Dihydrotestosterone sensitises LNCaP cells to death induced by epigallocatechin-3-Gallate (EGCG) or an IGF-I receptor inhibitor.
Compelling evidence has accumulated for chemopreventive effects for the active component of green tea Epigallocatechin-3-Gallate (EGCG) particularly for prostate cancer (CaP). We have assessed interactions between the effects of EGCG and two main regulators of prostate cell function, dihydrotestosterone (DHT) and insulin-like growth factor-1 (IGF-I). Using LNCaP (androgen-sensitive), PC3 and DU145 (androgen-resistant) CaP cell lines, we assessed the effect of EGCG alone on growth (0-200 microM) and on cell death (0-50 microM). EGCG decreased the proliferation of all the CaP cancer cells in a dose-dependent manner with an increase in apoptosis from 30 to 50 microM. With DU145 cells, a sub-apoptotic dose of EGCG (10-20 microM) reduced IGF-induced growth. With LNCaP cells, a sub-apoptotic dose of EGCG (8 microM) switched DHT from a growth promoter to a growth inhibitor. A similar reversal of DHT effect was seen in the presence of an IGF-I receptor inhibitor, AG1024 (1 microM). These responses appeared to be due to DHT sensitizing the cells to apoptosis by EGCG and AG1024 (P < 0.01 and P < 0.001 respectively). Our data suggests that both green tea and AG1024 are effective in inhibiting cell growth and inducing death in CaP cells but the effects of both are more effective in the presence of androgen.


Effects of combining erlotinib and RNA-interfered downregulation of focal adhesion kinase expression on gastric cancer.
To investigate the synergistic effects of combining erlotinib and RNA-interference downregulation of focal adhesion kinase (FAK) expression on the proliferation, apoptosis, invasion and migration of the human gastric adenocarcinoma cell line AGS. Cells were divided into five experimental groups: Group A, nontransfected control; Group B, transfected with empty vector; Group C, transfected with FAK-shRNA; Group D, erlotinib treatment; Group E, combination erlotinib treatment and transfected with FAK-shRNA. FAK protein levels were confirmed via Western blotting. Cell proliferation (CCK-8 assay, apoptosis (flow cytometry), cell invasion (transwell assay) and migration (scratch assay) were evaluated. RNA interference significantly decreased FAK protein levels. Cell proliferation, invasion and migration were significantly lower in Groups C, D and E compared with Group A, and significantly lower in Group E than in Groups C and D. RNA interference effectively silences FAK expression and inhibits malignant cell proliferation and invasion in gastric cancer cells. The effect of FAK inhibition is increased by co-treatment with erlotinib.


Establishment and characterization of a B-cell line from a patient with acute lymphoblastic leukemia.
A permanent lymphoblastoid cell line was established from the peripheral blood of a child with acute lymphoblastic leukemia. The cell line, designated SDK, grows in a stationary suspension culture, forming aggregates, in RPMI medium supplemented with 10% FCS, with a doubling time of 50-60 h. Immunologic markers and cytological features suggested that the SDK cells should be identified as being of B-cell origin. The cells failed to form rosettes with sheep erythrocytes, did not express T-cell antigens as defined by monoclonal antibodies, and exhibited surface and cytoplasmic immunoglobulin determinants. Chromosome analysis revealed the presence of three cell populations with (a) 46XY; (b) t(8q-; 14q+) or 2p-; 14q+) and (c) cells with unidentifiable markers. SDK demonstrated susceptibility to TPA-induced differentiation toward plasma cells.


Single-Molecule Force Spectroscopy of DNA-Based Reversible Polymer Bridges: Surface Robustness and Homogeneity.
Single-molecule force spectroscopy, as implemented in an atomic force microscope, provides a rarely-used method by which to monitor dynamic processes that occur near surfaces. Here, a methodology is presented and characterized that facilitates the study of polymer bridging across nanometer-sized gaps. The model system employed is that of DNA-based reversible polymers, and an automated procedure is introduced that allows the AFM tip-surface contact point to be automatically determined, and the distance d between opposing surfaces to be actively controlled. Using this methodology, the importance of several experimental parameters was systematically studied, e.g. the frequency of repeated tip/surface contacts, the area of the substrate surface sampled by the AFM, and the use of multiple AFM tips and substrates. Experiments revealed the surfaces to be robust throughout pulling experiments, so that multiple touches and pulls could be carried out on a single spot with no measurable affect on the results. Differences in observed bridging probabilities were observed, both on different spots on the same surface and, more dramatically, from one day to another. Data normalization via a reference measurement allows data from multiple days to be directly compared.


Idiopathic polyhydramnios: persistence across gestation and impact on pregnancy outcomes.
To investigate the likelihood of resolution of idiopathic polyhydramnios in pregnant women and compare outcomes between resolved and persistent cases. One hundred and sixty-three women with idiopathic polyhydramnios who delivered at two medical centers during a 3 year period (January 2012-January 2015) were included in the study. Exclusion criteria included congenital fetal anomalies, maternal diabetes, isoimmunization, fetal infection, placental tumors or anomalies, and multiple gestation. Polyhydramnios was defined as SDP≥8cm or AFI≥24cm. Resolved cases were defined as those with AFI and/or SDP falling and remaining below 24cm and 8cm respectively. Pregnancy outcomes were compared between resolved and persistent cases. Two-sample t-test or Wilcoxon rank-sum test was used for continuous variables while chi-square test or Fisher's exact test was used for categorical measures. Resolution was noted in 61 of 163 (37%) patients. There were no differences in maternal age, gravidity or parity between resolved and persistent cases. Mean gestational age at diagnosis of polyhydramnios and overall mean AFI were significantly lower in the cases that resolved (29.7±4.5 weeks vs 33.4±4.1 weeks, p<0.0001; 23.3±3.5cm vs 25.8 23.3±4.0cm, p=0.0002). Similar to AFI measurements, mean SDP was also lower in cases with resolution (p=0.002). There was no difference in induction rates, mode of delivery, amnioinfusion rates, meconium staining of amniotic fluid and fetal heart rate abnormalities influencing intrapartum management between the two groups. Induction of labor for fetal indication and rupture of membranes were significantly more common in the persistent group. Cesarean delivery for abnormal lie and fetal distress did not differ between the groups. There was an increased risk of macrosomia (>4000g) and preterm delivery (<37 weeks) in the persistent group (p<0.05). Resolution rate was approximately 37% and more likely in cases diagnosed earlier in pregnancy and with lower mean amniotic fluid volume. Preterm delivery and macrosomia were more common in cases that persisted across gestation.


A single-step immunization by sustained antigen release.
An inert polymer pellet less than 1 mm in diameter, implanted subcutaneously in mice, releases free antigen continuously and enhances antibody formation for over 6 months. The immune response stimulated by sustained antigen delivery is comparable to the secondary response induced by the same total dose of antigen emulsified in complete Freund's adjuvant. The sustained release polymer implants, using antigens over a wide molecular weight range, proved effective in eliciting prolonged antibody formation. The antigens tested included bovine serum albumin, gamma-globulin, and ribonuclease. Superior biocompatibility and release characteristics make the ethylene-vinyl acetate copolymer a promising method for the single-step induction of immunity.


Surgical findings during exploratory laparotomy are closely related to mortality in premature infants with necrotising enterocolitis.
This study investigated whether a correlation existed between surgical findings during the first laparotomy for necrotising enterocolitis (NEC) and death and, or, disease progression. We included infants admitted within one day of birth to our tertiary neonatal department at Rigshospitalet, Denmark, from 2006 to 2015, who underwent a laparotomy for acute NEC. They were classified according to the locality and extent of intestinal necrosis by a paediatric surgeon, based on the surgical findings. We correlated the surgical findings with postoperative outcomes, namely death and, or, progression of NEC. The first laparotomy showed that 48 infants had NEC, including 21 who demonstrated postoperative progression. Of these, six died before undergoing another laparotomy and 14 of the 15 infants who underwent relaparotomy also died. There was a significant association between surgical findings and NEC-related mortality (p = 0.03). The association between surgical findings and the progression of NEC was also significant (p < 0.0001). Surgical findings during laparotomy for NEC were strongly correlated with mortality, which was close to 100% after relaparotomy. Considering the discouraging outcome, further studies should focus on alternative surgical approaches, such as proximal diverting jejunostomy and the clip and drop technique for the treatment of severe NEC.


Obesity Doubles Mortality in Patients Hospitalized for Severe Acute Respiratory Syndrome Coronavirus 2 in Paris Hospitals, France: A Cohort Study on 5,795 Patients.
Preliminary data from different cohorts of small sample size or with short follow-up indicate poorer prognosis in people with obesity compared with other patients. This study aims to precisely describe the strength of association between obesity in patients hospitalized with coronavirus disease 2019 (COVID-19) and mortality and to clarify the risk according to usual cardiometabolic risk factors in a large cohort. This is a prospective cohort study including 5,795 patients aged 18 to 79 years hospitalized from February 1 to April 30, 2020, in the Paris area, with confirmed infection by severe acute respiratory syndrome coronavirus 2. Adjusted regression models were used to estimate the odds ratios (ORs) and 95% CIs for the mortality rate at 30 days across BMI classes, without and with imputation for missing BMI values. Eight hundred ninety-one deaths had occurred at 30 days. Mortality was significantly raised in people with obesity, with the following ORs for BMI of 30 to 35 kg/m2 , 35 to 40 kg/m2 , and >40 kg/m2 : 1.89 (95% CI: 1.45-2.47), 2.79 (95% CI: 1.95-3.97), and 2.55 (95% CI: 1.62-3.95), respectively (18.5-25 kg/m2 was used as the reference class). This increase holds for all age classes. Obesity doubles mortality in patients hospitalized with COVID-19.


The importance of right atrial pacing electrode position and pacing configuration for intra-atrial and inter-atrial conduction times.
Pace prevention of atrial tachyarrhythmias is based in part on the reduction of intra-atrial (IAA) and/or inter-atrial (IEA) conduction. We previously introduced a novel pacing mode using floating atrial ring electrodes on a VDD-lead (BIdirectional MO nophasic impulSe: BIMOS). The effects of BIMOS pacing on IAA and IEA conduction times has not been studied. In nine Merino sheep electrode catheters were placed at the His-Bundle (HBE), high right atrium (HRA), coronary sinus ostium (Cs-Os), and left lateral atrium (LLA). A VDD-lead was introduced with floating electrodes in the high and mid right atrium (Floating). IAA (S/P-HRA, S/P-Cs-Os, S/P-HBE, S/P-Floating), IEA conduction times (S/P-LLA), and P-wave duration (PD) were measured during sinus rhythm (S), during bipolar cathodal pacing (P) in the HRA, in the Cs-Os position, as well as during BIMOS floating pacing. The mean PD during S was significantly shorter than during HRA- (66. 6+/-12.8ms; vs. 116.2+/-11.1ms; p<0.05) and Cs-Os-P (66.6+/-12.8ms vs. 94.4+/-9.0ms; p<0.05). In comparison to HRA-P, BIMOS configuration lead to a significant reduction of the P-wave duration (116.2+/-11.1ms vs. 85. 4+/-8.8ms; p<0.05). During BIMOS pacing, the global atrial conduction time was significantly shorter than during pacing in the HRA and Cs-Os position. The results of this study demonstrate a clear reduction of IAA and IEA conduction times using BIMOS configurations compared to conventional HRA-P. Furthermore, BIMOS pacing produced a more homogeneous atrial activation when compared with conventional HRA- and Cs-Os-P.


Initial experience of using intracardiac echocardiography (ICE) for guiding balloon mitral valvuloplasty (BMV).
BMV is an established treatment for rheumatic mitral valve stenosis. The procedure is historically guided by fluoroscopy, and the role of intracardiac echocardiogram (ICE) guidance is not well defined. We report our initial experience of using ICE to guide BMV procedures. During BMV procedure, ICE catheter was inserted into the right atrium from the right femoral vein, and the septal puncture was monitored by ICE, as well as positioning of the balloon in the mitral valve. Comparisons were made between ICE, transthoracic echocardiography (TTE), and catheterization derived hemodynamic measurements (cath). Seventeen patients with mitral stenosis underwent the procedure. The mean age was 44.4 ± 21 years. The mean MV area increased from 0.9 ± 0.1 cm(2) to 1.7 ± 0.2 cm(2), P < 0.0001 and the mean gradient decreased from 12.6 ± 5.8 mmHg to 4.9 ± 1.8 mmHg, P < 0.001. Atrial septum puncture and guidance of the balloon into the MV apparatus were obtained in all patients under ICE guidance. Severe MR developed in one patient and was readily detected by ICE. ICE derived gradient measurements were comparable to those obtained by TTE, and cath. ICE guidance of BMV is feasible, and useful in monitoring safe septal puncture, optimizing balloon positioning, and in detecting complications. The hemodynamic measurements obtained were comparable to those obtained by TTE, and cath.


In vivo exposure to sodium fluoride does not modify the yield of viral tumour-induced periosteal bone nor of heterotopic bone induced by human tumour KB cells in mice.
In mice local or systemic administration of fluoride (5-25 mg NaF/kg body weight) during the proliferative phase of bone formation (up to 20 days) has no effect on the yield of bone formed either by local stimulation of periosteal membrane by Moloney sarcoma virus-induced tumour or on bone induced heterotopically by human KB cells. The lack of stimulatory activity of fluoride on rapidly induced osteogenesis in mice is in agreement with recent reports which show that fluoride is not a potent mitogen for human osteoblasts grown in vitro.


[The importance of Walthard inclusions from the stand point of differential diagnosis of the female genital tuberculosis (author's transl)].
Walthard inclusions are described, which are frequent found like small nodules or cysts on the serosa of fallopian tubes and uterine ligaments under routine pelviscopy. The importance of Walthard inclusions is emphasized for distinguishing from the tubercles. During the three years period the incidence of Walthard inclusions from the 574 pelviscopic examinations was 19.7%.


Clostridium difficile ribotype 078 cultured from post-surgical non-healing wound in a patient carrying ribotype 014 in the intestinal tract.
Extra-intestinal infections caused by Clostridium difficile are rare. The risk of extra-intestinal infections associated with C. difficile may be particularly relevant in environments contaminated with C. difficile spores. This paper describes the case of a non-diarrheic patient colonized with C. difficile ribotype 014 in the intestinal tract who developed a post-surgical wound infection by C. difficile ribotype 078. The infection responded to metronidazole administered first intravenously and then orally. This case indicates that C. difficile may not only be related to diarrheic diseases, but also to infections of non-healing wounds, especially in situations when C. difficile is the only isolated pathogen.


Individual differences effects on the psychological refractory period.
The goal of this study was to assess the impact of individual neuropsychological differences on the ability to share attention between concurrent tasks. Participants (n = 20) were trained on six single task practice sessions and dual-task was assessed with reaction time performance on a psychological refractory period (PRP) paradigm. Neuropsychological test scores were also acquired. Furthermore, one of the known variables that can influence performances on neuropsychological tests is gender, which was added as a potential predictor. Results show that the small PRP group was associated with better performances in processing speed, inhibition, flexibility and working memory on neuropsychological tests. Gender also had an impact on the PRP, males having a lower PRP than females. A multiple regression was performed to determine which variables explained the most PRP duration, which showed that 49.1% of the variance of the PRP length could be explained by gender, reaction times of the PRP practice trials at the sixth session, the denomination and flexibility conditions of the Modified Stroop Task as well as results on the Symbol Search Test. Gender was the variable that explained the PRP variance the most (23%). Processing speed also seemed to be a great determinant of the PRP as well as the ability to alternate between task-sets as assessed by the Flexibility condition of the Modified Stroop Task. Thus, this study reveals that good performances on certain neuropsychological tests could predict one's ease to manage two tasks simultaneously with a higher chance for males to perform better.


Determination of 1,2-dibromoethane, 1,4-dichlorobenzene and naphthalene residues in honey by gas chromatography--mass spectrometry using purge and trap thermal desorption extraction.
A highly sensitive method for the determination of 1,2-dibromoethane, 1,2-dichlorobenzene and naphthalene residues in honey was developed, using gas chromatography-mass spectrometry combined with a purge and trap thermal desorption system as the extraction technique. Optimal conditions for isolation and separation were established and calibration curves were constructed. Linearity was held between 2.4 and 300 microg kg(-1) honey for 1,2-dibromoethane, 0.5 and 300 microg kg(-1) for 1,4-dichlorobenzene and 0.125 and 3000 microg kg(-1) for naphthalene. The detection limits were found to be 0.8, 0.15 and 0.05 microg kg(-1) honey for 1,2-dibromoethane, 1,4-dichlorobenzene and naphthalene, respectively. The method was applied to the analysis of 25 Greek honey samples. 1,2-Dibromoethane was not found in the majority of the samples, while only one sample was found to contain both 1,4-dichlorobenzene and naphthalene residues at concentrations exceeding 10 microg kg(-1).


Curcumin ameliorates focal segmental glomerulosclerosis by inhibiting apoptosis and oxidative stress in podocytes.
Focal segmental glomerulosclerosis (FSGS), a podocyte disease, is the leading cause of end-stage renal disease (ESRD). Nevertheless, the current effective treatment for FSGS is deficient. Curcumin (CUR) is a principal curcuminoid of turmeric, which is a member of the ginger family. Previous studies have shown that CUR has renoprotective effects. However, the mechanism of CUR in anti-FSGS is not clear. This study aimed to explore the mechanism of CUR against FSGS through a combination of network pharmacological methods and verification of experiments. The analysis identified 98 shared targets of CUR against FSGS, and these 98 targets formed a network of protein-protein interactions (PPI). Of these 98 targets, AKT1, TNF, IL-6, VEGFA, STAT3, MAPK3, HIF1A, CASP3, IL1B, and JUN were identified as the hub targets. Molecular docking suggested that the best binding to CUR is MAPK3 and AKT1. Apoptotic process and cell proliferation were identified as the main biological processes of CUR against FSGS by gene ontology (GO) analysis. The most enriched signaling pathway in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was the PI3K-AKT signaling pathway. Western blots and flow cytometry showed that CUR could inhibit adriamycin (ADR) induced apoptosis, oxidative stress damage, and attenuate podocyte epithelial-mesenchymal transition (EMT) by repressing the AKT signaling pathway. Collectively, our study demonstrates that CUR can attenuate apoptosis, oxidative stress damage, and EMT in FSGS in vitro. These results supply a compelling basis for future studies of CUR for the clinical treatment of FSGS.


Surface protein distributions in cells isolated from solid tumours and their metastases.
Methods have been developed which isolate single viable cells from the primary growths of two tumour systems (a lymphosarcoma and a carcinoma) and their secondary deposits. Subsequent comparisons of the surface-membrane structure of pairs of these primary and secondary cells, using lactoperoxidase-catalysed radioiodination coupled with polyacrylamide-gel electrophoresis, suggest that their overall structures are qualitatively very similar. This latter picture is still maintained when the isolated cells are treated with trypsin or incubated in complete medium before radioiodination. Analysis of the incorporated label into defined sections of the electrophoretic patterns revealed small quantitative differences between primary and secondary cells. In particular, slightly reduced incorporation into certain surface components of secondary cell preparations was seen. However, these did not occur for all the animals investigated, and also they did not consistently occur if the isolated cells were incubated in complete medium. The most similar overall change observed for the two tumour systems was a slight reduction in the secondary cells of a 20K mol. wt surface component.


The Ethical Unjustifications of COVID-19 Triage Committees.
The ever-debated question of triage and allocating the life-saving ventilator during the COVID-19 pandemic has been repeatedly raised and challenged within the ethical community after shortages propelled doctors before life and death decisions (Anderson-Shaw and Zar 2020; Huxtable 2020; Jongepier 2020; Peterson, Largent, and Karlawish 2020). The British Medical Association's ethical guidance highlighted the possibility of an initial surge of patients that would outstrip the health system's ability to deliver care "to existing standards," where utilitarian measures have to be applied, and triage decisions need to maximize "overall benefit" (British Medical Association 2020, 3) In these emergency circumstances, triage that "grades according to their needs and the probable outcomes of intervention" will prioritize or eliminate patients for treatment, and health professionals may be faced with obligations to withhold or withdraw treatments to some patients in favour of others (British Medical Association 2020, 4). This piece is a response and extension to articles published on the manner of involvement for ethics and ethicists in pandemic triage decisions, particularly examining the ability and necessity of establishing triage committees to ameliorate scarce allocation decisions for physicians.


Draft Genome Sequence of the Efficient Bioflocculant-Producing Bacterium Paenibacillus sp. Strain A9.
Paenibacillus sp. strain A9 is an important bioflocculant-producing bacterium, isolated from a soil sample, and is pale pink-pigmented, aerobic, and Gram-positive. Here, we report the draft genome sequence and the initial findings from a preliminary analysis of strain A9, which is a novel species of Paenibacillus.


Historiographical approaches to biogeography: a critical review.
We performed a critical review of the historiographical studies on biogeography. We began with the pioneering works of Augustin and Alphonse de Candolle. Then, we analyzed the historical accounts of biogeography developed by (1) Martin Fichman and his history on the extensionism-permanentism debate; (2) Gareth Nelson and his critique of the Neo-Darwinian historiography of biogeography; (3) Ernst Mayr, with his dispersalist viewpoint; (4) Alan Richardson, who wrote a microhistory on the biogeographic model constructed by Darwin; (5) Michael Paul Kinch and the ideas discussed in the 19th century about the geographical distribution of living beings; (6) Janet Browne, who highlighted the importance of the pre-Darwinian naturalists; (7) Peter Bowler, who focused mainly on the influence of paleontology on biogeography; (8) James Larson, who looked into the practices of the naturalists of Northern Europe in the late 18th century; and (9) Malte Ebach, who like Larson, was more interested in analysing the practices rather than the ideas of naturalists who studied the geographical distribution of organisms. Finally, these works are compared with each other. There has not been a dominant paradigm in the construction of historical narratives of biogeography; however, they provide a useful context for understanding problems of biogeography that continue to be debated to this day.


Comparison of different real-time PCR chemistries and their suitability for detection and quantification of genetically modified organisms.
The real-time polymerase chain reaction is currently the method of choice for quantifying nucleic acids in different DNA based quantification applications. It is widely used also for detecting and quantifying genetically modified components in food and feed, predominantly employing TaqMan and SYBR Green real-time PCR chemistries. In our study four alternative chemistries: Lux, Plexor, Cycling Probe Technology and LNA were extensively evaluated and compared using TaqMan chemistry as a reference system. Amplicons were designed on the maize invertase gene and the 5'-junction of inserted transgene and plant genomic DNA in MON 810 event. Real-time assays were subsequently compared for their efficiency in PCR amplification, limits of detection and quantification, repeatability and accuracy to test the performance of the assays. Additionally, the specificity of established assays was checked on various transgenic and non-transgenic plant species. The overall applicability of the designed assays was evaluated, adding practicability and costs issues to the performance characteristics. Although none of the chemistries significantly outperformed the others, there are certain characteristics that suggest that LNA technology is an alternative to TaqMan when designing assays for quantitative analysis. Because LNA probes are much shorter they might be especially appropriate when high specificity is required and where the design of a common TaqMan probe is difficult or even impossible due to sequence characteristics. Plexor on the other hand might be a method of choice for qualitative analysis when sensitivity, low cost and simplicity of use prevail.


Trends in the utilization of specific health care services among older Manitobans: 1985 to 2000.
This paper examines 16-year trends in the utilization of hospital and physician services by Manitobans aged 75 and more, using data from the Manitoba Population Health Research Data Repository. Trends are examined across five measures of hospital services (separations, short-stay days, long-stay days, cataract surgeries, and hip/knee replacements) and two measures of physician care (overall visit rate, and proportion having seven or more visits). Results show changes in the utilization of these services among older adults living in Manitoba over time, with the extent of change varying with the service under consideration, age, and location of residence. Previously large utilization differentials are shown to be shrinking; for example, cataract surgery rates across regions and physician visit rates by age. For other services, such as the rates of hip or knee replacement surgery, the differences across regions are increasing. Findings indicate that global generalizations about the impact of older adults on the health care system are subject to question, as regional differences and differences between age groups (75-84, 85+) can be significant.


Diastolic orthostatic hypertension and cardiovascular prognosis in type 2 diabetes: a prospective cohort study.
In patients with type 2 diabetes, the prognostic impact of an orthostatic rise in blood pressure is not known. Therefore, the aim of this study was to determine the prognostic implications of the diastolic orthostatic blood pressure response in a cohort of patients with type 2 diabetes. We also evaluated associations between different orthostatic blood pressure responses and markers of subclinical cardiovascular organ damage. Office blood pressures were measured in the sitting and in the standing position in 749 patients with type 2 diabetes who participated in the CARDIPP study (Cardiovascular Risk factors in Patients with Diabetes-a Prospective study in Primary care). Diastolic orthostatic hypertension was defined as a rise of diastolic blood pressure ≥10 mmHg and diastolic orthostatic hypotension was defined as a drop of diastolic blood pressure ≥10 mmHg. Recruitment took place between the years 2005-2008, and patients were followed until any of the primary outcome events (cardiovascular death or hospitalization for either myocardial infarction or stroke) occurred or until December 31st, 2014. Measurements of aortic pulse wave velocity and of carotid intima-media thickness were performed at base-line. Diastolic orthostatic hypertension was found in 140 patients (18.7 %) and was associated with significantly lower risk of cardiovascular events (crude hazard ratio compared with patients with normal systolic and diastolic orthostatic blood pressure response: 0.450, 95 % C.I. 0.206-0.987, P = 0.046). Diastolic orthostatic hypotension was found in 31 patients (4.1 %) and was associated with higher values for aortic pulse wave velocity and carotid intima-media thickness, compared with patients with normal systolic and diastolic orthostatic blood pressure response. Diastolic orthostatic hypertension is common in patients with type 2 diabetes, and may be a novel marker for decreased cardiovascular risk in these patients.


Transcriptional regulation of bone sialoprotein gene expression by Osx.
Osteoporosis is the most common metabolic bone disease characterized by decreased bone mass, decreased bone strength, and increased risk of fracture. It is due to unbalance between bone formation and bone resorption. Bone formation is a complex process which involves the differentiation of mesenchymal stem cells to osteoblasts. Osteoblasts produce a characteristic extracellular collagenous matrix that subsequently becomes mineralized. Osterix (Osx) is an osteoblast-specific transcription factor required for osteoblast differentiation. Bone sialoprotein (Bsp) is a member of the SIBLING gene family. Expression of Bsp correlates with the differentiation of osteoblasts and the onset of mineralization. Our preliminary data showed that Bsp was abolished in Osx-null mice; however, the detailed mechanism of Osx regulation on Bsp is not fully understood. In this study, regulation of Bsp expression by Osx was further characterized. It was shown that overexpression of Osx led to Bsp upregulation. Inhibition of Osx by small interfering RNA resulted in Bsp downregulation in osteoblast. Transfection assay demonstrated that Osx was able to activate Bsp promoter reporter in a dose-dependent manner. To define minimal region of Bsp promoter activated by Osx, a series of deletion mutants of Bsp promoter were generated, and the minimal region was narrowed down to the proximal 100 bp. Point-mutagenesis studies showed that one GC-rich site was required for Bsp promoter activation by Osx. ChIP assays demonstrated that endogenous Osx associated with native Bsp promoter in primary osteoblasts. Our observations provide evidence that Osx targets Bsp expression directly.


Stabilization of swine wastes by anaerobic digestion.
The purpose of this study was to evaluate the efficiency of the anaerobic digestion to reduce the organic load of swine waste. Anaerobic digestion is a biological means of decomposition of manure in an oxygen-free environment, and has the advantage of producing a fuel gas (methane) and an odor-free residue rich in nutrients, which can be used as fertilizer. The experiments were carried out in laboratory scale batch digester, at temperature controlled at 35 +/- 2 degrees C, without agitation. The digester was fed with 5% and 15% manure slurry. Results showed an average reduction of TCOD of 58% and DCOD of 85%. The methane content of biogas varied from 55 to 65% and stabilized sludge presented characteristics for use in agriculture.


Targeted-cryosurgical ablation of the prostate with androgen deprivation therapy: quality of life in high-risk prostate cancer patients.
To present preliminary results on health-related quality of life (QoL), prostate-associated symptoms and therapeutic effects of targeted-cryosurgical ablation of the prostate (TCSAP) with androgen deprivation therapy (ADT) in high-risk prostate cancer (PCa) patients. Thirty-four men with high-risk PCa features underwent TCSAP, and ADT was added to improve the treatment outcomes. High-risk parameters were defined as either prostate-specific antigen (PSA) = or > 100ng/mL, or Gleason score = or > 8, or both. The Genito-Urinary Group of the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Core 30 (EORTC QLQ-C30) with prostate-cancer-specific module (QLQ-PR25) was used for evaluating morbidities and PSA levels were recorded every 3 months. PSA failure was defined as the inability to reach a nadir of 0.4 ng/mL or less. Although it was not statistically significant, the global health status scores increased after TCSAP with ADT. The scores for five functional scales also became higher after treatment. The most prominent symptom after treatment was sexual dysfunction, followed by treatment-related and irritative voiding symptoms. TCSAP with ADT appears to be minimally invasive with high QoL except for sexual dysfunction. Long-term follow-up of PSA data and survival is necessary before any conclusions can be made on the efficacy of this promising new therapeutic modality in the treatment of PCa.


Hepatic artery thrombosis-related risk factors after living donor liver transplantation: single-center experience from Turkey.
The purpose of this retrospective study is to evaluate the risk factors hepatic artery thrombosis (HAT) after orthotopic liver transplantation (OLT) in a consecutive series from a single center. Between January 2010 and May 2012, we performed 278 living donor liver transplantations, including 189 males and 89 females. We compared the risk factors between HAT and non-HAT groups according to the following variables: age, gender, body mass index (BMI), graft weight, use of graft, Child-Pugh and model for end stage liver disease score, level of hemoglobin, blood pressure, operation time, blood transfusion, presence of ascites, international normalized ratio (INR) level, and etiology. Eighteen patients, including 15 males and 3 female, had HAT after the operation (mean age, 45.1 years; age range, 22-60 years). There were no pediatric patients in the HAT group. HAT rate was 6.5% in our series. Graft loss and retransplantation due to HAT was 38.7% in a 2-year period. Biliary leakage was observed in 72 (25.8%) living donor liver transplantations; this rate was higher in patients with HAT (n = 8; 44.4%). The infection rate was 50% (n = 9) in the HAT group and was 32.7% (n = 91) in the non-HAT group. Mean INR value was 2.15 in the HAT group and 1.72 in the non-HAT group. When we compared the groups according to use of graft for anastomosis, biliary lekage, infection, and INR value, the differences were statistically significant (P < .05). Although the results of OLT have improved over the past years, HAT is still associated with substantial morbidity, high incidence of graft failure, and high mortality rates. The most important findings associated with HAT in our series were found as INR levels, bile leakage, and resistant infections. Use of vascular graft for hepatic artery anastomosis was found to increase HAT risk.


Role of squalene synthase in prostate cancer risk and the biological aggressiveness of human prostate cancer.
We previously conducted a genome-wide linkage analysis of Japanese nuclear families affected with prostate cancer and showed that the susceptibility to prostate cancer was closely linked to D8S550 at 8p23. The role of farnesyl diphosphate farnesyltransferase (FDFT1), which is located under the peak marker D8S550 at 8p23, and squalene synthase, the enzyme encoded by FDFT1, in prostate cancer was studied. The association among common variants of FDFT1 with prostate cancer risk, the promoter activities of FDFT1 with different genotypes and the effects of inhibition of squalene synthase were studied, and the FDFT1 transcript levels of human prostate samples were quantified. The A allele of rs2645429 was significantly associated with prostate cancer risk in a Japanese familial prostate cancer population. Rs2645429 was located in the promoter region of FDFT1, and the AA genotype showed significantly increased promoter activity. The knockdown of FDFT1 mRNA expression or squalene synthase inhibition led to a significant decrease in prostate cancer cell proliferation. Additionally, human prostate cancer specimens expressed significantly higher levels of FDFT1 mRNA compared with noncancerous specimens. Finally, aggressive cancers showed higher transcript levels. FDFT1 and its encoded enzyme, squalene synthase, may play an important role in prostate cancer development and its aggressive phenotypes.


Experimental investigations on increased resistance to infections with Candida albicans and Staphylococcus aureus Smith by 4-imino-1,4-diazobicyclo-(3.1.0)-hexane-2-on BM 06.002 (prop. INN imexon) in mice.
BM 06.002 increases the resistance of mice to experimentally induced chronic infection with Candida albicans. Furthermore, BM 06.002 leads to increased resistance in the case of experimentally induced infection with Staphylococcus aureus Smith, when a subtherapeutic dose of sulfadiazine is applied. In mice immunosuppressively pretreated with hydrocortisone, BM 06.002 effectuates immunorestauration.


Differences between diabetic and non-diabetic patients with community-acquired pneumonia in primary care in Spain.
Diabetes is one of the underlying risk factors for developing community-acquired pneumonia (CAP). The high prevalence of diabetes among population and the rising incidence of this illness, converts it as an important disease to better control and manage, to prevent its secondary consequences as CAP. The objective of this research is to describe the characteristics of the patients with diabetes and the differences with the no diabetes who have had an episode of CAP in the context of the primary care field. A retrospective, observational study in adult patients (> 18 years-old) who suffer from CAP and attended at primary care in Spain between 2009 and 2013 was developed using the Computerized Database for Pharmacoepidemiological Studies in Primary Care (BIFAP). We carried out a descriptive analysis of the first episodes of CAP, in patients with or without diabetes as comorbidity. Other morbidity (CVA, Anaemia, Arthritis, Asthma, Heart disease, Dementia, Depression, Dysphagia, Multiple sclerosis, Epilepsy, COPD, Liver disease, Arthrosis, Parkinson's disease, Kidney disease, HIV) and life-style factors were also included in the study. A total of 51,185 patients were included in the study as they suffer from the first episode of CAP. Of these, 8012 had diabetes as comorbidity. There were differences between sex and age in patients with diabetes. Patients without diabetes were younger, and had less comorbidities including those related to lifestyles such as smoking, alcoholism, social and dental problems than patients with diabetes. Patients who developed an episode of CAP with diabetes have more risk factors which could be reduced with an appropriate intervention, including vaccination to prevent successive CAP episodes and hospitalization. The burden of associated factors in these patients can produce an accumulation of risk. Health care professional should know this for treating and control these patients in order to avoid complications. Diabetes and those other risk factors associated could be reduced with an appropriate intervention, including vaccination to prevent the first and successive CAP episodes and the subsequent hospitalization in severe cases.


Estimates of child mortality reductions attributed to vitamin A supplementation in sub-Saharan Africa: scale up, scale back, or refocus?
Vitamin A supplementation (VAS) has been implemented in over 82 countries globally, primarily because of its beneficial effect in preventing child mortality. Secular reductions in child mortality and the implementation of alternative programs to promote vitamin A intake have led to questions on the need for national VAS programs. This study aimed to estimate child mortality changes related to VAS using current, scale-back, and scale-up coverage scenarios. Data related to demographic characteristics, fertility, intervention coverage, anthropometry, child mortality and cause-of-death structure were integrated into the Lives Saved Tool (LiST). We estimated the cause-specific (LiST model) and all-cause mortality reductions related to VAS based on evidence from recent meta-analyses. Between 2008 and 2018, VAS coverage declined in most sub-Saharan African (SSA) countries. In 2019 alone, 12% and 24% reductions in all-cause mortality related to VAS were expected to avert from 105,332 to 234,704 child deaths, respectively, in SSA; whereas the cause-specific mortality model (LiST) estimated that 141,670 child deaths were averted in 2019. Estimates of VAS-related child mortality reductions were highly variable among countries. Our scaling-back scenario led to highly variable country-level results, with expected increases in mortality rates, from a low of 0.04/1000 live births to as high as 49.3/1000 live births, suggesting that some countries could start considering scaling back, while others need to scale up. Excess child mortality that would be preventable by VAS has declined, but is still significant in many SSA countries. While scale-up of VAS is needed for most of the countries, scaling back can also be considered in some countries. Policy decisions, however, should be guided by more recent data on food consumption, vitamin A statuses, child health, and vitamin A fortification coverage.


[Meningeal metastasis of uterine leiomyosarcoma. Case report and literature review].
Brain metastases are the most commonly seen intracranial lesions in adults. What is more, meningiomas are the most common primary intracranial tumours after gliomas and their imaging characteristics are well known in both CT and MRI scans. However, there are lesions that can mimic meningiomas in imaging studies, including metastases of extracranial tumours, confronting us with a diagnostic and therapeutic challenge. We present the case of a patient with meningeal metastasis of a uterine leiomyosarcoma that was not known at the time of the surgical intervention.


[Cervical spine operation using coralline hydroxyapatite].
To study the result of using coralline hydroxyapatite (CHA) in cervical anterior fusion and posterior laminoplasty operation. 45 patients of cervical operation using CHA were followed up. Of these patients, 27 received anterior fusion and titanium plate fixation (34 levels fused). 18 with spinal stenosis at least at 3 levels, (posterior spinal process-splitting laminoplasty) had 79 split spinal processes fused by CHA. The average follow-up time was 25.9 months. The union rate of CHA was 85.3% in average by anterior approach. The height and angle of the fused level and the anterior salient were well maintained. The JOA score improvement rate was 58.7%. The CHA grafts were kept well between the split spinous processes with a union rate of 70.9%. After operation, the spinal cord moved dorsally, and the flat rate was improved from 0.19 to 0.41. No apparent posterior salient occurred. The JOA score improvement rate was 51%. The effect of CHA in cervical antero posterior operation is similar to that of autograft bone, and it is unnecessary to resect a piece of bone from a health region of the body.


Prevalence of antibodies against hepatitis C virus in a dialysis unit.
Anti-HCV was tested in 77 uremic patients, 48 on hemodialysis (HD), 29 on CAPD, by immunoenzymatic 1st and 2nd generation assays (ELISA I, II) and 4-antigen (4-RIBA) immublotting. The investigation was extended to the staff (n = 29) and to HCV-positive patients' families (n = 30). The prevalence using 2nd generation tests was double (21%) that in 1st generation tests (11%). A greater incidence in the HD than in the CAPD group (23 vs. 17%) and a highly significative correlation to dialytic age were observed. No one among the sanitary personnel and only 2 family members were found HCV positive, suggesting a low infectivity via the parenteral inevident route. Extracorporeal circulation and particularly the exposure time to the treatment seem to be the main risk factors.


Expression in Escherichia coli, purification, refolding and antifungal activity of an osmotin from Solanum nigrum.
Heterologous protein expression in microorganisms may contribute to identify and demonstrate antifungal activity of novel proteins. The Solanum nigrum osmotin-like protein (SnOLP) gene encodes a member of pathogenesis-related (PR) proteins, from the PR-5 sub-group, the last comprising several proteins with different functions, including antifungal activity. Based on deduced amino acid sequence of SnOLP, computer modeling produced a tertiary structure which is indicative of antifungal activity. To validate the potential antifungal activity of SnOLP, a hexahistidine-tagged mature SnOLP form was overexpressed in Escherichia coli M15 strain carried out by a pQE30 vector construction. The urea solubilized His6-tagged mature SnOLP protein was affinity-purified by immobilized-metal (Ni2+) affinity column chromatography. As SnOLP requires the correct formation of eight disulfide bonds, not correctly formed in bacterial cells, we adapted an in vitro method to refold the E. coli expressed SnOLP by using reduced:oxidized gluthatione redox buffer. This method generated biologically active conformations of the recombinant mature SnOLP, which exerted antifungal action towards plant pathogenic fungi (Fusarium solani f. sp.glycines, Colletotrichum spp., Macrophomina phaseolina) and oomycete (Phytophthora nicotiana var. parasitica) under in vitro conditions. Since SnOLP displays activity against economically important plant pathogenic fungi and oomycete, it represents a novel PR-5 protein with promising utility for biotechnological applications.


Avicularin alleviates acute liver failure by regulation of the TLR4/MyD88/NF-κB and Nrf2/HO-1/GPX4 pathways to reduce inflammation and ferroptosis.
Acute liver failure (ALF) is an inflammation-mediated hepatocyte death process associated with ferroptosis. Avicularin (AL), a Chinese herbal medicine, exerts anti-inflammatory and antioxidative effects. However, the protective effect of AL and the mechanism on ALF have not been reported. Our in vivo results suggest that AL significantly alleviated lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced hepatic pathological injury, liver enzymes, inflammatory cytokines, reactive oxygen species and iron levels and increased the antioxidant enzyme activities (malondialdehyde and glutathione). Our further in vitro experiments demonstrated that AL suppressed inflammatory response in LPS-stimulated RAW 264.7 cells via blocking the toll-like receptor 4 (TLR4)/myeloid differentiation protein-88 (MyD88)/nuclear factor kappa B (NF-κB) pathway. Moreover, AL attenuated ferroptosis in D-GalN-induced HepG2 cells by activating the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1)/glutathione peroxidase 4 (GPX4) pathway. Therefore, AL can alleviate inflammatory response and ferroptosis in LPS/D-GalN-induced ALF, and its protective effects are associated with blocking TLR4/MyD88/NF-κB pathway and activating Nrf2/HO-1/GPX4 pathway. Moreover, AL is a promising therapeutic option for ALF and should be clinically explored.


Effect of stress, adrenocorticotropin or corticosteroid treatment, adrenalectomy, or hypophysectomy on hypothalamic immunoreactive adrenocorticotropin concentrations.
Despite variations in plasma and/or pituitary ACTH concentrations, no changes in median eminence or medial basal hypothalamic immunoreactive ACTH-like concentrations were seen in rats studied 6 weeks posthypophysectomy, 12 h or 28 days after bilateral adrenalectomy, 6 days after implantation of corticosterone pellets (75 mg), after dexamethasone administration (300 micrograms/100 g BW, ip, twice daily for 4 days), or after chronic immobilization stress. The lack of concordant variation of brain, pituitary, and plasma ACTH concentrations further supports the suggestion of a nonpituitary, central nervous system origin of some portion of brain immunoreactive ACTH-like activity.


Population dynamics of the chestnut gall wasp, Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae) : V. Estimation of the effect of predation by spiders on the mortality of imaginal wasps based on the precipitin test.
During the course of a 10-year investigation on the population dynamics of Dryocosmus kuriphilus, a rapid decrease in the number of individuals was noticed in the adult stage. To detect the role of predation by spiders in causing this high adult mortality, spiders collected from the survey station were tested for their reaction to Drycosmus antiserum. About 20∼50% of the spiders collected during the emergence period of D. kuriphilus reacted positively, showing that the species of Thomisidae, Argiopidae, Agelenidae and Salticidae were the important predators. The predation ratio for the total adult wasp population was estimated at 8.1% in 1968, 20.2% in 1969 and much higher in 1970, increasing with decreasing population density of the wasps. However, the predation by spiders could not account for all of the adult mortality of D. kuriphilus.


Loss of matrix metalloproteinase-9 or matrix metalloproteinase-12 protects apolipoprotein E-deficient mice against atherosclerotic media destruction but differentially affects plaque growth.
Epidemiological and histological evidence implicates proteinases of the matrix metalloproteinase (MMP) family in atherosclerosis and aneurysm formation. We previously indicated a role for urokinase-type plasminogen activator in atherosclerotic media destruction by proteolytic activation of MMPs. However, the role of specific MMPs, such as MMP-9 and MMP-12, in atherosclerosis remains undefined. MMP-9- or MMP-12-deficient mice were crossed in the atherosclerosis-prone apolipoprotein E-deficient background and fed a cholesterol-rich diet. Mice were killed at 15 or 25 weeks of diet to study intermediate and advanced lesions, respectively. Loss of MMP-9 reduced atherosclerotic burden throughout the aorta and impaired macrophage infiltration and collagen deposition, while MMP-12 deficiency did not affect lesion growth. MMP-9 or MMP-12 deficiency conferred significant protection against transmedial elastin degradation and ectasia in the atherosclerotic media. This study is the first to provide direct genetic evidence for a significant involvement of MMP-9, but not of MMP-12, in atherosclerotic plaque growth. In addition, deficiency of MMP-9 or MMP-12 protected apolipoprotein E-deficient mice against atherosclerotic media destruction and ectasia, mechanisms that implicate the involvement of these MMPs in aneurysm formation.


The pharmacokinetics and metabolism of the anilide local anaesthetics in neonates. I. Lignocaine.
The pharmacokinetics and metabolism of lignocaine in premature neonates was studied after subcutaneous administration. The collection of serial urine together with a limited number of blood samples from neonates enabled simultaneous computer fitting of data to a pharmacokinetic model. The disposition kinetics of lignocaine in four neonates were compared with similar data reported for adults. Neonates had prolonged t1/2 (neonate mean: 3.16 h; adult mean: 1.80 h), and an increased total volume of distribution (neonate mean: 2.75 l/kg; adult mean: 1.11 l/kg) compared with adults. Total plasma clearance (Cltp) normalised on body weight showed no significant difference between neonates (mean: 0.610 l/h/kg) and adults (mean: 0.550 l/h/kg). The urinary excretion of lignocaine and several of its metabolites was studied in 8 neonates and 11 adults. Neonates were shown to excrete much more unchanged lignocaine (mean: 19.67%) compared with adults (mean: 4.27%) and the proportion of the dose excreted as 4-hydroxyxylidine is considerably reduced in neonates (neonate mean: 8.89%; adult mean: 63.78%). The use of the two pharmacokinetic parameters, t1/2 and Cltp, as indices of drug elimination ability are discussed.


Effects of tripeptides derived from milk proteins on polymorphonuclear oxidative and phosphoinositide metabolisms.
The tripeptide GLF (glycyl-leucyl-phenylalanine) was isolated from human milk proteins. This peptide increased phagocytosis by human and murine macrophages and protected mice against Klebsiella pneumoniae infection. Specific binding sites on human polymorphonuclear leukocytes (PMNs) have been demonstrated recently. The aim of the present research was to study the action of this peptide on rat and human PMN oxidative burst and to investigate the consequences of cell stimulation on polyphosphoinositide hydrolysis. A biphasic stimulating concentration-dependent effect of GLF on PMN chemiluminescence and superoxide anion generation was demonstrated. One of the peaks of the oxidative response occurred around 10(-9) M, which correlates with the Kd of high affinity receptors of GLF. The other maximum, around 10(-4) M, might be due to the hydrophobic nature of the tripeptide. O2- generation mimicked the phorbol myristate acetate response: after a lag period of 2-5 min, O2- release gradually increased for 10-15 min until a plateau was reached. Furthermore, GLF enhanced phosphoinositide breakdown with maximal IP3 production at 10(-7) M. Various analogs of GLF were synthesized in order to define the relative importance of the different amino acids and their position in the tripeptide molecule: glycyl-phenylalanine-leucine was devoid of biological properties but enhanced the activity of GLF on the metabolic burst at high concentrations; peptides leucyl-leucyl-phenylalanine and leucyl-leucyl-tyrosine, which displaced GLF from its specific membrane receptors, exerted stimulating effects on PMN oxidative and phosphoinositide metabolisms. It is quite conceivable that these short peptides, which may be generated in the newborn during digestion and which are able to stimulate phagocytic cells, are implicated in the defense of the neonate immature organism against infection.


[Psychological stress caused by work: burnout syndrome].
In a variety of helping professions emotional stress can lead to "burnout". The syndrome is characterised by emotional exhaustion, attitudinal hardening (loss of empathy) and a sense of decreased accomplishment. Longterm consequences of burnout can be mental problems such as depression or psychosomatic disorders. Affected employees are less productive and sick more often. They may even quit their jobs or need early retirement. In spite of this considerable loss of working force often neither those affected nor the employers are aware of the problem in time. Burnout seems to be caused by disproportionally high efforts (time, emotional involvement, empathy) and poor satisfaction (negative outcome) in addition to stressful working conditions (high demands). It affects mainly nurses, physicians, social workers, teachers and other similar professions. In order to prevent burnout, awareness of the problem must be promoted. Examples of job-specific measures for nurses and teachers are given. Their goal is to lower the professional stress and improve satisfaction. Social support and improved team cooperation can further protect against burnout.


Comparison of visual outcomes and subjective visual quality after bilateral implantation of a diffractive trifocal intraocular lens and blended implantation of apodized diffractive bifocal intraocular lenses.
To compare the visual acuity (VA) and quality of vision between bilateral implantation of a trifocal intraocular lens (IOL) and blended bifocal IOLs with an intermediate add in the dominant eye and a near add in the nondominant eye. Patients with either trifocal or blended bifocal IOLs implanted were recruited after surgery. Subjects returned for a single diagnostic visit between 3 and 24 months after surgery. VA was tested at various distances, including low-contrast acuity and acuity at their preferred reading distance. A binocular defocus curve was obtained, and subjective visual function and quality of vision were evaluated. Twenty-five trifocal subjects and 30 blended bifocal subjects were enrolled. There were no significant differences in low-contrast acuity, preferred reading distance, or acuity at that reading distance. Binocular vision at 4 m, 60 cm, and 40 cm was not statistically significantly different. The trifocal provided statistically significantly better visual acuity (P<0.05) at vergences from -0.5 to -1.5 D (from 2 m to 67 cm viewing distance, P<0.05). There was no statistically significant difference in the near vision subscale scores of the 39-question National Eye Institute Visual Function Questionnaire or the overall scores of the Quality of Vision questionnaire, though significantly more trifocal subjects reported that the observed visual disturbances were "bothersome" (P<0.05). Both lens modalities provided subjects with excellent binocular near and distance vision, with similar low rates of visual disturbances and good reported functional vision. The trifocal IOL provided significantly better intermediate VA in the viewing distance range of 2 m to 67 cm, corresponding to viewing things such as a car dashboard or grocery shelf. VA was similar between groups at viewing distances from 60 to 40 cm, corresponding to computer or reading distance.


Clinical and functional target validation using tissue and cell microarrays.
Expression levels of thousands of genes or proteins can be readily determined using microarray techniques. However, this represents only the first step in understanding the biological and medical significance of these molecules. New high-throughput techniques, such as tissue and cell microarrays, will facilitate clinical and functional analysis of molecular targets.


Screening for Fabry Disease in Japanese Patients with Young-Onset Stroke by Measuring α-Galactosidase A and Globotriaosylsphingosine.
Fabry disease is an X-linked lysosomal storage disorder caused by mutations in GLA, which encodes the enzyme α-galactosidase A (α-Gal A). Although the prevalence of Fabry disease in patients with stroke has been reported to range from 0% to 4%, few cohort studies have examined Japanese stroke patients. We aimed to clarify the prevalence of Fabry disease and the frequency of GLA mutations among patients with young-onset stroke in Japan. From April 2015 to December 2016, we enrolled patients with young-onset (≤60 years old) ischemic stroke or intracerebral hemorrhage. We measured α-Gal A activity and the concentration of globotriaosylsphingosine in plasma. Genetic evaluations were performed in patients with low α-Gal A activity or high concentrations of globotriaosylsphingosine. Overall, 516 patients (median age of onset, 52 years old; 120 women) were consecutively enrolled in this study. Five patients (4 men and 1 woman) had low α-Gal A activity, and no patients were detected with the screen for plasma globotriaosylsphingosine levels. The genetic analysis did not identify a causative mutation responsible for classic Fabry disease in any of the patients, but 2 patients (.4%) carried the p.E66Q in GLA. No patient with Fabry disease was detected in our young-onset stroke cohort.


Neo-modified Koyanagi technique for the single-stage repair of proximal hypospadias.
To retrospectively analyze the results of a modification of the Koyanagi hypospadias repair developed to reduce complications. Twelve boys underwent a modified Koyanagi repair over the past 3 years. The technique included the preservation of a wide vascular pedicle for each parameatal flap. The distal urethral plate was left intact and attached on each lateral side to the medial side of each distal Koyanagi pedicle flap. One boy (8%) developed a urethral fistula. There was no instance of meatal stenosis, sacculation or urethral stricture. Modifying the Koyanagi repair by preserving a wide vascular pedicle for the distal skin flaps and utilizing the distal urethral plate as the base for the distal neourethra has lowered the incidence of complications. This technique is a suitable alternative to a two-stage approach for the correction of proximal hypospadias.


Seizures in a 77-year-old-woman after a bolus dose of remifentanil.
Seizures in patients with no history of epilepsy has been associated with administration of remifentanil in two previous case reports. We present a case where a bolus dose of remifentanil (50 micrograms/kg) was followed by a self terminating generalised tonic-clonic seizure in a patient admitted for acute hip surgery, and no previous history of epilepsy.


Induction of endoplasmic reticulum stress is associated with the anti-tumor activity of monepantel across cancer types.
Monepantel is an anti-helminthic drug that also has anti-cancer properties. Despite several studies over the years, the molecular target of monepantel in mammalian cells is still unknown, and its mechanism-of-action is not fully understood, though effects on cell cycle, mTOR signalling and autophagy have been implicated. Viability assays were performed on >20 solid cancer cell cells, and apoptosis assays were performed on a subset of these, including 3D cultures. Genetic deletion of BAX/BAK and ATG were used to establish roles of apoptosis and autophagy in killing activity. RNA-sequencing was performed on four cell lines after monepantel treatment, and differentially regulated genes were confirmed by Western blotting. We showed that monepantel has anti-proliferative activity on a broad range of cancer cell lines. In some, this was associated with induction of apoptosis which was confirmed using a BAX/BAK-deficient cell line. However, proliferation is still inhibited in these cells following monepantel treatment, indicating cell-cycle disruption as the major anti-cancer effect. Previous studies have also indicated autophagic cell death occurs following monepantel treatment. We showed autophagy induction in multiple cell lines; however, deletion of a key autophagy regulator ATG7 had minimal impact on monepantel's anti-proliferative activity, suggesting autophagy is associated with, but not required for its anti-tumour effects. Transcriptomic analysis of four cell lines treated with monepantel revealed downregulation of many genes involved in the cell cycle, and upregulation of genes linked to ATF4-mediated ER stress responses, especially those involved in amino-acid metabolism and protein synthesis. As these outcomes are all associated with mTOR signalling, cell cycle and autophagy, we now provide a likely triggering mechanism for the anti-cancer activity of monepantel.


[Spontaneous and induced production of tumor necrosis factor (TNF alpha) by peripheral blood leukocytes in patients with acute leukemia type M 4 and M5 according to FAB].
Tumor necrosis factor (TNF) is a cytokine with pleiotropic biological activity. We have undertaken the present study to evaluate a possible ability of peripheral blood cells of healthy subjects and patients with acute monocytic leukemia to produce TNF alpha. We studied 17 leukemia patients and 8 control subjects. Spontaneous production of TNF alpha induced by LPS and Newcastle disease virus (NDV) were determined in blood cell cultures. Leukemic cells released markedly higher level of TNF alpha than control cells cultured in vitro in contrast to control cells producing TNF spontaneously. It seems that leukemic cells can produce TNF in response to very small amount of LPS; however, we cannot exclude that spontaneous TNF may be involved in autocrine regulation of proliferation and differentiation of bone marrow monocyte precursors.


Little evidence of transdifferentiation of bone marrow-derived cells into pancreatic beta cells.
Bone marrow cells contain at least two distinct types of stem cells which are haematopoietic stem cells and mesenchymal stem cells. Both cells have the ability to differentiate into a variety of cell types derived from all three germ layers. Thus, bone marrow stem cells could possibly be used to generate new pancreatic beta cells for the treatment of diabetes. In this study, we investigated the feasibility of bone marrow-derived cells to differentiate into beta cells in pancreas. Using green fluorescent protein transgenic mice as donors, the distribution of haematogenous cells in the pancreas was studied after bone marrow transplantation. In the pancreas of green fluorescent protein chimeric mice, green fluorescent protein-positive cells were found in the islets, but none of these cells expressed insulin. Previous data has suggested that tissue injury can recruit haematopoietic stem cells or their progeny to a non-haematopietic cell fate. Therefore, low-dose streptozotocin (30 or 50 mg/kg on five consecutive days) was injected into the mice 5 weeks after bone marrow transplantation, but no green fluorescent protein-positive cells expressing insulin were seen in the islets or around the ducts of the pancreas. Our data suggests that bone marrow-derived cells are a distinct cell population from islet cells and that transdifferentiation from bone marrow-derived cells to pancreatic beta cells is rarely observed.


Removal of Thermafil Obturators Using 2 Different Techniques: A Comparative Ex Vivo Study.
The aim of this ex vivo study was to compare 2 techniques for removal of Thermafil obturators from curved root canals in mandibular molars. Sixty mesial root canals in extracted mandibular molars were distributed into matched pairs according to degree and radius of curvature and were prepared to size 35/.04 and obturated with Thermafil obturators size 35. Each tooth was embedded in resin, mounted in a mannequin head, and isolated with rubber dam to simulate clinical conditions. Removal of obturators was undertaken under a dental operating microscope by an experienced endodontist using either the FragRemover loop device, or reciprocating NiTi instruments. Pre- and postoperative micro-CT scans were taken to compare the amount of remaining filling material and of removed dentin. Eventual problems during removal attempts were recorded. Using Reciproc, 24 carriers (80%) were removed, 16 in one piece, 8 in small pieces, and in 6 cases removal failed. The FragRemover removed 12 carriers (40%), all of these completely, but in 18 cases removal failed (P < .05). Reciproc left less remaining filling material than the FragRemover (P < .05) but removed significantly more dentin in the apical part of the root canal (P < .01). There was no significant difference concerning the total amount of removed dentin (P > .05) between both groups. Seven complications occurred in the Reciproc group. In 5 teeth, working length could not be reached; in 2 teeth, a Reciproc file R25 fractured. With the FragRemover, 64 complications occurred, in 36 cases, including second removal attempts, the loop slipped over the carrier, in 24 cases the top of the carrier was sheared off. In 9 cases, separation of the core occurred too far apically, not allowing further use of the FragRemover. In 4 cases, the wire loop tore. Reciproc was more effective in removal of Thermafil carriers, whereas when using the FragRemover less dentin was removed in the apical part of the root canal and more obturation material was left.


Cell wall anchoring of the 37-kilodalton oncofetal antigen by Lactobacillus plantarum for mucosal cancer vaccine delivery.
The 37-kDa oncofetal antigen (OFA), a tumor immunogen expressed on all mammalian cancers examined to date, was secreted and anchored to the cell wall of Lactobacillus plantarum using homologous signal peptides and LPxTG anchors. Orally administered L. plantarum expressing anchored OFA induced a specific immune response against OFA in mice.


Evaluating the causality between skin tanning, radiated disorders, and basal cell carcinoma: a multivariable Mendelian randomization analysis.
The causality of ease of skin tanning and radiation-related disorders of the skin and subcutaneous tissue with basal cell carcinoma (BCC) remains unclear. Our objective was to investigate whether ease of skin tanning and radiation-related disorders of the skin and subcutaneous tissue have a relation with the occurrence and development of BCCs. In this work, independent genetic variants strongly associated (P < 5e-08) with ease of skin tanning and radiation-related disorders of the skin and subcutaneous tissue were selected as instrumental variables from corresponding genome-wide association studies (GWASs). Summary-level data for BCC was obtained from the European Bioinformatics Institute (EBI). Two-sample univariable and multivariable Mendelian randomization (MR) were performed. Sensitivity analyses were preformed via MR-Egger regression, heterogeneity test, pleiotropy test, and leave-one-out sensitivity test. We observed positive causal effect both for ease of skin tanning [odds ratio (OR) = 2.102, 95% confidence interval (CI) = 1.915-2.306, P = 2.71e-55] and radiation-related disorders of the skin and subcutaneous (OR = 1.603, 95% CI = 1.483-1.734, P = 3.41e-32) on occurrence of BCCs based on univariable MR analyses. In the multivariable mendelian randomization (MVMR) analysis of BCC risk, we also observed a direct causal effect of ease of skin tanning (ORMVMR = 1.623, 95% CI = 1.445-1.824, PMVMR =3.41e-16) and radiation-related disorders of the skin and subcutaneous (ORMVMR = 1.208, 95% CI = 1.107-1.319, PMVMR = 2.46e-05) on BCCs. The findings suggest that the high risk of BCCs can be attributed to ease of skin tanning and radiation-related disorders of the skin and subcutaneous tissue.


Fractures of the ulnar coronoid process.
The ulnar coronoid process plays a central role in elbow stability due to its unique anatomic characteristics. A fracture of the coronoid, although uncommon, represents a serious injury that can adversely affect functional outcome if not treated appropriately. Several surgical interventions addressing different fracture patterns are being increasingly recognised as effective treatment options even for smaller fragments. A review of the literature was performed in order to evaluate different treatment strategies applied to clearly defined fracture configurations. 14 articles reporting data for the management of 236 coronoid fractures met our inclusion criteria and were subjected to critical analysis. The data suggest that recognition of specific coronoid fracture patterns, use of appropriate classification systems and application of staged surgical protocols can stabilise the elbow effectively and lead to favourable outcomes.


Artificial intelligence-based technology to make a three-dimensional pelvic model for preoperative simulation of rectal cancer surgery using MRI.
A new technique that allows visualization of whole pelvic organs with high accuracy and usability is needed for preoperative simulation in advanced rectal cancer surgery. In this study, we developed an automated algorithm to create a three-dimensional (3D) model from pelvic MRI using artificial intelligence (AI) technology. This study included a total of 143 patients who underwent 3D MRI in a preoperative examination for rectal cancer. The training dataset included 133 patients, in which ground truth labels were created for pelvic vessels, nerves, and bone. A 3D variant of U-net was used for the network architecture. Ten patients who underwent lateral lymph node dissection were used as a validation dataset. The correctness of the vascular labelling was assessed for pelvic vessels and the Dice similarity coefficients calculated for pelvic bone. An automatic segmentation algorithm that extracts the artery, vein, nerve, and pelvic bone was developed, automatically producing a 3D image of the entire pelvis. The total time needed for segmentation was 133 seconds. The success rate of the AI-based segmentation was 100% for the common and external iliac vessels, but the rates for the vesical vein (75%), superior gluteal vein (60%), or accessory obturator vein (63%) were suboptimal. Regarding pelvic bone, the average Dice similarity coefficient between manual and automatic segmentation was 0.97 (standard deviation 0.0043). Though there is room to improve the segmentation accuracy, the algorithm developed in this study can be utilized for surgical simulation in the treatment of advanced rectal cancer.


Autistic and Catatonic Spectrum Symptoms in Patients with Borderline Personality Disorder.
Recent literature has shown that a considerable percentage of patients with severe mental disorders can develop, over time, full-blown or subthreshold catatonia. Some studies corroborate the model of an illness trajectory in which different mental disorders would be arranged along a continuum of severity until the development of catatonia. In such an illness pathway, autistic traits (AT) and borderline personality disorder (BPD) may represent important steps. In order to further explore the association between AT, BPD, and catatonia, the aim of this study was to compare catatonic spectrum symptoms and AT among patients with major depressive disorder (MDD), BPD, and healthy controls (CTL), also evaluating possible predictive dimensions of the different diagnoses. A total of 90 adults affected by BPD, 90 adults with a diagnosis of MDD, and 90 CTL, homogeneous in terms of gender and age, were recruited from six Italian university departments of psychiatry and assessed with the SCID-5-RV, the Catatonia Spectrum (CS), and the Adult Autism Subthreshold Autism Spectrum (AdAS Spectrum). The total CS score was significantly higher in the BPD and MDD groups than in the CTL group, while the majority of CS domain scores were significantly higher in the BPD group than in the MDD group, which scored significantly higher than the CTL group. The total AdAS Spectrum score and the AdAS Spectrum domain scores were significantly higher in the BPD group than in the MDD group, which in turn scored significantly higher than the CTL group. The CS domains "psychomotor activity" and "impulsivity", and AdAS Spectrum domains "verbal communication", "empathy", and "hyper-/hyporeactivity to sensory input" were associated with the risk of presenting a diagnosis of BPD.


Prediction of serum anti-HSP27 antibody titers changes using a light gradient boosting machine (LightGBM) technique.
Previous studies have proposed that heat shock proteins 27 (HSP27) and its anti-HSP27 antibody titers may play a crucial role in several diseases including cardiovascular disease. However, available studies has been used simple analytical methods. This study aimed to determine the factors that associate serum anti-HSP27 antibody titers using ensemble machine learning methods and to demonstrate the magnitude and direction of the predictors using PFI and SHAP methods. The study employed Python 3 to apply various machine learning models, including LightGBM, CatBoost, XGBoost, AdaBoost, SVR, MLP, and MLR. The best models were selected using model evaluation metrics during the K-Fold cross-validation strategy. The LightGBM model (with RMSE: 0.1900 ± 0.0124; MAE: 0.1471 ± 0.0044; MAPE: 0.8027 ± 0.064 as the mean ± sd) and the SHAP method revealed that several factors, including pro-oxidant-antioxidant balance (PAB), physical activity level (PAL), platelet distribution width, mid-upper arm circumference, systolic blood pressure, age, red cell distribution width, waist-to-hip ratio, neutrophils to lymphocytes ratio, platelet count, serum glucose, serum cholesterol, red blood cells were associated with anti-HSP27, respectively. The study found that PAB and PAL were strongly associated with serum anti-HSP27 antibody titers, indicating a direct and indirect relationship, respectively. These findings can help improve our understanding of the factors that determine anti-HSP27 antibody titers and their potential role in disease development.


Toll-like receptor 2 (TLR2) engages endoplasmic reticulum stress sensor IRE1α to regulate retinal innate responses in Staphylococcus aureus endophthalmitis.
Endoplasmic reticulum (ER) stress response has been implicated in a variety of pathophysiological conditions, including infectious and inflammatory diseases. However, its contribution in ocular bacterial infections, such as endophthalmitis, which often cause blindness is not known. Here, using a mouse model of Staphylococcus (S.) aureus endophthalmitis, our study demonstrates the induction of inositol-requiring enzyme 1α (IRE1α) and splicing of X-box binding protein-1 (Xbp1) branch of the ER-stress pathway, but not the other classical ER stress sensors. Interestingly, S aureus-induced ER stress response was found to be dependent on Toll-like receptor 2 (TLR2), as evident by reduced expression of IRE1α and Xbp1 mRNA splicing in TLR2 knockout mouse retina. Pharmacological inhibition of IRE1α using 4µ8C or experiments utilizing IRE1α-/- macrophages revealed that IRE1α positively regulates S aureus-induced inflammatory responses. Moreover, IRE1α inhibition attenuated S aureus-triggered NF-κB, p38, and ERK pathways activation and cells treated with these pathway-specific inhibitors reduced Xbp1 splicing, suggesting a positive feedback inhibition. In vivo, inhibition of IRE1α diminished the intraocular inflammation and reduced PMN infiltration in mouse eyes, but, increased the bacterial burden and caused more retinal tissue damage. These results revealed a critical role of the IRE1α/XBP1 pathway as a regulator of TLR2-mediated protective innate immune responses in S aureus-induced endophthalmitis.


Mining of a novel reductase and its application for asymmetric reduction of p-methoxyacetophenone.
(R)-1-(4-methoxyphenyl) ethanol [(R)-1b] is an essential precursor for the synthesis of aryl propanoic acids' anti-inflammatatory drugs. Biocatalysts for (R)-1b preparation are limited and reductase has problems of low substrate concentration and low conversion rate. As a result, there is a constant need for discovering novel biocatalysts with excellent catalytic performances. In this study, a novel reductase LpSDR from Lacisediminihabitans profunda for the biocatalytic reduction of p-methoxyacetophenone (1a) to (R)-1b was obtained based on gene-mining technology, and some key reaction parameters were also investigated to improve the conversion rate of 1a using whole cells of recombinant Escherichia coli expressing reductase LpSDR as biocatalysts. It was found that the optimal concentration of isopropanol, ZnSO4·7H2O solution, 1a, and recombinant E. coli resting cells, the optimal reaction temperature, buffer pH, and reaction time were 1.95 mol l-1, 0.75 mmol l-1, 75 mmol l-1, 250 g (wet weight) l-1, 28°C, 7.0, and 21 h, respectively. Under the above conditions, a conversion rate of 99.5% and an enantiomeric excess of 99.6% were obtained, which were superior to the corresponding values previously reported. This study provides a novel reductase LpSDR, which is helpful in reducing 1a to (R)-1b.


Maternal obesity classes, preterm and post-term birth: a retrospective analysis of 479,864 births in England.
Preterm (< 37 weeks gestation) and post-term birth (≥42 weeks gestation) are associated with increased morbidity and mortality for mother and infant. Obesity (body mass index (BMI) ≥30 kg/m2) is increasing in women of reproductive age. Maternal obesity has been associated with adverse pregnancy outcomes including preterm and post-term birth. However, the effect sizes vary according to the subgroups of both maternal BMI and gestational age considered. The aim of this retrospective analysis was to determine the association between maternal obesity classes and gestational age at delivery. A secondary data analysis of 13 maternity units in England with information on 479,864 singleton live births between 1990 and 2007. BMI categories were: underweight (< 18.5 kg/m2), recommended weight (18.5-24.9 kg/m2), overweight (25.0-29.9 kg/m2) and obesity classes I (30.0-34.9 kg/m2), II (35.0-39.9 kg/m2), IIIa (40-49.9 kg/m2) and IIIb (≥50 kg/m2). Gestational age at delivery categories were: Gestational age at delivery (weeks): extreme preterm (20-27), very preterm (28-31), moderately preterm (32-36), early term (37, 38), full term (39-40), late term (41) and post-term (≥42). The adjusted odds of births in each gestational age category (compared to full-term birth), according to maternal BMI categories were estimated using multinomial logistic regression. Missing data were estimated using multiple imputation with chained equations. There was a J-shaped association between the absolute risk of extreme, very and moderate preterm birth and BMI category, with the greatest effect size for extreme preterm. The absolute risk of post-term birth increased monotonically as BMI category increased. The largest effect sizes were observed for class IIIb obesity and extreme preterm birth (adjusted OR 2.80, 95% CI 1.31-5.98). Women with class IIIb obesity have the greatest risks for inadequate gestational age. Combining obesity classes does not accurately represent risks for many women as it overestimates the risk of all preterm and post-term categories for women with class I obesity, and underestimates the risk for women in all other obesity classes.


Phase II study to assess the efficacy, safety and tolerability of the mitotic spindle kinesin inhibitor AZD4877 in patients with recurrent advanced urothelial cancer.
AZD4877 is a potent inhibitor of the mitotic spindle kinesin, Eg5. Early-phase clinical studies in a broad range of cancers showed that AZD4877 is well tolerated. This Phase II study evaluated the efficacy, safety and pharmacokinetics (Cmax) of AZD4877 in patients with previously treated advanced urothelial cancer (ClinicalTrials.gov identifier NCT00661609). AZD4877 25 mg was administered once-weekly for 3 weeks of each 4-week cycle until disease progression, death, unacceptable toxicity or withdrawal. The primary objective was to determine the objective response rate (RECIST). Recruitment was to be halted if ≤ 2 of the first 20 evaluable patients achieved an objective tumor response. Cmax was assessed on days 1 and 8 of cycle 1. None of the first 20 patients evaluable for efficacy achieved an objective response; enrollment was therefore halted. During this initial analysis, a further 21 patients were recruited. Overall, 39 patients were evaluable for efficacy, including one with confirmed partial response (PR) and seven patients with stable disease for ≥ 8 weeks (including one unconfirmed PR). The most commonly reported treatment-related adverse events (TRAEs) were neutropenia (22 patients), fatigue (12), leukopenia (7) and constipation (6); the most commonly reported grade ≥ 3 TRAE was neutropenia (21). Four patients had serious TRAEs. On days 1 and 8, the geometric mean Cmax of AZD4877 was 138 ng/ml (CV = 75 %) and 144 ng/ml (CV = 109 %), respectively. AZD4877 was generally tolerable in patients with advanced urothelial cancer. Given the limited clinical efficacy, further development of AZD4877 in urothelial cancer is not planned.


Obstructive sleep apnea is associated with depressed myocardial mechanoenergetics.
To evaluate the association between the myocardial mechanoenergetic efficiency index (MEEi) and the Apnea-Hypopnea Index (AHI) in the initial phase of obstructive sleep apnea (OSA) diagnosis. In this cohort study, we included a total of 382 eligible participants without cardiovascular disease in a tertiary outpatient clinic between January 2013 and January 2015. We recorded demographic, clinical, polysomnographic and echocardiographic variables of the patients. In addition, myocardial mechanoenergetic efficiency (MEE) and MEEi were calculated by an echocardiography-derived validated measurement. The mean (±SD) age of the participants was 48.47 ± 12.13, and male/female ratio was 287/95. Comparing with non-OSA, MEEi was significantly lower in OSA patients at all stages (0.35 ± 0.08 vs. 0.42 ± 0.05; p < .001). MEEi was negatively correlated with hypertension (r = -0.518, p < .001), body mass index (r = -0.382, p < .001), AHI (r = -0.656, p < .001), total apne (r = -0.525, p < .001), hypopnea (r = -0.415, p < .001), systolic pulmonary pressure (r = -0.318, p < .001), relative wall thickness (RWT; r = -0.415, p < .001), and positive correlated with left ventricular ejection fraction (r = 0.586, p < .001). According to multiple linear regression analysis AHI (β = -0.625, p < .001), total apnea (β = -0.402, p = .001), hypopnea (β = -0.395, p = .001), LV ejection fraction (β = 0.478, p < .001) and RWT (β = -0.279, p < .001) have an independent relationship with MEEi. MEEi was lower in OSA patients. A reduced MEEi may reflect a disturbance in energy use of the myocardium. Consequently, our results may provide insight into the mechanisms leading to structural cardiac diseases in OSA patients.


Oligonephropathy, developmental programming and nutritional management of low-gestation newborns.
Intrauterine growth retardation is associated with oligonephropathy and the risk of hypertension and renal disease in adulthood. Animal study data show that nephron density is directly related to protein nutrition status during nephrogenesis. Among infants born at low gestation, postnatal growth retardation and major deficits in energy and protein intake have been reported. Since nephrogenesis occurs mainly in late gestation, increasing protein intake during nephrogenesis could minimize the risk of long-term morbidities associated with oligonephropathy.


MetaMine--a tool to detect and analyse gene patterns in their environmental context.
Modern sequencing technologies allow rapid sequencing and bioinformatic analysis of genomes and metagenomes. With every new sequencing project a vast number of new proteins become available with many genes remaining functionally unclassified based on evidences from sequence similarities alone. Extending similarity searches with gene pattern approaches, defined as genes sharing a distinct genomic neighbourhood, have shown to significantly improve the number of functional assignments. Further functional evidences can be gained by correlating these gene patterns with prevailing environmental parameters. MetaMine was developed to approach the large pool of unclassified proteins by searching for recurrent gene patterns across habitats based on key genes. MetaMine is an interactive data mining tool which enables the detection of gene patterns in an environmental context. The gene pattern search starts with a user defined environmentally interesting key gene. With this gene a BLAST search is carried out against the Microbial Ecological Genomics DataBase (MEGDB) containing marine genomic and metagenomic sequences. This is followed by the determination of all neighbouring genes within a given distance and a search for functionally equivalent genes. In the final step a set of common genes present in a defined number of distinct genomes is determined. The gene patterns found are associated with their individual pattern instances describing gene order and directions. They are presented together with information about the sample and the habitat. MetaMine is implemented in Java and provided as a client/server application with a user-friendly graphical user interface. The system was evaluated with environmentally relevant genes related to the methane-cycle and carbon monoxide oxidation. MetaMine offers a targeted, semi-automatic search for gene patterns based on expert input. The graphical user interface of MetaMine provides a user-friendly overview of the computed gene patterns for further inspection in an ecological context. Prevailing biological processes associated with a key gene can be used to infer new annotations and shape hypotheses to guide further analyses. The use-cases demonstrate that meaningful gene patterns can be quickly detected using MetaMine.MetaMine is freely available for academic use from http://www.megx.net/metamine.


The glycoprotein nature of murine gamma interferon.
Synthesis of murine gamma interferon (MuIFN gamma) by phytohemagglutinin (PHA)-stimulated spleen cells was inhibited in a dose-dependent manner by graded concentrations of tunicamycin or 2-deoxy-glucose, both of which inhibit glycosylation. The homologous (murine) and heterologous (rat) antiviral activities of the MuIFN gamma preparations secreted in the presence of the various concentrations of either glycosylation inhibitor were reduced to similar degrees. MuIFN gamma synthesized in the presence of tunicamycin (tunicamycin-MuIFN gamma) exhibited a lower molecular weight (MW), a lack of binding to immobilized Concanavalin-A (Con A), and different charge properties when compared to MuIFN gamma produced in absence of inhibitor (control-MuIFN gamma). Potent rabbit and rat neutralizing antibodies raised against a control-MuIFN gamma subcomponent, isolated by its specific binding to a Con A affinity column, neutralized the antiviral activity of tunicamycin-MuIFN gamma to the same degree as the immunogen.


Depth and age-dependent distribution of keratocytes in healthy human corneas: a study using scanning-slit confocal microscopy in vivo.
To document keratocyte distribution and changes with age in the cellular network of the human cornea in vivo. Department of Ophthalmology, University of Rostock, Rostock, Germany. Forty-nine eyes of 31 healthy subjects of various ages were examined with a modified Microphthal scanning-slit confocal microscope (SSCM) (Hund) to document keratocyte distribution in the intact living cornea. Optical sections made by confocal microscopy were recorded on videotape, and the keratocyte density was determined for the total volume of the cornea and for the stromal sublayers. The highest cell density was in the anterior stroma of the cornea immediately posterior to Bowman's membrane (24 320 cells/mm(3) +/- 6740 [SD]), the lowest in the central area (11,610 +/- 4290 cells/mm(3)), and an intermediate density in the posterior stroma immediately adjacent to Descemet's membrane (18,850 +/- 4610 cells/mm(3)). The differences were statistically significant (P <.005). The keratocyte density was significantly lower in the anterior and posterior regions in the group older than 50 years: Cell density at 4% depth was 20,960 +/- 8200 cells/mm(3) and at 96%, 15 520 +/- 4290 cells/mm(3) (P <.05). In healthy living corneas, the keratocyte density was high in the areas adjacent to Bowman's and Descemet's membranes and was lower in patients older than 50 years than in those younger than 50 years. Further studies are needed to document the rate of change with age and to better understand the role and capacity of aging keratocytes in regenerative processes following corneal diseases or surgical procedures.


Are monoglyceride-lipase, triglyceride-lipase and phospholipase A of rat liver microsomes distinct protein entities?
1. Different extraction and purification techniques were employed for the separation of MG-lipase, TG-lipase and phospholipase A from rat liver microsomes. 2. Up to 60 per cent of the microsomal content of TG-lipase and phospholipase could be extracted with 1 M KCl or NaCl. MG-lipase was extracted more readily by detergents (e.g. emulphogen). 3. MG-lipase is more resistant to detergent and heat inactivation than TG-lipase and phospholipase A. It is retained, using the technique of affinity chromatography, on a column of CH-Sepharose coupled to monooleoylglycerol. In addition, MG-lipase was separated from TG-lipase by electrofocusing. 4. TG-lipase and phospholipase A were partially separated by gel filtration on Sephadex G-200 in the presence of 1 mM dithiothreitol and by chromatography on CH-Sepharose 4b. 5. On the basis of the present extraction and purification studies, it is concluded that mg-lipase is an enzyme protein distinct from TG-lipase and phospholipase A.


Exercise enhances the effectiveness of vitamin D therapy in rats with Alzheimer's disease: emphasis on oxidative stress and inflammation.
Alzheimer's disease (AD) is characterized by gradual loss of memory and cognitive functions which can affect anyone. Authors declared that there is a link between vitamin D and brain function. It has been proven that vitamin D plays an important role in improving AD cognitive functions. Researchers have found that exercise has many beneficial effects on humans. In addition to cardioprotection, it has been demonstrated that exercise provides an effective improvement in different brain functions. So in our study, we aimed to evaluate the effect of each of vitamin D and/ or exercise on AD and if they could be used as a potential line for treating AD. This study was conducted on fifty female white albino rats divided equally into 5 groups: control group, Alzheimer group induced by Lipopolysaccharide, Alzheimer group treated with vitamin D, Alzheimer group treated with exercise and Alzheimer group treated with both vitamin D and exercise. The following parameters were assessed in rat brain tissues: acetylcholine esterase (AChE) activity, levels of amyloid β 42 and tau proteins, dopamine brain neurotransmitter, BDNF and NGF by ELISA. Serum levels of IL-6 and IL-10 were assessed by ELISA. MDA, GSH and vitamin D levels were also estimated in addition to cognitive function tests and histopathological examination of rat brain tissues. In Alzheimer group, there was a significant increase in the proinflammatory cytokine IL-6, the lipid peroxidation marker MDA, amyloid β and tau proteins, levels. In addition to a significant increase in time consumed in T-maze test. Alzheimer group also showed a significant decrease in the anti-inflammatory cytokine IL-10, the anti-oxidative stress biomarker GSH, the neurotransmitters AChE and dopamine, and the growth factors BDNF and NGF as well as serum vitamin D levels. Treatment with either vitamin D or exercise significantly improved cognitive dysfunction and the histopathological picture of the brains of Alzheimer's rats with the best results in combined vitamin D and exercise treated group. The treated groups, especially combined vitamin D and exercise group, showed a significant decrease in IL-6, MDA, amyloid β and tau proteins levels, but on the other hand they showed a significant increase in IL-10, GSH, AChE, dopamine, BDNF and NGF. These data suggest that combined vitamin D and exercise could be considered as a potential and effective line for treating AD.


Number of examined lymph nodes and nodal status assessment in pancreaticoduodenectomy for pancreatic adenocarcinoma.
The accuracy of the assessment of the nodal status in resected cephalic pancreatic adenocarcinoma (PA) depends on the number of examined lymph nodes (NELN). This study assesses the impact of the NELN on N staging and survival and propose a minimal number of examined lymph nodes (MNELN) ensuring reliability of the pN status determination. 188 consecutive patients treated by pancreaticoduodenectomy (PD) for PA. Correlations between NELN and survivals of pN0 and pN1 groups and with the rate of pN1 patients were studied. A probability model based on the binomial law was built to estimate the MNELN able to detect pN1 patients with a sensitivity ≥ 95%. Overall and disease free 5-year survivals were 27.2% and 24.6% respectively. 135 patients (71.8%) were staged pN1. The median NELN was 17 (range 0-68). Overall and disease free survivals of pN1 patients were not related to NELN. The influence of NELN on survival in pN0 patients due to stage migration did not reach significance. The probability model showed that a MNELN of 16 nodes was required to detect pN1 patients with a sensitivity of 95%. A MNELN of 16 is required to assess pN status and should be considered as a quality criterion in future studies and trials on PD for PA.


Physical activity interventions are delivered consistently across hospitalized older adults but multimorbidity is associated with poorer rehabilitation outcomes: A population-based cohort study.
Older adults live with multimorbidity including frailty and cognitive impairment often requiring hospitalization. While physical activity interventions (PAIs) are a normal rehabilitative treatment, their clinical effect in hospitalized older adults is uncertain. To observe PAI dosing characteristics and determine their impact on clinical performance parameters. A single-site prospective observational cohort study in an older persons' unit. Seventy-five older persons' unit patients ≥65 years. PAI; therapeutic contact between physiotherapy clinician and patient. Parameters included changes in activities-of-daily-living (Barthel Index), handgrip strength, balance confidence, and gait velocity, measured between admission and discharge (episode). Dosing characteristics were PAI temporal initiation, frequency, and duration. Frailty/cognition status was dichotomized independently per participant yielding 4 subgroups: frail/nonfrail and cognitively-impaired/cognitively-unimpaired. Median (interquartile range) PAI initiation occurred after 2 days (1-4), frequency was 0.4 PAIs per day (0.3-0.5), and PAI duration per episode was 3.75 hours (1.8-7.2). All clinical parameters improved significantly across episodes: grip strength median (interquartile range) change, 2.0 kg (0.0-2.3) (P < .01); Barthel Index, 5 (3-8) (P < .01); gait velocity, 0.06 m.∙s-1 (0.06-0.16) (P < .01); and balance confidence, -3 (-6 to -1) (P < .01). Physical activity intervention dosing remained consistent within subgroups. While several moderate to large associations between amount of PAIs and change in clinical parameters were observed, most were within unimpaired subgroups. PAI dosing is consistent. However, while clinical changes during hospital episodes are positive, more favourable responses to PAIs occur if patients are nonfrail/cognitively-unimpaired. Therefore, to deliver a personalized rehabilitation approach, adaptation of PAI dose based on patient presentation is desirable.