Acute cytomegalovirus glomerulonephritis: an experimental model.
HA/ICR mice injected intraperitoneally with an LD(10) of murine cytomegalovirus develop a transiet acute glomerulonephritis characterized by cytomegalic intranuclear inclusions limited to mesangial cells. Viruria and proteinuria occur on the 3rd day and usually subside by the 10th day, accompanied by resolution of the glomerular lesion. By electron microscopy, virus synthesized within mesangial cell nuclei accumulates in intercelluar channels that course through the mesangial matrix. Virus and cellular debris are also found in channels surrounded by matrix in the juxtaglomerular area and between the epithelial cells of the macula densa. By immunofluorescence, viral antigen first concentrated in glomerular mesangial cells on the 5th day is cleared centripetally, appearing as fine granules in the juxtaglomerular area by the 7th day after infection. By the 10th day, changes are restricted to the juxtaglomerular region, and thereafter hyalinization at the glomerular vascular pole or cellular disarray of the juxtaglomerular zone with lymphocytic infiltration extending into the renal interstitium from few glomeruli occurs in approximately 40 per cent of the mice associated with persistence of viral antigen in these foci. These findings indicate that at least one route of transport resulting in viruria involves the glomerular mesangium, juxtaglomerular zone, and the distal convoluted tubule. Persistence of viral antigen or other noxious substances along this route may result in a unique juxtaglomerular lesion and interstital nephritis.


A longitudinal analysis of the fluctuation of food stores in Belo Horizonte, Minas Gerais, Brazil.
Changes in food environments have the potential to affect consumption, nutritional status, and health, and understanding these changes is of utmost importance. This study, therefore, aimed to examine the fluctuation of food stores that sell fruits and vegetables over five years in the health promotion service area of Primary Health Care (PHC) in Belo Horizonte, Minas Gerais, Brazil. This was an ecological study that used data from a food environment audit conducted in the realm of Brazilian PHC. Buffers of 1 mile (equivalent to 1600 m) were created around health promotion services to define food environments. All food stores and open-air food markets that sold fruits and vegetables (FV) within this buffer area were considered eligible. The data collection was performed during two periods: the baseline, in 2013, and after five years, in 2018. This study compares the fluctuation by the type of stores and according to the health vulnerability index (HVI). After 5 years, 35.2% of the stores were stable; 154 stores were closed, and 155 were opened. The stability was greater in low-vulnerability areas, and the fluctuation differed by type of store only for areas with high vulnerability. The number of supermarket decreased in high HVI territories; and local stores, showed greater stability when compared to specialized FV markets. The differences in store fluctuations according to the vulnerability of areas demonstrate the importance of food supply policies considering the local characteristics to reduce inequities of access to healthy foods.


Nutrition knowledge in urban Blacks.
A simple questionnaire has been presented to Blacks at Baragwanath Hospital and in Soweto to test their current knowledge of basic nutrition and some related attitudes. The survey of 400 persons included nurses, schoolchildren, and a group of others whose standard of general education was lower. The vast majority had received nutrition instruction, were aware of basic requirements, and, given a free choice, could select reasonable diets. Fatness is no longer considered a desirable attribute in these urban Blacks but reasons for this vary.


The effect of CYP2D6 drug-drug interactions on hydrocodone effectiveness.
The hepatic cytochrome 2D6 (CYP2D6) is a saturable enzyme responsible for metabolism of approximately 25% of known pharmaceuticals. CYP interactions can alter the efficacy of prescribed medications. Hydrocodone is largely dependent on CYP2D6 metabolism for analgesia, ondansetron is inactivated by CYP2D6, and oxycodone analgesia is largely independent of CYP2D6. The objective was to determine if CYP2D6 medication coingestion decreases the effectiveness of hydrocodone. This was a prospective observational study conducted in an academic U.S. emergency department (ED). Subjects were included if they had self-reported pain or nausea and were excluded if they were unable to speak English, were less than 18 years of age, had liver or renal failure, or carried diagnoses of chronic pain or cyclic vomiting. Detailed drug ingestion histories for the preceding 48 hours prior to the ED visit were obtained. The patient's pain and nausea were quantified using a 100-mm visual analog scale (VAS) at baseline prior to drug administration and following doses of hydrocodone, oxycodone, or ondansetron. We used a mixed model with random subject effect to determine the interaction between CYP2D6 drug ingestion and study drug effectiveness. Odds ratios (ORs) were calculated to compare clinically significant VAS changes between CYP2D6 users and nonusers. A total of 250 (49.8%) of the 502 subjects enrolled had taken at least one CYP2D6 substrate, inhibitor, or inducing pharmaceutical, supplement, or illicit drug in the 48 hours prior to ED presentation. CYP2D6 drug users were one-third as likely to respond to hydrocodone (OR = 0.33, 95% confidence interval [CI] = 0.1 to 0.8) and more than three times as likely as nonusers to respond to ondansetron (OR = 3.4, 95% CI = 1.3 to 9.1). There was no significant difference in oxycodone effectiveness between CYP2D6 users and nonusers (OR = 0.53, 95% CI = 0.3 to 1.1). CYP2D6 drug-drug interactions appear to change effectiveness of commonly prescribed drugs in the ED. Drug-drug interaction should be considered prior to prescribing CYP2D6 drugs.


Blast mines: physics, injury mechanisms and vehicle protection.
Since World War II, more vehicles have been lost to land mines than all other threats combined. Anti-vehicular (AV) mines are capable of disabling a heavy vehicle, or completely destroying a lighter vehicle. The most common form of AV mine is the blast mine, which uses a large amount of explosive to directly damage the target. In a conventional military setting, landmines are used as a defensive force-multiplier and to restrict the movements of the opposing force. They are relatively cheap to purchase and easy to acquire, hence landmines are also potent weapons in the insurgents' armamentarium. The stand-offnature of its design has allowed insurgents to cause significant injuries to security forces in current conflicts with little personal risk. As a result, AV mines and improvised explosive devices (IEDs) have become the most common cause of death and injury to Coalition and local security forces operating in Iraq and Afghanistan. Detonation of an AV mine causes an explosive, exothermic reaction which results in the formation of a shockwave followed by a rapid expansion of gases. The shockwave is mainly reflected by the soillair interface and fractures the soil cap overthe mine. The detonation products then vent through the voids in the soil, resulting in a hollow inverse cone which consists of the detonation gases surrounded by the soil ejecta. It is the combination of the detonation products and soil ejecta that interact with the target vehicle and cause injury to the vehicle occupants. A number of different strategies are required to mitigate the blast effects of an explosion. Primary blast effects can be reduced by increasing the standoff distance between the seat of the explosion and the crew compartment. Enhancement of armour on the base of the vehicle, as well as improvements in personal protection can prevent penetration of fragments. Mitigating tertiary effects can be achieved by altering the vehicle geometry and structure, increasing vehicle mass, as well as developing new strategies to reduce the transfer of the impulse through the vehicle to the occupants. Protection from thermal injury can be provided by incorporating fire resistant materials into the vehicle and in personal clothing. The challenge for the vehicle designer is the incorporation of these protective measures within an operationally effective platform.


Perineuronal nets ensheath fast spiking, parvalbumin-immunoreactive neurons in the medial septum/diagonal band complex.
Perineuronal nets, composed of extracellular matrix material, have previously been associated with parvalbumin-immunoreactive neurons in the medial septum/diagonal band (MS/DB) complex of the rat. The aim of this study was to correlate the presence of perineuronal nets with electrophysiological properties and parvalbumin immunoreactivity in MS/DB neurons. Intracellular recordings were made from cells in a brain slice preparation maintained in vitro, and neurons were characterized into four populations: (i) slow-firing neurons, (ii) burst-firing neurons, (iii) fast spiking neurons with narrow action potentials and a small degree of spike frequency adaptation, and (iv) regular spiking neurons with broader action potentials and a high degree of spike frequency adaptation. Following electrophysiological characterization, neurons were filled with biocytin, processed for parvalbumin immunoreactivity and stained for perineuronal nets using Wisteria floribunda lectin. The three substances were viewed with triple fluorescence. Fast spiking, nonadapting neurons, shown previously to contain parvalbumin immunoreactivity, were nearly all ensheathed by perineuronal nets. There was a population of small parvalbumin-immunoreactive neurons which did not possess perineuronal nets, and which were not encountered with the intracellular electrodes. The other three neuron types in the MS/DB did not contain parvalbumin immunoreactivity or perineuronal nets. In keeping with this neurochemical profile for electrophysiologically identified neurons, burst-firing neurons had action potential parameters more similar to those of regular spiking than of fast spiking neurons. We conclude that fast spiking neurons, presumed to be GABAergic septohippocampal projection neurons, are surrounded by supportive structures to enable the high level of neuronal discharge required for producing disinhibition of hippocampal pyramidal neurons.


A tumor-penetrating peptide enhances circulation-independent targeting of peritoneal carcinomatosis.
Peritoneal carcinomatosis is a major source of morbidity and mortality in patients with advanced abdominal neoplasms. Intraperitoneal chemotherapy (IPC) is an area of intense interest given its efficacy in ovarian cancer. However, IPC suffers from poor drug penetration into peritoneal tumors. As such, extensive cytoreductive surgery is required prior to IPC. Here, we explore the utility of iRGD, a tumor-penetrating peptide, for improved tumor-specific penetration of intraperitoneal compounds and enhanced IPC in mice. Intraperitoneally administered iRGD significantly enhanced penetration of an attached fluorescein into disseminated peritoneal tumor nodules. The penetration was tumor-specific, circulation-independent, and mediated by the neuropilin-binding RXXK tissue-penetration peptide motif of iRGD. Q-iRGD, which fluoresces upon cleavage, including the one that leads to RXXK activation, specifically labeled peritoneal metastases displaying different growth patterns in mice. Importantly, iRGD enhanced intratumoral entry of intraperitoneally co-injected dextran to approximately 300% and doxorubicin to 250%. Intraperitoneal iRGD/doxorubicin combination therapy inhibited the growth of bulky peritoneal tumors and reduced systemic drug toxicity. iRGD delivered attached fluorescein and co-applied nanoparticles deep into fresh human peritoneal metastasis explants. These results indicate that intraperitoneal iRGD co-administration serves as a simple and effective strategy to facilitate tumor detection and improve the therapeutic index of IPC for peritoneal carcinomatosis.


Discrimination and Sleep: Differential Effects by Type and Coping Strategy.
Discrimination has been posited as a contributor of sleep disparities for Latinxs. The strategy used to cope with discrimination may reduce or exacerbate its effects on sleep. This study examined whether different types of discrimination (everyday and major lifetime discrimination) were associated with sleep indices (quality, disturbances, efficiency) and whether coping strategy used moderated associations. Data of Latinx adults (N = 602; 51% women, 65% Dominican, Mage = 46.72 years) come from the Latino Health and Well-being Project, a community-based, cross-sectional study of Latinxs in Lawrence, MA. Multiple linear regressions were estimated separately for each sleep outcome. Everyday discrimination was significantly associated with poorer sleep quality and greater disturbances; major lifetime discrimination was significantly associated with worse sleep across the three sleep indices. Coping strategy moderated associations between discrimination and sleep. Compared with Latinxs who used passive coping, those who used passive-active coping strategies had poorer sleep quality the more they experienced everyday discrimination. Latinxs who used any active coping strategy, compared with passive coping, had greater sleep disturbances the more frequently they experienced major lifetime discrimination. Findings show that everyday discrimination and major lifetime discrimination are associated with different dimensions of sleep and suggest that coping with discrimination may require the use of different strategies depending on the type of discrimination experienced.


Crystal structure of metarhodopsin II.
G-protein-coupled receptors (GPCRs) are seven transmembrane helix (TM) proteins that transduce signals into living cells by binding extracellular ligands and coupling to intracellular heterotrimeric G proteins (Gαβγ). The photoreceptor rhodopsin couples to transducin and bears its ligand 11-cis-retinal covalently bound via a protonated Schiff base to the opsin apoprotein. Absorption of a photon causes retinal cis/trans isomerization and generates the agonist all-trans-retinal in situ. After early photoproducts, the active G-protein-binding intermediate metarhodopsin II (Meta II) is formed, in which the retinal Schiff base is still intact but deprotonated. Dissociation of the proton from the Schiff base breaks a major constraint in the protein and enables further activating steps, including an outward tilt of TM6 and formation of a large cytoplasmic crevice for uptake of the interacting C terminus of the Gα subunit. Owing to Schiff base hydrolysis, Meta II is short-lived and notoriously difficult to crystallize. We therefore soaked opsin crystals with all-trans-retinal to form Meta II, presuming that the crystal's high concentration of opsin in an active conformation (Ops*) may facilitate all-trans-retinal uptake and Schiff base formation. Here we present the 3.0 Å and 2.85 Å crystal structures, respectively, of Meta II alone or in complex with an 11-amino-acid C-terminal fragment derived from Gα (GαCT2). GαCT2 binds in a large crevice at the cytoplasmic side, akin to the binding of a similar Gα-derived peptide to Ops* (ref. 7). In the Meta II structures, the electron density from the retinal ligand seamlessly continues into the Lys 296 side chain, reflecting proper formation of the Schiff base linkage. The retinal is in a relaxed conformation and almost undistorted compared with pure crystalline all-trans-retinal. By comparison with early photoproducts we propose how retinal translocation and rotation induce the gross conformational changes characteristic for Meta II. The structures can now serve as models for the large GPCR family.


Retrospective reporting of the duration of post-partum amenorrhea: a survival analysis.
Duration of post-partum amenorrhoea (PPA) is the period from the end of pregnancy to begin of menstruation. It is a temporary infecundable period. The attainment of first menstruation after delivery is treated as the termination of PPA. The main aim of this paper is to investigate the differentials of the duration of PPA according to the characteristics of mothers and their child. The data are taken from a sample survey of Palpa and Rupandehi districts. A total of 1019 ever-married mothers were interviewed. 481 mothers were provided information on the duration of PPA to their last but one born child (retrospective status reporting). Survival analysis technique has been used. It was found that the distribution of PPA showed a heaping at the multiple of three months. Mean duration of PPA was found to be ten months and it was higher among mothers of higher parity, older mothers and higher age at birth of child. Younger mothers have lower duration than that of the older. Mean duration was increased with the increased parity of mothers. A strong positive association was observed between the duration of PPA and breastfeeding (BF). The increased duration of PPA was found with the increased birth interval. The survival status of child was found a strong consequence on amenorrheaic period. Inverse association was found with education as well as high socioeconomic status of the family. A significant association was found with residential as well as caste/ethnic groups. Mean duration of PPA was found to be 10.6 months. Amenorrheaic period was found higher among mothers of higher parity, older ages, and higher age at birth of the child. A positive relationship was found with BF whereas inverse association was found with the level of education. Duration of PPA was decreased with the increased level of socio-economic status.


Verapamil treatment induces cytoprotective autophagy by modulating cellular metabolism.
Verapamil, an L-type calcium channel blocker, has been used successfully to treat cardiovascular diseases. Interestingly, we have recently shown that treatment of cancer cells with verapamil causes an effect on autophagy. As autophagy is known to modulate chemotherapy responses, this prompted us to explore the impact of verapamil on autophagy and cell viability in greater detail. We report here that verapamil causes an induction of autophagic flux in a number or tumor cells and immortalized normal cells. Moreover, we found that inhibition of autophagy in COLO 205 cells, via treatment with the chloroquine (CQ) or by CRISPR/Cas9-mediated disruption of the autophagy genes Atg7 and Atg5, causes an upregulation of apoptotic markers in response to verapamil. In search of a mechanism for this effect and because autophagy can often mitigate metabolic stress, we examined the impact of verapamil on cellular metabolism. This revealed that in normal prostate cells, verapamil diminishes glucose and glycolytic intermediate levels leading to adenosine 5'-triphosphate (ATP) depletion. In contrast, in COLO 205 cells it enhances aerobic glycolysis and maintains ATP. Importantly, we found that the autophagic response in these cells is related to the activity of l-lactate dehydrogenase A (LDHA, EC 184.108.40.206), as inhibition of LDHA reduces both basal and verapamil-induced autophagy and consequently decreases cell viability. In summary, these findings not only identify a novel mechanism of cytoprotective autophagy induction but they also highlight the potential of using verapamil together with inhibitors of autophagy for the treatment of malignant disease. ENZYMES: l-lactate dehydrogenase (LDHA, EC 18.104.22.168).


Serological diagnosis of schistosomiasis. 1 Demonstration and significance of bilharzial antigenaemia.
Circulating bilharzial antigens were demonstrated in 39% out of 87 bilharzial patients via the IHA and CF techniques using hyperimmune rabbit anti - S.mansoni worm serum. Correlation between the percentage of positive cases and the age, duration of infection, clinical presentation, type of bilharzial infection, chemotherapeutic effect and the intradermal test (I.D.T.) were made. The percentage of positive cases was found to increase with age and with the duration of infection. More positivity was encountered in hepatosplenomegalic and in S.mansoni-infected cases. Most of the I.D positive cases were also IHA positive, but still some I.D. negative cases were IHA positive. Specific chemotherapy resulted in rise in the number of IHA positive cases which was followed by gradual fall in teh next six months until they became negative. On the other hand, using the CF technique, all cases turned negative after treatment with no phase of augmentation. However, antigenaemia reappeared in few cases. These cases relapsed later. Comparing the results of IHA and CF indicates that each test may be detecting a different group of antigens.


Strand exchange of telomeric DNA catalyzed by the Werner syndrome protein (WRN) is specifically stimulated by TRF2.
Werner syndrome (WS), caused by loss of function of the RecQ helicase WRN, is a hereditary disease characterized by premature aging and elevated cancer incidence. WRN has DNA binding, exonuclease, ATPase, helicase and strand annealing activities, suggesting possible roles in recombination-related processes. Evidence indicates that WRN deficiency causes telomeric abnormalities that likely underlie early onset of aging phenotypes in WS. Furthermore, TRF2, a protein essential for telomere protection, interacts with WRN and influences its basic helicase and exonuclease activities. However, these studies provided little insight into WRN's specific function at telomeres. Here, we explored the possibility that WRN and TRF2 cooperate during telomeric recombination processes. Our results indicate that TRF2, through its interactions with both WRN and telomeric DNA, stimulates WRN-mediated strand exchange specifically between telomeric substrates; TRF2's basic domain is particularly important for this stimulation. Although TRF1 binds telomeric DNA with similar affinity, it has minimal effects on WRN-mediated strand exchange of telomeric DNA. Moreover, TRF2 is displaced from telomeric DNA by WRN, independent of its ATPase and helicase activities. Together, these results suggest that TRF2 and WRN act coordinately during telomeric recombination processes, consistent with certain telomeric abnormalities associated with alteration of WRN function.


Analysis of a chimeric class-I patatin-GUS gene in transgenic potato plants: High-level expression in tubers and sucrose-inducible expression in cultured leaf and stem explants.
Patatin is a family of lipid acyl hydrolases that accounts for 30 to 40% of the total soluble protein in potato (Solanum tuberosum L.) tubers. To examine the regulation of the patatin genes, we constructed a chimeric gene containing 2.5 kb of 5' flanking sequence from the class I patatin genomic clone PS20 transcriptionally fused to β-glucuronidase (GUS) and introduced it into potato plants using an Agrobacterium tumefaciens Tiplasmid vector. While the chimeric gene was expressed at high levels in tubers and in stolons attached to developing tubers, it was not normally expressed in leaves, stems, roots, or in stolons before tuberizatization. However, the expression of the class I patatin-GUS construct was not "tuber-specific" since leaf and stem explants cultured on medium containing 300 to 400 mM sucrose showed GUS activity equal or greater than that of tubers. The sucrose induction of GUS activity in leaf and stem explants was accompanied by the accumulation of patatin protein and large amounts of starch, but not by the morphological changes that normally are associated with tuberization. In contrast, the GUS reporter gene under the control of the 35S promoter of cauliflower mosaic virus showed an essentially uniform pattern of expression in transgenic potato plants and was not induced by sucrose.


Pilot of "Families for Health": community-based family intervention for obesity.
To develop and evaluate "Families for Health", a new community based family intervention for childhood obesity. Programme development, pilot study and evaluation using intention-to-treat analysis. Coventry, England. 27 overweight or obese children aged 7-13 years (18 girls, 9 boys) and their parents, from 21 families. Families for Health is a 12-week programme with parallel groups for parents and children, addressing parenting, lifestyle change and social and emotional development. Change in baseline BMI z score at the end of the programme (3 months) and 9-month follow-up. Attendance, drop-out, parents' perception of the programme, child's quality of life and self-esteem, parental mental health, parent-child relationships and lifestyle changes were also measured. Attendance rate was 62%, with 18 of the 27 (67%) children completing the programme. For the 22 children with follow-up data (including four who dropped out), BMI z score was reduced by -0.18 (95% CI -0.30 to -0.05) at 3 months and -0.21 (-0.35 to -0.07) at 9 months. Statistically significant improvements were observed in children's quality of life and lifestyle (reduced sedentary behaviour, increased steps and reduced exposure to unhealthy foods), child-parent relationships and parents' mental health. Fruit and vegetable consumption, participation in moderate/vigorous exercise and children's self-esteem did not change significantly. Topics on parenting skills, activity and food were rated as helpful and used with confidence by most parents. Families for Health is a promising new childhood obesity intervention. Definitive evaluation of its clinical effectiveness by randomised controlled trial is now required.


High-Throughput Drug Screening System Based on Human Induced Pluripotent Stem Cell-Derived Atrial Myocytes ∼ A Novel Platform to Detect Cardiac Toxicity for Atrial Arrhythmias.
Evaluation of proarrhythmic properties is critical for drug discovery. In particular, QT prolongation in electrocardiograms has been utilized as a surrogate marker in many evaluation systems to assess the risk of torsade de pointes and lethal ventricular arrhythmia. Recently, new evaluation systems based on human iPS cell-derived cardiomyocytes have been established. On the other hand, in clinical situations, it has been reported that the incidence of atrial arrhythmias such as atrial fibrillation has been increasing every year, with the prediction of a persistent increase in the near future. As to the increased incidence of atrial arrhythmias, in addition to the increased population of geriatric patients, a wide variety of drug treatments may be related, as an experimental method to detect drug-induced atrial arrhythmia has not been established so far. In the present study, we characterized the atrial-like cardiomyocytes derived from human induced pluripotent stem cells and examined their potential for the evaluation of drug-induced atrial arrhythmia. Atrial-like cardiomyocytes were induced by adding retinoic acid (RA) during the process of myocardial differentiation, and their characteristics were compared to those of RA-free cardiomyocytes. Using gene expression and membrane potential analysis, it was confirmed that the cells with or without RA treatment have atrial or ventricular like cardiomyocytes, respectively. Using the ultra-rapid activating delayed rectifier potassium current (IKur) channel inhibitor, which is specific to atrial cardiomyocytes, Pulse width duration (PWD) 30cF prolongation was confirmed only in atrial-like cardiomyocytes. In addition, ventricular like cardiomyocytes exhibited an early after depolarization by treatment with rapidly activating delayed rectifier potassium current (IKr) channel inhibitor, which induces ventricular arrhythmia in clinical situations. Here, we have established a high-throughput drug evaluation system using human iPS cell-derived atrial-like cardiomyocytes. Based on the obtained data, the system might be a valuable platform to detect potential risks for drug-induced atrial arrhythmias.


Binaural response properties of low-frequency neurons in the gerbil dorsal nucleus of the lateral lemniscus.
Differences in intensity and arrival time of sounds at the two ears, interaural intensity and time differences (IID, ITD), are the chief cues for sound localization. Both cues are initially processed in the superior olivary complex (SOC), which projects to the dorsal nucleus of the lateral lemniscus (DNLL) and the auditory midbrain. Here we present basic response properties of low-frequency (< 2 kHz) DNLL neurons and their binaural sensitivity to ITDs and IIDs in the anesthetized gerbil. We found many neurons showing binaural properties similar to those reported for SOC neurons. IID-properties were similar to that of the contralateral lateral superior olive (LSO). A majority of cells had an ITD sensitivity resembling that of either the ipsilateral medial superior olive (MSO) or the contralateral LSO. A smaller number of cells displayed intermediate types of ITD sensitivity. In neurons with MSO-like response ITDs that evoked maximal discharges were mostly outside of the range of ITDs the gerbil naturally experiences. The maxima of the first derivative of their ITD-functions (steepest slope), however, were well within the physiological range of ITDs. This finding is consistent with the concept of a population rather than a place code for ITDs. Moreover, we describe several other binaural properties as well as physiological and anatomical evidence for a small but significant input from the contralateral MSO. The large number of ITD-sensitive low-frequency neurons implicates a substantial role for the DNLL in ITD processing and promotes this nucleus as a suitable model for further studies on ITD-coding.


Fusion-activated Ca2+ entry via vesicular P2X4 receptors promotes fusion pore opening and exocytotic content release in pneumocytes.
Ca(2+) is considered a key element in multiple steps during regulated exocytosis. During the postfusion phase, an elevated cytoplasmic Ca(2+) concentration ([Ca(2+)])(c) leads to fusion pore dilation. In neurons and neuroendocrine cells, this results from activation of voltage-gated Ca(2+) channels in the plasma membrane. However, these channels are activated in the prefusion stage, and little is known about Ca(2+) entry mechanisms during the postfusion stage. This may be particularly important for slow and nonexcitable secretory cells. We recently described a "fusion-activated" Ca(2+) entry (FACE) mechanism in alveolar type II (ATII) epithelial cells. FACE follows initial fusion pore opening with a delay of 200-500 ms. The site, molecular mechanisms, and functions of this mechanism remain unknown, however. Here we show that vesicle-associated Ca(2+) channels mediate FACE. Using RT-PCR, Western blot analysis, and immunofluorescence, we demonstrate that P2X(4) receptors are expressed on exocytotic vesicles known as lamellar bodies (LBs). Electrophysiological, pharmacological, and genetic data confirm that FACE is mediated via these vesicular P2X(4) receptors. Furthermore, analysis of fluorophore diffusion into and out of individual vesicles after exocytotic fusion provides evidence that FACE regulates postfusion events of LB exocytosis via P2X(4). Fusion pore dilation was clearly correlated with the amplitude of FACE, and content release from fused LBs was accelerated in fusions followed by FACE. Based on these findings, we propose a model for regulation of the exocytotic postfusion phase in nonexcitable cells in which Ca(2+) influx via vesicular Ca(2+) channels regulates fusion pore expansion and vesicle content release.


Molecular dynamics simulation of ion mobility in gases.
A force field molecular dynamics method is developed to directly simulate ion drift in buffer gases driven by an electric field. The ion mobility and collision cross sections (CCSs) with relevance to ion mobility spectrometry can be obtained from the simulated drift velocity in high-density buffer gases (pressure ∼50 bars) and high electric fields (∼107 V/m). Compared to trajectory methods, the advantage of the molecular dynamics method is that it can simultaneously sample the internal dynamic motions of the ion and the ion-gas collisions. For ions with less than 100 atoms, the simulated collision cross section values can be converged to within ±1%-2% by running a 100 ns simulation for 5-19 h using one computer core. By using a set of element-based Lennard-Jones parameters that are not tuned for different atomic types in different molecules, the simulated collision cross sections for 15 small molecular ions (number of atoms ranging from 17 to 85, mass ranging from 74.1 to 609.4 g/mol) are consistent with experimental values: the mean unsigned error is 2.6 Å2 for He buffer gas and 4.4 Å2 for N2 buffer gas. The sensitivity of the simulated CCS values to random diffusion, drift velocity, electric field strength, temperature, and buffer gas density is examined.


A genetic study of dentin growth in the mandibular second and third molars of male mice.
Although the influence of genetic and environmental factors on the completed crown size of mandibular molars in mice is well known, such influence on the growth process of the mandibular second and third molars of mice has not yet been clarified. The purpose of the present study is to examine the growth pattern of the dentinal formation and evaluate the relative contributions of both genetic and environmental factors to longitudinal dentin formation by the use of the partial diallel cross-genetic approach, the time-marking method and a computerized image measurement system. The following results were obtained: 1) the onset of dentinal formation in the second molar was on the 6-9th day after birth, while that of the third molar was on the 9-12th day after birth; 2) the best fitted growth curve of the dentinal formation for the first molar was the fifth order polynomial equation, while for the second molar the fourth order polynomial equation was best, and for the third molar the third order polynomial equation was best; 3) maternal effect strongly influenced the second and the third molar dentin growth at an early stage of postnatal growth. Thus, it could be concluded that the dentinal formation growth pattern differed among the three kinds of mouse molars while maternal environmental factors, in addition to genetic factors, played an important role in the mouse second and third molar dentin growth during the early stage of postnatal growth.


Sensitivity of cancer cells to Plk1 inhibitor GSK461364A is associated with loss of p53 function and chromosome instability.
Polo-like kinases are a family of serine threonine kinases that are critical regulators of cell cycle progression and DNA damage response. Predictive biomarkers for the Plk1-selective inhibitor GSK461364A were identified by comparing the genomics and genetics of a panel of human cancer cell lines with their response to a drug washout followed by an outgrowth assay. In this assay, cell lines that have lost p53 expression or carry mutations in the TP53 gene tended to be more sensitive to GSK461364A. These more sensitive cell lines also had increased levels of chromosome instability, a characteristic associated with loss of p53 function. Further mechanistic studies showed that p53 wild-type (WT) and not mutant cells can activate a postmitotic tetraploidy checkpoint and arrest at pseudo-G(1) state after GSK461364A treatment. RNA silencing of WT p53 increased the antiproliferative activity of GSK461364A. Furthermore, silencing of p53 or p21/CDKN1A weakened the tetraploidy checkpoint in cells that survived mitotic arrest and mitotic slippage. As many cancer therapies tend to be more effective in p53 WT patients, the higher sensitivity of p53-deficient tumors toward GSK461364A could potentially offer an opportunity to treat tumors that are refractory to other chemotherapies as well as early line therapy for these genotypes.


A kinetochore-based ATM/ATR-independent DNA damage checkpoint maintains genomic integrity in trypanosomes.
DNA damage-induced cell cycle checkpoints serve as surveillance mechanisms to maintain genomic stability, and are regulated by ATM/ATR-mediated signaling pathways that are conserved from yeast to humans. Trypanosoma brucei, an early divergent microbial eukaryote, lacks key components of the conventional DNA damage-induced G2/M cell cycle checkpoint and the spindle assembly checkpoint, and nothing is known about how T. brucei controls its cell cycle checkpoints. Here we discover a kinetochore-based, DNA damage-induced metaphase checkpoint in T. brucei. MMS-induced DNA damage triggers a metaphase arrest by modulating the abundance of the outer kinetochore protein KKIP5 in an Aurora B kinase- and kinetochore-dependent, but ATM/ATR-independent manner. Overexpression of KKIP5 arrests cells at metaphase through stabilizing the mitotic cyclin CYC6 and the cohesin subunit SCC1, mimicking DNA damage-induced metaphase arrest, whereas depletion of KKIP5 alleviates the DNA damage-induced metaphase arrest and causes chromosome mis-segregation and aneuploidy. These findings suggest that trypanosomes employ a novel DNA damage-induced metaphase checkpoint to maintain genomic integrity.


Giant Traumatic Diaphragmatic Hernia: A Report of Delayed Presentation.
Diaphragmatic rupture is an uncommon injury after blunt abdominal trauma. The diaphragmatic defect may not be obvious in imaging studies immediately after the initial injury. Patients may have delayed presentation when the diaphragmatic defect enlarges and allows abdominal content to herniate into the thoracic cavity. Here, we present the case of a 30-year-old man who presented with the emergency department complaining of shortness of breath at rest for two days duration. He reported having shortness of breath for the last five years, but he attributed it to his smoking. The shortness of breath was associated with cough productive and vague abdominal pain. The patient had an unremarkable relevant medical history. He reported having a motor vehicle accident five years ago that was severe but he did not sustain any significant injuries or fractures. Upon examination, the patient appeared in respiratory distress. Respiratory examination revealed diminished air entry on the left hemithorax and the abdominal examination revealed increased generalized tenderness with increased bowel sounds. The patient underwent a thoracic computed tomography scan, which unexpectedly demonstrated a huge left-sided diaphragmatic defect with bowel loops observed to occupy the left hemithorax completely. The patient was stabilized and shifted to emergency laparotomy during which the hernia content was reduced and the defect was closed with a mesh. The patient reported the resolution of his symptoms after the surgery. Intensive chest physiotherapy exercises were performed. After six months of follow-up, the patient remained asymptomatic with no active complaints. The diaphragmatic hernia may have delayed presentations after several years of blunt abdominal trauma. The case highlighted the importance of initial imaging studies after blunt trauma may not identify the diaphragmatic defect.


Metabolic amplifying pathway increases both phases of insulin secretion independently of beta-cell actin microfilaments.
Two pathways control glucose-induced insulin secretion (IS) by beta-cells. The triggering pathway involves ATP-sensitive potassium (K(ATP)) channel-dependent depolarization, Ca(2+) influx, and a rise in the cytosolic Ca(2+) concentration ([Ca(2+)](c)), which triggers exocytosis of insulin granules. The metabolic amplifying pathway augments IS without further increasing [Ca(2+)](c). The underlying mechanisms are unknown. Here, we tested the hypothesis that amplification implicates actin microfilaments. Mouse islets were treated with latrunculin B and cytochalasin B to depolymerize actin or jasplakinolide to polymerize actin. They were then perifused to measure [Ca(2+)](c) and IS. Metabolic amplification was studied during imposed steady elevation of [Ca(2+)](c) by tolbutamide or KCl or by comparing the magnitude of [Ca(2+)](c) and IS changes produced by glucose and tolbutamide. Both actin polymerization and depolymerization augmented IS triggered by all stimuli without increasing (sometimes decreasing) [Ca(2+)](c), which indicates a predominantly inhibitory function of microfilaments in exocytosis at a step distal to [Ca(2+)](c) increase. When [Ca(2+)](c) was elevated and controlled by KCl or tolbutamide, the amplifying action of glucose was facilitated by actin depolymerization and unaffected by polymerization. Both phases of IS were larger in response to high-glucose than to tolbutamide in low-glucose, although triggering [Ca(2+)](c) was lower. This difference in IS, due to amplification, persisted when the IS rate was doubled by actin depolymerization or polymerization. In conclusion, metabolic amplification is rapid and influences the first as well as the second phase of IS. It is a late step of stimulus-secretion coupling, which does not require functional actin microfilaments and could correspond to acceleration of the priming process conferring release competence to insulin granules.


A role for the androgen metabolite, 5alpha-androstane-3beta,17beta-diol, in modulating oestrogen receptor beta-mediated regulation of hormonal stress reactivity.
Activation of the hypothalamic-pituitary-adrenal (HPA) axis is a basic response of animals to environmental perturbations that threaten homeostasis. These responses are regulated by neurones in the paraventricular nucleus of the hypothalamus (PVN) that synthesise and secrete corticotrophin-releasing hormone (CRH). Other PVN neuropeptides, such as arginine vasopressin and oxytocin, can also modulate activity of CRH neurones in the PVN and enhance CRH secretagogue activity of the anterior pituitary gland. In rodents, sex differences in HPA reactivity are well established; females exhibit a more robust activation of the HPA axis after stress than do males. These sex differences primarily result from opposing actions of sex steroids, testosterone and oestrogen, on HPA function. Ostreogen enhances stress activated adrenocorticotrophic hormone (ACTH) and corticosterone (CORT) secretion, whereas testosterone decreases the gain of the HPA axis and inhibits ACTH and CORT responses to stress. Data show that androgens can act directly on PVN neurones in the male rat through a novel pathway involving oestrogen receptor (ER)beta, whereas oestrogen acts predominantly through ERalpha. Thus, we examined the hypothesis that, in males, testosterone suppresses HPA function via an androgen metabolite that binds ERbeta. Clues to the neurobiological mechanisms underlying such a novel action can be gleaned from studies showing extensive colocalisation of ERbeta in oxytocin-containing cells of the PVN. Hence, in this review, we address the possibility that testosterone inhibits HPA reactivity by metabolising to 5alpha-androstane-3beta,17beta-diol, a compound that binds ERbeta and regulates oxytocin containing neurones of the PVN. These findings suggest a re-evaluation of studies examining pathways for androgen receptor signalling.


Miniaturized Cultivation Profiling Returns Indolo-Sesquiterpene Michael Adducts from the Australian Soil-Derived Aspergillus terreus CMB-SWF012.
An integrated program of chemical profiling (GNPS) coupled with an expanded format 24-well-plate miniaturized cultivation profiling (MATRIX) utilizing traditional as well as grain/pulse and cereal media permitted rapid prioritization of Aspergillus terreus CMB-SWF012 as a source of unprecedented natural products. Scaled-up cultivation on rice and PDA yielded the rare tripeptides asterripeptides A-C (1-3), new indolo-sesquiterpene Michael adducts terreusides A and B (4 and 5), and known precursors asterresin A (6) and (+)-giluterrin (7). Structures for 1-7 were assigned by detailed spectroscopic and chemical analysis and biosynthetic considerations.


Successful treatment of aggressive HIV-associated non-Hodgkin's lymphoma with combination chemotherapy, biotherapy with rituximab and HAART: presentation of a therapeutic option.
The incidence of non-Hodgkin's lymphoma (NHL) in individuals infected with human immunodeficiency virus (HIV) is more than 60 times higher than in matched controls. In the vast majority of cases aggressive pathological subtypes and advanced stages prevail, extranodal sites are involved and systemic symptoms are present. The prognosis of HIV-NHL remains poor and the optimal therapeutic approach has yet to be defined. We report a 48-year-old Ethiopian woman with advanced-stage HIV infection, who developed diffuse large cell, immunoblastic type B-cell NHL and was treated with a modified CHOP-like chemotherapy combined with Rituximab and supported with growth factor. Highly active antiretroviral therapy (HAART) and opportunistic infections prophylaxis were administered concomitantly. The patient completed 6 cycles of therapy and currently, 76 weeks after diagnosis, is in complete clinical remission. Despite the fact that there was a transient decrease in the CD4-positive cell number and a 1.5 log increase in plasma viral load there were no opportunistic infections, nor was life-threatening toxicity seen. Rituximab seems a well-tolerable and advantageous adjunct to chemotherapy and HAART in the treatment of aggressive HIV-associated NHL andshould be investigated in large trials in the future.


The concept of nemesis in psychoneurosis.
A patient who has a nemesis fear as the basic process in his psychoneurosis feels that he is destined to repeat the life course of one of his parents to eventual long-term psychosis, or incapacitating physical illness or death by illness or accident. He feels that this will occur at about the same age as that at which his parent suffered his misfortune. The patient during his childhood and adolescence had a traumatic relationship with this parent, and is haunted by guilt feelings about it, and fears that avenging destiny, or nemesis, requires that he pay with a similar misfortune for that which he feels he caused. These patients improve much in psychotherapy, but the underlying nemesis fear, though much reduced in severity, is not entirely eliminated.


NorVold Abuse Questionnaire for men (m-NorAQ): validation of new measures of emotional, physical, and sexual abuse and abuse in health care in male patients.
There are far more prevalence studies on abuse of females than on males as subjects of abuse. The NorVold Abuse Questionnaire (NorAQ) measures emotional, physical, and sexual abuse, as well as abuse in health care, in women and men. The aim of this study was to test the concurrent validity and test-retest reliability of the questions in the version of NorAQ administered to men (m-NorAQ) against the interview model. The validation was tested in a subsample (n = 86) of a male patient sample who had filled out the m-NorAQ (N = 1667). Respondents completed m-NorAQ twice and were then interviewed. Before the interview, respondents were instructed to answer questions based on personal experience. The interview consisted of 4 open-ended questions about lifetime experiences of emotional, physical, and sexual abuse and abuse in health care. Results indicated that respondents in the subsample had discussed the experiences of abuse more often in both formal and informal settings than had subjects in the total patient sample. Measures of sensitivity for m-NorAQ were good to excellent (emotional abuse, 83%; physical abuse, 76%; sexual abuse, 68%; abuse in health care, 93%), as were those for specificity (emotional abuse, 72%; physical abuse, 92%; sexual abuse, 99%, abuse in health care, 90%); likelihood ratios were satisfactory (emotional abuse, 3; physical abuse, 9; sexual abuse, 46; abuse in health care, 9); and test-retest reliability measures were excellent (emotional abuse, 80%-95%; physical abuse, 77%-88%; sexual abuse, 91%-100%; abuse in health care, 84%-92%). m-NorAQ showed good to excellent concurrent validity for the different types of abuse and excellent reliability for all questions about abuse. In spite of methodological challenges, validation studies must be conducted as a minimum precaution to ensure that an instrument accurately measures abuse as intended.


[Glycaemia control in critically ill patients is justified and effective].
Hyperglycemia and insulin resistance develop in the majority of severe acute illness and/or injury. One of the main causes of hyperglycemia in critically ill patients is the release of counterregulatory stress hormones and proinflammatory cytokines, in addition to increased production of glucose along with its decreased utilization. Hyperglycemia plays an important role not only in influencing the cascade of inflammatory cytokines, but it also increases oxidative stress. In the past, stress hyperglycemia was thought to be an evolutionary protective, natural adaptive response of the body to current threat, which allows increased entry of glucose into the cells of non-insulin-tissues, thus improving chances for survival. At present, however, this state of insulin resistance, glucose intolerance and hyperglycemia is called "stress diabetes" or "diabetes of injury". Ever since the time of the breakthrough "Leuven" study, which brought significant reduction in morbidity and mortality in surgical critically ill patients with tight glycemic control, hospitals, particularly their intensive care units, have focused on the treatment of hyperglycemia. Although extensive observational data have shown a consistent, almost linear relationship between blood glucose concentrations seen in hospitalized patients and the incidence of adverse clinical results, there have been particular doubts concerning the universality of control, its safety, and pitfalls resulting from hypoglycemia. This controversial debate is currently enriched by the recent international trial - the NICE-SUGAR, whose post-hoc analyses are currently underway. Despite the controversy there is no doubt that the deliberate control of blood glucose control in critically ill patients is justified. It is the insulin application regimen--the insulin protocol per se--that remains the biggest problem in the implementation of glycemic control. Regarding targets, it is necessary to take into account that the best positive effects on outcomes can be anticipated in certain subgroups of critically ill patients, which is currently the subject of further study. Continued streamlining, achieving optimal blood glucose ranges in critically ill patients will allow us to develop and apply computer algorithms that greatly simplify and improve continuous monitoring of blood glucose. Procedures seeking optimal intensive control in critically ill patients are accepted in intensive care units. However, it is undoubtedly necessary to improve monitoring techniques and the quality of biosensors in order to ensure the safety and effectiveness of interventions aimed at reducing blood glucose levels while using advanced protocols. Automatic closed systems are a promise for the future.


Prognostic risk stratification of gliomas using deep learning in digital pathology images.
Evaluation of tumor-tissue images stained with hematoxylin and eosin (H&E) is pivotal in diagnosis, yet only a fraction of the rich phenotypic information is considered for clinical care. Here, we propose a survival deep learning (SDL) framework to extract this information to predict glioma survival. Digitized whole slide images were downloaded from The Cancer Genome Atlas (TCGA) for 766 diffuse glioma patients, including isocitrate dehydrogenase (IDH)-mutant/1p19q-codeleted oligodendroglioma, IDH-mutant/1p19q-intact astrocytoma, and IDH-wildtype astrocytoma/glioblastoma. Our SDL framework employs a residual convolutional neural network with a survival model to predict patient risk from H&E-stained whole-slide images. We used statistical sampling techniques and randomized the transformation of images to address challenges in learning from histology images. The SDL risk score was evaluated in traditional and recursive partitioning (RPA) survival models. The SDL risk score demonstrated substantial univariate prognostic power (median concordance index of 0.79 [se: 0.01]). After adjusting for age and World Health Organization 2016 subtype, the SDL risk score was significantly associated with overall survival (OS; hazard ratio = 2.45; 95% CI: 2.01 to 3.00). Four distinct survival risk groups were characterized by RPA based on SDL risk score, IDH status, and age with markedly different median OS ranging from 1.03 years to 14.14 years. The present study highlights the independent prognostic power of the SDL risk score for objective and accurate prediction of glioma outcomes. Further, we show that the RPA delineation of patient-specific risk scores and clinical prognostic factors can successfully demarcate the OS of glioma patients.


Hysterectomy and kidney cancer risk: a meta-analysis.
Recent cohort findings suggest that women who underwent a hysterectomy have an elevated relative risk of kidney cancer, although evidence from past studies has been inconsistent. We conducted a systematic review and meta-analysis of published cohort and case-control studies to summarize the epidemiologic evidence investigating hysterectomy and kidney cancer. Studies published from 1950 through 2012 were identified through a search of PubMed and of references from relevant publications. Meta-analyses were conducted using random-effects models to estimate summary relative risks (SRRs) and 95% confidence intervals (CIs) for hysterectomy, age at hysterectomy (<45, 45+ years) and time since hysterectomy (<10, 10+ years). The SRR for hysterectomy and kidney cancer for all published studies (seven cohort, six case-control) was 1.29 (95% CI, 1.16-1.43), with no evidence of between-study heterogeneity or publication bias. The summary effect was slightly weaker, although still significant, for cohorts (SRR, 1.26; 95% CI, 1.11-1.42) compared with case-control findings (1.37; 95% CI, 1.09-1.73) and was observed irrespective of age at hysterectomy, time since the procedure and model adjustment for body mass index, smoking status and hypertension. Women undergoing a hysterectomy have an approximate 30% increased relative risk of subsequent kidney cancer. Additional research is needed to elucidate the biological mechanisms underlying this association.


Automatic and motivational predictors of children's physical activity: integrating habit, the environment, and the Theory of Planned Behavior.
Physical activity determinant studies now often include both environmental and sociocognitive factors but few of them acknowledge and explore the mechanisms underlying relevant environmental influences. This study explored environmental correlates of children's self-reported physical activity and potential mediation through the Theory of Planned Behavior (TPB) and habit strength. Six hundred and twenty-one pupils aged 9-11 years were recruited from 4 primary schools in the UK. TPB variables, habit strength and environmental variables were assessed at baseline. Self-reported physical activity was assessed 1 week later. Mediation tests revealed that 43% of the association between convenient facilities and intention was mediated through subjective norms (17%) and habit (26%), while 15% of the association between convenient facilities and physical activity was mediated through habit strength alone. A significant direct effect of convenient facilities and resources in the home environment on physical activity was also found. The school environment was not significantly related to children's physical activity intentions or behavior. The results suggest that the environment influences children's physical activity both directly and indirectly and that habit strength seems to be the most important mediator for this association.


Psychosocial aspects and psychiatric disorders in young adult with thalassemia major.
Beta-thalassemia major (TM), a chronic, genetically determined hematological disorder, has received little investigation on the psychological aspects of the disease and the psychosocial adjustment of patients with this anemia. Unfortunately, the few psychological studies referred on the literature are generally limited to the investigation of the only children with thalassemia major. The study was planned to assess the self-image, the quality of life, the way of coping and to investigate the existence of psychiatric disorders in young adults with thalassemia major. 147 patients were included in the study. Patients were psychologically investigated by three interviews: the first connects some psychosocial information, the second submits patients to the psychological test and the third gives back the results obtained by the test. The psychological test consists of the ways of coping questionnaire (WCQ), the Machover's test, The short form 36-health survey questionnaire (SF-36) and symptom-check-list-90 revised (SCL-90-R) were performed on all patients. Vis-à-vis identity and self-image were found to be low with feeling of insufficiency and being exposed to vulnerability in 80% of patients with TM. Evaluation of mean values of symptomatological dimensions in these patients showed a personality characterized by somatization (SOM), depression (DEP) and obsessive-compulsive traits. The principal coping strategy used is escape-avoidance. No statistically significant differences occurred to relation to gender, age, level education and SCL-90-R and WCQ scores. Estimation of the SF-36 scores showed that the emotional role and social function values were considerably lower than in all of the domain. As a result, the study showed that most of the patients with TM had severe psychosocial problems. Relying on these data, it was concluded that medical therapy of these patients should be supported with psychological aid and psychiatric treatment.


COVID-19: changing patterns among neurosurgical patients from North India, efficacy of repeat testing, and inpatient prevalence.
COVID-19 has affected surgical practice globally. Treating neurosurgical patients with the restrictions imposed by the pandemic is challenging in institutions with shared patient areas. The present study was performed to assess the changing patterns of neurosurgical cases, the efficacy of repeated testing before surgery, and the prevalence of COVID-19 in asymptomatic neurosurgical inpatients. Cases of non-trauma-related neurosurgical patients treated at the Postgraduate Institute of Medical Education and Research (PGIMER) before and during the COVID-19 pandemic were reviewed. During the pandemic, all patients underwent a nasopharyngeal swab reverse transcription-polymerase chain reaction test to detect COVID-19 at admission. Patients who needed immediate intervention were surgically treated following a single COVID-19 test, while stable patients who initially tested negative for COVID-19 were subjected to repeated testing at least 5 days after the first test and within 48 hours prior to the planned surgery. The COVID-19 positivity rate was compared with the local period prevalence. The number of patients who tested positive at the second test, following a negative first test, was used to determine the probable number of people who could have become infected during the surgical procedure without second testing. Of the total 1769 non-trauma-related neurosurgical patients included in this study, a mean of 337.2 patients underwent surgery per month before COVID-19, while a mean of 184.2 patients (54.6% of pre-COVID-19 capacity) underwent surgery per month during the pandemic period, when COVID-19 cases were on the rise in India. There was a significant increase in the proportion of patients undergoing surgery for a ruptured aneurysm, stroke, hydrocephalus, and cerebellar tumors, while the number of patients seeking surgery for chronic benign diseases declined. At the first COVID-19 test, 4 patients (0.48%) tested were found to have the disease, a proportion 3.7 times greater than that found in the local community. An additional 5 patients tested positive at the time of the second COVID-19 test, resulting in an overall inpatient period prevalence of 1%, in contrast to a 0.2% national cumulative caseload. It is possible that COVID-19 was prevented in approximately 67.4 people every month by using double testing. COVID-19 has changed the pattern of neurosurgical procedures, with acute cases dominating the practice. Despite the fact that the pandemic has not yet reached its peak in India, COVID-19 has been detected 3.7 times more often in asymptomatic neurosurgical inpatients than in the local community, even with single testing. Double testing displays an incremental value by disclosing COVID-19 overall in 1 in 100 inpatients and thus averting its spread through neurosurgical services.


First isolation and genotyping of viruses from recent outbreaks of viral haemorrhagic septicaemia (VHS) in Slovenia.
In November and December 2007, the virus causing viral haemorrhagic septicaemia (VHS) was detected in rainbow trout Oncorhynchus mykiss from 2 fish farms in Slovenia. During 2008 and 2009 the infection spread only among rainbow trout farms and 4 new outbreaks were confirmed. High mortality and clinical signs of VHS were observed among the diseased fish. VHSV was confirmed by virus isolation, immunoperoxidase test, reverse transcriptase polymerase chain reaction (RT-PCR) and phylogenetic analysis. Based on 1 complete (1524 nucleotides [nt]) and 9 partial (600 nt) glycoprotein gene nucleotide sequences, 9 VHSV isolates from the 6 VHS outbreaks were genetically closely related (99 to 100% identity), and were classified into the Subgroup I-a of Genotype I, most closely related to the German isolates Dstg21-07, Dstg36-06, and Dstg54-1-07 (99 to 100% identity). Phylogenetic analysis and epidemiological investigations confirmed that the VHS virus had been (re)introduced with imported live fish, and that subsequent outbreaks were linked to the initial infection. Our study shows that direct nucleotide sequencing of RT-PCR products, amplified from the tissue of VHSV-infected fish, represents a reliable tool for fast routine genotyping in diagnostic laboratories. This is the first report of a natural epidemic associated with VHSV infection in Slovenia since the eradication of the disease in 1977.


Association between GH receptor polymorphism (exon 3 deletion), serum IGF1, semen quality, and reproductive hormone levels in 838 healthy young men.
GH activity may be involved in male reproductive function. A common genetic polymorphism in the gene encoding the GH receptor (GHR) results in deletion of the entire exon 3 sequence (GHRd3 isoform). The short GHRd3/d3 isoform seems more sensitive compared with full-length receptors (GHRfl/fl). TO INVESTIGATE THE ASSOCIATIONS BETWEEN GH ACTIVITY, EVALUATED BY EXON 3 GHR POLYMORPHISM, AND SERUM IGF1 VS REPRODUCTIVE HORMONES, SEMEN QUALITY, AND PRE- AND POSTNATAL GROWTH IN HEALTHY YOUNG MALES (N=838, MEAN AGE: 19.4 years). Compared with GHRfl/fl homozygous individuals (n=467) GHRd3/d3 homozygous individuals (n=69) tended to have larger semen volume (3.2 (2.4-4.3) vs 3.6 (2.6-4.7) ml, P=0.053) and higher serum inhibin-B levels (208 pg/ml (158-257) vs 227 pg/ml (185-264), P=0.050). Semen quality, levels of gonadotropins, testosterone, estradiol, sex hormone-binding globulin, and IGF1 were not associated with GHRd3 genotype. A twofold increase in serum IGF1 was associated with a 13% (4-23) increase in calculated free testosterone (P=0.004). By contrast IGF1 was inversely associated with serum inhibin-B (P=0.027), but showed no associations to semen quality. GHR genotype and serum IGF1 were not associated with size at birth or final height. GHRd3 polymorphism seemed only to have a weak influence on male reproductive function of borderline significance. The sensitive GHRd3/d3 genotype may slightly increase testicular function, as evaluated by semen volume and levels of inhibin-B, but does not seem to influence Leydig cell steroidogenesis. GHR genotype did not influence pre- and postnatal growth.


Relationship between hepatic blood flow and overall metabolism: the hepatic arterial buffer response.
The impact of hepatic blood flow on overall metabolism is not generally appreciated but is of major consequence for homeostasis. Endocrine glands produce and secrete hormones, but for effective and rapid control of hormonal levels, a rapid turnover is required. The liver is the principal organ of clearance for a wide variety of hormones and other endogenous substances. Altered total hepatic blood flow results in altered hepatic clearance rates. The main intrinsic control of hepatic blood flow is designed to hold total hepatic blood flow steady. The liver cannot control portal venous flow (which is simply the sum of outflows of the extrahepatic splanchnic organs); therefore, to hold total blood flow steady, the hepatic artery adjusts total flow in relation to alterations in portal blood flow. This control is referred to as the hepatic arterial buffer response and it functions to regulate blood flow per se rather than nutrient or oxygen regulation. The hepatic arterial flow does not change in response to altered hepatic metabolic activity as shown by the lack of expected vascular changes with altered oxygen content of blood, altered metabolic activity induced by enzyme induction or enzyme inhibition, and altered oxygen uptake or biliary secretion. The hepatic artery is not subservient to local metabolic needs of the liver but rather is controlled in a manner that subserves the homeostatic needs of the entire body. The mechanism of the arterial buffer response is not yet clear but various hypotheses are discussed.


Assessment of Respiratory Health Symptoms and Asthma in Children near a Drying Saline Lake.
Residents of the Imperial Valley, a rural, agricultural border region in California, have raised concerns over high rates of pediatric asthma symptoms. There is an urgent need to understand the influences and predictors of children's respiratory health in Imperial Valley. We assessed the impacts of sociodemographic, lifestyle, and household factors on children's respiratory health and asthma prevalence by administering a survey to parents of elementary school children (n = 357) in northern Imperial Valley. We observed an overall asthma prevalence of 22.4% and respiratory symptoms and allergies were widely reported, including wheezing (35.3%), allergies (36.1%), bronchitic symptoms (28.6%), and dry cough (33.3%). Asthmatics were significantly more likely to report respiratory symptoms, but high rates of wheezing, allergies, and dry cough were observed among nonasthmatics, suggesting the possibility for underdiagnosis of respiratory impairment in our school-age population. Having an asthmatic mother and exposure to environmental tobacco smoke were also associated with greater odds of asthma. Our findings provide evidence to support community concerns about children's respiratory health, while also suggesting that household and demographic characteristics have limited explanatory power for assessing asthma in this population. This work provides critical baseline data with which to evaluate local environmental factors and their influence on asthma and respiratory symptoms.


Boosting the performance of MALT lymphoma classification in patients with primary Sjögren's Syndrome through data augmentation: a case study.
Sjögren's Syndrome (SS) patients with mucosa associated lymphoid tissue lymphomas (MALTLs) and diffuse large B-cell lymphomas (DLBCLs) have 10-year survival rates of 80% and 40%, respectively. This highlights the unique biologic burden of the two histologic forms, as well as, the need for early detection and thorough monitoring of these patients. The lack of MALTL patients and the fact that most studies are single cohort and combine patients with different lymphoma subtypes narrow the understanding of MALTL progression. Here, we propose a data augmentation pipeline that utilizes an advanced synthetic data generator which is trained on a Pan European data hub with primary SS (pSS) patients to yield a high-quality synthetic data pool. The latter is used for the development of an enhanced MALTL classification model. Four scenarios were defined to assess the reliability of augmentation. Our results revealed an overall improvement in the accuracy, sensitivity, specificity, and AUC by 7%, 6.3%, 9%, and 6.3%, respectively. This is the first case study that utilizes data augmentation to reflect the progression of MALTL in pSS.


Associations of demographic and socioeconomic factors with stage at diagnosis of breast cancer.
Stage at diagnosis is one of the most important prognostic factors of breast cancer survival. Because in the breast cancer case this may vary with socioeconomic characteristics, this study was performed to recognize the relationship between demographic and socioeconomic factors with stage at diagnosis in Iran. This cross-sectional, descriptive study conducted on 526 patients suffering from breast cancer and registered in Cancer Research Center of Shahid Beheshti University of Medical Sciences from 2008 to 2013. A reliable and valid questionnaire about family levels of socioeconomic status filled in by interviewing the patients via phone. For analyzing the data, Multinomial logistic regression, Kendal tau-b correlation coefficient and Contingency Coefficient tests were executed by SPSS22. Economic status, educational attainment of patient and household head and/or a combination of these were considered as parameters for socioeconomic status. First, the relationship between stage at diagnosis and demographic and socioeconomic status was assessed in univariate analysis then these relationships assessed in two different models of multinomial logistic regression. The mean age of the patients was 48.3 (SD=11.4). According to the results of this study, there were significant relationships between stage at diagnosis of breast cancer with patient education (p=0.011), living place (p=0.044) and combined socioeconomic status (p=0.024). These relationships persisted in multiple multinomial logistic regressions. Other variables, however, had no significant correlation. Patient education, combined socioeconomic status and living place are important variables in stage at diagnosis of breast cancer in Iranian women. Interventions have to be applied with the aim of raising women's accessibility to diagnostic and medical facilities and also awareness in order to reducing delay in referring. In addition, covering breast cancer screening services by insurance is recommended.


The effect of step stool use and provider height on CPR quality during pediatric cardiac arrest: A simulation-based multicentre study.
We aimed to explore whether a) step stool use is associated with improved cardiopulmonary resuscitation (CPR) quality; b) provider adjusted height is associated with improved CPR quality; and if associations exist, c) determine whether just-in-time (JIT) CPR training and/or CPR visual feedback attenuates the effect of height and/or step stool use on CPR quality. We analysed data from a trial of simulated cardiac arrests with three study arms: No intervention; CPR visual feedback; and JIT CPR training. Step stool use was voluntary. We explored the association between 1) step stool use and CPR quality, and 2) provider adjusted height and CPR quality. Adjusted height was defined as provider height + 23 cm (if step stool was used). Below-average height participants were ≤ gender-specific average height; the remainder were above average height. We assessed for interaction between study arm and both adjusted height and step stool use. One hundred twenty-four subjects participated; 1,230 30-second epochs of CPR were analysed. Step stool use was associated with improved compression depth in below-average (female, p=0.007; male, p<0.001) and above-average (female, p=0.001; male, p<0.001) height providers. There is an association between adjusted height and compression depth (p<0.001). Visual feedback attenuated the effect of height (p=0.025) on compression depth; JIT training did not (p=0.918). Visual feedback and JIT training attenuated the effect of step stool use (p<0.001) on compression depth. Step stool use is associated with improved compression depth regardless of height. Increased provider height is associated with improved compression depth, with visual feedback attenuating the effects of height and step stool use.


Prevalence of anemia in Japanese adolescents: 30 years' experience in screening for anemia.
Iron deficiency is the most common cause of anemia in adolescents and is of continuing concern in Japan. We have conducted screening for anemia in adolescents in the Tokyo area since 1966. Screening for the prevalence of anemia is important both for identifying populations at risk and determining the appropriate treatment for individual patients. The number of junior and senior high school students enrolled in the screening program was 793 in 1966, which increased to more than 70,000 by 1989. A normal hemoglobin (Hb) level was present in 90% of students before 1981, and in 98% of boys and 95% of girls in 1990. The percentage of girls with a normal Hb level has decreased gradually since 1991. The high incidence of mild hypochromic microcytic anemia suggests an increase in the prevalence of iron deficiency. Dieting may be the cause of this undesirable phenomenon in adolescents.


Evaluation of the BinaxNOW Staphylococcus aureus test for rapid identification of Gram-positive cocci from VersaTREK blood culture bottles.
The ability of the rapid BinaxNOW Staphylococcus aureus (BNSA) immunochromatographic test (Alere Scarborough, Inc., ME) to accurately differentiate S. aureus from coagulase-negative staphylococci (CoNS) and other Gram-positive cocci (GPC) directly from VersaTREK blood culture bottles was evaluated. A total of 319 positive patient blood culture bottles with GPC seen in clusters with Gram staining were tested using the BNSA test and a direct tube coagulase test (DTCT). The BNSA test was accurate for the detection and differentiation of S. aureus from CoNS and other GPC within 30 min from the time of blood culture positivity and demonstrated a test sensitivity and specificity of 95.8% and 99.6%, respectively. BNSA test results were faxed to the antimicrobial stewardship pharmacist by noon each day in order to evaluate empirical antimicrobial therapy and facilitate more rapid changes or modifications if necessary. Same-day reporting of BNSA test results in conjunction with an antimicrobial stewardship program was more impactful in improving treatment for inpatients with documented S. aureus bacteremia than in reducing empirical vancomycin use in inpatients with CoNS during the first 24 h following reporting.


The population genetic structure of the facultatively sexual parasitic nematode Strongyloides ratti in wild rats.
We have investigated the population genetic structure of the parasitic nematode Strongyloides ratti in wild rats. In the UK, S. ratti reproduces predominantly by mitotic parthenogenesis, with sexual forms present at a rate of less than 1%. S. ratti was found to be a prevalent parasite and substantial genetic diversity was detected. Most rats were infected with a genotypic mixture of parasites. A hierarchical analysis of the genetic variation found in S. ratti sampled across Britain and Germany showed that 73.3% was explained by variation between parasites within individual hosts and 25.3% by variation between rats within sample sites. Only a small proportion (1.4%) of the total genetic variation was attributable to genetic subdivision between sample sites, suggesting that there is substantial gene flow between these sites. Most parasites sampled were found to exist in Hardy-Weinberg equilibrium and this population genetic structure is discussed in view of the virtual absence of sexual reproduction.


[Microsurgical management of petroclival meningiomas combined trans-subtemporal and suboccipital retrosigmoid keyhole approach].
With the development of modern skull base minimally invasive technology mature and neural radio surgery techniques, it is necessary to re-examine the therapeutic strategy for the treatment of petroclival meningiomas. To sum up the operative experience and methods in microsurgical resection of petroclival meningiomas by the combining trans-subtemporal and suboccipital retrosigmoid keyhole approach. To explore the minimally invasive operation approach of petroclival meningiomas, to raise the removal degree and to improve the postoperative result using this approach. The clinical data of the consecutive 21 patients with the petroclival meningiomas were reviewed retrospectively. The method, degree of tumor resection,techniques of the combining keyhole approach, Karnofsky performance score (KPS) before and after operation were also analyzed. The neuronavigation guided operation was performed in 9 cases, and 12 cases were operated in the neuroelectrophysiological monitoring. Total excision of the tumor resection (Simpson, I-II levels) was conducted in 18 cases (85.7%, 18/21), and 3 patients underwent close resection (Simpson III level, 14.3%, 3/21). Postoperative three-dimensional CT showed good lock bone flap restoration; Postoperative pathology confirmed meningioma. Postoperative cranial nerve dysfunction or new original nerve dysfunction were aggravated in 5 cases (23.8%) , including transient trochlear nerve (3 cases), abducent nerve (1 case), and the motor branch of trigeminal nerve paralysis (1 case). Abducent nerve paralysis (1 case) appeared, with hearing impairment. After the 3-month follow-up, 11 cases had the same KPS aspreoperation, 7 cases improved, and 3 cases not improved. The KPS score was 77.14±23.12 on average, and there was no statistically significant difference compared with that before operation (P>0.05). The postoperative follow-up for half a year showed fluent speaking and writing in 19 cases (KPS 70 or higher), and general recovery in 2 cases (KPS<70). The postoperative follow-up for 3-29 months showed no tumor recurrence or progress. The combining trans-subtemporal and suboccipital retrosigmoid keyhole approach is simple, safe, and minimally invasive, and an ideal operation approach of petroclival meningioma. To master the operation skills and the intraoperative matters needing attention in the operation, is favorable to improve the resection rate and curative effect.


[Variability of the level of thermoresistance as an indicator of the functional status of a cell].
Effect of acclimating temperature on the thermoresistance of isolated ciliated gill epithelium of Anodonta anatina was studied. One strip of gills from each mussel was used to determine the initial level of thermoresistance (survival time at 40 degrees C). Other strips were kept at 24 degrees C until their death, and their thermoresistance and fluorescence after Ethidium bromide staining were determined at different time intervals. After a short exposure at 24 degrees C the mean level of thermoresistance of the epithelium increased. The individual shift in the value and direction of cell thermoresistance in different mussels negatively correlated with its initial level. It resulted in narrowing the extent of individual variability of the level of thermoresistance. The prolongation of exposure at 24 degrees C decreased the mean level of thermoresistance, disturbed the negative correlation, and increased the variability of thermoresistance level. The value of cell fluorescence remained constant within 48 h. The increase in fluorescence was observed 12 h after reducing the mean thermoresistance level. Hence, the dynamics of variability in the thermoresistance level is a most sensitive indication of the cell functional state.


CycleGAN-Based Image to Image Translation for Realistic Surgical Training Phantoms.
Training in surgery is essential for surgeons to develop skill and dexterity. Physical training phantoms provide excellent haptic feedback and tissue properties for stitching and operating with authentic instruments and are easily available. However, they lack realistic traits and fail to reflect the complex environment of a surgical scene. Generative Adversarial Networks can be used for image-to-image translation, addressing the lack of realism in physical phantoms, by mapping patterns from the intraoperative domain onto the video stream captured during training with these surgical simulators. This work aims to achieve a successful I2I translation, from intra-operatory mitral valve surgery images onto a surgical simulator, using the CycleGAN model. Different experiments are performed - comparing the Mean Square Error Loss with the Binary Cross Entropy Loss; validating the Fréchet Inception Distance as a training and image quality metric; and studying the impact of input variability on the model performance. Differences between MSE and BCE are modest, with MSE being marginally more robust. The FID score proves to be very useful in identifying the best training epochs for the CycleGAN I2I translation architecture. Carefully selecting the input images can have a great impact in the end results. Using less style variability and input images with good feature details and clearly defined characteristics enables the network to achieve better results.Clinical Relevance- This work further contributes for the domain of realistic surgical training, successfully generating fake intra operatory images from a surgical simulator of the cardiac mitral valve.


White matter (dis)connections and gray matter (dys)functions in visual neglect: gaining insights into the brain networks of spatial awareness.
Seminal case reports collected during the middle part of the XX century, designated the parietal lobe as the principal area of damage in patients suffering from contralesional spatial neglect (Brain WC. Visual disorientation with special reference to lesions of the right cerebral hemisphere. Brain 1941;64:224-72; Paterson A, Zangwill O. Disorders of visual space perception associated with lesions of the right cerebral hemisphere. Brain 1944;67:331-58; McFie J, Piercy MF, Zangwill O. Visual spatial agnosia associated with lesions of the right hemisphere. Brain 1950;73:167-90). Based on this evidence, textbooks of neurology have traditionally referred to neglect as a "parietal sign". This view found complete accomplishment in the 1986 group study by Vallar and Perani, who confirmed that the inferior parietal lobe was the area most frequently involved in neglect patients with lesions confined to the cerebral cortex and lesions involving subcortical gray matter nuclei. In the same study, it was found that lesions limited to subcortical white matter were rarely associated with neglect. Here, we reconsider recent accumulating evidence, gathered from investigations in animals and human patients, supporting the partially different view that damage involvement of long-range white matter bundles connecting the parietal to the frontal lobe, importantly influence the occurrence and severity of spatial neglect. These findings do not dispute the role of the parietal and frontal cortex in spatial attention and space-related behaviour, but call for a reappraisal of the respective roles of disruption of white matter connections and damage of gray matter cortical modules in the pathophysiology of neglect. Disentangling the connectional and modular anatomical correlates of neglect may be crucial to better understand the pathophysiology of this syndrome, to explain the manifold clinical dissociations often encountered in clinical practice and to increase the impact of behavioural and pharmacological interventions. In this review, we focus on the role of within-hemisphere white-matter disconnection. The role of interhemispheric disconnection, perhaps the oldest connectionist theory of neglect (Geschwind N. Disconnexion syndromes in animals and man - part II. Brain 1965;88:585-644), was extensively treated elsewhere (Bartolomeo P, Thiebaut de Schotten M, Doricchi F. Left unilateral neglect as a disconnection syndrome, Cerebral Cortex 2007;45:3127-48). We first summarise the structure of long-range white matter connections within the cerebral hemispheres and sketch a brief historical review of the original findings suggesting the role of intrahemispheric disconnection to neglect. We then revisit some of the current functional interpretation of the neglect syndrome in the light of disconnectionist approach and review evidences favouring or disfavouring a purely disconnectionist interpretation of the syndrome. Finally, we address the issue of diagnostic criteria to be used in future anatomo-clinical studies aiming at investigating the role of white matter and gray matter dysfunctions in spatial neglect.


Social inequalities in tobacco-attributable mortality in Spain. The intersection between age, sex and educational level.
First study of social inequalities in tobacco-attributable mortality (TAM) in Spain considering the joint influence of sex, age, and education (intersectional perspective). Data on all deaths due to cancer, cardiometabolic and respiratory diseases among people aged ≥35 years in 2016 were obtained from the Spanish Statistical Office. TAM was calculated based on sex-, age- and education-specific smoking prevalence, and on sex-, age- and disease-specific relative risks of death for former and current smokers vs lifetime non-smokers. As inequality measures, the relative index of inequality (RII) and the slope index of inequality (SII) were calculated using Poisson regression. The RII is interpreted as the relative risk of mortality between the lowest and the highest educational level, and the SII as the absolute difference in mortality. The crude TAM rate was 55 and 334 per 100,000 in women and men, respectively. Half of these deaths occurred among people with the lowest educational level (27% of the population). The RII for total mortality was 0.39 (95%CI: 0.35-0.42) in women and 1.61 (95%CI: 1.55-1.67) in men. The SII was -41 and 111 deaths per 100,000, respectively. Less-educated women aged <55 years and men (all ages) showed an increased mortality risk; nonetheless, less educated women aged ≥55 had a reduced risk. TAM is inversely associated with educational level in men and younger women, and directly associated with education in older women. This could be explained by different smoking patterns. Appropriate tobacco control policies should aim to reduce social inequalities in TAM.


A 2-Protein Signature Predicting Clinical Outcome in High-Grade Serous Ovarian Cancer.
High-grade serous ovarian cancer (HGSOC) accounts for approximately 70% deaths in ovarian cancer. The overall survival (OS) of HGSOC is poor and still remains a clinical challenge. High-grade serous ovarian cancer can be divided into 4 molecular subtypes. The prognosis of different molecular subtypes is still unclear. We aimed to investigate the prognostic values of immunohistochemistry-based different molecular subtypes in patients with HGSOC. We analyzed the protein expression of representative biomarkers (CXCL11, HMGA2, and MUC16) of 3 different molecular subtypes in 110 formalin-fixed, paraffin-embedded HGSOC by tissue microarrays. High CXCL11 expression predicted worse OS, not disease-free survival (DFS; P = 0.028 for OS, P = 0.191 for DFS). High HMGA2 expression predicted worse OS and DFS (P = 0.037 for OS, P = 0.021 for DFS). MUC16 expression was not associated with OS or DFS (P = 0.919 for OS, P = 0.517 for DFS). Multivariate regression analysis showed that CXCL11 combined with HMGA2 signature was an independent predictor for OS and DFS in patients with HGSOC. CXCL11 combined with HMGA2 signature was a clinically applicable prognostic model that could precisely predict an HGSOC patient's OS and tumor recurrence. This model could serve as an important tool for risk assessment of HGSOC prognosis.


Association between attention-deficit/hyperactivity disorder and epilepsy in pediatric populations.
There is a bidirectional relationship between attention-deficit/hyperactivity disorder (ADHD) and epilepsy. ADHD increases seizure risk, while patients with epilepsy have an increased prevalence of ADHD. The reasons explaining this association are not fully understood. Proposed mechanisms include effects of antiepileptic medications, underlying neurodevelopmental vulnerability, the effects of chronic seizures and subclinical epileptiform activity on cognitive functions and adrenergic dysfunction. There may also be a common genetic defect underlying both disorders in some families. Antiepileptics associated with ADHD-like side effects include phenobarbital, gabapentin, vigabatrin and topiramate. Methylphenidate has been studied in a double-blind setting against placebo for treatment of ADHD comorbid with epilepsy, and has a good risk-benefit ratio. Amphetamine, atomoxetine, clonidine and guanfacine only have case series to support their use and bupropion should be avoided.


Identification of three protein kinases which phosphorylate threonyl-tRNA synthetase from rat liver.
Threonyl-tRNA synthetase is phosphorylated in Chinese hamster ovary cells labeled with 32Pi [(1984) J. Biol. Chem. 259, 11160-11161]. Phosphorylation of the purified synthetase from rat liver has been examined with five different protein kinases. Three of the enzymes phosphorylate the synthetase, protease activated kinase I, the cAMP-dependent protein kinase, and the Ca2+, phospholipid-dependent protein kinase. Phosphorylation occurs exclusively on seryl residues. Two-dimensional phosphopeptide maps of tryptic digests of the phosphorylated synthetase are distinct with each protein kinase. These data suggest that multiple phosphorylation of the synthetase may occur in vivo.


Soybean biodiesel methyl esters, free glycerin and acid number quantification by 1H nuclear magnetic resonance spectroscopy.
Production of alternative fuels, such as biodiesel, from transesterification of vegetable oil driven by heterogeneous catalysts is a promising alternative to fossil diesel. However, achieving a successful substitution for a new renewable fuel depends on several quality parameters. (1)H NMR spectroscopy was used to determine the amount of methyl esters, free glycerin and acid number in the transesterification of soybean oil with methanol in the presence of hydrotalcite-type catalyst to produce biodiesel. Reaction parameters, such as temperature and time, were used to evaluate soybean oil methyl esters rate conversion. Temperatures of 100 to 180 °C and times of 20 to 240 min were tested on a 1 : 12 molar ratio soybean oil/methanol reaction. At 180 °C/240 min conditions, a rate of 94.5 wt% of methyl esters was obtained, where free glycerin and free fatty acids were not detected.


Adapting an effective primary care provider STD/HIV prevention training programme.
Sexually acquired human immunodeficiency virus (HIV) infection continues to be the major source of HIV infection in the USA. Preventing sexual transmission of HIV can be accomplished through patient behaviour change. Such behaviour change can also decrease risk of other sexually transmitted diseases (STDs) and unwanted pregnancies, both far more common problems than HIV infection. Primary care physicians and other providers can increase patients' safe sex practices by conducting effective sexual risk assessment (RA) and risk reduction (RR) counselling, but physicians both infrequently and incompletely do sexual RA and RR. A programme was developed to improve primary care physicians' prevention practice using Simulated Patient Instructors (SPIs) and mailed educational materials. Programme evaluations showed improved sexual RA and RR practice both by self-report as well as by observation by Simulated Patient Evaluators (SPEs). This paper briefly reviews these findings and then presents adaptations made to improve the programme's content, decrease its cost and increase its availability for training many other care providers. Evaluation of the adapted programme indicates that content and training methods are highly regarded by a diverse array of trainees. To disseminate the modified programme beyond the local area, a Train-the-Trainer programme and manual have been developed, including discussion of recruiting, training and using SPIs for sexual risk reduction. Wider use of this training, as well as more effective and more readily available STD/HIV prevention training, are needed to attain national goals of provider clinical prevention practice.


Explicit representations of all refractive optical interfaces without spherical aberration.
The following explicit model, valid for high aperture refraction with homogenous and isotropic materials, encompasses all explicit solutions of the first-order nonlinear differential equation representing the perfect image-forming process of any axial object point into its axial image point. Solutions include well-known cases, such as flats, spheres, prolate ellipsoids, prolate hyperboloids, and other sections of nondegenerate Cartesian ovals of revolution, now classified according to the recurrent explicit solution introduced herein. We also present some series expansions, given in cylindrical coordinates z(r), for more efficient computation. Explicit solutions allow accurate and expedite thickness calculation as compared to the regular series, parametric, or implicit solutions commonly used. The results of this study are useful in the design of centered optical systems that are perfectly aligned.


Facile Synthesis of Graphene from Waste Tire/Silica Hybrid Additives and Optimization Study for the Fabrication of Thermally Enhanced Cement Grouts.
This work evaluates the effects of newly designed graphene/silica hybrid additives on the properties of cementitious grout. In the hybrid structure, graphene nanoplatelet (GNP) obtained from waste tire was used to improve the thermal conductivity and reduce the cost and environmental impacts by using recyclable sources. Additionally, functionalized silica nanoparticles were utilized to enhance the dispersion and solubility of carbon material and thus the hydrolyzable groups of silane coupling agent were attached to the silica surface. Then, the hybridization of GNP and functionalized silica was conducted to make proper bridges and develop hybrid structures by tailoring carbon/silica ratios. Afterwards, special grout formulations were studied by incorporating these hybrid additives at different loadings. As the amount of hybrid additive incorporated into grout suspension increased from 3 to 5 wt%, water uptake increased from 660 to 725 g resulting in the reduction of thermal conductivity by 20.6%. On the other hand, as the concentration of GNP in hybrid structure increased, water demand was reduced, and thus the enhancement in thermal conductivity was improved by approximately 29% at the same loading ratios of hybrids in the prepared grout mixes. Therefore, these developed hybrid additives showed noticeable potential as a thermal enhancement material in cement-based grouts.


A dynamic microscale mid-throughput fibrosis model to investigate the effects of different ratios of cardiomyocytes and fibroblasts.
Cardiac fibrosis is a maladaptive remodeling of the myocardium hallmarked by contraction impairment and excessive extracellular matrix deposition (ECM). The disease progression, nevertheless, remains poorly understood and present treatments are not capable of controlling the scarring process. This is partly due to the absence of physiologically relevant, easily operable, and low-cost in vitro models, which are of the utmost importance to uncover pathological mechanisms and highlight possible targets for anti-fibrotic therapies. In classic models, fibrotic features are usually obtained using substrates with scar mimicking stiffness and/or supplementation of morphogens such as transforming growth factor β1 (TGF-β1). Qualities such as the interplay between activated fibroblasts (FBs) and cardiomyocytes (CMs), or the mechanically active, three-dimensional (3D) environment, are, however, neglected or obtained at the expense of the number of experimental replicates achievable. To overcome these shortcomings, we engineered a micro-physiological system (MPS) where multiple 3D cardiac micro-tissues can be subjected to cyclical stretching simultaneously. Up to six different biologically independent samples are incorporated in a single device, increasing the experimental throughput and paving the way for higher yielding drug screening campaigns. The newly developed MPS was used to co-culture different ratios of neonatal rat CMs and FBs, investigating the role of CMs in the modulation of fibrosis traits, without the addition of morphogens, and in soft substrates. The expression of contractile stress fibers and of degradative enzymes, as well as the deposition of fibronectin and type I collagen were superior in microtissues with a low amount of CMs. Moreover, high CM-based microconstructs simulating a ratio similar to that of healthy tissues, even if subjected to both cyclic stretch and TGF-β1, did not show any of the investigated fibrotic signs, indicating a CM fibrosis modulating effect. Overall, this in vitro fibrosis model could help to uncover new pathological aspects studying, with mid-throughput and in a mechanically active, physiologically relevant environment, the crosstalk between the most abundant cell types involved in fibrosis.