoma arising from the vagus nerve and review the diagnostic strategies. PET scanning has poor sensitivity and specificity and does not therefore contribute to confirm the diagnosis which depends on exploring the mediastimun surgically.


Altered activity of the sodium-potassium pump in arteries of rats with steroid hypertension.
1. Ouabain-sensitive uptake of 86Rb, a measure of the Na+-K+ pump activity, was studied in tail arteries of rats made hypertensive with deoxycorticosterone and saline. 2. Decreased activity of the ouabain-sensitive Na+-K+ pump supports the hypothesis that the activity of Na+-K+ pump is suppressed in volume expanded hypertension.


The effects of vigour on measures of obesity across time.
Past studies expected measures of obesity to be positively associated with positive affects. However, this hypothesis was not tested in reference to a specific positive affect. We tested the hypothesized unidirectional effects of measures of obesity on vigour, representing a positive affect, and of vigour on measures of obesity. We used a longitudinal design, separately for men and women. Participants were 1,876 and 931 healthy men and women, respectively, examined at Time 1 (T1) and Time 2 (T2), about 2 years apart. Measures of obesity included body mass index, waist circumference, and waist-to-hip ratio. Vigour was assessed by the Shirom-Melamed Vigour Measure. We used structural equation modelling to test our hypotheses. We found that for both genders, T1 measures of obesity did not predict either T1 or T2 vigour. Among both genders, we found support for the effects of T1 vigour on T1 but not on T2 measures of obesity. To the extent that the 'Jolly fat hypothesis' refers to the effects of measures of obesity on positive affects, we failed to support it for vigour as a positive affect. Vigour has contemporaneous but not longitudinal effects on body weight.


High-Quality Genome Sequence of Bacillus anthracis Strain 14RA5914 Isolated during an Outbreak in Germany in 2014.
Bacillus anthracis is a zoonotic agent causing anthrax, a notifiable disease in animals. The last anthrax outbreak among cattle in Germany occurred in April 2014 in Saxony-Anhalt. Here we report a high-quality genome sequence of the Bacillus anthracis strain 14RA5914 Dobichau isolated from the spleen of a dead cow.


Normalization of blood loss in women with heavy menstrual bleeding treated with an oral contraceptive containing estradiol valerate/dienogest.
The study was conducted to assess the efficacy of estradiol valerate/dienogest (E₂V/DNG) administered using an estrogen step-down and progestogen step-up approach in a 28-day regimen in the treatment of heavy menstrual bleeding (HMB) using clinical end points allowing E₂V/DNG to be compared with other available medical therapies. This was a pooled analysis of data from two identically designed randomized, placebo-controlled, multiple center studies conducted in Europe, Australia and North America that assessed the effectiveness of E₂V/DNG in reducing menstrual blood loss (MBL) in women with HMB. Women aged ≥ 18 years with objectively confirmed HMB were randomized to E₂V/DNG (n=220) or placebo (n=135) for seven treatment cycles. Outcomes analyzed included absolute reduction in MBL from baseline, proportion of women successfully treated (defined as MBL below 80 mL and ≥ 50% reduction in MBL), proportion with MBL below 80 mL and proportion with ≥ 50% reduction in MBL from baseline. At study end, 63.6% and 11.9% of patients were successfully treated with E₂V/DNG and placebo, respectively, with 68.2% and 15.6% of women with MBL below 80 mL, and 70.0% and 17.0% with MBL reduction ≥ 50% (all p<.001). E₂V/DNG is highly effective for the treatment of HMB and is associated with a high rate of treatment success.


The impact of the COVID-19 pandemic on ectopic pregnancy presentation and treatment: A retrospective cohort study.
We examined ectopic pregnancy (EP) incidence, presentation and management, before and during the COVID-19 pandemic, and following initiation of vaccination against COVID-19. In a single-center retrospective cohort study, we compared incidence, presentation and management of EP, between 98 women who presented during the pandemic (March 1 2020 to August 31, 2021), and 94 women diagnosed earlier (March 1 2018 to August 31, 2019). Sub-periods before and after introduction of the vaccination were compared. Age and parity were similar between the periods. For the pandemic compared to the earlier period, the median gestational age at EP presentation was higher (6.24 ± 1.25 vs. 5.59 ± 1.24, P<0.001), and the proportions were higher of symptomatic women (42.9% vs. 27.7%, p = 0.035) and urgent laparoscopies (42.9% vs. 24.5%, p = 0.038). In a multivariable linear model, women who presented during the pandemic were more likely to undergo an urgent laparoscopy [OR 2.30, 95%CI (1.20-4.41)], P = 0.012. In urgent surgeries performed during the pandemic compared to the earlier period, the proportion of women with a hemoglobin drop >2 gr/dL was greater (60% vs. 30%, p = 0.024). Statistically significant differences were not found in sonographic or laboratory findings, in rupture or massive hemoperitoneum rates, or in the need for blood transfusion in urgent laparoscopy. Outcomes were similar before and after introduction of vaccinations. During the pandemic, and even after the introduction of vaccination, women with EP were more likely to undergo urgent surgery, and blood loss was greater. This is likely due to delayed diagnosis.


Pre-operative chemotherapy for stage IIIa (N2) non-small cell lung cancer.
From June 1990 to December 1993, 36 patients were enrolled in a phase II study, aimed at determining the feasibility of surgery, patterns of disease recurrence and survival after neoajuvant chemotherapy in non-small cell lung cancer (NSCLC) stage IIIA-N2. Twenty-seven patients underwent invasive staging procedures (i.e. mediastinoscopy or needle biopsy). Two CHT schedules were used. Cisplatin (P) 90 mg/mq, day 1, mitomycin (M) 6 mg/mq, day 1, and vindesine (V) 5 mg/mq, days 1, 8, 15, were administered every 3 weeks for 3 cycles in the first 20 patients. The last 16 patients were treated with cisplatin (P) 90 mg/mq, day 1, mitomycin (M) 6 mg/mq, day 1, and vinorelbina 20 mg/mq, days 1, 8, 15. Thoracotomy was performed 15-20 days after haematological recovery in the objective-responders. Thirty-two patients were evaluable for response to CHT. The overall objective response (OR) rate was 78.1%. There were three complete (CR) (9.4%) and 22 partial responses (PR) (68.7%). The 25 patients with OR underwent radical surgery (16 pneumonectomies, one bilobectomy, seven lobectomies and one wedge resection). The only morbidity reported was a late broncho-pleural fistula (on post-operative day 37). There were three post-operative deaths in patients who underwent pneumonectomy: two due to an empyema following a broncho-pleural in fistula and one by pulmonary embolism. Histology was negative for the three CRs. Six patients with residual nodal involvement at surgery underwent radiotherapy. Relapse occurred in seven resected patients. Presently 14 patients are alive, all but one being disease-free, with a median follow-up of 30.5 months (15-47). Median survival was 31 months (5-47). Actuarial 3-year survival rate is 49%. Our results confirm the high response rate of CHT, as well as the feasibility and the overall low complication rate of both treatments (CHT and surgery).


Confusional symptomatology distinguishes early- and late-onset Alzheimer's disease.
Symptoms of confusion were examined in 75 patients with Alzheimer's disease (AD). Mild confusion was found in 20, and mode-rate/severe confusion in 8 patients. Confusion was more frequent in the late-onset (26/44-59%) than in the early-onset AD group (2/31-6%) (p less than 0.0001), and patients with confusion were older (p less than 0.0001) than those without confusion. The frequency of confusion was higher in patients with ischemic heart disease (13/28-46%) than in patients without this vascular factor (10/47-21%) (p less than 0.05). An inverse relation was found between confusional symptomatology and parietal-lobe symptoms. The findings in this study suggest that a subgroup of AD patients, fulfilling the NINCDS-ADRDA criteria for probable AD, is characterized by a clinical picture of mild confusional symptomatology together with no or mild parietal-lobe symptomatology, higher age and higher frequency of ischemic heart disease. This group contrasts with the other subgroup of pure AD, which is characterized by a clinical picture of marked parietal-lobe symptomatology, almost no confusional symptomatology, lower age and lower frequency of ischemic heart disease.


Reducing perioperative blood loss with antifibrinolytics and antifibrinolytic-like agents for patients undergoing total hip and total knee arthroplasty.
Total hip and knee arthroplasties may be associated with a significant amount of perioperative blood loss. The severity of blood loss may be great enough to require the use of blood transfusions to treat perioperative anemia. Various methods of blood preservation have been studied. The use of antifibrinolytics and antifibrinolytic-like agents to reduce perioperative bleeding has been researched in orthopaedics and other surgical subspecialties. This review aims to evaluate the current evidence supporting the use of tranexamic acid, aminocaproic acid, fibrin tissue adhesive, and aprotinin in the reduction of perioperative blood loss in total hip and knee arthroplasties.


Integrating material properties from magnetic resonance elastography into subject-specific computational models for the human brain.
Advances in brain imaging and computational methods have facilitated the creation of subject-specific computational brain models that aid researchers in investigating brain trauma using simulated impacts. The emergence of magnetic resonance elastography (MRE) as a non-invasive mechanical neuroimaging tool has enabled in vivo estimation of material properties at low-strain, harmonic loading. An open question in the field has been how this data can be integrated into computational models. The goals of this study were to use a novel MRI dataset acquired in human volunteers to generate models with subject-specific anatomy and material properties, and then to compare simulated brain deformations to subject-specific brain deformation data under non-injurious loading. Models of five subjects were simulated with linear viscoelastic (LVE) material properties estimated directly from MRE data. Model predictions were compared to experimental brain deformation acquired in the same subjects using tagged MRI. Outcomes from the models matched the spatial distribution and magnitude of the measured peak strain components as well as the 95th percentile in-plane peak strains within 0.005 mm/mm and maximum principal strain within 0.012 mm/mm. Sensitivity to material heterogeneity was also investigated. Simulated brain deformations from a model with homogenous brain properties and a model with brain properties discretized with up to ten regions were very similar (a mean absolute difference less than 0.0015 mm/mm in peak strains). Incorporating material properties directly from MRE into a biofidelic subject-specific model is an important step toward future investigations of higher-order model features and simulations under more severe loading conditions.


Ubiquitin, cellular inclusions and their role in neurodegeneration.
Covalent binding of ubiquitin to proteins marks them for degradation by the ubiquitin/ATP-dependent pathway. This pathway plays a major role in the breakdown of abnormal proteins that result from oxidative stress, neurotoxicity and mutations. Failure to eliminate ubiquitinated proteins disrupts cellular homeostasis, causing degeneration. Inclusions containing ubiquitinated proteins are commonly detected in many neurological disorders. These aggregates are mostly cytosolic; nevertheless, ubiquitinated inclusions are found in endosomes/lysosomes in Alzheimer's disease and prion encephalopathies, and in nuclei in disorders associated with CAG/polyglutamine repeats, such as Huntington's disease and spinocerebellar ataxias. Ubiquitinated aggregates must result from a malfunction or overload of the ubiquitin/ATP-dependent pathway or from structural changes in the protein substrates, halting their degradation. Prevention of protein aggregation in these diseases might offer new therapeutic leads.


Birth stress and lateral preferences.
Conflicting evidence exists concerning the possible role of birth stress in the etiology of left-sided lateral preferences. In order to clarify this issue, associations among lateral preferences of hand, eye, and foot and eight indices of prenatal and perinatal stress were examined in the present study on a sample of 987 boys and girls who participated in the Philadelphia Collaborative Perinatal Project. Controls were instituted for some of the methodological and measurement problems encountered in past birth stress and laterality research. Results showed that subjects with different lateral preferences did not differ significantly in their distributions of all but one birth stress items. Hence, there was no substantial evidence for a link between birth stress and left-sided preferences. Alternative hypotheses for the etiology of left-sidedness should therefore be explored.


Genomic analyses of the metastasis-derived prostate cancer cell lines LNCaP, VCaP, and PC3-AR.
The lymph node metastasis-derived LNCaP, the bone metastasis-derived PC3 (skull), and VCaP (vertebral) cell lines are widely used as preclinical models of human prostate cancer (CaP) and have been described in more than 19,000 publications. Here, we report on short-read whole-genome sequencing and genomic analyses of LNCaP, VCaP, and PC3 cells stably transduced with WT AR (PC3-AR). LNCaP, VCaP, and PC3-AR cell lines were sequenced to an average depth of more than 30-fold using Illumina short-read sequencing. Using various computational methods, we identified and compared the single-nucleotide variants, copy-number profiles, and the structural variants observed in the three cell lines. LNCaP cells are composed of multiple subpopulations, which results in nonintegral copy number states and a high mutational load when the data is analyzed in bulk. All three cell lines contain pathogenic mutations and homozygous deletions in genes involved in DNA mismatch repair, along with deleterious mutations in cell-cycle, Wnt signaling, and other critical cellular processes. PC3-AR cells have a truncating mutation in TP53 and do not express the p53 protein. The VCaP cells contain a homozygous gain-of-function mutation in TP53 (p.R248W) that promotes cancer invasion, metastasis, and progression and has also been observed in prostate adenocarcinomas. In addition, we detect the signatures of chromothripsis of the q arms of chromosome 5 in both PC3-AR and VCaP cells, strengthening the association of TP53 inactivation with chromothripsis reported in other systems. Our work provides a resource for genetic, genomic, and biological studies employing these commonly-used prostate cancer cell lines.


[The Cryopre-servation: history and the ethical issue of storing embryos].
This review article deals with the technique which is called cryopreservation. Text describes the principle of this method, history and ethical problems in assisted reproduction.


Probing the nitrite and nitric oxide reductase activity of cbb3 oxidase: resonance Raman detection of a six-coordinate ferrous heme-nitrosyl species in the binuclear b3/CuB center.
In this work we report the first spectroscopic evidence demonstrating that cbb3 oxidase catalyzes the reduction of nitrite to nitrous oxide under reducing anaerobic conditions. The reaction proceeds through the formation of a ferrous six-coordinate heme b3-nitrosyl species that has been characterized by resonance Raman spectroscopy.


Fabrication of drug-loaded polymer microparticles with arbitrary geometries using a piezoelectric inkjet printing system.
Carrier geometry is a key parameter of drug delivery systems and has significant impact on the drug release rate and interaction with cells and tissues. Here we present a piezoelectric inkjet printing system as a simple and convenient approach for fabrication of drug-loaded polymer microparticles with well-defined and controlled shapes. The physical properties of paclitaxel (PTX)-loaded poly(lactic-co-glycolic acid) (PLGA) inks, such as volatility, viscosity and surface tension, were optimized for piezoelectric inkjet printing, and PTX-loaded PLGA microparticles were fabricated with various geometries, such as circles, grids, honeycombs, and rings. The resulting microparticles with 10% (w/w) PTX exhibited a fairly homogeneous shape and size. The microparticle fabrication by piezoelectric inkjet printing was precise, reproducible, and highly favorable for mass production. The microparticles exhibited a biphasic release profile with an initial burst due to diffusion and a subsequent, slow second phase due to degradation of PLGA. The release rate was dependent on the geometry, mainly the surface area, with a descending rate order of honeycomb>grid, ring>circle. The PTX-loaded microparticles showed a comparable activity in inhibiting the growth of HeLa cells. Our results demonstrate that a piezoelectric inkjet printing system would provide a new approach for large-scale manufacturing of drug carriers with a desired geometry.


Comparative studies on lipid and colony-stimulating factor-induced macrophage growth.
We previously reported that lipids such as cholesterol esters, triglycerides, and some phospholipids that constitute cell membranes or serum lipoproteins induced growth of mouse peritoneal macrophages in vitro. In this paper, we compared the macrophage growth-stimulating activity of cardiolipin (CL), an active phospholipid with that of CSF-1. Growth kinetics and maximal degree of growth of exudated macrophages induced by CL were similar to those of CSF-1. CL did not stimulate macrophages to release soluble macrophage growth factors. Also, the activity of CL was not blocked as much by anti-CSF-1, suggesting that most of the effect of CL was direct and not mediated by CSF-1 or other protein factors. There was no synergistic effect between CL and CSF-1. CL induced growth of both exudate and resident macrophages, whereas CSF-1 induced very little resident macrophage growth. Furthermore, although the growth-stimulating activities of both substances were inhibited by IFN-gamma and TNF, CL was more resistant to these inhibitory effects. These results suggest that the lipid has some different characters from CSF-1 and may induce the growth of resident macrophages in inflammations or tumors.


Application of cyclodextrin-modified gold nanoparticles in enantioselective monolith capillary electrochromatography.
β-cyclodextrin modified gold nanoparticles (CD-GNP) were employed as the stationary phase in monolith capillary electrochromatography (CEC) to facilitate enantioseparation. CD-GNP were covalently bound to the surface of the thiolated porous polymer monolithic column. The fabricated enantioselective monolithic column was characterized by a variety of spectroscopic methods. The column exhibited steady EOF mobility over pH values ranging from 4.6 to 9.7. Additionally, the column was stable under CEC separation conditions over 180 min. Moreover, the column exhibited good column-to-column reproducibility. The CD-GNP-modified monolithic column was employed in the efficient CEC separation of three pairs of drug enantiomers (chlorpheniramine, zopiclone and tropicamide). The results exhibit reproducible run-to-run enantioseparations and the monolith column can maintain its enantioselectivity for more than 1 month if the column is stored in a CD-GNP solution at 4°C.


Effect of drinks that are added as flavoring in oral midazolam premedication on sedation success.
Midazolam is one of the most frequently used agents for sedation in pediatric dentistry. The injectable form of midazolam can also be given orally. However, its bad taste has negative effects on ingestion of the drug. In this study, we aimed to evaluate the effect of drinks which were added to mask the bitter taste of midazolam for drug acceptance and sedation. In the laboratory; the pH values of 2.5 ml midazolam (15 mg x 3 ml(-1)) and the mixtures of Pepsi Cola, 10% sodium citrate, fresh pomegranate juice, and fresh grapefruit juice in equal volumes were measured. Seventy-five patients between 2 and 8 years of age who were assessed to have anxiety with the Frankl Behavior Scale and whose dental treatment was planned under sedation were randomly divided into five groups. Pepsi Cola (Group I), 10% sodium citrate (Group II), pomegranate juice (Group III), and grapefruit juice (Group IV) which were added to 0.75 mg x kg(-1) midazolam in equal volumes, and (Group V) 0.75 mg x kg(-1) midazolam given orally to children. The drug compliance of children was evaluated. After 15 min, 4-5 l x min(-1) of N(2)O : O(2) (40 : 60) inhalation was started via a nasal hood. During the procedure, heart rate, and SpO2 were monitored with pulse oximetry and sedation levels with the Ramsay Sedation Scale (RSS). Anesthetist, dentist, and parental satisfaction levels were recorded. The groups had similar demographics, drug ingestion was better in Groups I and II, but the mean RSS was the highest in Group II (P < 0.05). As well as making drug ingestion much simpler, the addition of sodium citrate to the midazolam administered orally to the children increased the effectiveness of sedation.


[Adenocarcinoma of the cervix--analysis of 363 cases].
Form 1959 through to May, 1993, 363 patients with adenocarcinoma of uterine cervix were initially treated which accounted for 2.5% of all initially treated cases of cervical carcinoma. Patients' mean age was 50.5 years. Forty-six percent of the patients were post-menopausal. The main symptom was irregular vaginal and/or discharge. Over 88% of the cases were in stage II and III of the disease. They were treated with radiotherapy alone. The overall 5-year survival rate was 51.8%. The recurrence rate was 17.4% with a mean time to recurrence of 33.3 months. Thirteen patients recurred beyond 5 years. Recurrence was accompanied with lung metastasis in 17 patients and supraclavicular eymph node metastases in 11. Among patients whose tumor recurred the 1- and 2-year survival rate were 35.2% and 19.2% respectively. The most significant prognostic factors were the stage of disease, the size of the tumor and the histologic subtypes


Removal of tetracycline from water by catalytic photodegradation combined with the microalga Scenedesmus obliquus and the responses of algal photosynthesis and transcription.
The antibiotic tetracycline (TC) and its degradation products (TDPs) in degradation solution present serious environmental problems, such as human health damage and ecological risk; thus further treatment is required before being released into the aquatic environment. Furthermore, their environmental impact on microalgae remains unclear. In this study, TC was degraded by photocatalysis using birnessite and UV irradiation, followed by biological purification using the microalga Scenedesmus obliquus. In addition, the photosynthetic activity and transcription of the microalgae were examined to evaluate the toxicity of TC and TDPs. The results show that photocatalytic degradation efficiency reached 92.7% after 30 min, and 11 intermediate products were detected. The microalgae achieved a high TC removal efficiency (99.7%) after 8 days. Exposure to the degraded TC solution (D) resulted in significantly lower (p < 0.05) biomass than the pure TC (T), and S. obliquus in the T treatment showed better resilience than the D treatment. Transcriptomic assays for different treatments revealed differential gene expression mainly involving the photosynthesis, ribosome, translation and peptide metabolic progresses. The up-regulation of photosynthesis-related genes and differential expression of chloroplast genes may be important for S. obliquus to acquire high photosynthetic efficiency and growth recovery when exposed to TC and TDPs. Our study provides a reference for TC removal using a combination of catalytic degradation and microalgal purification, and it is also helpful for understanding the environmental risk of TDPs in natural aquatic environments.


Purification and properties of aspartate transcarbamylase from Mycobacterium smegmatis.
Aspartate transcarbamylase (carbamoyl-phosphate: L-aspartate carbamoyltransferase, EC 220.127.116.11) has been purified from Mycobacterium smegmatis TMC 1546 using streptomycin sulphate precipitation, ammonium sulphate precipitation, DE-52 chromatography, second ammonium sulphate precipitation, Sephadex G-200 gel filtration, and aspartate-linked CNBr-activated Sepharose 4B affinity chromatography in successive order. The enzyme was purified 231.6-fold, and the preparation was found to be homogeneous on column chromatography and polyacrylamide gel electrophoresis. The purified enzyme had a molecular weight of 246,000 and was composed of two asymmetrical subunits. The kinetic and regulatory properties of aspartate transcarbamylase from M. smegmatis were also studied. The enzyme was found to be an allosteric in nature with carbamyl phosphate showing positive cooperativity and UMP exhibiting a negative cooperativity. CTP was found to be the most potent inhibitor among nucleotides. Phosphate acted as a non-competitive product inhibitor with respect to aspartate. Succinate and maleate exerted a competitive inhibition when aspartate was the variable substrate.


Characterization of biotin interference in 21 Vitros 5600 immunoassays and risk mitigation for patient safety at a large academic medical center.
Biotin and streptavidin are commonly used reagents in clinical immunoassays. Several cases of biotin interference with immunoassay testing for patients taking biotin supplements have been reported, yet, not all analytes and platforms susceptible to biotin interference have been characterized. The objectives of this study are to characterize biotin interference with 21 immunoassays using the Ortho Clinical Diagnostics Vitros 5600, evaluate a biotin-depletion method, and apply risk mitigation strategies for biotin interference during routine clinical testing at our institution. Residual serum without and with increasing concentrations of exogenous biotin were used to evaluate biotin interference with 21 immunoassays using the Vitros 5600. Biotin-depletion was evaluated by comparing measured analyte concentrations in serum with and without exogenous biotin and streptavidin-microparticle pretreatment. Focused education for healthcare professionals about biotin interference was performed in February 2018. Samples with suspected biotin interference were investigated using this biotin-depletion method, and analyte testing by alternate methodology for select samples. Exogenous biotin in serum caused dose-dependent negative biases in 15 immunometric assays, and dose-dependent positive biases in 6 competitive immunoassays. Streptavidin-microparticle pretreatment of serum containing exogenous biotin demonstrated recoveries 100 ± 15% of expected values for all 21 analytes. Physicians identified 21 samples suspicious for biotin interference over 11 months, and streptavidin-microparticle pretreatment verified 11 cases of biotin interference. Analytical bias caused by biotin interference is dependent on biotin concentration but independent of analyte concentration for immunometric methods using the Vitros 5600, and dependent on both biotin and analyte concentration for competitive immunoassays. Multi-disciplinary education and a lab streptavidin-microparticle pretreatment method help mitigate risk of erroneous results due to biotin interference for patient safety.


Demonstration of a pontine-hippocampal projection containing a ranatensin-like peptide.
Previous studies have demonstrated a neuronal system containing an immunoreactive ranatensin-like peptide (irRT) in rat brain. The results of the present study describe the ascending projections of a prominent irRT cell group in the pons. A major pontine irRT fiber system was shown to innervate the hippocampus but not the mammillary body.


Impact of sociocultural factors on hospital length of stay in children with nephrotic syndrome in Japan.
Hospital length of stay (LOS) of Japanese inpatients is extraordinarily long, partly because of the nature of the health insurance system in Japan. We investigated factors that affect the LOS in Japan, taking pediatric nephrosis as an example. The characteristics of 78 hospitalized children with steroid-sensitive nephrotic syndrome were studied. The median LOS of the patients was 47 days. Most of the patients' mothers (92.3%) stayed in hospital during admission to care for their children. Using multivariate Cox's regression analysis with time-dependent covariates, non-working mother and smaller number of siblings were found to be factors significantly associated with prolonged LOS. These factors are presumed to allow the mothers to care for their hospitalized children for a long period. No medical or clinical factors played a significant role in the prolonged LOS. These results suggest that sociocultural factors have a greater influence on the LOS in Japan than do medical factors such as disease course, coexisting illness, complications, and treatment response.


Comparative performance of various human papillomavirus assays available in Korea for detecting cervical intraepithelial neoplasia.
The aim of this study was to evaluate the clinical performance for detecting cervical intraepithelial neoplasia (CIN) 2 or higher lesions among available human papillomavirus infection (HPV) genotyping tests in Korea. Eligible patients visited 13 tertiary hospitals for colposcopic biopsy following cervical cytology and HPV genotyping test between January and December 2018. Baseline characteristics including age, body mass index (BMI), and parity were collected from 3798 patients. The performance of the Roche Cobas HPV 4800 was evaluated against other domestic HPV assays to detect CIN2 or higher. A total of seven types of HPV genotyping tests were analyzed in the research institutes. A total of 1358 patients (35.8%) tested Anyplex II HPV 28 and 701 patients (18.5%) tested Cobas 4800 HPV. The overall sensitivity in the detection of CIN2 or higher was 41.5% (38.9-44.1) in patients positive for HPV 16/18. The Cobas test for HPV 16/18 was concordant with other assays evaluated for detection of CIN2 or higher and showed sensitivity of 46.6%, which was not significantly different from other assays. Although Anyplex II HPV28 (Seegene) showed slightly decreased sensitivity for detecting CIN2 or higher lesion with HPV 16/18 positive (39.8%, p < 0.05) compared to Cobas 4800, in aspect of high-risk HPV positive, Anyplex II HPV28 showed increased sensitivity (96.9%, p < 0.05). The performance of the HPV genotype test that were commonly used in Korea was concordant with Cobas HPV test. Further studies are needed to evaluate the safety, efficiency, and cost-effectiveness of the various commercially available domestic HPV assays.


Neurophysiological responses to unpleasant stimuli (acute electrical stimulations and emotional pictures) are increased in patients with schizophrenia.
Patients with schizophrenia have often been described as insensitive to nociceptive signals, but objective evidence is sparse. We address this question by combining subjective behavioral and objective neurochemical and neurophysiological measures. The present study involved 21 stabilized and mildly symptomatic patients with schizophrenia and 21 control subjects. We applied electrical stimulations below the pain threshold and assessed sensations of pain and unpleasantness with rating scales, and Somatosensory Evoked Potentials (SEPs/EEG). We also measured attention, two neurochemical stress indices (ACTH/cortisol), and subjective VEPs/EEG responses to visual emotional stimuli. Our results revealed that, subjectively, patients' evaluations do not differ from controls. However, the amplitude of EEG evoked potentials was greater in patients than controls as early as 50 ms after electrical stimulations and beyond one second after visual processing of emotional pictures. Such responses could not be linked to the stress induced by the stimulations, since stress hormone levels were stable. Nor was there a difference between patients and controls in respect of attention performance and tactile sensitivity. Taken together, all indices measured in patients in our study were either heightened or equivalent relative to healthy volunteers.


Challenges faced by cancer patients in Uganda: Implications for health systems strengthening in resource limited settings.
Uganda Cancer Institute (UCI), the only comprehensive cancer treatment center in Uganda, registers about 4000 new cancer patients a year. However, many cancer patients in Uganda never receive treatment due to a variety of challenges. We therefore conducted a study to identify and assess the challenges faced by cancer patients in Uganda. A cross-sectional study conducted in April-May 2017 among adult cancer patients. 359 participants participated in an interviewer-administered survey. We used stratified random sampling to select the study participants. Data was analyzed in SPSS Statistics 24. 35 % of the patients delayed initiating cancer treatment and 41 % missed medical appointments along their care journey. Delayed and missed appointments were mainly due to lack of money for cancer medicines, transportation and accommodation. Patients also expressed challenges with side effects of cancer treatment: 52 % sought help from health workers when they experienced side effects; 14 % used alternative medicine; and 21 % did not inform anyone. In addition, 55 % of the participants had limited knowledge about their disease and treatment. Other challenges when at UCI included: being hungry and thirsty throughout the day, long waiting hours, not having a resting place, not understanding what comes next, and having their records lost by hospital staff. Challenges faced by cancer patients in Uganda result in enormous delays in initiation and continuation of cancer treatment. These challenges are often a result of the poor social-economic status of the patients; inadequate infrastructure for cancer care; and inefficiencies in the health care system. To improve the experience of patients, the National Cancer Control Plan should consider establishing regional cancer centers; creating a reliable supply of cancer medicines; and integrating navigation programmes into cancer care. Strengthening the whole health system, in relation to cancer service delivery, should remain a top priority for Uganda and other resource limited settings.


Reduced wave amplitudes of brainstem auditory response in high-risk babies born at 28-32week gestation.
To examine brainstem auditory electrophysiology in high-risk babies born at 28-32week gestation by analysing the amplitudes of wave components in maximum length sequence brainstem auditory evoked response (MLS BAER). 94 preterm babies, ranging in gestation 28-32weeks, with perinatal problems (high-risk) were recruited. The amplitudes of MLS BAER wave components were studied at term age (37-42weeks postconceptional age). Compared with normal term controls, the amplitude in the high-risk preterm babies was significantly smaller at the highest click rate 910/s for wave I (p<0.01), at all 91-910/s for wave III (all p<0.01) and at 455 and 910/s (p<0.05 and 0.01) for wave V. Compared with age-matched low-risk preterm controls, the amplitude was significantly smaller at 455 and 910/s for wave I (p<0.05 and 0.05), 91-910/s for wave III (p<0.05-0.001), and 227-910/s (p<0.05 and 0.01) for wave V. No differences in the V/I and V/III amplitude ratios were found between the high-risk preterm babies and the controls. The amplitudes of MLS BAER wave components, mainly more central components, were reduced in the high-risk preterm babies born at 28-32week gestation. Electrophysiological activity of the brainstem auditory neuron in such babies is depressed, mainly attributed to or related to the associated perinatal problems.


The development of drug resistance mutations K103N Y181C and G190A in long term Nevirapine-containing antiviral therapy.
We built a cohort study of HIV patients taking long-term first-line Antiretroviral Therapy in 2003. In this assay, we focused on the development of primary drug resistance mutations against Non-Nucleoside Reverse Transcriptase Inhibitor (NNRTI), K103N, Y181C and G190A. The cohort study was built in Henan province, China. We used Single Genome Amplification (SGA) to analyze the frequency of K103N, Y181C and G190A in serial plasma samples of three individual patients. We also performed standard genotype HIV drug resistance assay in 204 patients of this cohort study to analyze the frequency of these mutations. In the SGA sequences, the K103N decreased and vanished, while the frequency of Y181C and G190A increased in individual patient receiving long-term Antiretroviral Therapy (ART). In the sequences of standard genotype HIV drug resistance assay, the frequency of K103N, Y181C and G190A had the similar pattern with that in SGA sequences. Among these patients, the viral suppression were still sufficient after receiving ART for 72 months, and 78.6% (160/204) patients could have their CD4 count over than 200cells/ul. In some patients, first-line ART had the possibility to provide sufficient treatment effect for over than 72 months, but in long-term treatment, the dominant NNRTI drug resistance mutation K103N could reduced, while the proportion of variants with mutation Y181C or G190A may increased. This result was not similar with that in vitro study, which state that variant with K103N or Y181C had an equal viral fitness with wild type.


Prediction of protein structural classes using hybrid properties.
In this paper, amino acid compositions are combined with some protein sequence properties (physiochemical properties) to predict protein structural classes. We are able to predict protein structural classes using a mathematical model that combines the nearest neighbor algorithm (NNA), mRMR (minimum redundancy, maximum relevance), and feature forward searching strategy. Jackknife cross-validation is used to evaluate the prediction accuracy. As a result, the prediction success rate improves to 68.8%, which is better than the 62.2% obtained when using only amino acid compositions. Therefore, we conclude that the physiochemical properties are factors that contribute to the protein folding phenomena and the most contributing features are found to be the amino acid composition. We expect that prediction accuracy will improve further as more sequence information comes to light. A web server for predicting the protein structural classes is available at http://app3.biosino.org:8080/liwenjin/index.jsp.


Feasibility and usability of a non-immersive virtual reality tele-cognitive app in cognitive rehabilitation of patients affected by Parkinson's disease.
Cognitive impairment is one of the most common non-motor features of Parkinson's disease (PD). The aim of the present study was to evaluate the feasibility and acceptability/usability of a protocol using a non-immersive virtual reality tele-cognitive app, performed remotely in a sample of Italian patients with PD. Non-demented patients with mild PD were included in the study. Patients performed the cognitive rehabilitation in a remote way, at home (three training sessions lasting 20 min/week for 6 weeks) using the NeuroNation app, downloaded for free on the patients' smartphones. The usability and feasibility of the tele-cognitive rehabilitation program were assessed with the System Usability Scale (SUS) and the Goal Attainment Scaling (GAS). Sixteen patients (9 men and 7 women; mean age 58.4 ± 8.3 years; mean disease duration 4.6 ± 2.1 years) were included in the study. At the end of the study, the mean SUS was 83.4 ± 11.5. The GAS score recorded at the end of the study (65.6 ± 4.2) was significantly higher than at baseline (38.5 ± 2.4; P-value <0.001). In our sample, good feasibility and usability were observed for a 6-week cognitive rehabilitation protocol based on the non-immersive virtual reality tele-cognitive app NeuroNation. Our data support the usefulness of cognitive rehabilitation performed in a remote way in PD patients.


The priority areas and possible pathways for health cooperation in BRICS countries.
As one of the largest alliances of middle-income countries, the BRICS, known as an acronym for five countries including "Brazil, Russia, India, China, and South Africa", represents half of the global population. The health cooperation among BRICS countries will benefit their populations and other middle- and low-income countries. This study aims to summarize the current status of health cooperation in BRICS countries and identify opportunities to strengthen BRICS participation in global health governance. A literature review was conducted to analyze the status, progress, and challenges of BRICS' health cooperation. Content analysis was used to review the 2011-2021 annual joint declarations of the BRICS Health Ministers Meetings. The priority health areas were identified through segmental frequency analysis. Our research suggested that communicable diseases, access to medicine, and universal health coverage appeared most frequently in the content of declarations, indicating the possible top health priorities among BRICS' health collaboration. These priority areas align with the primary health challenges of each country, including the threats of double burden of diseases, as well as the need for improving health systems and access to medicines. Respective external cooperation, inter-BRICS health cooperation, and unified external cooperation are the main forms of health cooperation among BRICS countries. However, challenges such as the lack of a unified image and precise position, lack of practical impact, and weak discourse power have impeded the impact of BRICS on health governance. This study suggests that the BRICS countries should recognize their positioning, improve their unified image, and establish cooperative entities; at the same time, they should increase their practical strength, promote non-governmental cooperation, and expand the cooperation space through the "BRICS Plus" mechanism with countries with similar interests to join.


Increased expression of miR-126 and miR-10a predict prolonged relapse-free time of primary oestrogen receptor-positive breast cancer following tamoxifen treatment.
Adjuvant tamoxifen is a valid treatment option for women with oestrogen receptor (ER)-positive breast cancer. However, up to 40% of patients experience distant or local recurrence or die. MicroRNAs have been suggested to be important prognosticators in breast cancer. This study aims to identify microRNAs with the potential to predict tamoxifen response. We performed a global microRNA screen (1105 human microRNAs) in primary tumours of six matched pairs of postmenopausal, ER-positive breast cancer patients treated with tamoxifen, who were either recurrence free or had developed a recurrence (median follow up: 8.84 years; range: 1.28-12.7 years). Patients of this discovery set and the 81 patients of the validation set (median follow up: 8.64 years; range: 0.21-19.85 years) were treated at the Robert Bosch Hospital, Stuttgart, Germany, between 1986 and 2005. Out of the top 20 deregulated microRNAs (12 up-regulated, eight down-regulated) miR-126 (Hazard Ratio (HR) = 0.56, 95% confidence interval (CI): 0.38-0.83; Holm-adj. P = 0.022) and miR-10a (HR = 0.53, 95% CI: 0.33-0.85; Holm-adj. P = 0.031) were identified as significant predictors of tamoxifen outcome by multivariate Cox regression analysis in the independent validation set of 81 postmenopausal, ER-positive patients. Kaplan-Meier survival analyses based on cut-offs determined by receiver operating characteristics curves confirmed that a higher expression of miR-126 and miR-10a in the patients tumour was associated with longer relapse-free time (log-rank P = 0.037, P<0.0001, respectively). Our data suggest that miR-126 and miR-10a are independent predictors for tumour relapse in early postmenopausal breast cancer patients treated with adjuvant tamoxifen.


Multiple lumbar arachnoid cysts. Case report.
Arachnoid cysts are a rare cause of compression of the contents of the lumbar spinal canal; in the literature only about 100 cases are reported. The various methods of diagnosis are discussed in the light of a recent case observed by the authors.


Self-reported utilization of preventive health services by retired employees age 65 and older.
Increased utilization of preventive services among the aging has been associated with improved health status and decreased medical costs. We sought to examine the use of the Health Risk Appraisal (HRA) in benchmarking compliance and characterizing those retired employees who met preventive service guidelines. A retrospective cohort study of retired employees age 65 and older. Nation-wide health promotion program offered by General Motors Corporation. 59,670 retired General Motors employees age 65 and older who participated in a nationwide mailed HRA health promotion program. Preventive health services compliance was measured using selected HRA questions. Gender, HRA participation patterns, overall health risk status, medical plan selection and disease status were examined as predictors of increased compliance. Multivariate logistic regression models were developed to test the relative contributions of participant characteristics to increased utilization. The self-reported HRA data indicated that compliance levels were higher than national averages. The Healthy People 2000 goals for the preventive services studied were met and exceeded (with the exception of tetanus immunization). Higher compliance was associated with being male, younger than 70 years, multiple-year HRA participation, overall low risk status and HMO insurance plan selection. The results from the HRA indicated that this population participated at a higher level than a comparable national sample exceeding goals set by Healthy People 2000.


Tumour necrosis factor alpha induces only minor inflammatory changes in the central nervous system, but augments experimental meningitis.
Although tumour necrosis factor alpha is said to play a key role in bacterial meningitis and other CNS diseases, the effects of this pro-inflammatory cytokine have only been studied in part and are incompletely understood. In a rat model, we investigated the effect of intracisternal injection of recombinant rat-specific tumour necrosis factor alpha (5, 35, 70 and 280 microg tumour necrosis factor alpha) (i) alone, (ii) combined with pneumococcal cell wall components, on regional cerebral blood flow, intracranial pressure, white blood cell count in the cerebrospinal fluid, and brain water content. Tumour necrosis factor a dose-dependently caused an increase in regional cerebral blood flow (up to 221 +/- 43% of baseline values) over the six hour observation period and mild cerebrospinal fluid leukocytosis; intracranial pressure and brain water content were unchanged. Hypothesizing that regional cerebral blood flow changes are dependent on nitric oxide, tumour necrosis factor alpha-induced regional cerebral blood flow increase was abolished by Aminoguanidine, a selective inhibitor of inducible nitric oxide synthase. Combination of the lowest tumour necrosis factor alpha dose and a low dose pneumococcal cell wall preparation magnified the inflammatory effect of both. We conclude that intrathecally injected tumour necrosis factor alpha alone results in only minor inflammatory changes, whereas it dramatically augments experimental meningitis.


Circulating differentiated and senescent lymphocyte subsets and incident diabetes risk in older adults: The Cardiovascular Health Study.
Cellular senescence is a feature of aging implicated in the pathophysiology of diabetes mellitus (DM). Whether senescent lymphocytes are associated with the future occurrence of DM is uncertain. We used cryopreserved peripheral blood mononuclear cells collected from 1860 Cardiovascular Health Study participants (average age 80.2 years) and flow cytometry immunophenotyping to evaluate the longitudinal relationships of naive (CD45RA+ ), memory (CD45RO+ ), senescent (CD28- ), and T effector memory RA+ (TEMRA) (CD28- CD57+ CD45RA+ ) CD4+ and CD8+ T cells, and memory B cells (CD19+ CD27+ ), with the risk of incident DM. In exploratory analyses we evaluated the relationships of 13 additional innate lymphocyte and CD4+ and CD8+ subsets with incident DM risk. Over a median follow-up time of 8.9 years, 155 cases of incident DM occurred. In Cox models adjusted for demographic variables (age, sex, race, study site and flow cytometry analytical batch) or diabetes risk factors (demographic variables plus education, body mass index, smoking status, alcohol use, systolic blood pressure, hypertension medication use and physical activity), no significant associations were observed for any CD4+ , CD8+ or CD19+ cell phenotypes with incident DM. These results suggest the frequencies of naive, memory and senescent T cells and memory B cells are not strongly associated with incident DM risk in older adults.


Mitochondrial dysfunction in bipolar disorder: evidence from magnetic resonance spectroscopy research.
Magnetic resonance spectroscopy (MRS) affords a noninvasive window on in vivo brain chemistry and, as such, provides a unique opportunity to gain insight into the biochemical pathology of bipolar disorder. Studies utilizing proton ((1)H) MRS have identified changes in cerebral concentrations of N-acetyl aspartate, glutamate/glutamine, choline-containing compounds, myo-inositol, and lactate in bipolar subjects compared to normal controls, while studies using phosphorus ((31)P) MRS have examined additional alterations in levels of phosphocreatine, phosphomonoesters, and intracellular pH. We hypothesize that the majority of MRS findings in bipolar subjects can be fit into a more cohesive bioenergetic and neurochemical model of bipolar illness that is both novel and yet in concordance with findings from complementary methodological approaches. In this review, we propose a hypothesis of mitochondrial dysfunction in bipolar disorder that involves impaired oxidative phosphorylation, a resultant shift toward glycolytic energy production, a decrease in total energy production and/or substrate availability, and altered phospholipid metabolism.


Analysis of immune markers in human cardiac allograft recipients and association with coronary artery vasculopathy.
Because coronary artery vasculopathy (CAV) is a common cause of late cardiac allograft loss in humans, there is a need to develop and test non-invasive surrogate markers capable of detecting and predicting this disease entity. We performed a cross-sectional analysis of immune-based surrogate markers in 65 primary cardiac allograft recipients with or without angiographically documented CAV. Anti-donor cellular immunity was determined by interferon gamma (IFN)-gamma enzyme-linked immunosorbent spot (ELISPOT) assays using donor HLA-derived peptides (indirect pathway), and anti-donor alloantibodies were detected by flow cytometry using HLA-coated beads. Anti-donor cellular and humoral immunity were detected more frequently in patients with CAV (17 of 32, 53.1%) than in controls (4 of 33, 12.1%) (p < 0.001). Anti-donor cellular and humoral immunity were detected in different sub-groups of CAV patients; peripheral blood lymphocytes (PBLs) from only 1 of 32 CAV patients reacted to donor peptides with simultaneous detection of peripheral anti-donor alloantibodies. Immune reactivity in cardiac transplant recipients with CAV differs significantly from those without CAV and the detected responses are heterogeneous in nature. Serial assessments of anti-donor immunity using different methods will be required to detect and possibly predict outcome in cardiac transplant recipients.


Cognitive deficits and biases for food and body in bulimia: investigation using an affective shifting task.
Studies suggest that attentional deficits and biases play a role in the development and maintenance of eating disorders. Many of these studies have methodological limitations and their results are difficult to interpret. In this study, we examine attentional deficits and biases in bulimia. 18 bulimic participants and 18 controls performed an adaptation of the go/no-go affective shifting task. That task allows the investigation of attention, inhibitory control and mental flexibility for stimuli related to the body and food. Bulimic participants tended to react faster than controls in the go/no-go affective task. They also had poorer discrimination ability than controls and showed inhibition problems, particularly when the targets were related to food. The magnitude of these effects ranged from moderate to large. No difference between groups was found concerning mental flexibility. These results suggest that bulimics present cognitive deficits and are more impulsive, especially with food-related stimuli. These cognitive deficits and biases may be at least partially responsible for the development and maintenance of bulimia.


Postpartum metabolic syndrome and high-sensitivity C-reactive protein after gestational hypertension and pre-eclampsia.
To evaluate the association between metabolic syndrome (MetS) and high-sensitivity C-reactive protein (hsCRP), a biomarker of chronic inflammation and an independent predictor for cardiovascular disease overall and in subgroups of women with/without pre-eclampsia and gestational hypertension (GHT). A prospective cohort study was conducted in Nairobi, Kenya. Women with pre-eclampsia or GHT and normotensive women within 12 weeks postpartum underwent physical, anthropometric, fasting lipid profile, plasma glucose, and hsCRP measurements at 6 months postpartum. A generalized linear regression model with Poisson distribution adjusted for body mass index and age was used to estimate the association between elevated hsCRP and MetS overall and stratified by pre-eclampsia or GHT. In the 171 women included in the study, risk of elevated hsCRP (>3 mg/L) was greater among women with compared to those without MetS (adjusted relative risk [ARR] 1.70, 95% confidence interval [CI] 1.05-2.73, P=0.03) and was statistically significantly higher in the hypertensive (ARR 2.16 95% CI 1.01-4.62, P=0.04) but not in the normotensive (ARR 1.46, 95% CI 0.93-2.28) group. Increased risk of elevated hsCRP postpartum can guide longitudinal mechanistic and intervention studies to reduce postpartum cardiovascular morbidity in women with MetS, especially after pre-eclampsia or GHT.