Treatment success rates among tuberculosis patients notified from the private sector in West Bengal, India.
In India, an estimated 50% of tuberculosis (TB) patients are diagnosed and managed by private providers (PPs). However, there is limited information on treatment outcomes among those treated by PPs largely because of a lack of systems for patient follow-up and documentation of the outcomes. Tuberculosis Health Action Learning Initiative (THALI) project implemented in six districts of West Bengal, supported PPs in managing TB patients and systematically documented the TB treatment outcomes of such patients. This provided us an opportunity to describe the treatment outcomes and assess factors associated with 'unsuccessful outcomes' among TB patients notified by PPs from January to April 2018. This was a retrospective cohort study using routinely collected data. During the study reference period, 2347 patients were notified from 389 PPs. The patient's mean (SD) age was 39.9 (17.2) years and 61% were males. Of the total, about 86% had pulmonary TB, 95% were new cases, and 23% were microbiologically confirmed (either on sputum smear microscopy or Xpert assay). Among 2347 patients, 19% received free drugs from the National Tuberculosis Program (NTP) under the supervision of PPs while the rest purchased anti-TB drugs from the private pharmacies. The 'successful outcomes' (cured and treatment completed) were seen in 86.1% (95% confidence interval (CI), 84.6%-87.4%) patients and 8.6% were 'not evaluated'. The patients who received free NTP drugs (adjusted relative risk-4.0 (95% CI: 3.1-5.0)) had a higher risk of 'unsuccessful outcomes' than those who availed of drugs from private pharmacies. The high 'successful outcomes' among TB patients treated by PPs are promising. However, higher 'unsuccessful outcomes' among patients on free NTP drugs need to be explored, and suitable corrective actions have to be taken.


Contribution of different sources to the pollution of wet weather flows in combined sewers.
Experiments performed on "Marais" catchment, in central Paris, aimed to follow up the quality of wet weather flows from the entry to the exit of a combined sewer network. SS, VSS, COD, BOD5, Cd, Cu, Pb, Zn concentrations were measured for an important number of rain events in roof, yard, street runoff, as well as in dry and wet weather flows at the catchment outlet. Mass entry-exit totals, at the scale of the catchment, were calculated over 31 rain events in order to evaluate the contribution of different types of runoff, of sanitary sewage and of sewer sediments to the total wet weather pollutant loads at the catchment outlet. The erosion of in-sewer pollutant stocks was found to be the main source of particles and of organic matter in wet weather flows, whereas heavy metal loads mainly originated from roof runoff, due to the corrosion of metallic roofs. Particles eroded inside the sewer during rain events were found to be quite different from the particles constituting the main part of sewer sediments: they are organic and biodegradable, with rather important settling velocities and seem to accumulate during dry weather periods. A change of the chemical form of heavy metals was noticed during the transport in the sewer and it is suspected that a fraction of the dissolved metals from the runoff is adsorbed on sewer sediments.


Pepstatin inhibition of rat myocardial acid protease.
The protease inhibitor, pepstatin, inhibited acid protease in rat heart homogenates with a Ki of 7 x 10(-11) M by a noncompetitive mechanism, but it had no effect on myocardial acid protease activity when given in high doses to intact animals. Exposure of heart slices to 10(-7) M pepstatin for two hours also failed to produce demonstrable acid protease inhibition, suggesting that myocardial acid protease activity is unaffected by this potent inhibitor unless the integrity of the cell is disturbed.


[Kidney transplant in patients with HIV infection].
Until recently, human immunodeficiency virus (HIV) infection was an absolute contraindication to solid organ transplantation because it was feared that the anti-rejection therapy could result in accelerated HIV disease. At the end of the 1990s it became clear that HIV infection, once deemed a fatal disease, could be effectively turned into a chronic condition by the use of highly active antiretroviral therapy. Since then, the mortality rate from opportunistic infections has decreased dramatically, while liver and renal insufficiency have become the major causes of morbidity and mortality in these patients in the long term. A growing number of HIV patients develop end-stage renal disease secondary to immune-mediated glomerulonephritis, HIV-associated nephropathy, nephrotoxic effects induced by antiretroviral medication, or diabetic and vascular nephropathy, and therefore need maintenance dialysis. For this reason we have to reconsider kidney transplant as a possible treatment option. During the last decade, the results of many studies have shown that transplantation can be safe and effective as long as the HIV infection is effectively controlled by antiretroviral therapy. The short- and medium-term patient and graft survival rates in HIV-positive transplant recipients are comparable with those of the overall transplant population, but the incidence of acute rejection episodes is higher. The main clinical problem in the management of HIV-positive transplant recipients originates from the interference between immunosuppressive regimens and antiretroviral drugs. Thus, a close collaboration between infectious disease specialists and nephrologists is mandatory in order to optimize transplantation programs in these patients.


Epicardial Fat Volume Is Associated with Endothelial Dysfunction, but not with Coronary Calcification: From the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil).
The increase in epicardial fat volume (EFV) is related to coronary artery disease (CAD), independent of visceral or subcutaneous fat. The mechanism underlying this association is unclear. Coronary artery calcium (CAC) score and endothelial dysfunction are related to coronary events, but whether EFV is related to these markers needs further clarification. To evaluate the association between automatically measured EFV, cardiovascular risk factors, CAC, and endothelial function. In 470 participants from the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil) with measures of EFV, CAC score and endothelial function, we performed multivariable models to evaluate the relation between cardiovascular risk factors and EFV (response variable), and between EFV (explanatory variable) and endothelial function variables or CAC score. Two-sided p <0.05 was considered statistically significant. Mean age was 55 ± 8 years, 52.3% of patients were men. Mean EFV was 111mL (IQ 86-144), and the prevalence of CAC score=0 was 55%. In the multivariable analyses, increased EFV was related to female sex, older age, waist circumference, and triglycerides (p<0.001 for all). Higher EFV was associated with worse endothelial function: as compared with the first quartile, the odds ratio for basal pulse amplitude were (q2=1.22, 95%CI 1.07-1.40; q3=1.50, 95%CI 1.30-1.74; q4=1.50, 95%CI 1.28-1.79) and for peripheral arterial tonometry ratio were (q2=0.87, 95%CI 0.81-0.95; q3=0.86, 95%CI 0.79-0.94; q4=0.80, 95%CI 0.73-0.89), but not with CAC score>0. Higher EFV was associated with impaired endothelial function, but not with CAC. The results suggest that EFV is related to the development of CAD through a pathway different from the CAC pathway, possibly through aggravation of endothelial dysfunction and microvascular disease.


Targeted next generation sequencing in SPAST-negative hereditary spastic paraplegia.
Molecular characterization is important for an accurate diagnosis in hereditary spastic paraplegia (HSP). Mutations in the gene SPAST (SPG4) are the most common cause of autosomal dominant forms. We performed targeted next generation sequencing (NGS) in a SPAST-negative HSP sample. Forty-four consecutive HSP patients were recruited from an adult neurogenetics clinic in Sydney, Australia. SPAST mutations were confirmed in 17 subjects, and therefore 27 SPAST-negative patients were entered into this study. Patients were screened according to mode of inheritance using a PCR-based library and NGS (Roche Junior 454 sequencing platform). The screening panel included ten autosomal dominant (AD) and nine autosomal recessive (AR) HSP-causing genes. A genetic cause for HSP was identified in 25.9 % (7/27) of patients, including 1/12 classified as AD and 6/15 as AR or sporadic inheritance. Several forms of HSP were identified, including one patient with SPG31, four with SPG7 (with one novel SPG7 mutation) and two with SPG5 (including two novel CYP7B1 frameshift mutations). Additional clinical features were noted, including optic atrophy and ataxia for patients with SPG5 and ataxia and a chronic progressive external ophthalmoplegia-like phenotype for SPG7. This protocol enabled the identification of a genetic cause in approximately 25 % of patients in whom one of the most common genetic forms of HSP (SPG4) was excluded. Targeted NGS may be a useful method to screen for mutations in multiple genes associated with HSP. More studies are warranted to determine the optimal approach to achieve a genetic diagnosis in this condition.


Transgenic tobacco plants overexpressing chitinases of fungal origin show enhanced resistance to biotic and abiotic stress agents.
Genes encoding defense-related proteins have been used to alter the resistance of plants to pathogens and other environmental challenges, but no single fungal gene overexpression has produced broad-spectrum stress resistance in transgenic lines. We have generated transgenic tobacco (Nicotiana tabacum) lines that overexpress the endochitinases CHIT33 and CHIT42 from the mycoparasitic fungus Trichoderma harzianum and have evaluated their tolerance to biotic and abiotic stress. Both CHIT33 and CHIT42, individually, conferred broad resistance to fungal and bacterial pathogens, salinity, and heavy metals. Such broad-range protective effects came off with no obvious detrimental effect on the growth of tobacco plants.


Galangin Inhibits Thrombin-Induced MMP-9 Expression in SK-N-SH Cells via Protein Kinase-Dependent NF-κB Phosphorylation.
Galangin, a member of the flavonol compounds of the flavonoids, could exert anti-inflammatory effects in various cell types. It has been used for the treatment of arthritis, airway inflammation, stroke, and cognitive impairment. Thrombin, one of the regulators of matrix metalloproteinase (MMPs), has been known as a vital factor of physiological and pathological processes, including cell migration, the blood⁻brain barrier breakdown, brain edema formation, neuroinflammation, and neuronal death. MMP-9 especially may contribute to neurodegenerative diseases. However, the effect of galangin in combating thrombin-induced MMP-9 expression is not well understood in neurons. Therefore, we attempted to explore the molecular mechanisms by which galangin inhibited MMP-9 expression and cell migration induced by thrombin in SK-N-SH cells (a human neuroblastoma cell line). Gelatin zymography, western blot, real-time PCR, and cell migration assay were used to elucidate the inhibitory effects of galangin on the thrmbin-mediated responses. The results showed that galangin markedly attenuated the thrombin-stimulated phosphorylation of proto-oncogene tyrosine-protein kinase (c-Src), proline-rich tyrosine kinase 2 (Pyk2), protein kinase C (PKC)α/β/δ, protein kinase B (Akt), mammalian target of rapamycin (mTOR), p42/p44 mitogen-activated protein kinase (MAPK), Jun amino-terminal kinases (JNK)1/2, p38 MAPK, forkhead box protein O1 (FoxO1), p65, and c-Jun and suppressed MMP-9 expression and cell migration in SK-N-SH cells. Our results concluded that galangin blocked the thrombin-induced MMP-9 expression in SK-N-SH cells via inhibiting c-Src, Pyk2, PKCα/βII/δ, Akt, mTOR, p42/p44 MAPK, JNK1/2, p38 MAPK, FoxO1, c-Jun, and p65 phosphorylation and ultimately attenuated cell migration. Therefore, galangin may be a potential candidate for the management of brain inflammatory diseases.


Laboratory experiments to determine if crayfish can communicate chemically in a flow-through system.
The importance of chemical cues for transmitting information concerning sexual identity, agonistic state, and stress-related condition in the crayfishProcambarus clarkii (Girard) was examined in a flow-through system. Experiments tested the effects of "conditioned water" from stimulus tanks on the behavior of solitary male or female crayfish. Twenty males and 20 females were subjected to a random sequence of five treatments: unconditioned water (control), conditioned water flowing through tanks containing a solitary male or female, and conditioned water from tanks holding either two males or two females. Durations of the following behaviors were recorded: chelae up, chela(e) in baffle hole, chela waving, climbing, digging, grooming, gross body movement, and meral spread. Results indicated that crayfish chemically detected another animal within 0.25 m without additional visual or tactile stimuli; however, crayfish apparently did not "communicate" information on sexual identity, agonistic state, or stress condition, nor does this detection necessarily imply discrimination between stimuli from crayfish and other taxa (e.g., fish). Our conclusions are contrasted with the two previous reports on chemical communication in crayfish in which experimental animals were tested in static systems. We suggest that a temporal separation of molting and copulation and a long reproductive receptivity period for females (which would allow abundant intersexual encounters) could account for a lack of selective pressure to evolve long-distance sex pheromones.


Studying the mechanism of CD47-SIRPα interactions on red blood cells by single molecule force spectroscopy.
The interaction forces and binding kinetics between SIRPα and CD47 were investigated by single-molecule force spectroscopy (SMFS) on both fresh and experimentally aged human red blood cells (hRBCs). We found that CD47 experienced a conformation change after oxidation, which influenced the interaction force and the position of the energy barrier between SIRPα and CD47. Our results are significant for understanding the mechanism of phagocytosis of red blood cells at the single molecule level.


Safety and efficacy of catheter ablation of atrial fibrillation in the very elderly (≥80 years old): Insights from the UC San Diego AF Ablation Registry.
Catheter ablation improves outcomes in symptomatic atrial fibrillation (AF) patients. However, its safety and efficacy in the very elderly (≥80 years old) is not well described. Ablation of AF in the very elderly is safe and effective. We performed a retrospective study of all patients who underwent catheter ablation enrolled in the University of California, San Diego AF Ablation Registry. The primary outcome was freedom from atrial arrhythmias on or off antiarrhythmic drugs (AADs). Of 847 patients, 42 (5.0%) were 80 years of age or greater with a median age of 81.5 (80-82.3) and 805 (95.0%) were less than 80 years of age with a median age of 64.4 (57.6-70.2). Among those who were ≥80 years old, 29 were undergoing de novo ablation (69.0%), whereas in the younger cohort, 518 (64.5%) were undergoing de novo ablation (p = .548). There were no statistically significant differences in fluoroscopy (p = .406) or total procedure times (p = .076), AAD use (p = .611), or procedural complications (p = .500) between groups. After multivariable adjustment, there were no statistically significant differences in recurrence of any atrial arrhythmias on or off AAD (adjusted hazard ratio [AHR]: 0.75; 95% confidence interval [CI]: 0.45-1.23; p = .252), all-cause hospitalizations (AHR: 0.86; 95% CI: 0.46-1.60; p = .626), or all-cause mortality (AHR: 4.48; 95% CI: 0.59-34.07; p = .147) between the very elderly and the younger cohort. In this registry analysis, catheter ablation of AF appears similarly effective and safe in patients 80 years or older when compared to a younger cohort.


Adverse events in cancer patients with sickle cell trait or disease: case reports.
Given the relatively high prevalence of sickle cell trait and disease among African Americans and established racial disparities in cancer outcomes, we reviewed the literature regarding adverse events in cancer patients with these hematologic genotypes. Erythrocyte sickling can result from extreme hypoxia and other physiologic stressors, as might occur during cancer therapy. Further, tumoral hypoxia, a poor prognostic and predictive factor, could lead to a cycle of local sickling and increased hypoxia. A search of PubMed produced 150 publications, most of which were excluded because of incidental relevance. Eleven case reports of patients diagnosed from 1993 to 2013 were reviewed. Two reports of patients with sickle cell trait describe an abundance of sickled erythrocytes within tumors, and a third report describes sickling-related events requiring multiday hospitalization. Eight reports of patients with sickle cell disease delineated multiorgan failure, vaso-occlusive crises, and rapid renal deterioration. Hypothesized triggers are delayed clearance of anticancer agents attributable to baseline kidney damage, activation of vasoadherent neutrophils from treatment to counter chemotherapy-induced neutropenia, hypoxia from general anesthesia, and intratumoral hypoxia. Clinical implications include pretreatment genotyping for prophylaxis, dose adjustment, and enhanced patient monitoring. With the current lack of high-quality evidence, however, the scope of poor outcomes remains unknown.


Antiangiogenic effects of the novel camptothecin ST1481 (gimatecan) in human tumor xenografts.
ST1481 (gimatecan) is a novel lipophilic camptothecin with a promising preclinical pharmacological profile. On the basis of its high antitumor efficacy when delivered by the oral route, the compound is suitable for prolonged administration. This schedule of treatment has been reported as the most appropriate to exploit the antiangiogenic effects of cytotoxic drugs. The aim of the study was to investigate the antiangiogenic and antitumor effects of oral ST1481 in human tumor xenografts. In spite of a marginal drug effect against the s.c. growing A549 lung carcinoma following administration with an intermittent schedule (q4dx4 times, maximum tolerated dose: 2 mg/kg), tumor growth was strongly inhibited by a daily low-dose (0.5 mg/kg) prolonged administration. Immunohistochemical analysis showed a reduced number of microvessels in tumors of both treated groups versus controls and a significantly higher reduction in the daily versus the q4dx4-treated tumors (P < 0.0001, by Student's t test). In our experimental model, the relation between microvessel density and tumor size (r = 0.738, by the Spearman rank test) suggests a role of inhibition of tumor vasculature in tumor response. Significant inhibition of tumor angiogenesis (P < 0.0001 versus control tumors) was observed even with a very low drug dose (0.06 mg/kg) in the orthotopically implanted (i.d.) MeWo melanoma, under conditions causing minimal tumor growth inhibition. Additional evidences of the antiangiogenic activity of ST1481 were provided by antimotility effects on endothelial cells, in vivo inhibition of vascularization in the Matrigel assay, and down-regulation of the expression of the proangiogenic basic fibroblast growth factor in A549 tumor cells associated with inhibition of the pathway involving Akt. In conclusion, the available results support the possibility that the antiangiogenic properties of ST1481 contribute to its antitumor potential and that this effect might be enhanced by the continuous low-dose treatment.


Mogroside V ameliorates broiler pulmonary inflammation via modulating lung microbiota and rectifying Th17/Treg dysregulation in lipopolysaccharides-induced lung injury.
The dysbiosis of lung microbiota and inflammatory factors play a crucial role in the occurrence of lipopolysaccharides (LPS)-induced lung injury. Recently, mogroside V (MGV) has received increasing attention due to its potential health benefits in pneumonia, but its complex mechanism needs further experimental elucidation. In this study, we established an LPS-induced chicken lung injury model to investigate the protective effect of MGV on LPS-induced acute lung injury in broiler and its related mechanisms. A total of 192 one-day-old white-finned broilers were randomly assigned into 4 groups with 6 replicates: 1) control group: basal diet (d 1-44), saline (d 43); 2) LPS group: basal diet (d 1-44), LPS (d 43); 3) MGV group: basal diet + 0.2% MGV (d 1-44), saline (d 43); 4) MGV-LPS group: basal diet + 0.2% MGV (d 1-44), LPS (d 43). The results showed that pathological examination showed that lung tissue inflammation infiltration was reduced after MGV treatment. In addition, MGV can promote the balance of Th17 and Treg cell cytokines, significantly inhibit the expression of proinflammatory cytokines (IL-1β (P < 0.01), IL-6 (P < 0.001), IL-17F (P < 0.05)), and decrease immunosuppressive target expression (PD-L1 (P < 0.01), PD-1 (P < 0.001), RORα (P < 0.001)), activating the immune system. Furthermore, 16S rRNA sequencing analysis showed that MGV treatment could increase the abundance of beneficial bacteria in the lung and reduce the abundance of bacteria associated with inflammation. Generally, MGV intervention has a preventive effect on the pathological damage induced by lipopolysaccharides. Its mechanism is related to inhibiting the inflammatory response, regulating the Th17/Treg balance, and maintaining the stability of lung microbiota.


Free and esterified fatty acid and cholesterol synthesis in adult males and its effect on the doubly-labelled water method.
The purpose of the present study was to estimate whole-body fatty acid and cholesterol synthesis in weight-stable adults and to determine the likely effect on the doubly-labelled water (DLW) method for measuring energy expenditure. Synthesis was measured by 2H incorporation over 14 d in six adult males in approximate energy balance following noradrenaline infusion to maximize mobilization of free fatty acid from adipose tissue. The inter-individual variation in synthesis rates was large and in one subject the proportion of free fatty acid synthesized was ten times that of the mean of the rest of the group; the fasting concentration of esterified fatty acid in this subject was five times that of the rest of the group indicating likely violation of the assumptions underlying the calculation of whole-body synthesis. After 14 d of labelling in the other five subjects, 0.9 (SEM 0.3)% of the circulating free fatty acid, 9.3 (SEM 3.0)% of the esterified fatty acid, 14.6 (SEM 2.4)% of the free cholesterol and 28.3 (SEM 3.7)% of esterified cholesterol had been synthesized de novo. A high rate of synthesis correlated with a low pre-dose 2H abundance both within and between lipid classes suggesting that natural 2H abundance variations in some lipid classes may be used to determine their metabolic origin. Whole-body synthetic rates were 8 g/d for fatty acid and 0.3-0.5 g/d for cholesterol. These values correspond to very small errors on DLW-derived estimates of CO2 production; -2.5 litres/d for fatty acid and -0.1 to -0.2 litres/d for cholesterol. These results, obtained in subjects typically consuming a diet with a lower fat and cholesterol content that the typical Western diet, suggest that the DLW method is unlikely to be affected by fatty acid and cholesterol synthesis in subjects in energy balance consuming a typical Western diet.


Waiting for markets to change me-High-stakeholders' views of antibiotic use and antibiotic resistance in pig production in Brazil.
Overuse of veterinary antibiotics is a risk factor for antimicrobial resistance (AMR), which is a global public health emergency. More than 70% of the antibiotics consumed worldwide are used in farm animals, mainly in poultry and pig herds. Brazil is the fourth largest pork producer globally and the second-largest user of antibiotics in animals. Qualitative research can help understand the complexities around antibiotic use (AMU) in Brazilian pig herds and identify stakeholders' attitudes concerning the rational AMU and AMR in the production chain. This study aimed to explore the knowledge and attitudes of high-level professionals in the animal production chain about AMU and AMR in pig farming, the relationship with pig welfare and AMU in Brazil. We conducted 32 in-depth interviews with individuals active in the pig industry. The majority of the participants considered AMU excessive and inappropriate in pig farms in Brazil. However, attitudes toward a restrictive AMU scenario in Brazilian pig farms were predominantly negative, justified by economic, sanitary and social barriers. These included unsatisfactory management and biosecurity conditions in pig farms that, in their opinion, justify AMU to prevent diseases; issues surrounding prescription and acquisition of veterinary drugs; and employment and income relationships arising from the sale of antibiotics. The views of high-level professionals in the Brazilian livestock chain reveal antibiotics as a structural element that enables pig production. Antibiotics were viewed as essential resources for producing cheap food. Foreign markets were considered the most relevant driver of change in AMU practices rather than pressure from Brazilian consumers. A common belief expressed was that AMR is more associated with the inappropriate AMU in human medicine than in the livestock sector. Resistance to change in these stakeholders may hinder the implementation of future public policies to restrict the use of antibiotics in Brazil. Our findings suggest that successful measures to deal with the AMU/AMR challenges in the pig chain shall not be rooted in personal behavior change. Instead, honest interdisciplinary dialogues and structural changes are needed to define common grounds and a way forward to break the cycle perpetuating antibiotics as structural commodities.


Arthroscopic stabilization in posterior or multidirectional instability of the shoulder.
The diagnosis and treatment of posterior or multidirectional instability of the shoulder can be difficult. Normal laxity must be differentiated from pathologic instability. Once clinically diagnosed, there are several likely factors that contribute to pathologic instability. Although many patients with posterior or multidirectional instability of the shoulder respond well to nonsurgical treatment, when surgical intervention is required, the success rate has not been as favourable as that for repairing isolated anterior instability of the shoulder.


Effects of dietary menhaden oil, soy, and a cyclooxygenase inhibitor on human breast cancer cell growth and metastasis in nude mice.
The purpose of Study 1 was to examine the effect of dietary soy on the progression of MDA-MB-435 human breast cancer cell solid tumors in nude mice. When toasted soy chips were fed at levels of 5%, 10%, or 20% (wt/wt) in a high-fat, linoleic acid-rich diet for 12 weeks, there was a trend for larger mammary fat pad tumors to occur with increasing soy intake. However, compared with the controls the severity of macroscopic lung metastasis was reduced significantly in the groups fed 10% and 20% soy. Study 2 compared the effects of diets containing 23% corn oil (CO), 18% menhaden oil (MO) + 5% CO, 18% MO + 5% CO + 10% soy chips, and MO or soy-supplemented diets + indomethacin treatment in the same animal model. Feeding the 18% MO diet without soy or indomethacin reduced primary tumor growth; statistically significant effects were not observed in any of the other groups. All three of the groups with MO supplementation showed a reduction in the occurrence and severity of macroscopic lung metastases, together with the expected decreases in tumor prostaglandin E levels. These effects were most pronounced when MO was combined with indomethacin treatment. When indomethacin was given with dietary soy, the previously reported suppressive effect of the cyclooxygenase inhibitor on MDA-MB-435 cell tumor progression was lost, despite reductions in tumor prostaglandin E concentrations.


Aspergillus diversity in the environments of nosocomial infection cases at a university hospital.
Aspergillus species (sp.) that causes opportunistic infections have been increasingly found in human mainly immunosuppressive patients around the world every year. The main objective was to use a rapid and cheap molecular method for monitoring Aspergillus infections and epidemiological approaches. In order to identity Aspergilli species (spp.), a number of molecular methods including restriction fragment length polymorphism (RFLP) have been employed in accordance with ribosomal RNA amplification. The focus of this study - a group of hospitalized patients with clinical and subclinical signs of infection. All of the collected clinical specimens were transported to the medical mycology lab and examined for Aspergillus identification. The environmental specimens were collected from air and surfaces inspected for the Aspergillus within the hospital sources. At first, growth characteristics and microscopic features on mycological media for the identification of Aspergillus sp. were performed. For the confirmation of Aspergillus isolates which similarly found in clinical and environmental sources, molecular method polymerase chain reaction/restriction fragment length polymorphism was carried out. From the mentioned specimens, 102 fungal isolates included Candida spp., Aspergillus spp. and other fungi. Aspergillus flavus (47%), Aspergillus fumigatus (29.4%) and Aspergillus niger (23.5%) all were found as the most common clinical isolates. In addition, Aspergillus isolates from environmental were Aspergillus niger (43.7%), Aspergillus flavus (41.7%), Aspergillus fumigatus (14.6%). Therefore, polymerase chain reaction-restriction fragment length polymorphism with a single restriction enzyme can be very useful in the identification of Aspergillus spp., because of its facility in use, speed, robust, and high sensitivity of diagnosis.


e-Mental Health Program Usage Patterns in Randomized Controlled Trials and in the General Public to Inform External Validity Considerations: Sample Groupings Using Cluster Analyses.
Randomized controlled trials (RCTs) with vigorous study designs are vital for determining the efficacy of treatments. Despite the high internal validity attributed to RCTs, external validity concerns limit the generalizability of results to the general population. Bias can be introduced, for example, when study participants who self-select into a trial are more motivated to comply with study conditions than are other individuals. These external validity considerations extend to e-mental health (eMH) research, especially when eMH tools are designed for public access and provide minimal or no supervision. Clustering techniques were employed to identify engagement profiles of RCT participants and community users of a self-guided eMH program. This exploratory approach inspected actual, not theorized, RCT participant and community user engagement patterns. Both samples had access to the eMH program over the same time period and received identical usage recommendations on the eMH program website. The aim of this study is to help gauge expectations of similarities and differences in usage behaviors of an eMH tool across evaluation and naturalistic contexts. Australian adults signed up to myCompass, a self-guided online treatment program created to reduce mild to moderate symptoms of negative emotions. They did so either by being part of an RCT onboarding (160/231, 69.6% female) or by accessing the program freely on the internet (5563/8391, 66.30% female) between October 2011 and October 2012. During registration, RCT participants and community users provided basic demographic information. Usage metrics (number of logins, trackings, and learning activities) were recorded by the system. Samples at sign-up differed significantly in age (P=.003), with community users being on average 3 years older (mean 41.78, SD 13.64) than RCT participants (mean 38.79, SD 10.73). Furthermore, frequency of program use was higher for RCT participants on all usage metrics compared to community users through the first 49 days after registration (all P values <.001). Two-step cluster analyses revealed 3 user groups in the RCT sample (Nonstarters, 10-Timers, and 30+-Timers) and 2 user groups in the community samples (2-Timers and 20-Timers). Groups seemed comparable in patterns of use but differed in magnitude, with RCT participant usage groups showing more frequent engagement than community usage groups. Only the high-usage group among RCT participants approached myCompass usage recommendations. Findings suggested that external validity concerns of RCT designs may arise with regards to the predicted magnitude of eMH program use rather than overall usage styles. Following up RCT nonstarters may help provide unique insights into why individuals choose not to engage with an eMH program despite generally being willing to participate in an eMH evaluation study. Overestimating frequency of engagement with eMH tools may have theoretical implications and potentially impact economic considerations for plans to disseminate these tools to the general public.


Changes in Hechtian strands in cold-hardened cells measured by optical microsurgery.
Optical microsurgical techniques were employed to investigate the mechanical properties of Hechtian strands in tobacco (Nicotiana tabacum) and Ginkgo biloba callus cells. Using optical tweezers, a 1. 5-microm diameter microsphere coated with concanavalin A was inserted though an ablated hole in the cell wall of a plasmolyzed cell and attached to a Hechtian strand. By displacing the adhered microsphere from equilibrium using the optical trapping force, the tensions of individual strands were determined. Measurements were made using both normal and cold-hardened cells, and in both cases, tensions were on the order of 10(-12) N. Significant differences were found in the binding strengths of cold-hardened and normal cultured cells. An increased number density of strands in cold-hardened G. biloba compared with normal cultured cells was also observed. Although no Hechtian strands were detected in any Arabidopsis callus cells, strands were present in leaf epidermal cells. Finally, the movement of attached microspheres was monitored along the outside of a strand while cycling the osmotic pressure.


Hypoplastic left heart syndrome: from fetus to fontan.
The care of children with hypoplastic left heart syndrome is constantly evolving. Prenatal diagnosis of hypoplastic left heart syndrome will aid in counselling of parents, and selected fetuses may be candidates for in utero intervention. Following birth, palliation can be undertaken through staged operations: Norwood (or hybrid) in the 1st week of life, superior cavopulmonary connection at 4-6 months of life, and finally total cavopulmonary connection (Fontan) at 2-4 years of age. Children with hypoplastic left heart syndrome are at risk of circulatory failure their entire life, and selected patients may undergo heart transplantation. In this review article, we summarise recent advances in the critical care management of patients with hypoplastic left heart syndrome as were discussed in a focused session at the 12th International Conference of the Paediatric Cardiac Intensive Care Society held on 9 December, 2016, in Miami Beach, Florida.


Characterization of T and B antigen-binding cells for beta-galactosidase. III. Independence of antigen-binding cells in normal animals from antigenic stimulation.
Antigen-binding cells to beta-galactosidase were enumerated in thymus and spleen of mice of different ages and found to remain at a constant frequency throughout life. Thymic beta-galactosidase-binding cells (ZBCs),6 representing T-binding cells, showed no appreciable fluctuation, whereas splenic ZBCs, a mixture of T- and B-binding cells, were slightly depressed at birth but found at normal frequencies by 1 week of age. In addition, germfree mice, both within the 1st week after birth and after maturity, had nearly as many binding cells as did conventionally reared age-matched mice. These results considered together suggest that the ability of cells of both T and B origin to recognize antigen, as revealed by their ability to bind beta-galactosidase, arise independently of specific or nonspecific antigenic stimulation, as part of the normal ontogenic sequence of steps of differentiation. This is consistent with the theory of clonal precommitment of T and B cells.


Oviposition on and mining in bolls of Bt and non-Bt cotton by resistant and susceptible pink bollworm (Lepidoptera: Gelechiidae).
Transgenic cotton that produces insecticidal crystal protein Cry1Ac of Bacillus thuringiensis (Bt) has been effective in controlling pink bollworm, Pectinophora gossypiella (Saunders). We compared responses to bolls of Bt cotton and non-Bt cotton by adult females and neonates from susceptible and Cry1Ac-resistant strains of pink bollworm. In choice tests on caged cotton plants in the greenhouse, neither susceptible nor resistant females laid fewer eggs on Bt cotton bolls than on non-Bt cotton bolls, indicating that the Bt toxin did not deter oviposition. Multiple regression revealed that the number of eggs laid per boll was negatively associated with boll age and positively associated with boll diameter. Females also laid more eggs per boll on plants with more bolls. The distribution of eggs among bolls of Bt cotton and non-Bt cotton was clumped, indicating that boll quality rather than avoidance of previously laid eggs was a primary factor in oviposition preference. Parallel to the results from oviposition experiments, in laboratory no-choice tests with 10 neonates per boll, the number of entrance holes per boll did not differ between Bt cotton and non-Bt cotton for susceptible and resistant neonates. Also, like females, neonates preferred younger bolls and larger bolls. Thus, acceptance of bolls by females for oviposition and by neonates for mining was affected by boll age and diameter, but not by Bt toxin in bolls. The lack of discrimination between Bt and non-Bt cotton bolls by pink bollworm from susceptible and resistant strains indicates that oviposition and mining initiation are independent of susceptibility to Cry1Ac.


Accelerating joint relaxation-diffusion MRI by integrating time division multiplexing and simultaneous multi-slice (TDM-SMS) strategies.
To accelerate the acquisition of relaxation-diffusion imaging by integrating time-division multiplexing (TDM) with simultaneous multi-slice (SMS) for EPI and evaluate imaging quality and diffusion measures. The time-division multiplexing (TDM) technique and SMS method were integrated to achieve a high slice-acceleration (e.g., 6×) factor for acquiring relaxation-diffusion MRI. Two variants of the sequence, referred to as TDM3e-SMS and TDM2s-SMS, were developed to simultaneously acquire slice groups with three distinct TEs and two slice groups with the same TE, respectively. Both sequences were evaluated on a 3T scanner with in vivo human brains and compared with standard single-band (SB) -EPI and SMS-EPI using diffusion measures and tractography results. Experimental results showed that the TDM3e-SMS sequence with total slice acceleration of 6 (multiplexing factor (MP) = 3 × multi-band factor (MB) = 2) provided similar image intensity and microstructure measures compared to standard SMS-EPI with MB = 2, and yielded less bias in intensity compared to standard SMS-EPI with MB = 4. The three sequences showed a similar positive correlation between TE and mean kurtosis (MK) and a negative correlation between TE and mean diffusivity (MD) in white matter. Multi-fiber tractography also shows consistency of results in TE-dependent measures between different sequences. The TDM2s-SMS sequence (MP = 2, MB = 2) also provided imaging measures similar to standard SMS-EPI sequences (MB = 2) for single-TE diffusion imaging. The TDM-SMS sequence can provide additional 2× to 3× acceleration to SMS without degrading imaging quality. With the significant reduction in scan time, TDM-SMS makes joint relaxation-diffusion MRI a feasible technique in neuroimaging research to investigate new markers of brain disorders.


Determinants of nursing home costs in Florida: policy implications and support in national research findings.
Descriptive and econometric analysis of the major nonquality determinants of nursing home costs for Florida shows that mean costs, size, and occupancy rate increased between 1971 and 1976, that per diem costs and occupancy rate were inversely related, and that the per diem cost was lower in rural than in urban areas. Regression of the data shows that--next to inflation, as expressed by the Consumer Price Index--the occupancy rate accounts for most of the variation in per diem costs, followed by size, urban-rural location, and by type of control. The hypothetical "optimal," defined as lowest cost-size range, was calculated to be more than 350 beds. Recent research substantiates most of these findings. Medicaid Cost Reports from Florida's nursing homes were the source of the information analyzed; by 1976, the sixth year of the study, the data base covered nearly 9 of 10 licensed beds in the State. Some policy implications can be drawn from the analysis. Reductions in per diem costs could be achieved by higher occupancy rates, especially in the larger nursing homes, and a reduction in the rate of inflation would reduce the rate of increase in nursing home costs.


Prognostic or predictive value of circulating cytokines and angiogenic factors for initial treatment of multiple myeloma in the GIMEMA MM0305 randomized controlled trial.
Several new drugs are approved for treatment of patients with multiple myeloma (MM), but no validated biomarkers are available for the prediction of a clinical outcome. We aimed to establish whether pretreatment blood and bone marrow plasma concentrations of major cytokines and angiogenic factors (CAFs) of patients from a phase 3 trial of a MM treatment could have a prognostic and predictive value in terms of response to therapy and progression-free and overall survival and whether these patients could be stratified for their prognosis. Blood and bone marrow plasma levels of Ang-2, FGF-2, HGF, VEGF, PDGF-β, IL-8, TNF-α, TIMP-1, and TIMP-2 were determined at diagnosis in MM patients enrolled in the GIMEMA MM0305 randomized controlled trial by an enzyme-linked immunosorbent assay (ELISA). These levels were correlated both reciprocally and with the type of therapy and patients' characteristics and with a group of non-MM patients as controls. No significant differences were detected between the blood and bone marrow plasma levels of angiogenic cytokines. A cutoff for each CAF was established. The therapeutic response of patients with blood plasma levels of CAFs lower than the cutoff was better than the response of those with higher levels in terms of percentage of responding patients and quality of response. FGF-2, HGF, VEGF, and PDGF-β plasma levels at diagnosis have predictive significance for response to treatment. The stratification of patients based on the levels of CAFs at diagnosis and their variations after therapy is useful to characterize different risk groups concerning outcome and response to therapy. Clinical trial information can be found at the following link: NCT01063179.


Utility of the microcolony method for evaluation of multidrug-resistant tuberculosis patients in Karachi, Pakistan.
OBJECTIVE: To compare the microcolony method (MCM) with the reference culture method to evaluate culture conversion in multidrug-resistant tuberculosis (MDR-TB) patients.MATERIAL AND METHODS: Adult patients with Mycobacterium tuberculosis culture-positive MDR-TB undergoing second-line anti-tuberculosis treatment were recruited from two tertiary care chest clinics from January 2013 to October 2014. The MCM was performed in addition to MGIT™ and Löwenstein-Jensen medium (reference method) on sputum samples submitted on a monthly basis.RESULTS: Of 140 patients, culture conversion could be evaluated in 95 patients. The MCM showed 100% agreement with the reference M. tuberculosis culture in 83 of 95 patients who achieved culture conversion. In smear-positive and smear-negative cases, the mean time to positivity was 9.1 and 11.4 days for the MCM and 16.1 and 23.2 days for the reference M. tuberculosis culture respectively. The contamination rate for the MCM was 4.5% in comparison with 6.1% for the reference M. tuberculosis culture. The cost of MCM was estimated to be 30% that of the reference method.CONCLUSION: The MCM can be used in non-urban laboratories as a safe, rapid and cost-effective substitute for the reference M. tuberculosis culture to assess culture conversion in MDR-TB patients.Note: Abstract has been published in International Journal of Mycobacteriology 2015; 4: 159-160.


Liver transplantation for primary and metastatic liver cancers.
Liver transplantation for hepatic malignancies has emerged as a well-documented and proven treatment modality. However, early unsatisfactory results emphasized that only a highly selected patient population would benefit from transplantation. Currently, 15% of all liver transplants performed are for hepatocellular carcinoma (HCC). There is no controversy about the fact that liver transplantation for HCC in the adult population yields good results for patients whose tumour masses do not exceed the Milan criteria. It remains to be determined whether patients with more extensive tumours can be reliably selected to benefit from the procedure. In patients with small HCC at an early stage and preserved liver function, liver resection provides an alternative to transplant. Liver resection may offer similar survival results to orthotopic liver transplantation (OLT) in the short term, and does not carry the long-term effects of immunosuppression; however, long-term and disease-free survival favours liver transplantation. Very promising results have been obtained for cholangiocarcinoma treated by aggressive combination therapies, including chemo- and radiotherapy followed by OLT. Survival rate in these selected patients can approach that of patients with cholestatic liver disease, and the role of transplantation now requires re-evaluation. Similarly, hepatoblastoma is an excellent indication in paediatric patients with unresectable or recurrent tumours. Epithelioid hemangioendothelioma is also an appropriate indication for liver transplantation, even in the presence of extrahepatic metastases, unlike angiosarcoma which is associated with a very poor survival and considered as a contraindication. And finally for metastatic liver disease from neuroendocrine tumours, liver transplantation can result in long-term survival and even cure in well selected patients. Conversely, the value of transplantation for colorectal liver metastases (currently a contraindication) requires further evaluation by well-designed trials.


Leupaxin is a critical adaptor protein in the adhesion zone of the osteoclast.
Leupaxin is a cytoskeleton adaptor protein that was first identified in human macrophages and was found to share homology with the focal adhesion protein, paxillin. Leupaxin possesses several protein-binding domains that have been implicated in targeting proteins such as focal adhesion kinase (pp125FAK) to focal adhesions. Leupaxin can be detected in monocytes and osteoclasts, both cells of hematopoietic origin. We have identified leupaxin to be a component of the osteoclast podosomal signaling complex. We have found that leupaxin in murine osteoclasts is associated with both PYK2 and pp125FAK in the osteoclast. Treatment of osteoclasts with TNF-alpha and soluble osteopontin were found to stimulate tyrosine phosphorylation of both leupaxin and leupaxin-associated PYK2. Leupaxin was found to co-immunoprecipitate with the protein tyrosine phosphatase PTP-PEST. The cellular distribution of leupaxin, PYK2, and protein tyrosine phosphorylation-PEST co-localized at or near the osteoclast podosomal complex. Leupaxin was also found to associate with the ARF-GTPase-activating protein, paxillin kinase linker p95PKL, thereby providing a link to regulators of cytoskeletal dynamics in the osteoclast. Overexpression of leupaxin by transduction into osteoclasts evoked numerous cytoplasmic projections at the leading edge of the cell, resembling a motile phenotype. Finally, in vitro inhibition of leupaxin expression in the osteoclast led to a decrease in resorptive capacity. Our data suggest that leupaxin may be a critical nucleating component of the osteoclast podosomal signaling complex.


Intestinal infusion of endotoxin in adrenalectomized calves.
Four adrenalectomized calves, 3 to 4 months old, were allowed to develop adrenocorticosteroid insufficiency and then were given 50 mg of Escherichia coli endotoxin by infusion via cannula into the duodenum. Several physiologic variables were studied to detect changes indicative of endotoxemia. It is concluded that adrenocorticosteroid insufficiency in calves is not an independent factor allowing an intestinal source of endotoxin to cause the development of clinical signs and physiologic changes characteristic of endotoxemia.


Pharmacokinetics of mitomycin C in patients receiving the drug alone or in combination.
We have characterized plasma pharmacokinetics in 30 patients receiving mitomycin C (mitomycin) in doses ranging from 6 to 20 mg/m2 by iv bolus injection, either alone or in combination with other chemotherapeutic drugs. Plasma elimination of the drug was described by a two-compartment model in all but three cases, giving mean values for alpha-half-life of 8.2 mins, beta-half-life of 51.8 mins, and total body clearance of 332.9 ml/min/m2. The mean urinary excretion was 8.1% of the total dose administered. We did not detect alterations in pharmacologic parameters related to liver dysfunction, dose of mitomycin, and concomitant administration of other cytotoxic drugs.


Experiences from a prospective early rheumatoid arthritis study in southern Sweden.
Characteristics of an ongoing longitudinal observational study of patients with established rheumatoid arthritis (RA) are described. The sample comprised 183 patients enrolled from 1985 to 1989. Disease duration at inclusion was 1 year. Patients were referred from primary care and were included in the study irrespective of disease severity, with evaluation at a team care unit every 6 to 12 months. The assessment comprised a broad spectrum of standardized measures that covered the potentially most important disease outcomes. After 10 years the dropout rate was only 3%. Results to date from this study are summarized. Findings include disease course and remission rate, physical function, radiographic progression, hip involvement, mortality, work disability, and economic consequences.


Nutritional supplementation acutely increases albumin fractional synthetic rate in chronic hemodialysis patients.
Uremic malnutrition is associated with increased risk of hospitalization and death in chronic hemodialysis (CHD) patients. Most nutritional intervention studies in CHD patients traditionally have used concentrations of serum albumin as the primary outcome measure and showed slight or no significant improvements. A recent study showed that intradialytic parenteral nutrition (IDPN) improves whole-body protein synthesis in CHD patients. On the basis of this observation, it was hypothesized that the anabolic effects of IDPN are associated with increases in the fractional synthetic rate of albumin, a direct estimate of acute changes in hepatic albumin synthesis. Seven CHD patients were studied during two hemodialysis (HD) sessions, with and without IDPN, using primed-constant infusion of (13C) leucine 2 h before, during, and 2 h after HD. Plasma enrichments of (13C) leucine and (13C) ketoisocaproate were examined to determine the fractional synthetic rate of albumin. The results indicate that administration of IDPN significantly improves the fractional synthetic rate of albumin during HD (16.2 +/- 1.5%/d versus 12.8 +/- 1.7%/d; P < 0.05) in CHD patients in parallel with significant improvements in whole-body protein synthesis (5.05 +/- 1.3 mg/kg fat-free mass/min versus 3.22 +/- 0.3 mg/kg fat-free mass/min; P < 0.05). IDPN is protein anabolic in the acute setting in CHD patients, as evidenced by significant concomitant increases in the fractional synthetic rate of albumin and whole-body protein synthesis.


Use of Smartphones for Clinical and Medical Education.
Smartphone use in clinical settings and in medical education has been on the rise, benefiting both health care and health care providers. Studies have shown, however, that some health care facilities and providers are reluctant to switch to smartphones due to the threat of mixing personal apps with clinical care applications and the possibility that distraction created by smartphone use could lead to medication errors and errors linked to procedures, treatments, or tests. The purpose of this research was to examine the effects of smartphones in a clinical setting and for medical education, to determine their overall impact. The methodology for this qualitative study was a literature review, conducted over five electronic databases. The search was limited to articles published in English, between 2010 and 2016. Forty-one sources that focused on the implementation of and the barriers to use of smartphones in clinical and medical education environments were referenced. These studies revealed that smartphones have more positive than negative effects on the ability to enhance patient care and medical education. Smartphone use is clearly an effective and efficient method of enhancing patient care and medical education in the health care industry. Access to health care as well is enhanced by the use of this tool.


Development of an institutional review board preapproval process for Doctor of Nursing Practice students: process and outcome.
As Doctor of Nursing Practice (DNP) programs proliferate, effective collaboration with institutional review boards (IRBs) is important to protect human subjects. It is particularly important that faculty and students recognize which DNP students' projects should be considered as "human subjects research" or "quality improvement." The former require IRB review, whereas the latter may be eligible for expedited review or may be considered exempt. We report outcomes following implementation of a combination of didactic training, one-to-one consultation, and a decision support protocol to improve preparation for and collaboration with the IRB at a large university. In the first year of using this protocol, 53% of projects were deemed human subjects research and received IRB review. The other 47% were deemed quality improvement projects and did not require IRB review. We offer our experience as an approach for teaching students how to protect the subjects included in their quality improvement activities.


Direct three-dimensional characterization and multiscale visualization of wheat straw deconstruction by white rot fungus.
Microbial degradation of lignocellulose for resource and energy recovery has received increasing interest. Despite its obvious importance, the mechanism behind the biodegradation, especially the changes of morphological structure and surface characteristics, has not been fully understood. Here, we used three-dimensional (3D) characterization and multiscale visualization methods, in combination with chemical compositional analyses, to elucidate the degradation process of wheat straw by a white rot fungus, Phanerochaete chrysosporium. It was found that the fungal attack initiated from stomata. Lignin of the straw decayed in both size and quantity, and heterogeneity in the biodegradation was observed. After treatment with the fungus, the straw surface turned from hydrophobic to hydrophilic, and the adhesion of the straw surface increased in the fungal degradation. The morphology of the straw outer layer became heterogeneous and loose with the formation of many holes with various sizes. The wasp-tunnels-like structure of the collenchyma and parenchyma of the straw as well as the fungal hyphae interspersed inside the straw structure were clearly visualized in the 3D reconstruction structure. This work offers a new insight into the mechanism of lignocellulose biodegradation and demonstrates that multiscale visualization methods could be a useful tool to explore such complex processes.


Inducible Fibril Formation of Silk-Elastin Diblocks.
Silk-elastin block copolymers have such physical and biological properties that make them attractive biomaterials for applications ranging from tissue regeneration to drug delivery. Silk-elastin block copolymers that only assemble into fibrils at high concentrations can be used for a template-induced fibril assembly. This can be achieved by additionally including template-binding blocks that promote high local concentrations of polymers on the template, leading to a template-induced fibril assembly. We hypothesize that template-inducible silk-fibril formation, and hence high critical concentrations for fibril formation, requires careful tuning of the block lengths, to be close to a critical set of block lengths that separates fibril forming from nonfibril forming polymer architectures. Therefore, we explore herein the impact of tuning block lengths for silk-elastin diblock polypeptides on fibril formation. For silk-elastin diblocks ES m -SQ n , in which the elastin pentamer repeat is ES = GSGVP and the crystallizable silk octamer repeat is SQ = GAGAGAGQ, we find that no fibril formation occurs for n = 6 but that the n = 10 and 14 diblocks do show concentration-dependent fibril formation. For n = 14 diblocks, no effect is observed of the length m (with m = 40, 60, 80) of the amorphous block on the lengths of the fibrils. In contrast, for the n = 10 diblocks that are closest to the critical boundary for fibril formation, we find that long amorphous blocks (m = 80) oppose the growth of fibrils at low concentrations, making them suitable for engineering template-inducible fibril formation.


A transitional stage in the commitment of mesoderm to hematopoiesis requiring the transcription factor SCL/tal-1.
In this report, we describe the identification and characterization of an early embryoid body-derived colony, termed the transitional colony, which contains cell populations undergoing the commitment of mesoderm to the hematopoietic and endothelial lineages. Analysis of individual transitional colonies indicated that they express Brachyury as well as flk-1, SCL/tal-1, GATA-1, (beta)H1 and (beta)major reflecting the combination of mesodermal, hematopoietic and endothelial populations. This pattern differs from that found in the previously described hemangioblast-derived blast cell colonies in that they typically lacked Brachyury expression, consistent with their post-mesodermal stage of development (Kennedy, M., Firpo, M., Choi, K., Wall, C., Robertson, S., Kabrun, N. and Keller, G. (1997) Nature 386, 488-493). Replating studies demonstrated that transitional colonies contain low numbers of primitive erythroid precursors as well as a subset of precursors associated with early stage definitive hematopoiesis. Blast cell colonies contain higher numbers and a broader spectrum of definitive precursors than found in the transitional colonies. ES cells homozygous null for the SCL/tal-1 gene, a transcription factor known to be essential for development of the primitive and definitive hematopoietic systems, were not able to form blast colonies but did form transitional colonies. Together these findings suggest that the transitional colony represents a stage of development earlier than the blast cell colony and one that uniquely defines the requirement for a functional SCL/tal-1 gene for the progression to hematopoietic commitment.


X-ray spectroscopy characterization of self-assembled monolayers of nitrile-substituted oligo(phenylene ethynylene)s with variable chain length.
Self-assembled monolayers (SAMs) of nitrile-substituted oligo(phenylene ethynylene) thiols (NC-OPEn) with a variable chain length n (n ranging from one to three structural units) on Au(111) were studied by synchrotron-based high-resolution X-ray photoelectron spectroscopy and near-edge absorption fine-structure spectroscopy. The experimental data suggest that the NC-OPEn molecules form well-defined SAMs on Au(111), with all the molecules bound to the substrate through the gold-thiolate anchor and the nitrile tail groups located at the SAM-ambient interface. The packing density in these SAMs was found to be close to that of alkanethiolate monolayers on Au(111), independent of the chain length. Similar behavior was found for the molecular inclination, with an average tilt angle of ~33-36° for all the target systems. In contrast, the average twist of the OPEn backbone (planar conformation) was found to depend on the molecular length, being close to 45° for the films comprising the short OPE chains and ~53.5° for the long chains. Analysis of the data suggests that the attachment of the nitrile moiety, which served as a spectroscopic marker group, to the OPEn backbone did not significantly affect the molecular orientation in the SAMs.


Investigation of clinically acceptable agreement between two methods of automatic measurement of limb occlusion pressure: a randomised trial.
Development of automatic, pneumatic tourniquet technology and use of personalised tourniquet pressures has improved the safety and accuracy of surgical tourniquet systems. Personalisation of tourniquet pressure requires accurate measurement of limb occlusion pressure (LOP), which can be measured automatically through two different methods. The 'embedded LOP' method measures LOP using a dual-purpose tourniquet cuff acting as both patient sensor and pneumatic effector. The 'distal LOP' method measures LOP using a distal sensor applied to the patient's finger or toe of the operating limb, using photoplethysmography to detect volumetric changes in peripheral blood circulation. The distal LOP method has been used clinically for many years; the embedded LOP method was developed recently with several advantages over the distal LOP method. While both methods have clinically acceptable accuracy in comparison to LOP measured using the manual Doppler ultrasound method, these two automatic methods have not been directly compared. The purpose of this study is to investigate if the embedded and distal methods of LOP measurement have clinically acceptable agreement. The differences in pairs of LOP measurement in the upper and lower limbs of 81 healthy individuals were compared using modified Bland and Altman analysis. In surgery, it is common for cuff pressure to deviate from the pressure setpoint due to limb manipulation. Surgical tourniquet systems utilise a ± 15 mmHg pressure alarm window, whereby if the cuff pressure deviates from the pressure setpoint by > 15 mmHg, an audiovisual alarm is triggered. Therefore, if the difference (bias) ± SE, 95% CI of the bias and SD of differences ± SE in LOP measurement between the embedded and distal methods were all within ±15 mmHg, this would demonstrate that the two methods have clinically acceptable agreement. LOP measurement using the embedded LOP method was - 0.81 ± 0.75 mmHg (bias ± standard error) lower than the distal LOP method. The 95% confidence interval of the bias was - 2.29 to 0.66 mmHg. The standard deviation of the differences ± standard error was 10.35 ± 0.49 mmHg. These results show that the embedded and distal methods of LOP measurement demonstrate clinically acceptable agreement. The findings of this study demonstrate clinically acceptable agreement between the embedded and distal methods of LOP measurement. The findings support the use of the embedded LOP method of automatic LOP measurement using dual-purpose tourniquet cuffs to enable accurate, effective and simple prescription of personalised tourniquet cuff pressures in a clinical setting.


Cosmetic mastoidectomy for the combined supra/infratentorial transtemporal approach. Technical note.
The authors describe a cosmetic mastoidectomy technique for use when performing a combined supra/infratentorial craniotomy and transtemporal exposure. The technique involves a single temporal suboccipital bone flap and cosmetic mastoidectomy, removing the outer table of bone for later replacement. Replacement of the outer table of mastoid bone enables tamponade of a fat graft against the dura to reduce the risk of postoperative cerebrospinal fluid leaks. The technique has been performed in eight patients treated for petroclival meningiomas with excellent cosmetic results.


Symposium on microbiology update: old friends and new enemies. Bacillus cereus.
Bacillus cereus is an environmentally ubiquitous, Gram-positive, spore-forming bacillus responsible for 2 distinct foodborne disease syndromes as well as other manifestations of pathogenicity. The rapid-onset, "emetic," foodborne-disease syndrome is associated with an emetic toxin; the delayed-onset, "diarrheal" syndrome is associated with elaboration of enterotoxin. The majority of methods for detection of these toxins have relied on in vivo testing. More recent work on purification of enterotoxin facilitated the development of a rapid, specific, fluorescent immunodot assay and a tissue culture screening assay for enterotoxin. Work on characterization and detection of emetic toxin is ongoing.


Nuclear factor κB-dependent regulation of angiogenesis, and metastasis in an in vivo model of thyroid cancer is associated with secreted interleukin-8.
Development of novel strategies in the treatment of advanced thyroid cancer are needed. Our laboratory has previously identified a role for nuclear factor κB (NF-κB) signaling in human thyroid cancer cell growth, survival, and invasion. Our goal was to establish the role of NF-κB signaling on thyroid cancer growth and metastases in vivo and to begin to dissect mechanisms regulating this effect. We examined tumor formation of five thyroid cancer cell lines in an in vivo model of thyroid cancer and observed tumor establishment in two of the cell lines (8505C and BCPAP). Inhibition of NF-κB signaling by overexpression of a dominant-negative IκBα (mIκBα) significantly inhibited thyroid tumor growth in tumors derived from both cell lines. Further studies in an experimental metastasis model demonstrated that NF-κB inhibition impaired growth of tumor metastasis and prolonged mouse survival. Proliferation (mitotic index) was decreased in 8505C tumors, but not in BCPAP tumors, while in vitro angiogenesis and in vivo tumor vascularity were significantly inhibited by mIkBα only in the BCPAP cells. Cytokine antibody array analysis demonstrated that IL-8 secretion was blocked by mIκBα expression. Interestingly, basal NF-κB activity and IL-8 levels were significantly higher in the two tumorigenic cell lines compared with the nontumorigenic lines. Furthermore, IL-8 transcript levels were elevated in high-risk human tumors, suggesting that NF-κB and IL-8 are associated with more aggressive tumor behavior. These studies suggest that NF-κB signaling is a key regulator of angiogenesis and growth of primary and metastatic thyroid cancer, and that IL-8 may be an important downstream mediator of NF-κB signaling in advanced thyroid cancer growth and progression.


Preclinical evaluation of Som230 as a radiation mitigator in a mouse model: postexposure time window and mechanisms of action.
The somatostatin analog SOM230 has potent radioprophylactic and radiation mitigating properties that are unrelated to cytoprotection but appear to be due to suppression of secretion of pancreatic enzymes into the intestinal lumen. To determine the maximal postirradiation time window for administration, male CD2F1 mice were exposed to 8.5-11 Gy total-body radiation; SOM230 (0.5, 2 or 5 mg/kg) or vehicle was given by twice daily subcutaneous injections for 14 days, beginning 24-72 h after irradiation, and 30-day animal survival was recorded. The contribution of the gut to systemic cytokine levels was estimated by analyzing plasma samples obtained simultaneously from the portal vein and carotid artery. The effect of SOM230 on cell trypsin secretion was assessed in vitro and intestinal proteolytic activity was measured in vivo. SOM230 was associated with a 40-60% absolute improvement in overall postirradiation survival when treatment was started 48 h after irradiation and even exhibited a statistically significant survival benefit when started at 72 h. SOM230 ameliorated the radiation-induced decrease in chemokine (C-X-C motif) ligand 9 (CXCL9). SOM230 inhibited pancreatic acinar cell trypsin secretion in vitro in a dose-dependent fashion and reduced intraluminal and intestinal tissue proteolytic activity in vivo. SOM230 is an excellent radiation mitigator with a postirradiation time window in excess of 48 h. The mechanism likely involves preservation of intestinal barrier function due to decreased secretion of pancreatic enzymes into the bowel lumen.


Autoimmune hepatitides--update 2004.
This update reports is primarily made to update papers published in 2002 and 2003. A selection had to be realized since a search of the literature yielded more than 1600 journal articles. The prevailing opinion of an international group is especially emphasized. Autoimmune hepatitides (AIH)--as the name already suggests it--are no disease entity. We prefer to classify them under the terms of typical and atypical autoimmune hepatitides. Typical autoimmune hepatitis can be subdivided into a type 1 (with antibodies against nuclear material and/or smooth muscle) and a type 2 (with antibodies against endoplasmic reticulum). In the case of atypical autoimmune hepatitides, the detected autoantibodies would have to be stated. The essential feature of an autoimmune hepatitis is the presence of various high-titer autoantibodies circulating in serum, especially such of the IgG class. Special mention is made of the very rare liver ribosomal antibodies (LRA), which are mainly directed against 80S liver ribosomes and point to a so-called virus-associated autoimmune hepatitis. Also discussed are a genetic disposition, which is still speculative in some points, and the existence of so-called autogenes (physiological, pathological/pathogenic and so-called regulator genes). Various noxae, e.g. viruses, might also be "autogenous". Further research in the field of the autoimmune hepatitides (and also autoimmunity and autoimmune diseases as such) is therefore likely to concentrate on viruses and genes and their interrelations.


Hypothalamic dysfunction of the thrombospondin receptor α2δ-1 underlies the overeating and obesity triggered by brain-derived neurotrophic factor deficiency.
Brain-derived neurotrophic factor (BDNF) and its receptor, TrkB, are critical components of the neural circuitry controlling appetite and body weight. Diminished BDNF signaling in mice results in severe hyperphagia and obesity. In humans, BDNF haploinsufficiency and the functional Bdnf Val66Met polymorphism have been linked to elevated food intake and body weight. The mechanisms underlying this dysfunction are poorly defined. We demonstrate a chief role of α2δ-1, a calcium channel subunit and thrombospondin receptor, in triggering overeating in mice with central BDNF depletion. We show reduced α2δ-1 cell-surface expression in the BDNF mutant ventromedial hypothalamus (VMH), an energy balance-regulating center. This deficit contributes to the hyperphagia exhibited by BDNF mutant mice because selective inhibition of α2δ-1 by gabapentin infusion into wild-type VMH significantly increases feeding and body weight gain. Importantly, viral-mediated α2δ-1 rescue in BDNF mutant VMH significantly mitigates their hyperphagia, obesity, and liver steatosis and normalizes deficits in glucose homeostasis. Whole-cell recordings in BDNF mutant VMH neurons revealed normal calcium currents but reduced frequency of EPSCs. These results suggest calcium channel-independent effects of α2δ-1 on feeding and implicate α2δ-1-thrombospondin interactions known to facilitate excitatory synapse assembly. Our findings identify a central mechanism mediating the inhibitory effects of BDNF on feeding. They also demonstrate a novel and critical role for α2δ-1 in appetite control and suggest a mechanism underlying weight gain in humans treated with gabapentinoid drugs.


Neural factors in the development of renal function: effect of neonatal central catecholaminergic lesions with 6-hydroxydopamine.
Peripheral sympathetic neurons are thought to provide trophic regulatory signals for development of their target tissues. In the current study, we investigated the role of sympathetic tone in the functional development of the kidney in rats, using neonatal intracisternal administration of 6-hydroxydopamine (6-OHDA). This treatment destroys central catecholaminergic pathways and permanently reduces sympathetic activity without ablating peripheral nerve terminals. Renal function was evaluated over the first two postnatal weeks, when glomerular and tubular function undergo rapid development. Although basal renal clearance and the homeostatic response to fluid deprivation developed normally in the lesioned rats, the response to a maximally-effective dose of desmopressin acetate (DDAVP), a vasopressin analog, became deficient by the end of the second week. After weaning, the lesioned animals were unable to survive a chronic salt load, which requires sustained water reabsorption but high output of sodium. These data indicate that normal sympathetic tone is required for appropriate development of the responsiveness of the renal tubule to vasopressin.


Short-term efficacy and tolerability of venlafaxine extended release in adults with generalized anxiety disorder without depression: A meta-analysis.
Although efficacy of venlafaxine extended release (XR) for generalized anxiety disorder (GAD) has been reported in previous analyses in 2002 and 2004, the sample size was rather small and estimate of safety or tolerability was not clear. The present analysis had the advantage of large sample size and provided evidence for tolerability. Literature databases were searched, including Pubmed, Embase, Cochrane Central Register of Controlled Trials, Web of science and clinical trials. 10 eligible articles were finally selected and data was extracted and logged into the Review Manager 5.3 by two independent authors. The risk of bias was evaluated by the Cochrane Collaboration's Risk of Bias Tool and the stability of the results was assessed by sensitivity analysis. The publication bias was assessed by funnel plot and Egger's/Begg's test using Stata Version 12.0 software. In the current meta-analysis, 10 articles (14 studies) satisfying the inclusion criteria were analyzed. As efficacy outcomes, our findings indicated venlafaxine XR was significantly more effective than placebo according to mean change of the Hamilton Rating Scale for Anxiety total scores [mean difference = 3.31, 95% confidence interval(CI) 1.44-5.18, P = 0.0005], response [odds ratio(OR) = 1.83, 95%CI 1.58-2.12, P<0.00001], and remission (OR = 2.55, 95%CI 1.36-4.78, P = 0.003). In terms of tolerability, the most frequently reported treatment-emergent adverse events were nausea, dry mouth, dizziness, insomnia, somnolence, and headache. In addition, discontinuation due to all-cause (OR = 1.17, 95%CI 0.92-1.49, P = 0.19) was not significantly different between the two groups, whereas discontinuation due to adverse events was statistically higher in the venlafaxine XR group compared with the placebo treatment (OR = 2.80, 95%CI 2.21-3.54, P<0.00001) and discontinuation due to inefficacy was lower in venlafaxine than placebo treatment (OR = 0.26, 95%CI 0.17-0.40, P<0.00001). There was no significant publication bias and sensitivity analysis showed that our analysis exhibited high stability. We concluded that venlafaxine XR (75-225 mg/day) is an effective and well-tolerated pharmacological treatment option for adult patients with GAD.


Decreased expression of synaptic vesicle protein 2A, the binding site for levetiracetam, during epileptogenesis and chronic epilepsy.
We previously showed that gene expression of synaptic vesicle protein 2A (SV2A), the binding site for the antiepileptic drug levetiracetam, is reduced during epileptogenesis in the rat. Since absence of SV2A has been associated with increased epileptogenicity, changes in expression of SV2A could have consequences for the progression of epilepsy. Therefore we investigated hippocampal SV2A protein expression of temporal lobe epilepsy (TLE) patients and in rats during epileptogenesis and in the chronic epileptic phase. SV2A immunocytochemistry and Western blot analysis were performed on the hippocampus of autopsy controls, patients that died from status epilepticus (SE), and pharmacoresistant TLE patients. In addition, in epileptic rats, SV2A expression was determined after SE during the acute, latent, and chronic epileptic phase. In control tissue, presynaptic SV2A was expressed in all hippocampal subfields, with strongest expression in mossy fiber terminals. SV2A positive puncta were distributed in a patchy pattern over the somata and dendrites of neurons. SV2A decreased throughout the hippocampus of TLE patients with hippocampal sclerosis (HS), compared to autopsy control, SE, and non-HS tissue. In most rats, SV2A was already decreased in the latent period especially in the inner molecular layer and stratum lucidum. Similarly as in humans, SV2A was also decreased throughout the hippocampus of chronic epileptic rats, specifically in rats with a progressive form of epilepsy. These data support previous findings that reduced expression of SV2A could contribute to the increased epileptogenicity. Whether this affects the effectiveness of levetiracetam needs to be further investigated.


Organosulphur and related ligands in Suzuki-Miyaura C-C coupling.
Suzuki-Miyaura C-C cross coupling (SMC), an important synthetic strategy for many organic molecules, has several advantages such as mild reaction conditions, high tolerance toward various functional groups and ease in isolation of the product. Palladium(II) ligated with phosphines (particularly bulky and electron-rich) and N-heterocyclic carbenes (NHCs) has been found to be efficient in the catalysis of SMC. The drawback with many of these catalysts is their air/moisture sensitivity. Since 2000, palladium(II) complexes of organosulphur and related ligands have emerged as viable alternatives to palladium-phosphine/carbene complexes as they have sufficient thermal stability, air and moisture insensitivity. Moreover synthesis of complexes of such ligands is easy. In this perspective Suzuki-Miyaura C-C coupling reactions catalyzed with palladium(II)-complexes of organosulphur ligands have been reviewed. Catalysis of SMC with palladium(II) complexes of organoselenium and tellurium ligands, studied much less in comparison to those of organosulphur ligands, is also included.


[Assessment of injuries to the neck and dentomandibular system complicated by the infectious process].
The objective of the present work was to consider problems pertaining to forensic medical examination of injuries to the neck and dentition system complicated by pyoinflammatory processes. Two clinical observations are reported to illustrate significant difficulties encountered by forensic medical experts during assessment of the severity of harm inflicted to human health in the cases requiring differential characteristic of injuries, evaluation of opportuneness and completeness of the provided medical aid, and the role of these factors in the development of fatal pyoseptic complications. Special emphasis is laid on the analysis of possibilities to attain these goals based on the "Medical criteria for the assessment of severity of harm inflicted to human health" (2008).


Effectiveness of a web-based solution-focused brief chat treatment for depressed adolescents and young adults: randomized controlled trial.
Up to 9% of young people suffer from depression. Unfortunately, many in need of help remain untreated. The Internet offers anonymous ways to help depressed youth, especially those who are reluctant to search for help because of fear of stigma. Our goal was to evaluate the effectiveness of an individual chat treatment based on Solution-Focused Brief Therapy (SFBT) to young individuals aged 12-22 years with depressive symptoms by comparing it to a waiting list control group. For this study, 263 young people with depressive symptoms were randomized to the Web-based SFBT intervention, PratenOnline, or to a waiting list control condition. The chat treatment was delivered by trained professionals. Groups were compared on depressive complaints as measured by the Center for Epidemiologic Studies Depression Scale (CES-D) after 9 weeks and 4.5 months. For the chat group only, changes in depressive symptoms at 7.5 months after baseline were explored. The experimental SFBT condition (n=131) showed significantly greater improvement than the waiting list condition (n=132) in depressive symptoms at 9 weeks and 4.5 months on the CES-D, with a small between group effect size at 9 weeks (d=0.18, 95% CI -0.10 to 0.47) and a large effect size at 4.5 months (d=0.79, 95% CI 0.45-1.08). The percentage of participants showing a reliable and clinically significant change in depression was significantly larger for the SFBT intervention at 4.5 months only (28.2% vs 11.4% for the waiting list, P<.001, number needed to treat=6). At 7.5 months, the SFBT group showed further improvements. However, results have to be considered carefully because of high attrition rates. The Web-based SFBT chat intervention of PratenOnline was more effective than a waiting list control group in reducing depressive symptoms, and effects were larger at follow-up then at post-treatment. More studies are needed to find out if outcomes will be replicated, especially for those younger than 18 year old. Netherlands Trial Register: NTR 1696; http://www.trialregister.nl/trialreg/admin/rctview.asp?TC=1696 (Archived by WebCite at http://www.webcitation.org/6DspeYWrJ).


Effect of Cucumis sativus on Dysfunctional 3T3-L1 Adipocytes.
Obesity is caused by lipid accumulation in adipose tissues inducing adipocyte dysfunction, characterized by insulin resistance, increased lipolysis, oxidative stress, and inflammation, leading to increased levels of adipokines. Herein the capacity of the subfractions (SFs) SF1, SF2, and SF3 from the Cucumis sativus aqueous fraction and their combinations (M) to control adipocyte dysfunction in vitro, in 3T3-L1 adipocytes was studied. Adipocytes, previously treated with dexamethasone or IL-1 to induce dysfunction, were incubated with different concentrations of the subfractions for 24 h. 2-deoxyglucose consumption and glycerol release were evaluated, and a surface model was constructed to determine the most effective SF concentrations to improve both parameters. Effective SF combinations were assessed in their capacity to control metabolic, pro-oxidative, and pro-inflammatory conditions. SF1, SF2 (40 μg/ml each) and SF3 (20 μg/ml) improved 2-deoxyglucose consumption by 87%, 57%, and 87%, respectively. SF1 and SF2 (5 μg/ml each) and SF3 (40 μg/mL) increased glycerol secretion by 10.6%, 18.9%, and 11.8%, respectively. Among five combinations tested, only M4 (SF1 40 μg/ml:SF2 60 μg/ml:SF3 30 μg/ml) and M5 (SF1 40 μg/ml:SF2 60 μg/mL:SF3 10 μg/ml) controlled effectively the metabolic, pro-oxidative, and proinflammatory conditions studied. Glycine, asparagine, and arginine were the main components in these SFs.


Practice paper of the Academy of Nutrition and Dietetics abstract: the role of nutrition in health promotion and chronic disease prevention.
Food intake, lifestyle behaviors, and obesity are linked to the development of chronic diseases such as type 2 diabetes, certain cancers, and cardiovascular diseases. It is recognized that physical and social environment influences individuals' behaviors, and some population subgroups such as racial/ethnic minorities and individuals with low socioeconomic status or limited literacy or language abilities seem to be especially vulnerable to disparities in disease risk factors, disease prevalence, or health outcomes. Certain life cycle phases appear to be especially important for health promotion and disease prevention as the development of chronic diseases can take several decades. Such complex health issues often require system-wide, multifactorial, and multidisciplinary solutions. Social ecological models, with approaches spanning from individual level to macro policy level, can provide registered dietitians (RDs) and dietetic technicians, registered (DTRs) with a comprehensive framework to promote health and to prevent chronic diseases. Furthermore, the Nutrition Care Process can be utilized in carrying out the health promotion and disease prevention efforts. RDs and DTRs have the training and requisite skills to be leaders and active members of multidisciplinary teams to promote health and prevent chronic diseases across the life span. The position of the Academy of Nutrition and Dietetics states that primary prevention is the most effective, affordable method to prevent chronic disease, and that dietary intervention positively impacts health outcomes across the life span. RDs and DTRs are critical members of health care teams and are essential to delivering nutrition-focused preventive services in clinical and community settings, advocating for policy and programmatic initiatives, and leading research in disease prevention and health promotion. In concordance with the Academy's position, this practice paper provides an overview of practice examples, effective program components, and a comprehensive range of health promotion and chronic disease prevention strategies for RDs and DTRs. This paper supports the "Position of the Academy of Nutrition and Dietetics: The Role of Nutrition in Health Promotion and Chronic Disease Prevention" published in the July 2013 Journal of the Academy of Nutrition and Dietetics.


Baseline integrase drug resistance mutations and conserved regions across HIV-1 clades in Cameroon: implications for transition to dolutegravir in resource-limited settings.
Transition to dolutegravir-based regimens in resource-limited settings (RLS) requires prior understanding of HIV-1 integrase variants and conserved regions. Therefore, we evaluated integrase drug resistance mutations (DRMs) and conserved regions amongst integrase strand transfer inhibitor (INSTI)-naive patients harbouring diverse HIV-1 clades in Cameroon. A cross-sectional study was conducted amongst 918 INSTI-naive patients from Cameroon (89 ART-naive and 829 ART-experienced patients). HIV-1 sequences were interpreted regarding INSTI-DRMs using the Stanford HIVdb v8.9-1 and the 2019 IAS-USA list. Amino acid positions with <1% variability were considered as highly conserved. Subtyping was performed by phylogeny. Overall prevalence (95% CI) of INSTI-DRMs was 0.8% (0.4-1.7), with 0.0% (0.0-4.0) amongst ART-naive versus 0.9% (0.5-1.9) amongst ART-experienced patients; P = 0.44. Accessory mutations (95% CI) were found in 33.8% (30.9-37.0), with 38.2% (28.1-49.1) amongst ART-naive versus 33.4% (30.4-36.7) amongst ART-experienced patients; P = 0.21. Of 288 HIV-1 integrase amino acid positions, 58.3% were highly conserved across subtypes in the following major regions: V75-G82, E85-P90, H114-G118, K127-W132, E138-G149, Q168-L172, T174-V180, W235-A239 and L241-D253. Wide genetic diversity was found (37 clades), including groups M (92.3%), N (1.4%), O (6.2%) and P (0.1%). Amongst group M, CRF02_AG was predominant (47.4%), with a significantly higher frequency (95% CI) of accessory mutations compared with non-AG [41.4% (36.8-46.0) versus 27.1% (23.3-31.2) respectively; P < 0.001]. The low baseline of INSTI-DRMs (<1%) in Cameroon suggests effectiveness of dolutegravir-based regimens. In spite of high conservation across clades, the variability of accessory mutations between major circulating strains underscores the need for monitoring the selection of INSTI-DRMs while scaling up dolutegravir-based regimens in RLS.


We may not have it all together, but together we have it all. The importance of intra- and interdisciplinary collaboration.
The tremendous development of knowledge, the accumulation of clinical experience, and the enriching of everyday practice with the results of basic and clinical research created the need for splitting medicine in specialities and sub-specialities. Although this evolution is inevitable in order to meet the complex needs and demands of patients, we have to be aware of the risk of neglecting the holistic approach of a person in distress. Indeed, the organ-focused and pathology-centred approach of a patient may not exempt us of the rudiments of care such as the notion that a human being is more than the sum of its parts. This involves the absolute obligation for intra and inter-disciplinary cooperation. Recently, the European leading societies in Gynaecology, the European Board Obstetricians and Gynaecologists (EBCOG), the European Society of Human reproduction and Embryology (ESHRE), the European Society for Gynaecological Endoscopy (ESGE) and the European Academy for Gynaecological Surgery (+he Academy), have established some common impressive initiatives to improve women's health care. As a result of an intensive research programme coordinated by +he Academy with a multidisciplinary team of gynaecologists and surgeons a unique structured training and certification programme in endoscopic surgery is established. This project is a fine example making the dream of multidisciplinary approach reality and achieving some most impressive results. It is without any doubt a major step forwards in the improvement of the quality -assurance in endoscopic surgery in benefit of the patients care and has been accepted by our leading European and national societies. Recently, ESGE and ESHRE developed a new classification on female genital anomalies. An evidence-based -updated new classification published in the official journal of both societies was the ended result. THIS CREATIVE, SUCCESSFUL AND UNEVENTFUL COLLABORATION UNDERLINES THE NEED FOR RESPECTING SOME CRUCIAL POINTS: to clearly outline the topic of common action, to find the working platform, to define the method and to respect -independence of each society by clarifying the borders and the responsibilities. Keeping the window of the collaboration open is crucial for keeping the window to medicine of the future unlocked.


Mathematical models for HIV transmission dynamics: tools for social and behavioral science research.
HIV researchers have long appreciated the need to understand the social and behavioral determinants of HIV-related risk behavior, but the cumulative impact of individual behaviors on population-level HIV outcomes can be subtle and counterintuitive, and the methods for studying this are rarely part of a traditional social science or epidemiology training program. Mathematical models provide a way to examine the potential effects of the proximate biologic and behavioral determinants of HIV transmission dynamics, alone and in combination. The purpose of this article is to show how mathematical modeling studies have contributed to our understanding of the dynamics and disparities in the global spread of HIV. Our aims are to demonstrate the value that these analytic tools have for social and behavioral sciences in HIV prevention research, to identify gaps in the current literature, and to suggest directions for future research.


Correlates of Meeting the Physical Activity, Sedentary Behavior, and Sleep Guidelines for the Early Years among Belgian Preschool Children: The ToyBox-Study.
Physical activity, sedentary behavior, and sleep guidelines for preschool children were already established and integrated into the 24 h movement behavior guidelines in 2017. The aim of the current study was to investigate correlates of meeting or not meeting the physical activity, sedentary behavior, and sleep guidelines in Belgian preschool children. In total, 595 preschool children (53.3% boys, 46.7% girls, mean age: 4.2 years) provided complete data for the three behaviors and potentially associated correlates. Physical activity was objectively measured with accelerometers. Screen time, sleep duration, and correlates were reported by parents with the use of a questionnaire. Backward logistic regression was used to identify factors associated with meeting all guidelines for weekdays and weekend days. In the final model, older preschoolers (OR = 1.89), having a normal weight compared to being underweight (OR = 0.30), having parents that do not watch a lot of television (OR = 0.99), and having a father that attained higher education (OR = 1.91) were associated with meeting all guidelines on weekdays. For weekend days, a significant association was found for attending a sports club (OR = 1.08). Overall, only a few factors were associated with meeting the guidelines. A more comprehensive measurement of preschool children's potential correlates of physical activity, sedentary behavior, and sleep is warranted.


Di-(2-ethylhexyl) phthalate (DEHP) inhibits steroidogenesis and induces mitochondria-ROS mediated apoptosis in rat ovarian granulosa cells.
Increased oxidative stress (OS) due to ubiquitous exposure to di-(2-ethylhexyl) phthalate (DEHP) can affect the quality of oocytes by inducing apoptosis and hampering granulosa cell mediated steroidogenesis. This study was carried out to investigate whether DEHP induced OS affects steroidogenesis and induces apoptosis in rat ovarian granulosa cells. OS was induced by exposing granulosa cells to various concentrations of DEHP (0.0, 100, 200, 400 and 800 μM) for 72 h in vitro. Intracellular reactive oxygen species (ROS), oxidative stress (OS), mitochondrial membrane potential, cellular senescence, apoptosis, steroid hormones (estradiol & progesterone) and gene expression were analyzed. The results showed that an effective dose of DEHP (400 μg) significantly increased OS by elevating the ROS level, mitochondrial membrane potential, and β-galactosidase activity with higher mRNA expression levels of apoptotic genes (Bax, cytochrome-c and caspase3) and a lower level of an anti-apoptotic gene (Bcl2) as compared to the control. Further, DEHP significantly (P > 0.05) decreased the level of steroid hormones (estradiol and progesterone) in a conditioned medium and this effect was reciprocated with a lower expression (P > 0.05) of steroidogenic responsive genes (Cyp11a1, Cyp19A1, Star, ERβ1) in treated granulosa cells. Furthermore, co-treatment with N-Acetyl-Cysteine (NAC) rescues the effects of DEHP on OS, ROS, β-galactosidase levels and gene expression activities. Altogether, these results suggest that DEHP induces oxidative stress via ROS generation and inhibits steroid synthesis via modulating steroidogenic responsive genes, which leads to the induction of apoptosis through the activation of Bax/Bcl-2-cytochrome-c and the caspase-3-mediated mitochondrial apoptotic pathway in rat granulosa cells.


Evidence for a critical role of progesterone in the regulation of the midcycle gonadotropin surge and ovulation.
Serum concentrations of progesterone begin to rise just before the midcycle gonadotropin surge that leads to ovulation. To examine the role of progesterone in the regulation of these events, we evaluated the effects of a low dose (1 mg/day, orally) of the antiprogesterone RU 486 on the timing of the gonadotropin surge and ovulation in normally cycling women. The drug or a placebo was given for 5 or 15 days, starting when the dominant follicle reached 14-16 mm. RU 486 consistently delayed the timing of the midcycle gonadotropin surge and the subsequent collapse of the dominant follicle, despite rising estradiol concentrations and normal follicular development. Unexpectedly, RU 486 also delayed the emergence of the periovulatory progesterone rise. The addition of progesterone (5-10 mg/day, im, for 2 days) to a 5-day course of RU 486 after the emergence of a mature follicle readily induced LH and FSH surges and completely reversed the effects of RU 486 at midcycle. Our results suggest that RU 486 delays the midcycle gonadotropin surge and ovulation by suppressing or antagonizing an ovarian progestational signal. Progesterone may, thus, represent the ultimate ovarian signal to the estrogen-primed hypothalamic-pituitary unit to trigger the gonadotropin surge that leads to ovulation.


Perioperative Complications Curriculum for the OB/GYN Resident: A Pilot Study.
This article was migrated. The article was marked as recommended. Objectives: Our objective is to describe a process for development and implementation of a web-based curriculum addressing gynecologic perioperative complications. Methods: Residents, Fellows, and Faculty completed a needs assessment survey addressing satisfaction with baseline perioperative complications curriculum. Residents completed a knowledge pretest. Residents received weekly emails with links to developed topic-specific materials over 4 weeks. Residents completed a post-implementation survey to assess satisfaction and to retest knowledge. Results: 75% (21/28) of Residents and 47% (40/86) Fellows/Faculty completed the needs assessment. 95% (20/21) of Residents and 90% (36/40) Fellows/Faculty reported dissatisfaction with baseline perioperative complications curriculum. Ureteral injury, bowel injury, vaginal cuff dehiscence, and bladder injury were prioritized topics, while assigned readings, interactive web-based case modules, and intraoperative instruction were prioritized modalities for curriculum development. Resident pretest mean score was 72% (40-90%, SD = 15). Weekly curriculum emails were successfully distributed all Resident learners. Eighteen percent of Residents completed the post-implementation survey, with 100% reporting satisfaction with the web-based case modules. Post-test knowledge mean score was 84% (60-100%, SD = 15). Conclusion: Low satisfaction rate with baseline curriculum was reported. Materials were developed and distributed weekly to all Residents, who successfully and consistently accessed the developed web-based curriculum.