Physical-chemical stability and in vitro digestibility of hybrid nanoparticles based on the layer-by-layer assembly of lactoferrin and BSA on liposomes.
Hybrid nanoparticles were fabricated by the electrostatic deposition of positive bovine serum albumin (BSA) and negative lactoferrin (LF) onto the surface of anionic nanoliposomes (NLs). The resulting particles had a cumulative size of 156.27 ± 11.0 nm and decreased in negative charge. Transmission electron microscopy (TEM) revealed that the hybrid particles formed a smooth and spherical polyelectrolyte complex after globular protein deposition. Observations in size distribution and surface charge after heat treatment, pH alteration and long-term storage found that the particles coated with layers of polyelectrolytes, BSA and LF, had obviously better stability than the bare liposomes. In an in vitro gastrointestinal digestion study, monolayer coated NLs (LF-NLs) and double-layer coated NLs (BSA-LF-NLs) had similar changes in microstructure (TEM) and the release rate of model cargos (calcein), which were superior to the uncoated NLs. These results indicated that hybrid nanoparticles coated with the polyelectrolytes of BSA and LF on the surface of liposomes by electrostatic interaction may improve liposomal stability, and showed some implications for the fabrication of functional molecular delivery systems to control physical-chemical and digestion stability in food and nutrition areas.


The morphology of the femur in elderly Japanese females: Analysis using 3D-CT.
While various implants are currently used for the treatment of femoral trochanteric fractures in Japan, 'mismatches' have been known to occur between patient and implant type. The purpose of this study is to identify morphological characteristics of elderly Japanese females, both in terms of the shape of proximal femur and its anterior curvature in order to evaluate the suitability of implants for treatment of femoral trochanteric fractures using 3D-computed tomography (CT) scanning. This study used CT scan data taken from 50 elderly females (aged 70 years or older). Neck-shaft (NS) angle and height of the greater trochanter (GT) were measured under defined conditions (TPI: theoretical plane to place implant). Anterior curvature of the femoral shaft was also measured. On average, the NS angle was found to be 128.1° and GT height was 62.4 mm. Average curvature radius was 1040 mm. As for the NS angle, the centrum-collum-diaphysis angles of existing implants (125° or 130°) are reasonable. In elderly Japanese patients, the proximal femur shows specific characteristics. As GT height is very short, the implant may easily interfere with the lateral cortex. Additionally, the curvature radius calculated (1040 mm) was smaller than that of past reports, indicating strong curvature in the Japanese elderly. Our measurement results are potentially useful in the avoidance of intraoperative trouble caused by mismatch of implants.


Comparative evaluation of root canal disinfection by conventional method and laser: an in vivo study.
The aim of this study was to comparatively evaluate in vivo the disinfecting ability of conventional method and lasers in root canals. Study criteria included 60 single rooted teeth, which were indicated for root canal therapy followed to dental caries and trauma with intact crowns. Such selected patients were randomly divided into 2 groups, namely, Group A (30 teeth) and Group B (30 teeth). All clinical procedures were carried out under strict aseptic precautions. The teeth in Group A were subjected to biomechanical preparation followed by the treatment with the help of diode laser containing the gallium aluminum and arsenic, which emitted 980 nm wavelengths. The teeth in Group B were treated with routine method of biomechanical preparation along with irrigation using sodium hypochlorite and hydrogen peroxide. The microbiological samples were taken immediately after the access preparation and after the completion of the root canal disinfection and were sent for microbiological analysis. The teeth in Group A showed presence of common strains of bacteria ranging from Streptococci, Staphylococci, Klebsiella, and Pseudomonas. Reduction in the growth of microorganisms was found for all types of microorganisms. Only 8 samples exhibited the growth after treatment with laser. Results of Group B also showed the presence of common strains of anaerobic and aerobic bacteria as shown in earlier studies, predominantly Staphylococcus, Streptococci, and Pseudomonas. Statistical analysis showed non-significant P values for the microorganisms; however, only 3 samples showed the growth after treatment with conventional technique using sodium hypochlorite and hydrogen peroxide. Conventional method by using sodium hypochlorite and hydrogen peroxide as irrigating solutions is highly effective in disinfecting the root canal. Lasers when used can also reduce the bacterial load of the infected root canal.


Metabolic activation of intrahepatic CD8+ T cells and NKT cells causes nonalcoholic steatohepatitis and liver cancer via cross-talk with hepatocytes.
Hepatocellular carcinoma (HCC), the fastest rising cancer in the United States and increasing in Europe, often occurs with nonalcoholic steatohepatitis (NASH). Mechanisms underlying NASH and NASH-induced HCC are largely unknown. We developed a mouse model recapitulating key features of human metabolic syndrome, NASH, and HCC by long-term feeding of a choline-deficient high-fat diet. This induced activated intrahepatic CD8(+) T cells, NKT cells, and inflammatory cytokines, similar to NASH patients. CD8(+) T cells and NKT cells but not myeloid cells promote NASH and HCC through interactions with hepatocytes. NKT cells primarily cause steatosis via secreted LIGHT, while CD8(+) and NKT cells cooperatively induce liver damage. Hepatocellular LTβR and canonical NF-κB signaling facilitate NASH-to-HCC transition, demonstrating that distinct molecular mechanisms determine NASH and HCC development.


A systematic survey of randomised trials that stopped early for reasons of futility.
Randomised trial protocols may incorporate interim analyses, with the potential to stop the study for futility if early data show insufficient promise of a treatment benefit. Previously, we have shown that this approach will theoretically lead to mis-estimation of the treatment effect. We now wished to ascertain the importance of this phenomenon in practice. We reviewed the methods and results in a set of trials that had stopped for futility, identified through an extensive literature search. We recorded clinical areas, interventions, study design, outcomes, trial setting, sponsorship, planned and actual treatment effects, sample sizes; power; and if there was a data safety monitoring board, or a published protocol. We identified: if interim analyses were pre-specified, and how many analyses actually occurred; what pre-specified criteria might define futility; if a futility analysis formed the basis for stopping; who made the decision to stop; and the conditional power of each study, i.e. the probability of statistically significant results if the study were to continue to its complete sample size. We identified 52 eligible trials, covering many clinical areas. Most trials had multiple centres, tested drugs, and 40% were industry sponsored. There were 75% where at least one interim analysis was planned a priori; a majority had only one interim analysis, typically with about half the target total sample size. A majority of trials did not pre-define a stopping rule, and a variety of reasons were given for stopping. Few studies calculated and reported low conditional power to justify the early stop. When conditional power could be calculated, it was typically low, especially under the current trend hypothesis. However, under the original design hypothesis, a few studies had relatively high conditional power. Data collection often continued after the interim analysis. Although other factors will typically be involved, we conclude that, from the perspective of conditional power, stopping early for futility was probably reasonable in most cases, but documentation of the basis for stopping was often missing or vague. Interpretation of truncated trials would be enhanced by improved reporting of stopping protocols, and of their actual execution.


No relationship between fear of childbirth and pregnancy-/delivery-outcome in a low-risk Dutch pregnancy cohort delivering at home or in hospital.
To examine the relationship of fear of childbirth (FOC), general anxiety and depression during pregnancy and postpartum with birth complications. For this prospective cohort study 105 healthy women with low-risk pregnancies (until at least 30 weeks gestation) completed the Wijma Delivery Expectancy/Experience Questionnaire (W-DEQ) and the Hospital Anxiety and Depression Scale (HADS) at 30 weeks gestation and 6 weeks postpartum. These results were related with delivery characteristics. FOC during pregnancy was not related to complications during labour and delivery. In a regression analysis, both multiparity and medical interventions were predictors for higher postpartum FOC. A positive correlation was found between FOC during pregnancy and FOC at six weeks postpartum, corrected for complications during childbirth (r = 0.45, p < 0.001). The birth giving process was not related to FOC during pregnancy, but the pre-partum level of FOC certainly is predictive of the level of postpartum FOC, suggesting that FOC as measured during gestation may influence the interpretation of the birth experience itself. We did find a positive relationship between both parity and medical interventions during childbirth and FOC postpartum.


Modulation of murine B16F10 melanoma plasminogen activator production by a synthetic peptide derived from the laminin A chain.
Laminin is a large multidomain protein with diverse biological activities. We previously demonstrated that intact laminin as well as an A chain synthetic peptide (LamA2091-2108) stimulate tissue plasminogen activator (t-PA)-catalyzed plasminogen activation. Here we report that LamA2091-2108 increases t-PA production by the highly metastatic murine melanoma cell line B16F10, with no effect on the parental B16F1 line, which has a low metastatic capacity. Incubation of plasminogen with B16F10-conditioned medium results in direct activation of the zymogen to plasmin. Furthermore, following incubation of B16F10 cells with plasminogen, plasmin is eluted from the cell surface, suggesting that these cells contain binding sites for plasminogen/plasmin in close proximity to t-PA binding sites. Quantitation of t-PA activity using the synthetic substrate Val-Leu-Lys-p-nitroanilide indicates a minimal 10-fold increase in t-PA in the conditioned medium of B16F10 cells grown in the presence of LamA2091-2108, with no increased t-PA activity observed in B16F1-conditioned medium. Similar results were obtained in immunocapture experiments which are specific for t-PA antigen. In addition, B16F10 melanoma-associated t-PA catalyzes the plasminogen-dependent hydrolysis of laminin. Together these data suggest that degradation of basement membrane proteins by metastatic melanoma cells may release fragments (such as LamA2091-2108) which stimulate both the production and activity of metastasis-associated proteinases such as t-PA, providing a mechanism for augmentation of the metastatic capacity of B16F10 melanoma cells.


Disease chronicity and activity predict subclinical left ventricular systolic dysfunction in patients with systemic lupus erythematosus.
This study investigates parameters that could predict subclinical cardiac dysfunction in systemic lupus erythematosus (SLE) in the absence of valvular, clinical coronary artery and pericardial disease. A case-control trial. Rheumatology clinic, a university teaching hospital. Eighty-two female SLE patients (49 (SD 9) years) and 82 female normal subjects (49 (13) years) matched for age, body mass index, blood pressure and heart rate. All underwent standard echocardiography and tissue Doppler imaging. Twenty-two (27%) patients had evidence of impaired left ventricular (LV) long-axis function with mean myocardial peak systolic velocity (Sm) of basal six segments <4.4 cm/s and also subnormal stress-corrected midwall fractional shortening. Thirty-four (42%) patients demonstrated impaired right ventricular (RV) long-axis function. These occurred in the presence of comparable normal LV ejection fraction, cardiac index, and RV fractional area change to the control group. Patients with subnormal mean Sm were older (49 (8) vs 44 (9); p = 0.043) and had a higher prevalence of hypertension (46% vs 22%; p = 0.034), longer disease duration >10 years (82% vs 50%, p = 0.01), higher disease activity score (73% vs 48% for Systemic Lupus Erythematosus Disease Activity Index (SLEDAI)> or =1, p = 0.049) and end-organ damage index (64% vs 47% with Systemic Lupus International Collaborating Clinics Damage Index (SLICC)> or =1, p = 0.049) than those with normal values. Disease duration >10 years, disease activity index and increased arterial stiffness provided additional incremental predictive value of LV long-axis function. SLE patients have subclinical long and short-axis dysfunctions. Regular monitoring of cardiac function by tissue Doppler echocardiography may be indicated for patients who had SLE for >10 years, frequent flare or when arterial stiffening is demonstrated.


Band gaps in jagged and straight graphene nanoribbons tunable by an external electric field.
Band gap control by an external field is useful in various optical, infrared and THz applications. However, widely tunable band gaps are still not practical due to a variety of reasons. Using the orthogonal tight-binding method for π-electrons, we have investigated the effect of the external electric field on a subclass of monolayer chevron-type graphene nanoribbons that can be referred to as jagged graphene nanoribbons. A classification of these ribbons was proposed and band gaps for applied fields up to the SiO2 breakdown strength (1 V nm(-1)) were calculated. According to the tight-binding model, band gap opening (or closing) takes place for some types of jagged graphene nanoribbons in the external electric field that lies on the plane of the structure and perpendicular to its longitudinal axis. Tunability of the band gap up to 0.6 eV is attainable for narrow ribbons. In the case of jagged ribbons with armchair edges larger jags forming a chevron pattern of the ribbon enhance the controllability of the band gap. For jagged ribbons with zigzag and armchair edges regions of linear and quadratic dependence of the band gap on the external electric field can be found that are useful in devices with controllable modulation of the band gap.


Absence of Healing Impairment in Osteotomies Prepared via Osseodensification Drilling.
This study sought to qualitatively and quantitatively evaluate the effect of osteotomy preparation by conventional (control group) or OD (OD group) instrumentation on osteotomy healing. An incision of 10 cm was made in the anteroposterior direction over the hip in five sheep, and 15 osteotomies were prepared in the left ilium of the sheep (n = 3/sheep). Three different instrumentation techniques were utilized: (1) conventional/regular drilling (R [recommended by manufacturer]) in a 3-step series of a 2-mm pilot, 3.2-mm, and 3.8-mm twist drills; (2) OD clockwise (OD-CW) drilling with Densah Bur (Versah) 2.0-mm pilot, 2.8-mm, and 3.8-mm multi-fluted tapered burs; and (3) OD counterclockwise (OD-CCW) drilling with Densah Bur 2.0-mm pilot, 2.8-mm, and 3.8-mm multi-fluted tapered burs. Drilling was performed at 1,100 rpm with saline irrigation. Qualitative histomorphometric analysis of the osteotomies after 6 weeks did not show any healing impairment due to the instrumentation. Histologic analysis shows bone remodeling and growth in all samples, irrespective of osteotomy preparation technique, with the presence of bone chips observed along the length of the osteotomy wall in sites subjected to osseodensification drilling.


Development of segment specificity in identified lineages of the grasshopper CNS.
The purpose of this study was to determine the factors underlying differences in population size and composition between segmentally homologous neuronal lineages. The segmental median neuroblasts (MNBs) of grasshoppers are identified stem cells that each produce a midline group of neurons. We traced the embryonic development of the group in two disparate segments, counting MNB progeny and profiles of dying cells in fixed and stained preparations of staged embryos. In the metathoracic segment (T3), about 95 MNB progeny survive embryonic development, whereas in the next posterior segment, the first abdominal (A1), only about 60 survive. In T3, the MNB arises at 29% of embryogenesis and dies at 78%, whereas in A1 the MNB arises at 30% and dies at 73%. In T3, the number of MNB progeny initially increases at a steady rate, 10 cells being added per 5% of embryogenesis. Between 70% and 78% growth tapers off; although the T3 MNB continues to divide, cells die at the same time, specifically removing last-born progeny. By contrast, in A1 the MNB progeny increase in two phases, one from 30% to 45% and the other from 60% to 73%, again at the rate of 10 cells per 5%. Between the two phases, the number of A1 progeny is stable. The A1 MNB continues to divide, but cells die at the same time, specifically removing earlier-born progeny. The episodes of cell death in A1 and T3 coincide with embryonic molts, and thus may be hormonally triggered. Cell death is greater in A1 than T3, accounting for most of the difference in population size. The difference in MNB longevity makes a lesser contribution. The present data, together with corollary anatomical data (Thompson and Siegler, 1991), support the hypothesis that progeny fated to become certain neuronal types are selectively removed from the two MNB lineages: intersegmental interneurons from T3 and efferent neurons and local interneurons from A1.


Muscle-Related Polymorphisms (MSTN rs1805086 and ACTN3 rs1815739) Are Not Associated with Exceptional Longevity in Japanese Centenarians.
Myostatin (MSTN) and α-actinin-3 (ACTN3) genes are potentially associated with preservation of muscle mass and oxidative capacity, respectively. To explore the possible role of these genes in exceptional longevity (EL), the allele/genotype frequency distribution of two polymorphisms in MSTN (rs1805086, K153R) and ACTN3 (rs1815739, R577X) was studied in Japanese centenarians of both sexes (n = 742) and healthy controls (n = 814). The rs1805086 R-allele (theoretically associated with muscle mass preservation at the expense of oxidative capacity) was virtually absent in the two groups, where genotype distributions were virtually identical. Likewise, no differences in allele (p = 0.838 (women); p = 0.193 (men); p = 0.587 (both sexes)) or genotype distribution were found between groups for ACTN3 rs1815739 (p = 0.975 (women), p = 0.136 (men), p = 0.752 (both sexes)). Of note, however, the frequency of the rs1805086 R-allele observed here is the lowest been reported to date whereas that of the 'highly oxidative/efficient' rs1815739 XX genotype in Japanese male centenarians (33.3%) or supercentenarians of both sexes (≥110 years) are the highest (32.6%), for a non-American population. No definite conclusions can be inferred in relation to EL owing to its lack of association with both rs1815739 and rs1805086. However, it cannot be excluded that these gene variants could eventually be related to a "healthy" metabolic phenotype in the Japanese population. Further research might determine if such metabolic profile is among the factors that can potentially predispose these individuals to live longer than Caucasians and what genetic variants might be actually involved.


Which jump variables should be used to assess explosive leg muscle function?
The main purpose of this study was to determine the relationships between countermovement jump (CMJ) variables and acceleration and maximum speed performance. Twenty-three elite Australian football players were tested on a CMJ, which yielded several kinematic and kinetic variables describing leg muscle function. A 40 m sprint was also conducted to assess acceleration (10 m time) and an estimate of maximum speed (flying 20 m time). Players from one Australian Football League (AFL) club were tested and Pearson correlations for CMJ variables and sprint performance were calculated. Jump height, peak velocity, peak force, and peak power had less than 50% common variance, and therefore represented independent expressions of CMJ performance. Generally, the correlations between CMJ variables and sprinting performance were stronger for maximum speed (small to large effect sizes) than for acceleration (trivial to moderate sizes). The variable that produced the strongest correlation with acceleration was jump height (r = -0.430, P = .041) and with maximum speed was peak power/weight (r = -0.649, P = .001). The results indicate that if an integrated system comprising a position transducer and a force platform is available for CMJ assessment, jump height and peak power/weight are useful variables to describe leg muscle explosive function for athletes who perform sprints.


Connexin 43 expression predicts poor progression-free survival in patients with non-muscle invasive urothelial bladder cancer.
To evaluate the protein expression of connexin 43 (Cx43) in primary urothelial bladder cancer and test its association with the histopathological characteristics and clinical outcome. A tissue microarray containing 348 tissue samples from 174 patients with primary urothelial carcinomas of the bladder was immunohistochemically stained for Cx43. The intensity of staining was semiquantitatively evaluated (score 0, 1+, 2+), and the association with clinicopathological features was assessed. Univariable and multivariable analyses were performed to identify predictors for progression-free survival (PFS). Membranous Cx43 immunoreactivity was detected in 118 (67.8%) of 174 analysable urothelial carcinomas, of which 31 (17.8%) showed even a strong (score 2+) and mainly homogeneous staining. Strong expression levels of Cx43 (score 2+) were associated with higher tumour grade, multiplicity and increased proliferation (all p<0.05). In the subgroup of patients with stage pTa and pT1 bladder tumours (n=158), strong Cx43 expression (p<0.001), solid growth pattern (p<0.001) and increased Ki-67 proliferation fraction (p<0.05) were significantly associated with shorter PFS in an univariable Cox regression analysis. In multivariable Cox regression models, Cx43 immunoreactivity and histological growth pattern remained highly significant and adverse risk factors for PFS. The expression levels of Cx43 are frequent in non-muscle invasive bladder cancer (NMIBC), with high expression levels being associated with poor prognosis. Routine assessment of Cx43 expression may improve the identification of high-risk NMIBC.


The genetic variants of solute carrier family 11 member 2 gene and risk of developing type-2 diabetes.
Type-2 diabetes (T2DM) is a metabolic disorder characterized by long-terminsulin resistance, impaired insulin secretion from β-cells, and loss of beta cell mass and function. Inflammation and oxidative stress play a key role in the development of diabetes and are associated with insulin resistance. Notably, recent studies have demonstrated an association between body iron stores, insulin resistance and T2DM. Free iron, a powerful pro-oxidant molecule, is involved in oxidative stress, lipid peroxidation and endothelial dysfunction via its ability to generate free radicals. Specifically, the accumulation of iron in beta cells triggers oxidative stress and DNA damage, which have been reported to be associated with β-cell death and apoptosis. Solute carrier family-11 member-2 (SLC11A2) functions to transport ferrous iron and some divalent metal ions throughout the plasmamembrane and across endosomalmembranes. Functional polymorphisms in the SLC11A2 gene have been reported to cause excess storage of iron, resulting in iron overload. In this study, we evaluated the association between T2DM and SLC11A2 gene variants IVS4+44C/A, 1303C/A and 1254T/C by performing PCR-RFLP analysis on 100 T2DM patients and 100 healthy subjects. PCR products were digested with MnlI, MboI and SfanI restriction endonucleases and the products were then separated by 3% agarose gel electrophoresis. The genotype frequencies of the 1254T/C and 1303C/A SLC11A2 gene variants did not differ between healthy controls and T2DM patients (P > 0.05). But, inrecessive model (P = 0.037) and homozygous CC genotype (P = 0.030) for IVS4+44C/A showed significant correlation with T2DM risk. It is thought that presence of C allele of IVS4+44C/A plays pathological roles.


Disrupted Network Topology in Patients with Stable and Progressive Mild Cognitive Impairment and Alzheimer's Disease.
Recent findings suggest that Alzheimer's disease (AD) is a disconnection syndrome characterized by abnormalities in large-scale networks. However, the alterations that occur in network topology during the prodromal stages of AD, particularly in patients with stable mild cognitive impairment (MCI) and those that show a slow or faster progression to dementia, are still poorly understood. In this study, we used graph theory to assess the organization of structural MRI networks in stable MCI (sMCI) subjects, late MCI converters (lMCIc), early MCI converters (eMCIc), and AD patients from 2 large multicenter cohorts: ADNI and AddNeuroMed. Our findings showed an abnormal global network organization in all patient groups, as reflected by an increased path length, reduced transitivity, and increased modularity compared with controls. In addition, lMCIc, eMCIc, and AD patients showed a decreased path length and mean clustering compared with the sMCI group. At the local level, there were nodal clustering decreases mostly in AD patients, while the nodal closeness centrality detected abnormalities across all patient groups, showing overlapping changes in the hippocampi and amygdala and nonoverlapping changes in parietal, entorhinal, and orbitofrontal regions. These findings suggest that the prodromal and clinical stages of AD are associated with an abnormal network topology.


Comparison of carbohydrate and milk-based beverages on muscle damage and glycogen following exercise.
This study examined effects of carbohydrate (CHO), milk-based carbohydrate-protein (CHO-PRO), or placebo (P) beverages on glycogen resynthesis, muscle damage, inflammation, and muscle function following eccentric resistance exercise. Untrained males performed a cycling exercise to reduce muscle glycogen 12 hours prior to performance of 100 eccentric quadriceps contractions at 120% of 1-RM (day 1) and drank CHO (n = 8), CHO-PRO (n = 9; 5 kcal/kg), or P (n = 9) immediately and 2 hours post-exercise. At 3 hours post-eccentric exercise, serum insulin was four times higher for CHO-PRO and CHO than P (p < .05). Serum creatine kinase (CK) increased for all groups in the 6 hours post-eccentric exercise (p < .01), with the increase tending to be lowest for CHO-PRO (p < .08) during this period. Glycogen was low post-exercise (33+/-3.7 mmol/kg ww), increased 225% at 24 hours, and tripled by 72 hours, with no group differences. The eccentric exercise increased muscle protein breakdown as indicated by urinary 3-methylhistidine and increased IL-6 with no effect of beverage. Quadriceps isokinetic peak torque was depressed similarly for all groups by 24% 24 hours post-exercise and remained 21% lower at 72 hours (p < .01). In summary, there were no influences of any post-exercise beverage on muscle glycogen replacement, inflammation, or muscle function.


GIPR Agonism Inhibits PYY-Induced Nausea-Like Behavior.
The induction of nausea and emesis is a major barrier to maximizing the weight loss profile of obesity medications, and therefore, identifying mechanisms that improve tolerability could result in added therapeutic benefit. The development of peptide YY (PYY)-based approaches to treat obesity are no exception, as PYY receptor agonism is often accompanied by nausea and vomiting. Here, we sought to determine whether glucose-dependent insulinotropic polypeptide (GIP) receptor (GIPR) agonism reduces PYY-induced nausea-like behavior in mice. We found that central and peripheral administration of a GIPR agonist reduced conditioned taste avoidance (CTA) without affecting hypophagia mediated by a PYY analog. The receptors for GIP and PYY (Gipr and Npy2r) were found to be expressed by the same neurons in the area postrema (AP), a brainstem nucleus involved in detecting aversive stimuli. Peripheral administration of a GIPR agonist induced neuronal activation (cFos) in the AP. Further, whole-brain cFos analyses indicated that PYY-induced CTA was associated with augmented neuronal activity in the parabrachial nucleus (PBN), a brainstem nucleus that relays aversive/emetic signals to brain regions that control feeding behavior. Importantly, GIPR agonism reduced PYY-mediated neuronal activity in the PBN, providing a potential mechanistic explanation for how GIPR agonist treatment reduces PYY-induced nausea-like behavior. Together, the results of our study indicate a novel mechanism by which GIP-based therapeutics may have benefit in improving the tolerability of weight loss agents.


Exploring the conformational transitions of biomolecular systems using a simple two-state anisotropic network model.
Biomolecular conformational transitions are essential to biological functions. Most experimental methods report on the long-lived functional states of biomolecules, but information about the transition pathways between these stable states is generally scarce. Such transitions involve short-lived conformational states that are difficult to detect experimentally. For this reason, computational methods are needed to produce plausible hypothetical transition pathways that can then be probed experimentally. Here we propose a simple and computationally efficient method, called ANMPathway, for constructing a physically reasonable pathway between two endpoints of a conformational transition. We adopt a coarse-grained representation of the protein and construct a two-state potential by combining two elastic network models (ENMs) representative of the experimental structures resolved for the endpoints. The two-state potential has a cusp hypersurface in the configuration space where the energies from both the ENMs are equal. We first search for the minimum energy structure on the cusp hypersurface and then treat it as the transition state. The continuous pathway is subsequently constructed by following the steepest descent energy minimization trajectories starting from the transition state on each side of the cusp hypersurface. Application to several systems of broad biological interest such as adenylate kinase, ATP-driven calcium pump SERCA, leucine transporter and glutamate transporter shows that ANMPathway yields results in good agreement with those from other similar methods and with data obtained from all-atom molecular dynamics simulations, in support of the utility of this simple and efficient approach. Notably the method provides experimentally testable predictions, including the formation of non-native contacts during the transition which we were able to detect in two of the systems we studied. An open-access web server has been created to deliver ANMPathway results.


Engineering Synthetic Lipopeptide Antigen for Specific Detection of Mycobacterium avium subsp. paratuberculosis Infection.
Unlike other MAC members, Mycobacterium avium subsp. paratuberculosis (MAP) does not produce glycopeptidolipids (GPL) on the surface of the cell wall but a lipopentapeptide called L5P (also termed Lipopeptide-I or Para-LP-01) characterized in C-type (bovine) strains. This lipopeptide antigen contains a pentapeptide core, D-Phenylalanine-N-methyl-L-Valine-L-Isoleucine-L-Phenylalanine-L-Alanine, in which the N-terminal D-Phenylalanine is amido-linked with a fatty acid (C18-C20). The molecular and genetic characterization of this antigen demonstrated that L5P is unique to MAP. Knowledge of the structure of L5P enabled synthetic production of this lipopeptide in large quantities for immunological evaluation. Various studies described the immune response directed against L5P and confirmed its capability for detection of MAP infection. However, the hydrophobic nature of lipopeptide antigens make their handling and use in organic solvents unsuitable for industrial processes. The objectives of this study were to produce, by chemical synthesis, a water-soluble variant of L5P and to evaluate these compounds for the serological diagnosis of MAP using well-defined serum banks. The native L5P antigen and its hydrosoluble analog were synthesized on solid phase. The pure compounds were evaluated on collections of extensively characterized sera from infected and non-infected cattle. ROC analysis showed that L5P and also its water-soluble derivative are suitable for the development of a serological test for Johne's disease at a population level. However, these compounds used alone in ELISA have lower sensitivity (Se 82% for L5P and Se 62% for the water-soluble variant of L5P) compared to the Se 98% of a commercial test. Advantageously, these pure synthetic MAP specific antigens can be easily produced in non-limiting quantities at low cost and in standardized batches for robust studies. The fact that L5P has not been validated in the context of ovine paratuberculosis highlights the need to better characterize the antigens expressed from the different genetic lineages of MAP to discover new diagnostic antigens. In the context of infections due to other mycobacteria such as M. bovis or the more closely related species M. avium subsp. hominissuis, the L5P did not cross react and therefore may be a valuable antigen to solve ambiguous results in other tests.


Unmet Needs for Ancillary Services Among Men Who Have Sex with Men and Who Are Receiving HIV Medical Care - United States, 2013-2014.
Gay, bisexual, and other men who have sex with men (MSM) are disproportionately affected by human immunodeficiency virus (HIV) in the United States (1). Ancillary services, defined as services that support retention in HIV medical care and assist with day-to-day living, can improve the health of HIV-infected MSM and help them achieve viral suppression (2). To assess the unmet needs for ancillary services among MSM receiving outpatient HIV medical care during 2013-2014, CDC used data from the Medical Monitoring Project (MMP), a surveillance system designed to assess clinical and behavioral characteristics of adults receiving HIV care, to obtain nationally representative estimates of, and identify reasons for, unmet needs (3). Based on self-reported needs of persons responding to the MMP survey, the most prevalent unmet needs were for non-HIV medical care services: approximately 23% had an unmet need for dental care, and 19% had an unmet need for eye or vision care. Unmet needs were most prevalent among young, non-Hispanic black, and Hispanic/Latino MSM. State and local health departments, community-based organizations, and health care providers might improve the health of MSM living with HIV by promoting access to ancillary services using strategies that increase patient awareness of how to obtain these services, especially among young, non-Hispanic black, and Hispanic/Latino MSM.


Nutritional lipid supply can control the heat shock response of B16 melanoma cells in culture.
The in vitro culture of cells offers an extremely valuable method for probing biochemical questions and many commonly-used protocols are available. For mammalian cells a source of lipid is usually provided in the serum component. In this study we examined the question as to whether the nature of the lipid could become limiting at high cell densities and, therefore, prospectively influence the metabolism and physiology of the cells themselves. When B16 mouse melanoma cells were cultured, we noted a marked decrease in the proportions of n-3 and n-6 polyunsaturated fatty acids (PUFAs) with increasing cell density. This was despite considerable quantities of these PUFAs still remaining in the culture medium and seemed to reflect the preferential uptake of unesterified PUFA rather than other lipid classes from the media. The reduction in B16 total PUFA was reflected in changes in about 70% of the molecular species of membrane phosphoglycerides which were analysed by mass spectrometry. The importance of this finding lies in the need for n-3 and n-6 PUFA in mammalian cells (which cannot synthesize their own). Although the cholesterol content of cells was unchanged the amount of cholesterol enrichment in membrane rafts (as assessed by fluorescence) was severely decreased, simultaneous with a reduced heat shock response following exposure to 42°C. These data emphasize the pivotal role of nutrient supply (in this case for PUFAs) in modifying responses to stress and highlight the need for the careful control of culture conditions when assessing cellular responses in vitro.


Assessing healthcare providers' knowledge and practices relating to insecticide-treated nets and the prevention of malaria in Ghana, Laos, Senegal and Tanzania.
Research evidence is not always being disseminated to healthcare providers who need it to inform their clinical practice. This can result in the provision of ineffective services and an inefficient use of resources, the implications of which might be felt particularly acutely in low- and middle-income countries. Malaria prevention is a particularly compelling domain to study evidence/practice gaps given the proven efficacy, cost-effectiveness and disappointing utilization of insecticide-treated nets (ITNs). This study compares what is known about ITNs to the related knowledge and practices of healthcare providers in four low- and middle-income countries. A new questionnaire was developed, pilot tested, translated and administered to 497 healthcare providers in Ghana (140), Laos (136), Senegal (100) and Tanzania (121). Ten questions tested participants' knowledge and clinical practice related to malaria prevention. Additional questions addressed their individual characteristics, working context and research-related activities. Ordinal logistic regressions with knowledge and practices as the dependent variable were conducted in addition to descriptive statistics. The survey achieved a 75% response rate (372/497) across Ghana (107/140), Laos (136/136), Senegal (51/100) and Tanzania (78/121). Few participating healthcare providers correctly answered all five knowledge questions about ITNs (13%) or self-reported performing all five clinical practices according to established evidence (2%). Statistically significant factors associated with higher knowledge within each country included: 1) training in acquiring systematic reviews through the Cochrane Library (OR 2.48, 95% CI 1.30-4.73); and 2) ability to read and write English well or very well (OR 1.69, 95% CI 1.05-2.70). Statistically significant factors associated with better clinical practices within each country include: 1) reading scientific journals from their own country (OR 1.67, 95% CI 1.10-2.54); 2) working with researchers to improve their clinical practice or quality of working life (OR 1.44, 95% CI 1.04-1.98); 3) training on malaria prevention since their last degree (OR 1.68, 95% CI 1.17-2.39); and 4) easy access to the internet (OR 1.52, 95% CI 1.08-2.14). Improving healthcare providers' knowledge and practices is an untapped opportunity for expanding ITN utilization and preventing malaria. This study points to several strategies that may help bridge the gap between what is known from research evidence and the knowledge and practices of healthcare providers. Training on acquiring systematic reviews and facilitating internet access may be particularly helpful.


A multinational, drug utilization study to investigate the use of dexmedetomidine (Dexdor®) in clinical practice in the EU.
Dexmedetomidine (dexdor®) is approved in the European Union (EU) for sedation of adults in the intensive care unit (ICU). The present observational, retrospective study was requested by the European Medicines Agency to investigate dexmedetomidine use in clinical practice, with a particular focus on off-label use, including the paediatric population. Study countries and sites were chosen from those with highest dexmedetomidine use, based on sales. Site selection (blind) was conducted by a multispecialist, independent group. Anonymized data on demographics, treatment indication, dexmedetomidine dosing, concomitant medications and treatment effectiveness were collected retrospectively from records of all dexmedetomidine-treated patients at the site during the enrolment period. Informed consent was waived, to avoid influencing the prescribing of dexmedetomidine. Recruitment was completed within 18 months of first site initiation. Data from 2000 patients were collected from 16 hospitals in four EU countries (Finland 750, Poland 505, Germany 470, Austria 275). The median age was 62 years, with more males (70.2%) than females. Dexmedetomidine was primarily used in the adult ICU (86.0%) for ICU sedation (78.6%) and mostly dosed according the product label. The intended sedative effect was obtained in 84.9% of administrations. Paediatric use (5.9% of patients, mostly in Austria and Finland) occurred mainly in the adult or paediatric ICU (75.6%) for sedation (67.2%). Overall, most patients were treated with dexmedetomidine according to the product labelling. Use in children was limited but significant and similar in scope to that in adults. Administrations not fully according to the product labelling usually occurred in an ICU environment and reflected extensively investigated clinical uses of dexmedetomidine.


Delivery of small interfering RNA by peptide-targeted mesoporous silica nanoparticle-supported lipid bilayers.
The therapeutic potential of small interfering RNAs (siRNAs) is severely limited by the availability of delivery platforms that protect siRNA from degradation, deliver it to the target cell with high specificity and efficiency, and promote its endosomal escape and cytosolic dispersion. Here we report that mesoporous silica nanoparticle-supported lipid bilayers (or "protocells") exhibit multiple properties that overcome many of the limitations of existing delivery platforms. Protocells have a 10- to 100-fold greater capacity for siRNA than corresponding lipid nanoparticles and are markedly more stable when incubated under physiological conditions. Protocells loaded with a cocktail of siRNAs bind to cells in a manner dependent on the presence of an appropriate targeting peptide and, through an endocytic pathway followed by endosomal disruption, promote delivery of the silencing nucleotides to the cytoplasm. The expression of each of the genes targeted by the siRNAs was shown to be repressed at the protein level, resulting in a potent induction of growth arrest and apoptosis. Incubation of control cells that lack expression of the antigen recognized by the targeting peptide with siRNA-loaded protocells induced neither repression of protein expression nor apoptosis, indicating the precise specificity of cytotoxic activity. In terms of loading capacity, targeting capabilities, and potency of action, protocells provide unique attributes as a delivery platform for therapeutic oligonucleotides.


A novel decatenation assay for DNA topoisomerases using a singly-linked catenated substrate.
Decatenation is a crucial in vivo reaction of DNA topoisomerases in DNA replication and is frequently used in in vitro drug screening. Usually this reaction is monitored using kinetoplast DNA as a substrate, although this assay has several limitations. Here we have engineered a substrate for Tn3 resolvase that generates a singly-linked catenane that can readily be purified from the DNA substrate after restriction enzyme digestion and centrifugation. We show that this catenated substrate can be used with high sensitivity in topoisomerase assays and drug-inhibition assays.


Identifying Behaviors Predicting Early Morning Emotions by Observing Permanent Supportive Housing Residents: An Ecological Momentary Assessment.
Behavior and emotions are closely intertwined. The relationship between behavior and emotions might be particularly important in populations of underserved people, such as people with physical or mental health issues. We used ecological momentary assessment (EMA) to examine the relationship between emotional state and other characteristics among people with a history of chronic homelessness who were participating in a health coaching program. The goal of this study was to identify relationships between daily emotional states (valence and arousal) shortly after waking and behavioral variables such as physical activity, diet, social interaction, medication compliance, and tobacco usage the prior day, controlling for demographic characteristics. Participants in m.chat, a technology-assisted health coaching program, were recruited from housing agencies in Fort Worth, Texas, United States. All participants had a history of chronic homelessness and reported at least one mental health condition. We asked a subset of participants to complete daily EMAs of emotions and other behaviors. From the circumplex model of affect, the EMA included 9 questions related to the current emotional state of the participant (happy, frustrated, sad, worried, restless, excited, calm, bored, and sluggish). The responses were used to calculate two composite scores for valence and arousal. Nonwhites reported higher scores for both valence and arousal, but not at a statistically significant level after correcting for multiple testing. Among momentary predictors, greater time spent in one-on-one interactions, greater time spent in physical activities, a greater number of servings of fruits and vegetables, greater time spent interacting in a one-on-one setting as well as adherence to prescribed medication the previous day were generally associated with higher scores for both valence and arousal, and statistical significance was achieved in most cases. Number of cigarettes smoked the previous day was generally associated with lower scores on both valence and arousal, although statistical significance was achieved for valence only when correcting for multiple testing. This study provides an important glimpse into factors that predict morning emotions among people with mental health issues and a history of chronic homelessness. Behaviors considered to be positive (eg, physical activity and consumption of fruits and vegetables) generally enhanced positive affect and restrained negative affect the following morning. The opposite was true for behaviors such as smoking, which are considered to be negative.


Featured Article: Comparison of Diabetes Management Trajectories in Hispanic versus White Non-Hispanic Youth with Type 1 Diabetes across Early Adolescence.
Ethnic minority youth with type 1 diabetes (T1D) often have poorer glycemic control and lower rates of adherence compared to White Non-Hispanic (WNH) youth. Variables such as family conflict, autonomy support, and youth regimen responsibility have been shown to change over adolescence and impact diabetes management. However, these factors have been investigated in predominantly White samples. Few studies have examined potential differences in these variables and their trajectories for Hispanic youth over early adolescence. Youth with T1D (178 WNH and 33 Hispanic youth participants), as well as their maternal caregivers (174 WNH and 32 Hispanic maternal caregivers), completed measures of diabetes-specific autonomy support, diabetes-related family conflict, regimen responsibility, and blood glucose monitoring frequency at 4 timepoints over a 3-year period. At baseline, Hispanic youth had significantly poorer glycemic control, more family conflict, and fewer blood glucose checks on average compared to WNH youth. Similar to WNH youth, Hispanic youth have increasing independence for regimen tasks and decreasing parent autonomy support during this developmental period. However, while Hispanic youth had worsening diabetes management during early adolescence (as did WNH youth), Hispanic parents reported a more gradual change in youth's diabetes management over early adolescence. This study presents an important contribution to the existing literature on youth with T1D. Findings suggest potential strengths and targets for Hispanic youth navigating diabetes management during the adolescent period. It is important to continue to investigate the trajectories of ethnic minority youth with diabetes.


Functionalised iron oxide nanoparticles for multimodal optoacoustic and magnetic resonance imaging.
The novel attachment of the optoacoustic (OA) molecules indocyanine green (ICG) and Flamma®774 to the core of an iron oxide (Fe3O4) nanoparticle has resulted in the facile synthesis of a multimodal imaging probe for both multispectral optoacoustic tomography (MSOT) imaging and magnetic resonance imaging (MRI). The nanoparticles have been analysed structurally, optically and magnetically to demonstrate the multimodal characteristics. The OA analysis of the dyes ICG and Flamma®774 showed that they have absorbance at the near IR wavelengths of 790 and 780 nm, respectively, when conjugated to an iron oxide core. These wavelengths are ideal for spectral unmixing of the probe intensity from any endogenous contrast, such as oxy-(HbO2) and deoxy-hemoglobin (Hb). MRI showed that citrate capped Fe3O4 exhibited a good r2 contrast of 230 mM-1 s-1, which is in line with literature values. Upon optoacoustic dye modification, the r2 relaxivity coefficient is comparable with that of Flamma®774 iron oxide nanoparticles (FeO-774) with r2 = 212 mM-1 s-1, showing that an OA dye attachment can have little to no effect on the MRI contrast. Indocyanine green functionalised iron oxide (FeO-ICG) nanoparticles showed an r2 contrast that was dramatically reduced with r2 = 5 mM-1 s-1. These results indicate that the facile synthesis of an effective dual modality MRI-MSOT probe can be developed using an iron oxide core and simple ligand coordination chemistry using an optoacoustic dye.


Frequency-specific electrocochleography indicates that presynaptic and postsynaptic mechanisms of auditory neuropathy exist.
The physiological mechanisms underlying auditory neuropathy (AN) remain unclear and it is likely that the multiple disruptions are classified under the broadly defined term. Cochlear implantation is being more widely used in this population to bypass the suspected site-of-lesion although a number of cases have been identified within the Sydney Cochlear Implant Centre where this management strategy has been unsuccessful. It is likely that this relates to the different physiological mechanisms underlying AN. To investigate the site-of-lesion in AN, frequency-specific round window electrocochleography (ECochG) was used to assess local hair-cell, dendritic, and axonal currents generated within the cochlea in 14 subjects with AN and compared with responses from two normally hearing subjects. ECochG results were then compared with electrically evoked auditory brain stem response (EABR) measured after cochlear implantation. The results of this study demonstrate that two dominant patterns of ECochG waveforms (produced by a high-frequency alternating tone burst) can be identified in this population of AN subjects: (a) gross waveform showing a prolonged summating potential (SP) latency that, in most cases, is followed by a small compound action potential; and (b) gross waveform showing a normal latency SP waveform followed by a broad negative potential [assumed to reflect the dendritic potential (DP) identified in anaesthetized guinea-pigs]. This study demonstrates that in most subjects (n = 7) with a prolonged latency SP but no DP, normal morphology EABR waveforms were elicited for all electrode channels. On the other hand, all subjects (n = 7) who showed a normal latency SP followed by a broad negative DP, showed EABR waveforms that were absent or having poor wave V morphology. The authors' interpretation of this is that ECochG results may provide a classification of AN into pre- and postsynaptic lesions. We suggest that a presynaptic and postsynaptic type of AN exist that may have implications for the fitting of cochlear implants.


Ganetespib blocks HIF-1 activity and inhibits tumor growth, vascularization, stem cell maintenance, invasion, and metastasis in orthotopic mouse models of triple-negative breast cancer.
Targeted therapy against triple-negative breast cancers, which lack expression of the estrogen, progesterone, and HER2 receptors, is not available and the overall response to cytotoxic chemotherapy is poor. One of the molecular hallmarks of triple-negative breast cancers is increased expression of genes that are transcriptionally activated by hypoxia-inducible factors (HIFs), which are implicated in many critical aspects of cancer progression including metabolism, angiogenesis, invasion, metastasis, and stem cell maintenance. Ganetespib is a second-generation inhibitor of heat shock protein 90 (HSP90), a molecular chaperone that is essential for the stability and function of multiple client proteins in cancer cells including HIF-1α. In this study, human MDA-MB-231 and MDA-MB-435 triple-negative breast cancer cells were injected into the mammary fat pad of immunodeficient mice that received weekly intravenous injections of ganetespib or vehicle following the development of palpable tumors. Ganetespib treatment markedly impaired primary tumor growth and vascularization, and eliminated local tissue invasion and distant metastasis to regional lymph nodes and lungs. Ganetespib treatment also significantly reduced the number of Aldefluor-positive cancer stem cells in the primary tumor. Primary tumors of ganetespib-treated mice had significantly reduced levels of HIF-1α (but not HIF-2α) protein and of HIF-1 target gene mRNAs encoding proteins that play key roles in angiogenesis, metabolism, invasion, and metastasis, thereby providing a molecular basis for observed effects of the drug on the growth and metastasis of triple-negative breast cancer. Triple-negative breast cancers (TNBCs) respond poorly to available chemotherapy. TNBCs overexpress genes regulated by hypoxia-inducible factors (HIFs). Ganetespib induces degradation of HSP90 client proteins, including HIF-1α. Ganetespib inhibited TNBC orthotopic tumor growth, invasion, and metastasis. Ganetespib inhibited expression of HIF-1 target genes involved in TNBC progression.


Conservation of pregnancy-specific glycoprotein (PSG) N domains following independent expansions of the gene families in rodents and primates.
Rodent and primate pregnancy-specific glycoprotein (PSG) gene families have expanded independently from a common ancestor and are expressed virtually exclusively in placental trophoblasts. However, within each species, it is unknown whether multiple paralogs have been selected for diversification of function, or for increased dosage of monofunctional PSG. We analysed the evolution of the mouse PSG sequences, and compared them to rat, human and baboon PSGs to attempt to understand the evolution of this complex gene family. Phylogenetic tree analyses indicate that the primate N domains and the rodent N1 domains exhibit a higher degree of conservation than that observed in a comparison of the mouse N1 and N2 domains, or mouse N1 and N3 domains. Compared to human and baboon PSG N domain exons, mouse and rat PSG N domain exons have undergone less sequence homogenisation. The high non-synonymous substitution rates observed in the CFG face of the mouse N1 domain, within a context of overall conservation, suggests divergence of function of mouse PSGs. The rat PSG family appears to have undergone less expansion than the mouse, exhibits lower divergence rates and increased sequence homogenisation in the CFG face of the N1 domain. In contrast to most primate PSG N domains, rodent PSG N1 domains do not contain an RGD tri-peptide motif, but do contain RGD-like sequences, which are not conserved in rodent N2 and N3 domains. Relative conservation of primate N domains and rodent N1 domains suggests that, despite independent gene family expansions and structural diversification, mouse and human PSGs retain conserved functions. Human PSG gene family expansion and homogenisation suggests that evolution occurred in a concerted manner that maintains similar functions of PSGs, whilst increasing gene dosage of the family as a whole. In the mouse, gene family expansion, coupled with local diversification of the CFG face, suggests selection both for increased gene dosage and diversification of function. Partial conservation of RGD and RGD-like tri-peptides in primate and rodent N and N1 domains, respectively, supports a role for these motifs in PSG function.


Reimagining the peer-review system for translational health science journals.
Retractions of coronavirus disease 2019 (COVID-19) papers in high impact journals, such as The Lancet and the New England Journal of Medicine, have been panned as major scientific fraud in public media. The initial reaction to this news was to seek out scapegoats and blame individual authors, peer-reviewers, editors, and journals for wrong doing. This paper suggests that scapegoating a few individuals for faulty science is a myopic approach to the more profound problem with peer-review. Peer-review in its current limited form cannot be expected to adequately address the scope and complexity of large interdisciplinary science research collaboration, which is central in translational research. In addition, empirical studies on the effectiveness of traditional peer-review reveal its very real potential for bias and groupthink; as such, expectations regarding the capacity and effectiveness of the current peer review process are unrealistic. This paper proposes a new vision of peer-review in translational science that, on the one hand, would allow for early release of a manuscript to ensure expediency, whereas also creating a forum or a collective of various experts to actively comment, scrutinize, and even build on the research under review. The aim would be to not only generate open discussion and oversight respecting the quality and limitations of the research, but also to assess the extent and the means for that knowledge to translate into social benefit.


Bovine herpesvirus 1 productive infection stimulates inflammasome formation and caspase 1 activity.
Bovine herpesvirus 1 (BoHV-1), a significant viral pathogen of cattle, causes inflammation in affected tissue during acute infection. Consequently, we tested whether productively infected bovine cells stimulate inflammasome formation. Expression of two components required for inflammasome formation, the DNA sensor IFI16 (gamma-interferon-inducible protein 16) and NLRP3 (NOD-like receptor family, pyrin domain containing 3), were induced in bovine kidney cells by eight hours after infection. IFI16 was detected in punctate granules localized to the cytoplasm and nucleus. During productive infection, more than ten times more cells were caspase 1 positive, which is activated following inflammasome formation. Two caspase 1 inhibitors had no effect on productive infection. Conversely, another caspase 1 inhibitor, glyburide, significantly inhibited virus infection suggesting it had off-target effects on related enzymes or interfered with infection via non-enzymatic mechanisms. Collectively, these studies demonstrated that BoHV-1 infection stimulated inflammasome formation, which we predict is important for clinical symptoms in cattle.


Measurement of erythrocyte aggregation in a microchip stirring system by light transmission.
In the analysis of red blood cell (RBC) aggregation using optical detection, various shearing methods have been used to disperse RBCs in confined geometries. This study investigated RBC aggregation measurement in a microchip-stirring system by analysis of light transmission. A stirring-aided disaggregation mechanism in a microchip, consisting of a flat-cylindrical test chamber (D=4 mm, H=0.3 mm) and a magnetic stirrer (d=0.14 mm, l=2.2 mm), was used to generate a given shear which was large enough to disperse RBC aggregates, but not large enough to cause any mechanical hemolysis of cells. After stirring for 10 s followed by an abrupt halt of the stirring, the intensity of the light transmitted through a microchip was measured with respect to time and analyzed. A comparative study was conducted with varying test chamber height and hematocrit. The AI and t1/2 as typical aggregation indices obtained by analysis of transmitted light, which showed a good reproducibility (coefficient of variation (CV)<2.8%, n=10), also were found to be nearly independent of the chamber dimensions (CV<3.4%). The present aggregometry also showed the similar results of aggregation indices with varying hematocrits compared to those obtained using a laser-assisted optical rotational cell analyzer (LORCA). The essential feature of the present design is the adoption of a disposable microchip requiring a minimum blood sample volume as small as 6 mul, which enables it to be used easily in a clinical setting.


Insulin resistance and inflammation as precursors of frailty: the Cardiovascular Health Study.
Our research group has previously shown that the geriatric syndrome of frailty is associated with features of the metabolic syndrome (MetS) on cross-sectional analysis. To test whether MetS and its physiologic determinants-insulin resistance as measured by homeostasis model assessment score (IR-HOMA), increased inflammation and coagulation factor levels, and elevated blood pressure-are associated with incident frailty, we studied a subcohort of participants from the Cardiovascular Health Study observed from 1989/1990 through 1998/1999: 3141 community-dwelling adults, aged 69 to 74 years, without frailty and illnesses that increase inflammation markers or mimic frailty. The association of baseline MetS, IR-HOMA, levels of inflammation and coagulation factors, and systolic blood pressure (SBP) with time to onset of frailty was adjusted for demographic and psychosocial factors and incident events. Our main outcome measure was incident frailty. Metabolic syndrome was not significantly associated with incident frailty (hazard ratio, 1.16 (95% confidence interval [CI], 0.85-1.57). On the other hand, IR-HOMA and C-reactive protein levels were associated with incident frailty: for every standard deviation increment the hazard ratio for frailty was 1.15 (95% CI, 1.02-1.31) and 1.16 (95% CI, 1.02-1.32), respectively. The white blood cell count and factor VIIIc levels had a borderline association. Elevated systolic blood pressure had no association. Similar trends were found for incident prefrailty, a condition that precedes frailty. Two physiologic components of MetS- IR-HOMA and inflammation-are associated with incident frailty. Based on these results, IR-HOMA can be considered part of a larger process that leads to generalized decline.


Cardiolipin-protein complexes and initiation of complement activation after coronary artery occlusion.
Specific rabbit anti-cardiolipin (anti-CL) antibodies were used to investigate the hypothesis that cardiolipin, associated with mitochondrial membrane proteins, binds C1 and facilitates activation of the complement cascade following reperfusion of ischemic myocardium. By immunoelectron microscopy, anti-CL localized to subsarcolemmal mitochondria, emerging through breaks in membranes of damaged cardiac myocytes. Anti-CL reacted with > 15 mitochondrial constituents, most of which comigrated with the proteins that bind C1q in transblots of subsarcolemmal mitochondria, fractionated by polyacrylamide gel electrophoresis under reducing conditions in the presence of sodium dodecyl sulfate. A subset of the C1q-binding proteins > 24 to 37 kDa served as stable sites for assembly of C3, C5, and C9. Cardiac lymph, collected during the first hour after reperfusion of ischemic myocardium, contained proteins of diverse size that reacted with both anti-CL and C1q. Cardiac lymph, collected before occlusion and 4 to 5 hours after reperfusion, in comparison, had few if any C1q or anti-CL reactive proteins. Treatment with phospholipase suppressed the C1q-binding activity and anti-CL reactivity of the proteins in reperfusion lymph and those with similar properties in mitochondrial extracts. Our data suggest that during ischemia, mitochondria, extruded through breaks in the sarcolemma, unfold and release membrane fragments in which cardiolipin and protein are intimately associated. By binding C1 and supplying sites for the assembly of later-acting complement components, these fragments provide the means to disseminate the complement-mediated inflammatory response to ischemic injury.


A foreign invader or a reclusive native? DNA bar coding reveals a distinct European lineage of the zoonotic parasite Schistosoma turkestanicum (syn. Orientobilharzia turkestanicum ()).
Natural foci of Schistosoma turkestanicum (syn. Orientobilharzia turkestanicum) has been identified in the Gemenc Forest regions of Hungary utilising red deer as the definitive host. In order to identify the origins of this parasite in Europe standard DNA bar coding techniques were employed to sequence fragments of the cytochrome oxidase 1 (cox1) and the nuclear ribosomal internal transcribed region (ITS) from 10 individual adult male worms. Phylogenetic reconstruction using maximum likelihood phylogenetic reconstruction and haplotype networks of the cox1 showed all the worms to be of a distinct unique Hungarian lineage although some ITS haplotypes were shared with worms from populations in China and Iran. Molecular clock analysis suggests an early divergence event around 270,000years before present (YBP) between all S. turkestanicum populations giving rise to the Chinese, Iranian and Hungarian lineages. However, divergence of the sequences within the Hungarian population appears to have occurred approximately 63,000 YBP suggesting a long established population of S. turkestanicum in Europe. This suggests that the Hungarian population of S. turkestanicum has been native since the Ice Age and probably established itself during the last interglacial period as red deer moved into Europe from North Africa and the Middle East. This may also indicate that the parasite may have unknown populations established in several other countries in Eastern, Central and Southern Europe.


Impacts of the Cretaceous Terrestrial Revolution and KPg extinction on mammal diversification.
Previous analyses of relations, divergence times, and diversification patterns among extant mammalian families have relied on supertree methods and local molecular clocks. We constructed a molecular supermatrix for mammalian families and analyzed these data with likelihood-based methods and relaxed molecular clocks. Phylogenetic analyses resulted in a robust phylogeny with better resolution than phylogenies from supertree methods. Relaxed clock analyses support the long-fuse model of diversification and highlight the importance of including multiple fossil calibrations that are spread across the tree. Molecular time trees and diversification analyses suggest important roles for the Cretaceous Terrestrial Revolution and Cretaceous-Paleogene (KPg) mass extinction in opening up ecospace that promoted interordinal and intraordinal diversification, respectively. By contrast, diversification analyses provide no support for the hypothesis concerning the delayed rise of present-day mammals during the Eocene Period.


Big concerns with small projects: Evaluating the socio-ecological impacts of small hydropower projects in India.
Although Small Hydropower Projects (SHPs) are encouraged as sources of clean and green energy, there is a paucity of research examining their socio-ecological impacts. We assessed the perceived socio-ecological impacts of 4 SHPs within the Western Ghats in India by conducting semi-structured interviews with local respondents. Primary interview data were sequentially validated with secondary data, and respondent perceptions were subsequently compared against the expected baseline of assured impacts. We evaluated the level of awareness about SHPs, their perceived socio-economic impacts, influence on resource access and impacts on human-elephant interactions. The general level of awareness about SHPs was low, and assurances of local electricity and employment generation remained largely unfulfilled. Additionally most respondents faced numerous unanticipated adverse impacts. We found a strong relationship between SHP construction and increasing levels of human-elephant conflict. Based on the disparity between assured and actual social impacts, we suggest that policies regarding SHPs be suitably revised.


Melatonin in humans.
Melatonin, the hormone of the pineal gland, received a great deal of attention in the last decade because of its availability as over-the-counter drug or food supplement in some countries and suggested role in many vital physiological processes. Melatonin secretion is not restricted to mammals but is also produced in nonmammalian vertebrates, in some invertebrates, and in many plants, with the same molecular structure. The synthesis of melatonin is strictly controlled by lighting conditions and shows a clear circadian rhythm with low values during the daytime and significant increase at night. In this survey the basic data on melatonin significance in human physiology and in pathological processes as well as its possible therapeutic significance are reviewed and discussed.


Nucleotide excision repair impairment by nodularin in CHO cell lines due to ERCC1/XPF inactivation.
The problem of toxicity of cyanobacterial toxins is of increasing concern, as the incidence of such blooms grows. Among the toxins, the most abundant in the environment are hepatotoxins known as nodularins and microcystins. These toxins are responsible for almost all known cases of fresh and brackish water intoxication and are responsible for recurrent episodes of human and animal illness and death. Moreover, they are believed to be potent tumor promoters and initiators. However, the mechanisms by which these toxins induce liver cancer are not well understood. The aim of the present study was to determine the effect of nodularin on the kinetics of nucleotide excision repair (NER) in Chinese hamster ovary (CHO) cells exposed to UV radiation. The first set of experiments was performed to define the optimal treatment conditions for nodularin to avoid the possibility of encountering false positive signals in the comet assay due to the apoptogenic activity of nodularin. Based on the analysis of apoptosis, the 6-h treatment time of cells with nodularin (1mug/ml, 10mug/ml and 20mug/ml) was chosen for the alkaline comet assay. The kinetics of NER was determined in CHO cell lines: AA8 (wild-type) and mutant cell lines: UV135 (XPG(-)), UV41 (XPF(-)) and UV20 (ERCC1(-)) exposed to 20J/m(2) UV radiation. The micronucleus assay was performed to determine a residual DNA damage in four cell lines treated with nodularin (10mug/ml) and exposed to equitoxic doses UV radiation. Radiation doses of UV producing 50% of survival for AA8, UV135, UV20 and UV41 cell lines were calculated from UV survival curves. The results show that nodularin impairs the incision/excision step of NER in CHO cells by the ERCC1/XPF inactivation and leads to an increased level of UV-induced cytogenetic DNA damage.


Pediatric cystic nephromas: distinctive features and frequent DICER1 mutations.
Cystic nephromas (CNs) are uncommon benign renal neoplasms that present with a bimodal age distribution, affecting either infants/young children or adult females. Although differences between these age groups have been suggested, large studies of pediatric CN have not been conducted. As a result, the nomenclature and diagnostic criteria for these lesions remain controversial. In addition, the morphological overlap seen between CN and cystic partially differentiated nephroblastoma (CPDN) can result in diagnostic dilemmas. This study reviews the morphologic and radiographic features of 44 pediatric CN prospectively enrolled on a Children's Oncology Group protocol from 2007 to 2013. Although the typical multicystic architecture with thin septa described in adult CN was present in all of our pediatric cases, differences were also identified. We report distinctive features that add to the morphological spectrum of CN in children. Of the 44 cases, 16 had been previously analyzed and reported for DICER1 mutation, and either loss of function or missense mutations or both were identified in 15 of 16. In contrast, we analyzed 10 cases of adult CN, and all were negative for DICER1 mutations; similarly, 6 CPDNs previously analyzed and reported were negative for DICER1 mutations. Therefore, the clinical, morphological, and genetic differences between pediatric and adult CN, as well as between CN and CPDN, suggest that these 3 lesions represent distinct entities.


Comparison of slow larval and fast adult muscle innervated by the same motor neurone.
1. Muscles innervated by an identified set of motor neurones were compared between larval and adult stages. 2. The structure of the larval muscle is typically tonic: long sarcomeres, irregular Z-bands, and 10-12 thin filaments around each thick filament. The structure of the adult muscle is phasic: 3-4 micrometers sarcomeres, regular Z-bands, 6-8 thin filaments around each thick filament, and large mitochondrial volume. 3. The tensions produced by these muscles were correspondingly different. The larval twitch was about 7 times slower and the tetanus/twitch ratio 10 times greater than those of the adult. 4. No structural or physiological differences were observed in the neuromuscular junctions of the two stages. 5. The relatively unchanging functional relationship of a single motor neurone with two different muscle fibre types during two developmental stages is compared with the converse situation in which it has been reported that implantation of a different type of motor nerve into a muscle modifies contractile properties.


The decrease of soluble RAGE levels in rheumatoid arthritis patients following hormone replacement therapy is associated with increased bone mineral density and diminished bone/cartilage turnover: a randomized controlled trial.
The aim of the study was to prospectively investigate the effects of HRT on serum soluble receptor for advanced glycation end product (sRAGE) levels in RA patients and to determine whether sRAGE production is related to bone/cartilage metabolism. Eighty-eight post-menopausal RA patients were randomized to receive vitamin D3 and calcium supplementation with or without HRT (oestradiol plus noretisterone acetate). The levels of total sRAGE in sera were measured before, 1 and 2 years after treatment initiation. Potential associations between sRAGE levels, bone/cartilage metabolic markers and BMD were investigated. Patients receiving HRT displayed significantly decreased levels of serum sRAGE at 1 and 2 years as compared with levels at study entry. The increase in serum oestradiol was associated with the decline in sRAGE levels. Importantly, sRAGE levels at baseline significantly correlated with bone/cartilage turnover markers including C-terminal propeptide of type I procollagen, carboxyterminal telopeptide of type I collagen and cartilage oligomeric matrix protein, and the decrease of sRAGE levels paralleled with diminished concentration of these molecules. BMD in hip and femoral neck and progression of Larsen score at 1 year were associated with baseline sRAGE levels. The decline in sRAGE levels significantly correlated with an increase in total BMD following 2 years of treatment in patients receiving HRT but not in the control group. Our findings suggest that HRT decreases the levels of endogenous sRAGE in post-menopausal RA patients implicating its role in sRAGE regulation. In addition, serum sRAGE was associated with BMD and markers of bone/cartilage metabolism. These data suggest that sRAGE is involved directly or indirectly in bone metabolism. Trial registration. Current Controlled Trials, ISRCTN46523456, http://www.controlled-trials.com/isrctn/search.html?srch=ISRCTN46523456.


Gulf of Mexico hypoxia: exploring increasing sensitivity to nitrogen loads.
Hypoxia is a critical issue in the Gulf of Mexico that has challenged management efforts in recent years by an increase in hypoxia sensitivity to nitrogen loads. Several mechanisms have been proposed to explain the recent increase in sensitivity. Two commonly cited mechanisms are bottom-water reducing conditions preventing nitrification and thus denitrification, leading to more N recycling and production of oxygen-consuming organic matter, and carryover of organic matter from previous years increasing oxygen demand, making the system more sensitive. We use models informed by these mechanisms and fit with Bayesian inference to explore changes in Gulf of Mexico hypoxia sensitivity. We show that a model including an annually fit parameter representing variation in the fraction of nutrient loading and recycling contributing to bottom water oxygen demand provides a good fit to observations and is not improved by explicit inclusion of organic matter carryover to subsequent years. Both models support two stepwise increases in system sensitivity during the period of record. This change in sensitivity has greatly increased the nutrient reduction needed to achieve the established hypoxia goal. If the Gulf remains at the current state of sensitivity, our analysis suggests a roughly 70% reduction of spring TN loads from the 1988-1996 average of 6083 ton/day may be required.


Healthful and less-healthful foods and drinks from storefront and non-storefront businesses: implications for 'food deserts', 'food swamps' and food-source disparities.
Conceptualisations of 'food deserts' (areas lacking healthful food/drink) and 'food swamps' (areas overwhelm by less-healthful fare) may be both inaccurate and incomplete. Our objective was to more accurately and completely characterise food/drink availability in urban areas. Cross-sectional assessment of select healthful and less-healthful food/drink offerings from storefront businesses (stores, restaurants) and non-storefront businesses (street vendors). Two areas of New York City: the Bronx (higher-poverty, mostly minority) and the Upper East Side (UES; wealthier, predominantly white). All businesses on 63 street segments in the Bronx (n 662) and on 46 street segments in the UES (n 330). Greater percentages of businesses offered any, any healthful, and only less-healthful food/drink in the Bronx (42·0 %, 37·5 %, 4·4 %, respectively) than in the UES (30 %, 27·9 %, 2·1 %, respectively). Differences were driven mostly by businesses (e.g. newsstands, gyms, laundromats) not primarily focused on selling food/drink - 'other storefront businesses' (OSBs). OSBs accounted for 36·0 % of all food/drink-offering businesses in the Bronx (more numerous than restaurants or so-called 'food stores') and 18·2 % in the UES (more numerous than 'food stores'). Differences also related to street vendors in both the Bronx and the UES. If street vendors and OSBs were not captured, the missed percentages of street segments offering food/drink would be 14·5 % in the Bronx and 21·9 % in the UES. Of businesses offering food/drink in communities, OSBs and street vendors can represent substantial percentages. Focusing on only 'food stores' and restaurants may miss or mischaracterise 'food deserts', 'food swamps', and food/drink-source disparities between communities.


Simplified steps to heterologous prime-boost HIV vaccine development?
The RV 144 HIV vaccine efficacy study showed a reduction in HIV-1 infection risk in Thai volunteers who received two priming vaccinations of vCP1521 ALVAC (attenuated recombinant canarypox virus expressing HIV group-specific antigen, polymerase, and envelope genes) followed by two additional ALVAC vaccinations and coadministration of purified bivalent gp120 proteins (AIDSVAX B/E). In this issue of the JCI, Rouphael et al. build on these results by substituting a DNA plasmid cocktail expressing HIV-1 subtype C group-specific antigen, polymerase, and envelope antigen genes (DNA-HIV-PT123) for ALVAC in a phase 1b safety and immunogenicity study. The results indicate that the vaccine regimen is safe, elicits promising cross-subtype humoral and cellular responses, and opens up potentially simplified approaches to HIV-1 vaccine development.


Viroid Prevalence in Tunisian Citrus.
The citrus industry in Tunisia is based mainly on the production of local cultivars of sweet orange (Citrus sinensis), common mandarin (C. reticulata), clementine (C. clementina), and lemon (C. limon). Sour orange (C. aurantium) is the only rootstock presently being used in the major growing area located at Cap Bon where 80% of citrus is being produced. The presence of tristeza disease in the Mediterranean basin is a threat to the Tunisian citrus industry, and new rootstocks giving tristeza tolerant rootstock/scion combinations are urgently needed as an alternative to sour orange. Since some promising rootstocks are known to be sensitive to viroids (1), a survey was conducted to determine if the cultivars grown presently in Tunisia were infected with viroids. Following a preliminary report (2), an extensive survey was conducted from 1995-2001 that included 174 symptomless sources being grown at Cap Bon: 26 Maltaise demi-sanguine, 9 Maltaise sanguine, 20 Maltaise blonde, 4 orange doublefine, 16 Washington navel, 12 Valencia late, 29 common mandarin, 42 Cassar clementine, 5 Lunari lemon, and 11 Eureka lemon. These sources were graft-inoculated into Etrog citron that subsequently developed symptoms characteristic of viroid infection. Sequential polyacrylamide gel electrophoresis analysis and molecular hybridization using viroid-specific probes (1) revealed that all sources were infected with at least two viroids. Citrus exocortis viroid (CEVd), Hop stunt viroid (HSVd), and Citrus viroid III (CVd-III) were widespread and accounted for 68.4, 67.8, and 81.0% of the sources tested, respectively. Citrus bent leaf viroid (CBLVd) and Citrus viroid IV (CVd-IV) were only found in 32.7 and 2.3% of the sources. The most frequent viroid combinations were CEVd+HSVd+CVd-III (17.8%) and CEVd+CVd-III (17,2%), whereas HSVd+CVd-IV and CEVd+CBLVd+CVd-III+CVd-IV were found in a single source (0.6%). References: (1) N. Duran-Vila and J. S. Semancik. Pages 178-194 in: Viroids. CSIRO Publishing, Australia, 2003. (2) A. Najar et al. Pages 398-400 in: Proc. 15th Conf. Int. Org. Citrus Virol, 2002.


Reconfigurable optical logic unit with a terahertz optical asymmetric demultiplexer and electro-optic switches.
A reconfigurable optical logic unit that can execute any binary logic and arithmetic operation on the same hardware with different configurations is proposed. The design, based on cascaded terahertz optical asymmetric demultiplexer switches, introduces reconfigurability with the help of electro-optic switches. The model is simple and practicable.


Molecular Transformer: A Model for Uncertainty-Calibrated Chemical Reaction Prediction.
Organic synthesis is one of the key stumbling blocks in medicinal chemistry. A necessary yet unsolved step in planning synthesis is solving the forward problem: Given reactants and reagents, predict the products. Similar to other work, we treat reaction prediction as a machine translation problem between simplified molecular-input line-entry system (SMILES) strings (a text-based representation) of reactants, reagents, and the products. We show that a multihead attention Molecular Transformer model outperforms all algorithms in the literature, achieving a top-1 accuracy above 90% on a common benchmark data set. Molecular Transformer makes predictions by inferring the correlations between the presence and absence of chemical motifs in the reactant, reagent, and product present in the data set. Our model requires no handcrafted rules and accurately predicts subtle chemical transformations. Crucially, our model can accurately estimate its own uncertainty, with an uncertainty score that is 89% accurate in terms of classifying whether a prediction is correct. Furthermore, we show that the model is able to handle inputs without a reactant-reagent split and including stereochemistry, which makes our method universally applicable.


Frequency of embryo multinucleation detected by time-lapse system and its impact on pregnancy outcome.
To compare the detection rate of multinucleation with the time-lapse system and conventional control timing proposed by European Society of Human Reproduction and Embryology (ESHRE) consensus and evaluate its impact on pregnancy rates. Retrospective study. A private IVF center. A total of 686 embryos from 511 intracytoplasmic sperm injection (ICSI) cycles. None. A time-lapse system was used to acquire embryo images until ET; the stored data were reviewed for the presence and persistence of multinucleation. The detection rate of multinucleation was compared with ESHRE/ALPHA consensus-proposed embryo evaluating times (23 ± 1, 26 ± 1, 44 ± 1 hours). Morphokinetic characteristics of multinucleated embryos and the effect of multinucleation on pregnancy rate were researched. Multinucleation was detected in 159 embryos of 145 ICSI cycles. Using ESHRE/ALPHA consensus embryo evaluating times, only 44 (27.6%) out of 159 multinucleated embryos could be identified. In cycles with multinucleated ETs compared with cycles with no multinucleated embryos, clinical pregnancy rates (respectively, 23.4 vs. 44) and implantation rates (respectively, 23.3 vs. 43.6) were significantly lower. Time to 2-cell, 4-cell, and 6-cell stage was significantly longer in multinucleated embryos. Patient age (odds ratio [OR], 0.95; confidence interval [CI], 0.92-0.98) and presence of multinucleation (OR, 0.37; CI, 0.24-0.56) were the only significant predictors of clinical pregnancy rate. The time-lapse monitoring system seems to be a valuable tool to identify all cases with multinucleation. We conclude that the detection of multinucleation by time-lapse monitoring is associated with lower implantation and clinical pregnancy rates.


Ocular cicatricial pemphigoid.
Cicatricial pemphigoid is an inflammatory disease of presumed autoimmune etiology. It most commonly affects the conjunctiva and oral mucosa and less commonly the skin. The ocular manifestations of the disease include bilateral conjunctival shrinkage, xerosis, and corneal opacification. The progression of cicatricial pemphigoid is variable and can be interrupted by periods of remission or by periods of rapid exacerbation. Ocular surgery and topical pharmaceuticals may contribute to the exacerbation of this disease. Current treatment consists of systemic immunosuppressants and systemic corticosteroids.


Role of cephalosporins in the era of Clostridium difficile infection.
The incidence of Clostridium difficile infection (CDI) in Europe has increased markedly since 2000. Previous meta-analyses have suggested a strong association between cephalosporin use and CDI, and many national programmes on CDI control have focused on reducing cephalosporin usage. Despite reductions in cephalosporin use, however, rates of CDI have continued to rise. This review examines the potential association of CDI with cephalosporins, and considers other factors that influence CDI risk. EUCLID (the EUropean, multicentre, prospective biannual point prevalence study of CLostridium difficile Infection in hospitalized patients with Diarrhoea) reported an increase in the annual incidence of CDI from 6.6 to 7.3 cases per 10 000 patient bed-days from 2011-12 to 2012-13, respectively. While CDI incidence and cephalosporin usage varied widely across countries studied, there was no clear association between overall cephalosporin prescribing (or the use of any particular cephalosporin) and CDI incidence. Moreover, variations in the pharmacokinetic and pharmacodynamic properties of cephalosporins of the same generation make categorization by generation insufficient for predicting impact on gut microbiota. A multitude of additional factors can affect the risk of CDI. Antibiotic choice is an important consideration; however, CDI risk is associated with a range of antibiotic classes. Prescription of multiple antibiotics and a long duration of treatment are key risk factors for CDI, and risk also differs across patient populations. We propose that all of these are factors that should be taken into account when selecting an antibiotic, rather than focusing on the exclusion of individual drug classes.


Isolation and Antibiogram of Clostridium tetani from Clinically Diagnosed Tetanus Patients.
Clostridium tetani, the etiologic agent of tetanus, produces a toxin that causes spastic paralysis in humans and other vertebrates. This study was aimed for isolation, identification, and determination of antimicrobial susceptibility of C. tetani from clinically diagnosed tetanus patients. Isolation was done from deep-punctured tissues of the foot and arm injuries of 80 clinically diagnosed tetanus patients from the Pakistan Institute of Medical Sciences hospital. We successfully screened out five C. tetani isolates out of 80 samples based on the strain-specific characteristics confirmed through biochemical testing and toxin production. A disc diffusion method was used for antimicrobial susceptibilities and C. tetani isolates showed susceptibility to cefoperazone, chloramphenicol, metronidazole, penicillin G, and tetracycline, but were found to be resistant to erythromycin and ofloxacin. During animal testing, all the infected mice developed symptoms of tetanus. The results showed that identification of C. tetani is possible using biochemical and molecular tools and that the strains of C. tetani isolated had not developed resistance against the antibiotics most often used for the treatment of tetanus.


Brain-derived neurotrophic factor Val⁶⁶Met polymorphism affects resting regional cerebral blood flow and functional connectivity differentially in women versus men.
The human Val⁶⁶Met single nucleotide polymorphism in the brain-derived neurotrophic factor (BDNF) gene impacts BDNF signaling at the cellular level. At the neural-systems level, it is associated with differences in prefrontal cortex (PFC) and hippocampal function during performance of cognitive and affective tasks. Because the impact of this variant on basal prefrontal and hippocampal activity is not known but may be relevant to understanding the function of this gene in health and disease, we studied 94 healthy individuals with H₂ ¹⁵O PET to assess regional cerebral blood flow (rCBF) during rest and tested for between-genotype differences. Because BDNF and gonadal steroid hormones conjointly influence neuronal growth, survival, and plasticity in hippocampus and PFC, we also tested for sex × genotype interactions. Finally, in light of the known impact of BDNF on plasticity and dendritic arborization, we complimented direct rCBF comparisons with connectivity analyses to determine how activity in hippocampal and prefrontal regions showing between-genotype group differences covaries with rCBF in other nodes throughout the brain in a genotype- or sex-dependent manner. Compared with Val homozygotes, Met carriers had higher rCBF in prefrontal (BA25 extending into BA10) and hippocampal/parahippocampal regions. Moreover, there were significant sex × genotype interactions in regions (including frontal, parahippocampal, and lateral temporal cortex) in which Val homozygotes showed higher rCBF in females than males, but Met carriers showed the opposite relationship. Functional connectivity analysis demonstrated that correlations of BA25, hippocampus, and parahippocampus with frontal and temporal networks were positive for Val homozygotes and negative for Met carriers. In addition, sex × genotype analysis of functional connectivity revealed that genotype affected directionality of the inter-regional correlations differentially in men versus women. Our data indicate that BDNF allelic variation and sex interactively affect basal prefrontal and hippocampal function.


Hyperthermic intraperitoneal chemotherapy in interval debulking surgery for advanced epithelial ovarian cancer: A single-center, real-life experience.
An improvement in survival without increasing perioperative morbidity in patients with advanced epithelial ovarian cancer treated with hyperthermic intraperitoneal chemotherapy (HIPEC) after interval debulking surgery (IDS) has been recently demonstrated in a randomized controlled trial. This study was aimed at assessing the feasibility and perioperative outcomes of the use of HIPEC after IDS at a referral cancer center. Over the study period, 149 IDSs were performed. Patients who had at least International Federation of Gynecology and Obstetrics stage III disease, with <2.5 mm of residual disease (RD) at the end of surgery and were not participating in clinical trials received HIPEC. Moreover, specific exclusion criteria were considered. These patients were compared with 51 patients with similar clinical characteristics at the same institution and within the same timeframe who did not receive HIPEC. No differences in patient or disease characteristics with the exception of the type of neoadjuvant chemotherapy (P = .002) were found between the 2 groups. As for surgical characteristics, significant differences were found in RD after IDS (P = .007) and in the duration of surgery (P < .001), whereas the bowel resection and diversion rates (P = .583 and P = .213, respectively) and the postoperative intensive care unit and hospital stays (P = .567 and P = .727, respectively) were comparable. The times to start adjuvant chemotherapy were also similar (P = .998). Equally, the rates of any grade of both intraoperative complications (P = .189) and early postoperative complications (P = .238) were superimposable. In the authors' experience, the addition of HIPEC to IDS is feasible in 35% for the population. This value might increase with changes in the inclusion/exclusion criteria. HIPEC does not increase perioperative complications and does not affect a patient's recovery or time to start adjuvant chemotherapy. HIPEC should be offered to select patients listed for IDS.


Antioxidant activities of dioscorin, the storage protein of yam (Dioscorea batatas Decne) tuber.
Dioscorin, the storage protein of yam (Dioscorea batatas Decne) tuber (which is different from dioscorine found in tubers of Dioscorea hirsuta), was purified to homogeneity after DE-52 ion exchange column according to the methods of Hou et al. (J. Agric. Food Chem. 1999, 47, 2168-2172). A single band of 32 kDa dioscorin was obtained on a sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel with 2-mercaptoethanol treatment. This purified dioscorin was shown by spectrophotometric method to have scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical in a pH-dependent manner. There is a positive correlation between scavenging effects against DPPH (8-46%) and amounts of 32 kDa dioscorin (5.97-47.80 nmol) added in Tris-HCl buffer (pH 7.9), which are comparable to those of glutathione at the same concentrations. Using electron paramagnetic resonance (EPR) spectrometry for DPPH radical detection, it was found that the intensities of the EPR signal were decreased by 28.6 and 57 nmol of 32 kDa dioscorin in Tris-HCl buffer (pH 7.9) more than in distilled water compared to controls. EPR spectrometry was also used for hydroxyl radical detection. It was found that 32 kDa dioscorin could capture hydroxyl radical, and the intensities of the EPR signal were significantly decreased dose-dependently by 1.79-14.32 nmol of 32 kDa dioscorin (r = 0.975) compared to the control. It is suggested that 32 kDa dioscorin, the storage protein of yam tuber, may play a role as antioxidant in tubers and may be beneficial for health when people take it as a food additive or consume yam tubers.


Transmission potential and severity of COVID-19 in South Korea.
Since the first case of 2019 novel coronavirus (COVID-19) identified on Jan 20, 2020, in South Korea, the number of cases rapidly increased, resulting in 6284 cases including 42 deaths as of Mar 6, 2020. To examine the growth rate of the outbreak, we present the first study to report the reproduction number of COVID-19 in South Korea. The daily confirmed cases of COVID-19 in South Korea were extracted from publicly available sources. By using the empirical reporting delay distribution and simulating the generalized growth model, we estimated the effective reproduction number based on the discretized probability distribution of the generation interval. We identified four major clusters and estimated the reproduction number at 1.5 (95% CI: 1.4-1.6). In addition, the intrinsic growth rate was estimated at 0.6 (95% CI: 0.6, 0.7), and the scaling of growth parameter was estimated at 0.8 (95% CI: 0.7, 0.8), indicating sub-exponential growth dynamics of COVID-19. The crude case fatality rate is higher among males (1.1%) compared to females (0.4%) and increases with older age. Our results indicate an early sustained transmission of COVID-19 in South Korea and support the implementation of social distancing measures to rapidly control the outbreak.