Auricular leishmaniasis mimicking squamous cell carcinoma.
We report a rare case of auricular involvement by leishmaniasis, in order to demonstrate the importance of thorough investigation of cutaneous head and neck lesions, and also the importance of inclusion of infections such as leishmaniasis in the differential diagnosis of auricular lesions, especially in endemic areas. A 42-year-old man with multiple lesions on his head, neck and hands was referred to our centre. He had the following lesions: a painful, crusted, 8 x 8 cm plaque with indurated margins on the left parotid region and auricle; a red papule on the right temporal region; an ulcerative lesion on the skin overlying the proximal interphalangeal joint of the fifth finger of the right hand; and a bluish papule on the neck. Although histopathological examination of the Geimsa-stained specimen was misleading, a direct smear prepared from biopsies showed amastigotes, and therapy resulted in complete recovery. Leishmaniasis can be both under- or over-diagnosed. Especially in endemic areas, parasitic causes of chronic infections should always be kept in mind.


COVID-19 Infection and Death Rates Varied Widely Among U.S. States.
Socioeconomic and political reasons were cited for the differing outcomes.


Structure-directed combinatorial library design.
In recent years pharmaceutical companies have utilized structure-based drug design and combinatorial library design techniques to speed up their drug discovery efforts. Both approaches are routinely used in the lead discovery and lead optimization stages of the drug discovery process. Fragment-based drug design, a new power tool in the drug design toolbox, is also gaining acceptance across the pharmaceutical industry. This review will focus on the interplay between these three design techniques and recent developments in computational methodologies that enhance their integration. Examples of successful synergistic applications of these three techniques will be highlighted. Opinion regarding possible future directions of the field will be given.


Visualization of trans-interactions of a protocadherin-α between processes originating from single neurons.
Clustered protocadherin (Pcdh), a cell adhesion protein, is involved in the self-recognition and non-self-discrimination of neurons by conferring diversity on the cell surface. Although the roles of Pcdh in neurons have been elucidated, it has been challenging to visualize its adhesion activity in neurons, which is a molecular function of Pcdh. Here, we present fluorescent indicators, named IPADs, which visualize the interaction of protocadherin-α4 isoform (α4). IPADs successfully visualize not only homophilic α4 trans-interactions, but also combinatorial homophilic interactions between cells. The reversible nature of IPADs overcomes a drawback of the split-GFP technique and allows for monitoring the dissociation of α4 trans-interactions. Specially designed IPADs for self-recognition are able to monitor the formation and disruption of α4 trans-interactions between processes originating from the same neurons. We expect that IPADs will be useful tools for obtaining spatiotemporal information on Pcdh interactions in neuronal self-recognition and non-self-discrimination processes.


Molecular dynamics simulations of the spike trimeric ectodomain of the SARS-CoV-2 Omicron variant: structural relationships with infectivity, evasion to immune system and transmissibility.
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron is currently the most prevalent SARS-CoV-2 variant worldwide. Herein, we calculated molecular dynamics simulations of the trimeric spikeWT and SpikeBA.1 for 300 ns. Our results show that SpikeBA.1 has more conformational flexibility than SpikeWT. Our principal component analysis (PCA) allowed us to observe a broader spectrum of different conformations for SpikeBA.1, mainly at N-terminal domain (NTD) and receptor-binding domain (RBD). Such increased flexibility could contribute to decreased neutralizing antibody recognition of this variant. Our molecular dynamics data show that the RBDBA.1 easily visits an up-conformational state and the prevalent D614G mutation is pivotal to explain molecular dynamics results for this variant because to lost hydrogen bonding interactions between the residue pairs K854SC/D614SC, Y837MC/D614MC, K835SC/D614SC, T859SC/D614SC. In addition, SpikeBA.1 residues near the furin cleavage site are more flexible than in SpikeWT, probably due to P681H and D614G substitutions. Finally, dynamical cross-correlation matrix (DCCM) analysis reveals that D614G and P681H may allosterically affect the cleavage site S1/S2. Conversely, S2' site may be influenced by residues located between NTD and RBD of a neighboring protomer of the SpikeWT. Such communication may be lost in SpikeBA.1, explaining the changes of the cell tropism in the viral infection. In addition, the movements of the NTDWT and NTDBA.1 may modulate the RBD conformation through allosteric effects. Taken together, our results explain how the structural aspects may explain the observed gains in infectivity, immune system evasion and transmissibility of the Omicron variant.Communicated by Ramaswamy H. Sarma.


Thrombin: implications for intratumor therapy against metastasis.
The effect of an intratumor injection of thrombin as a potential cancer therapeutic agent was examined in two transplantable solid tumor models: a renal adenocarcinoma implanted beneath the kidney capsule in Wistar-Lewis rats and a rhabdomyosarcoma implanted similarly in Wag-Rij rats. For each tumor type, a group of test animals received a single dose of thrombin injected directly into the central tumor mass 6 weeks following tumor implantation. Corresponding groups of tumor-implanted controls consisted of animals that received an equal volume injection of saline instead of thrombin and animals that were not injected. The tumor-implanted animals receiving thrombin, saline, or no injection were handled identically. The saline-injected and noninjected control animals died of metastasis to the lung within the same well-defined period of time. All animals receiving thrombin showed significantly increased longevity, and the degree of increase was related to variations in tumor size at the time of injection. In every case, animal death was due to pulmonary metastasis verified at autopsy, but animals receiving thrombin therapy lived about 85% longer following tumor implantation than animals not receiving therapy.


Seq-ing the mechanisms of migraine.
The trigeminal ganglia (TG) play a crucial role in migraine pathophysiology. In this issue of Neuron, Yang et al. developed a single-cell atlas profiling the transcriptome and epigenome of mouse and human TG, thus providing a roadmap for therapeutic targeting.


Effect of vasopressin-like peptides on consumption of ethanol by the rat.
Rats were trained to accept ethanol in their drinking water, by successive small increments or decrements in alcohol concentration in response to the individual consumption of each rat. Those injected with desglycinamide9-lysine8-vasopressin (DGLVP), 1--4 microgram SC every second day, attained almost twice as high a final acceptance concentration (FAC) and mean daily ethanol intake (g/kg) as vehicle-treated controls. Hypophysectomized animals initially accepted the same alcohol concentrations as intact rats, but drank much larger volumes and correspondingly higher daily g/kg intakes. However, this was rapidly succeeded by rejection of all but very low concentrations, which was unaffected by DGLVP. During a subsequent free-choice period (water vs. ethanol at individual FACs), the groups maintained their relative positions with respect to ethanol intake. This was not altered by injection of vasopressin in the hypophysectomized rats, but was overcome by raising the alcohol concentration. The results suggest that vasopressin-like peptides facilitate acquisition of alcohol drinking behavior.


All-Arthroscopic Refixation of an Osteochondral Femoral Avulsion of the Anterior Cruciate Ligament with a Laxity-Minimizing Suture.
Torn anterior cruciate ligaments in children are rare injuries. The incidence of concomitant avulsion fracture in the skeletally immature patient is high. Reports of arthroscopic reconstruction in the literature are sparse. We present the case of a femoral avulsion fracture of the anterior cruciate ligament in a seven-year-old girl. The patient underwent arthroscopic refixation with DynacordTM Suture and Suture Button. At a follow-up of 8 weeks, the patient had free range of motion and a stable knee, with a negative Lachman- and Pivot-Shift-test in particular. Refixation of femoral avulsion fracture can lead to good results. Open physes and size of the knee should be taken into account. Anatomic reconstruction is compulsory to receive bone on bone healing.


Non-invasively measured central and peripheral factors of oxygen uptake differ between patients with chronic heart failure and healthy controls.
Maximum oxygen uptake is an established measurement of diagnosing chronic heart failure and underlies various central and peripheral factors. However, central and peripheral factors are little investigated, because they are usually measured invasively. The aim of this study was to compare non-invasively measured central and peripheral factors of oxygen uptake between patients with chronic heart failure and healthy controls. Ten male patients with heart failure with reduced ejection fraction (62 ± 4 years; body mass index: 27.7 ± 1.8 kg/m2; ejection fraction: 30 ± 4%) and ten male healthy controls (59 ± 3 years; body mass index: 27.7 ± 1.3 kg/m2) were tested for blood pressure, heart rate, stroke volume, cardiac output, and cardiac power output (central factors) as well as muscle oxygen saturation of the vastus lateralis and biceps brachii muscle (peripheral factors) during an incremental cycling test. Stroke volume and muscle oxygen saturation were non-invasively measured by a bioreactance analysis and near-infrared spectroscopy, respectively. Additionally, a maximum isometric strength test of the knee extensors was conducted. Magnitude-based inferences were computed for statistical analyses. Patients had a likely to most likely lower oxygen uptake, mean arterial pressure, and heart rate at maximum load as well as very likely lower isometric peak torque. Contrary, patients had a possibly to likely higher stroke volume and muscle oxygen saturation of the vastus lateralis muscle at maximum load. Differences in cardiac output, cardiac power output, and muscle oxygen saturation of the biceps brachii muscle at maximum load were unclear. Non-invasively measured central and peripheral factors of oxygen uptake differ between patients with chronic heart failure and healthy controls. Therefore, it is promising to measure both types of factors in patients with chronic heart failure to optimize the diagnosis and therapy.


Insecticidal potential of an endophytic Cladosporium velox against Spodoptera litura mediated through inhibition of alpha glycosidases.
Alpha glucosidase inhibitory activity was exhibited by partially purified fractions obtained from an endophytic Cladosporium velox, isolated from Tinospora cordifolia. Taking into account the increasing importance of digestive enzyme inhibitors as insecticidal agents, the entomopathogenic potential of the fractions obtained was evaluated against Spodoptera litura (Fab.), a polyphagous pest. Considerable mortality was obtained when the larvae were fed on diet supplemented with the partially purified extract. All the concentrations of the extract significantly prolonged the overall developmental period of S. litura. At higher concentrations, the extract influenced the longevity of females as well as their reproductive potential. Phytochemical analysis revealed the presence of phenolic compounds in the active fraction. The phenolic compound responsible for the bioactivities was purified and identified to be chlorogenic acid using HPLC and MS analysis. The content of chlorogenic acid in the extract was quantified to be 250μg/ml. The purified compound also demonstrated inhibition of alpha glycosidases in vivo. The present study indicates that the endophyte imparted resistance to the insects in the plants could be mediated through chlorogenic acid targeting the alpha glycosidases present in the gut of the insect. The isolate obtained can be exploited for the production of chlorogenic acid, which has the potential to be exploited as a biocontrol agent against S. litura.


Risk factors for outcome and complications of dorsal foraminotomy in cervical disc herniation.
Dorsal foraminotomy is a standard operative procedure for lateral cervical disc herniation. Factors associated with surgical complications and clinical outcome in dorsal foraminotomy of cervical disc herniation were evaluated in a retrospective cohort study. Thirty-nine patients were operated upon for unilateral, monosegmental, mediolateral cervical disc herniation (+/- associated spondylosis) from 1997 to 1999. Preoperative radiologic imaging and surgical reports were analyzed. Motor disfunction, neck irritation, and radicular pain were evaluated. Outcome was ranked according to modified Odom's criteria at 6 weeks and 1 year postoperatively. Six weeks after injury 7 of 39 patients (18%) showed neck irritation. No new neurologic deficit was seen. All patients with preoperative paresis improved; two had early relapses of a medial soft disc prolapse (2/39). Residual radicular pain was seen in 3 of 39 patients (8%) within 30 days postoperatively, necessitating surgical revision. Factors of surgical failure were associated spondylosis (2/3) and residual mediolateral disc protrusion (1/3). In one patient with associated spondylosis, local pain due to a symptomatic fracture of the lateral process of D1 resolved after revision. Duration of preoperative radicular pain was identified as a risk factor for unfavorable outcome. In lateral cervical disc herniation, associated spondylosis or medial disc protrusion poses a significant risk of surgical failure and complications of dorsal foraminotomy. Reducing the radicular failure rate by enlarging the bony decompression may lead to local failure. In well-selected patients with a lateral cervical free disc fragment, dorsolateral foraminotomy is successful and safe.


Quantitative assessment of regional variation in tissue clearing efficiency using optical coherence tomography (OCT) and magnetic resonance imaging (MRI): A feasibility study.
Tissue clearing has gained attention as a pioneering research tool for imaging of large tissue samples. This technique improves light transmission by reducing light scattering within tissues, either by removing lipids or by replacing water with a high refractive index solution. Although various clearing techniques have been developed, quantitative assessments on clearing efficacy depending on tissue properties are rare. In this study, we developed the quantitative mapping of regional clearing efficacy using mean free path in optical coherence tomography (OCT) and proton density in magnetic resonance imaging (MRI), and demonstrated its feasibility in the brain sample with four representative clearing techniques (benzyl alcohol and benzyl benzoate [BABB], ClearT, Scale, and passive CLARITY technique [PACT]). BABB (solvent-based clearing), involving both refractive index matching and lipid removal, exhibited best optical clearing performance with the highest proton density reduction both in gray and white matter. Lipid-removing techniques such as Scale (aqueous hyperhydration) and PACT (hydrogel embedding) showed higher clearing efficiency in white matter than gray matter in accordance with larger proton density increase in white matter. For ClearT (aqueous-based simple immersion), we observed lowest clearing efficiency in the white matter as well as poor lipid removal reflected in low proton density reduction. Our results showed the feasibility of the regional mapping of clearing efficacy and correlating optical transparency and proton density changes using OCT and MRI from existing tissue clearing techniques. This novel quantitative mapping of clearing efficacy depending on tissue types and clearing methods may be helpful in the development of optimized clearing methods for different biological samples.


The relationship between nurse characteristics and perceptions of psychotropic medications in residential facilities for the retarded.
A total of 227 nurses were surveyed regarding their attitudes, perceptions, and knowledge about the use of psychotropic drugs. In general, greater age, male sex, status, and higher qualifications tended to covary and were associated with perception of greater involvement by senior nurses in drug-related decisions, more satisfaction with the level of in-service training, awareness of certain side effects, perception of interdisciplinary decision making as a clinical reality, and the view that psychologists were not influential in medication-related decisions. Those without these characteristics (e.g., females, younger nurses) more often advocated greater use of medical and laboratory tests, ongoing training, and alternatives to drugs. Nurses working in nonambulatory units more often saw senior nursing staff as influential in decisions to stop drug treatment and more often recommended that alternatives to pharmacology be employed. Some tentative conclusions about the dynamics surrounding medication use in these facilities were offered to account for these patterns.


Maternal butyrate supplementation affects the lipid metabolism and fatty acid composition in the skeletal muscle of offspring piglets.
Maternal sodium butyrate (SB) intake has important effects on offspring growth and development. This study aimed to investigate the impacts of maternal SB supplementation during gestation and lactation on fatty acid composition and lipid metabolism in the offspring skeletal muscle of pigs. Twenty sows (Yorkshire, parity 2 to 3) were assigned to the control group (diets without SB, n = 10) and SB group (diets with 0.1% SB, n = 10). The results showed maternal SB supplementation throughout gestation and lactation increased (P < 0.05) body weight of offspring piglets at weaning. The concentrations of triglyceride in plasma and milk were enhanced (P < 0.05). Maternal SB induced (P < 0.05) lipid accumulation with increased expression of peroxisome proliferator activated receptor γ (PPARγ) by enrichment of the acetylation of H3 acetylation K27 (H3K27) in offspring skeletal muscle. Meanwhile, the concentrations of C18:2n-6, C18:3n-3, total polyunsaturated fatty acid (PUFA), n-6 PUFA and n-3 PUFA decreased (P < 0.05) in skeletal muscle of weaning piglets derived from SB sows. Together, these results showed that maternal SB supplementation could influence offspring growth performance, lipid metabolism and fatty acid composition of the skeletal muscle.


[Endophthalmos caused by intravitreal caterpillar hairs. Treatment by direct photocoagulation with argon laser].
We describe an infrequent case of endophthalmitis due to intraocular caterpillar hairs. We report a notable decrease of inflammation after Argon laser photocoagulation of the caterpillar hairs. We also present optical and electronic microscope photographs of one of the hairs extracted from the patient's conjunctiva.


Cleavage patterns, cell-lineages and cell specification are clues to phyletic lineages in Spiralia.
Embryos of molluscs, annelids, nemerteans and platyhelminthes show remarkable intra- and interphyletic resemblances and differences in mesentoblast, dorso-ventral axis and trochoblast specification. These variations have been used to investigate their evolutionary relationship. In molluscs and annelids a heterochronic shift parallels evolutionary relations based on adult characters. Nemerteans and platyhelminthes lack trochal cells and differ in the specification of the mesodermal precursor cell. Nemerteans also differ fundamentally with respect to axis specification related to the first cleavage. Therefore, close phylogenetic relations exist between molluscs and annelids, whereas nemerteans and platyhelminthes are only remotely related with each other and with molluscs and annelids.


Stem Cell Factor/Kit Signal Insufficiency Contributes to Hypoxia-Induced Intestinal Motility Dysfunctions in Neonatal Mice.
Gastrointestinal (GI) motility disorders represent a group of problems that more constantly encountered in preterm infants. However, whether hypoxia exposure contributes to the GI dysfunctions is still unclear. Newborn mice were exposed to hypoxia (10%) from P1 to P7. Intestinal motilities were examined by a strain gauge transducer. The proliferation of ICCs was detected by using immunostaining for BrdU, Ki67, Kit, Ano1, and insulin-like growth factor 1 receptor (IGF-1R+). Smooth muscle cells and enteric neurons were revealed by immunostaining for α-SMA and NF200, respectively. Apoptosis was assessed by TUNEL assay. Kit signal pathway was examined by western blot and qPCR. Intestinal motilities were found weakened significantly in the hypoxic small intestines as compared to controls on P8. Kit+ or Ano1+ interstitial cells of Cajal (ICCs) were found obviously decreased in the myenteric ICCs (ICC-MY) of neonatal mice after exposed to hypoxia. A large number of ICC progenitors (IGF-1R+) were found highly mitotic (BrdU+ Ki67+) to populate ICC during early postnatal development in the normoxic mice. We found the ICC proliferation was significantly inhibited upon hypoxia exposure, without increasing apoptosis (TUNEL+). We next identified that Kit phosphorylation was inhibited 3 days after hypoxia exposure. The inhibition of Kit signaling was largely due to decreased the expression of the ligand of Kit receptor, stem cell factor (SCF), in the intestinal walls. Exposure to imatinib, a Kit receptor inhibitor, for 3 days from P4 phenocopied the effect of hypoxia on the neonatal pups that resulted in inhibited intestinal motilities and decreased Kit+ ICC numbers. All together, our findings indicate the SCF/Kit signaling insufficiency may contribute to the underdevelopment of ICCs and intestinal motility dysfunction upon hypoxia exposure. The decease in ICC density is likely due to the cell cycle arrest of ICC progenitor cells.


Characteristics and diagnostic approaches in the detection of nonapparent bronchogenic carcinoma.
Bronchogenic carcinoma without parenchymal changes, located within normal hilum and mediastinum, does not show in routine chest X-ray. The value of tomography is demonstrated.


Effect of maternal NG-nitro-l-arginine administration on fetal growth and hypoxia-induced changes in newborn rats.
Nitric oxide (NO) inhibition with NG-nitro-l-arginine methyl ester (l-NAME) in the last trimester of pregnancy caused intrauterine growth retardation and hind-limb disruptions in rats. In the present study, the effect of maternal NO inhibition with NG-nitro-l-arginine (l-NNA) on hypoxic newborn rats was investigated. Timed-pregnant rats were obtained on gestational day 17. Four groups of rats were used: control, hypoxic, l-NNA and l-NNA + hypoxic groups. In the last two groups, l-NNA (2 mg/kg bolus, i.p.) was administered to the mothers of pups antenatally on 3 consecutive days. Hypoxia was induced in newborn rats by breathing of a mixture of 8% oxygen and 92% nitrogen for 3 h. Pups were then allowed to inhale normal atmospheric air for 30 min. All newborn rats were decapitated on the first day of life after hypoxia and reoxygenation. Brain, heart, lung, liver, kidney and intestinal tissues were studied biochemically. Hypoxia-induced biochemical changes were determined by measuring lipid peroxidation. Histopathologic examination of lung tissue was performed. Nitric oxide synthase inhibition in pregnancy did not cause fetal growth retardation. Hypoxia increased lipid peroxidation in all tissues except the heart; this increase was decreased by maternal l-NNA administration in brain, lung, liver and kidney tissues. However, lipid peroxidation was increased by NO synthase inhibition in the intestines. In the lungs, pulmonary hemorrhage was observed in the hypoxic group. Minimal pulmonary hemorrhage was detected in the l-NNA and l-NNA + hypoxic groups. These data suggest that antenatal administration of an NO synthase inhibitor acts as both a destructive and protective agent in hypoxic newborn rats.


Does vitamin blends supplementation affect the animal performance, carcass traits, and nutrient digestibility of young Nellore finishing bulls?
This study was realized to evaluate the effects of supplementation with blends of water and fat-soluble vitamins on animal performance and carcass traits of young Nellore bulls. Forty-three Nellore bulls, with an initial weight of 261±27.3 kg and a mean age of 8±1.0 months, were used. Five animals were slaughtered at the beginning of the experiment (reference group), to determine the initial empty body weight of the bulls that remained in the experiment. The remaining 38 bulls were fed ad libitum and distributed in a completely randomized design in a 2×2 factorial scheme, with or without supplementation of water-soluble (B-blend+ or B-blend-) and fat-soluble (ADE+ or ADE-) vitamin blends. Diets were isonitrogenous (120 g of crude protein/kg dry matter [DM] of total mixed ration) and consisted of a roughage:concentrate rate of 30:70 based on total DM of diet. The experiment lasted 170 days, with 30 days of adaptation and 140 days for data collection. At the beginning and end of the experimental period, the bulls were weighed to determine the average daily gain. To estimate the apparent digestibility of nutrients and microbial efficiency, spot collections of feces and urine were performed for five consecutive days. DM, ashes, organic matter, crude protein, ethereal extract, neutral detergent fiber corrected for residual ash and residual nitrogenous, and N intake and apparent digestibility were not influenced by vitamin supplementation, but total digestible nutrients intake and non-fibrous carbohydrates digestibility were influenced by B complex vitamin supplementation. Nitrogen balance, microbial efficiency, and performance data were not influenced (p>0.05) by vitamin supplementation. Vitamin supplementation (a blend of water-soluble and fat-soluble vitamins or their combinations) does not influence the animal performance and carcass traits of young Nellore bulls.


[The participation of the proximal convoluted tubule in the formation of the parietal layer of the renal corpuscle in the mouse (Mus musculus)].
The epithelium of the proximal convoluted tubule participates in the formation of the parietal layer in the area of the urinary pole, as can already be observed in the first renal corpuscles of the mouse fetus. Sexual hormones do not seem to have any influence on this structural characteristic, neither during prenatal nor postnatal life.


Impact assessment of crude oil mix, electricity generation mix, and vehicle technology on road freight emission reduction in China.
To achieve net zero emissions, the global transportation sector needs to reduce emissions by 90% from 2020 to 2050, and road freight has a significant potential to reduce emissions. In this context, emission reduction paths should be explored for road freight over the fuel life cycle. Based on panel data from 2015 to 2020 in China, China's version of the GREET model was established to evaluate the impact of crude oil mix, electricity mix, and vehicle technology on China's reduction in road freight emissions. The results show that the import share of China's crude oil has increased from 2015 to 2020, resulting in an increase in the greenhouse gas (GHG) emission intensity of ICETs in the well-to-tank (WTT) stage by 7.3% in 2020 compared with 2015. Second, the share of China's coal-fired electricity in the electricity mix decreased from 2015 to 2020, reducing the GHG emission intensity of battery electric trucks (BETs), by approximately 6.5% in 2020 compared to 2015. Third, different vehicle classes and types of BETs and fuel cell electric trucks (FCETs) have different emission reduction effects, and their potentials for energy-saving and emission reduction at various stages of the fuel life cycle are different. In addition, in a comparative study of vehicle technology, the results show that (1) for medium-duty trucks (MDTs) and heavy-duty trucks (HDTs), FCETs have lower GHG emission intensity than BETs, and replacing diesel-ICETs can significantly reduce GHG emissions from road freight; (2) for light-duty trucks (LDTs), BETs and FCETs have the highest GHG emission reduction potential; thus, improving technologies such as electricity generation, hydrogen fuel production, hydrogen fuel storage, and transportation will help to improve the emission reduction capabilities of BETs and FCETs. Therefore, policymakers should develop emission standards for road freight based on vehicle class, type, and technology.


Comparison of two devices for wound measurement.
Sixty leg ulcers were measured with two techniques, a common paper tape measure and a new technique, a plastic grid device. The plastic grid proved to be a reliable, valid, and feasible method to quickly assess wounds in a busy outpatient setting. The grid was superior to the tape measure in assessing large and/or irregular ulcers.


In-hospital blood collection increases the rate of indeterminate results in interferon-gamma release assays.
The interferon (IFN)-γ release assay (IGRA) has recently been established as a method to evaluate the infection status of tuberculosis instead of the tuberculin skin test. However, indeterminate results can create challenges to interpretation. The IGRA has been available in Japan since 2005, including the recently launched QuantiFERON-TB Gold Plus (QFT-plus) assay. The aim of this study was to investigate the clinical features and predictors of indeterminate results by the QFT-plus test in routine practice. This was a cross-sectional study of 1258 patients. Multivariate logistic regression models were employed to investigate the clinical factors related to indeterminate results by the QFT-plus. Overall, 91.8% of results were found to be conclusive and 8.2% were indeterminate. The QFT-plus indeterminate results were predominantly due to a low level of IFN-γ production by mitogens. Multivariate analysis indicated that an indeterminate result was significantly associated with age, sex, corticosteroid use, autoimmune disease, and inpatient setting. Certain types of individuals are at higher risk of an indeterminate IGRA result. The QFT-plus test for hospitalized patients should be avoided as much as possible, and it is better to perform the test for those patients in outpatient settings.


Comparison of short-term oncologic outcome of robotic thyroid surgery using dynamic risk stratification: A propensity score-matched comparison study.
The long-term oncologic outcome of robotic surgery for thyroid cancer is not well established. The aim of this study was to predict the long-term oncologic outcome of robotic surgery by using dynamic risk stratification in classic papillary thyroid carcinoma. A total of 444 propensity score-matched pairs of patients with papillary thyroid carcinoma treated with robotic surgery and conventional open surgery were classified into 4 response-to-therapy categories. The results were compared between the robotic surgery and open surgery groups. The median follow-up duration was 60 months. After propensity score matching, the robotic surgery group showed less extensive thyroid surgery and lymph node dissection and a higher proportion of patients who underwent radioactive iodine remnant ablation than the open surgery group; however, the dynamic risk stratification did not differ between the 2 groups (P = .086). The long-term oncologic outcome of robotic surgery is expected to be comparable with that of open surgery based on the dynamic risk stratification.


Antibacterial effects of hybrid tooth colored restorative materials against Streptococcus mutans: An in vitro analysis.
The objective of this study was to evaluate the antibacterial effect of two hybrid restoratives, namely resin modified glass ionomer cement (GC Fuji II™ LC, GC Corporation, Tokyo, Japan) and giomer (Beautifil-II, Shofu Inc., Kyoto, Japan) against Streptococcus mutans [Microbial Type Culture Collection (MTCC), 890]. The antibacterial effect was evaluated using an agar diffusion test. The prepared wells in petri dishes were completely filled with chlorhexidine (positive control group), resin modified glass ionomer cement and giomer respectively. Prepared bacterial suspension was poured over the petri dish and was spread evenly using the plate spreader. The culture plates were placed in the incubator for 24 h at 37°C. The antibacterial activity was evaluated after 24 h, 48 h, and 7 days for each group in triplicates. The results of the antibacterial effect of the tested materials were collected, statistically analyzed using the ANOVA test to determine the difference between the mean diameters of the inhibition zone produced. The mean zone of bacterial inhibition was found to be more with the giomer specimens at all time periods. However, this inhibitory activity showed a gradual decrease over a period of 7 days and the maximum inhibition was evident after 24 h with both the test materials.


Parity-based assessment of anemia and iron deficiency in pregnant women.
This study aimed to separately evaluate the prevalence of anemia and iron deficiency in nulliparous and multiparous women. We retrospectively examined data of women who delivered in our clinic from January 2016 to December 2018. Inclusion criteria were delivery occurring at ≥36 weeks and singleton pregnancy. Pregnant women with severe medical disorders were excluded. We estimated complete blood count (CBC) and serum ferritin (SF) in the first trimester and only CBC in the late second trimester. Data of nulliparas and multiparas were analyzed separately. Statistically significance was set at p < 0.05. Totally, 481 nulliparas and 603 and multiparas were enrolled. Mean hemoglobin values in the first trimester were 12.6 ± 1.0 and 12.4 ± 1.0 g/dl (p < 0.001), while median SF values were 42.7 (12.2, 108.2) and 27.7 (8.0, 72.6) ng/ml (p < 0.001) in nulliparas and multiparas, respectively. Hemoglobin in the late second trimester was 11.2 ± 0.9 and 10.7 ± 1.0 g/dl (p < 0.001) in nulliparas and multiparas, respectively. Low ferritin levels (SF < 12 ng/ml) were more frequently found in multiparas than in nulliparas (111/603 vs. 46/481, p < 0.001, Odds ratio [OR] = 2.13). Anemia in the first trimester (hemoglobin<11.0 g/dl) was found in 3.5% (17/481) and 8.8% (53/603) (p < 0.001; OR, 2.63), while that in late second trimester (hemoglobin<10.5) was observed in 21.0% (101/481) and 36.3% (219/603) (p < 0.001, OR = 2.15) nulliparas and multiparas, respectively. Non-anemic women (hemoglobin level ≥11.0) with low ferritin levels (SF < 12 ng/ml) in the first trimester showed higher rate of anemia development in the second trimester than those with both normal hemoglobin and ferritin levels, irrespective of parity (51.3% [19/37] vs. 16.2% [69/427], p < 0.001 in nulliparas and 76.9% [60/78] vs. 26.5% [125/472], p < 0.001 in multiparas]. Anemia and low SF levels occurred more commonly in multiparous than in nulliparas. Further, low SF was a risk factor for anemia development in later pregnancy.


A novel and facile synthesis of 3-(2-benzofuroyl)- and 3,6-bis(2-benzofuroyl)carbazole derivatives.
A facile synthesis of hitherto unreported 3-(2-benzofuroyl)carbazoles 3a-k, 3,6-bis(2-benzofuroyl)carbazoles 5a-k, and naphtho[2,1-b]furoylcarbazoles 3l and 5l is described. The synthesis mainly relies on the ultrasound-assisted Rap-Stoermer reaction of 3-chloroacetyl- (1) or 3,6-dichloroacetyl-9-ethyl-9H-carbazole (4) with various salicylaldehydes 2a-k as well as 2-hydroxy-1-naphthaldehyde (2l) in CH(3)CN with the presence of PEG-400 as catalyst. The procedure offers easy access to benzofuroylcarbazoles in short reaction times and the products are obtained in moderate to good yields.


Efficacy and safety of glecaprevir and pibrentasvir treatment for 8 or 12 weeks in patients with recurrent hepatitis C after liver transplantation: a Japanese multicenter experience.
Efficacy of 8-week regimen with direct-acting antivirals (DAA) for patients with hepatitis C after liver transplantation has not been clarified. This study aimed to clarify the efficacy and safety of glecaprevir and pibrentasvir therapy for 8 and 12 weeks in Japanese patients with recurrent hepatitis C after liver transplantation. A cohort study of liver transplant recipients with recurrent hepatitis C treated with glecaprevir (300 mg/day) and pibrentasvir (120 mg/day) was performed at nine liver transplant centers in Japan. Twenty-five patients with hepatitis C after liver transplantation were treated with glecaprevir and pibrentasvir. Twenty-four patients completed the treatment protocol; treatment was discontinued in one patient who had nausea at 3 days after the initiation of treatment. All the 24 patients who completed the 8- or 12-week treatment protocol achieved a sustained virological response 12 weeks after completion of treatment (SVR12). The SVR12 rates in patients with HCV genotype 1 and 2 were 100% (21 of 21 patients) and 75% (3 of 4 patients), respectively. All patients with prior DAA therapy failure (n = 6), jaundice (n = 4), and liver cirrhosis (n = 4) achieved SVR12. Seven of 8 patients (88%) with severe renal impairment achieved SVR12. Adverse events occurred in 6 of 25 patients (24%), including serious adverse events in 2 patients (8%). Treatment-related adverse events were nausea, pruritus, and mild renal dysfunction. Eight- or 12-week regimen of glecaprevir and pibrentasvir is efficacious and safe in patients with recurrent HCV infection after liver transplantation, even in difficult-to-treat populations, including patients with severe renal impairment, prior DAA experience, liver cirrhosis, or jaundice after liver transplantation.


Visual and auditory anomalies in Chediak-Higashi syndrome.
Albinism is correlated with misrouting of decussating retinal fibers in the brain. There is also evidence of anomalies of decussating auditory pathways in albinos. The Chediak-Higashi syndrome (CHS) is a rare form of partial albinism which includes increased susceptibility to infections, a hemorrhagic tendency and peripheral polyneuropathies. Binocular and monocular pattern-onset visually evoked potentials (VEPs) and monaural auditory brain stem responses (ABRs) were recorded from 4 subjects with CHS. Three of the CHS demonstrated asymmetric monocular VEPs and failed the Titmus stereovision test. All 4 CHS produced asymmetric ABRs similar to those reported for albinos. Although the hair, skin and irises are relatively well pigmented in CHS, these individuals apparently have anomalies of their central visual and auditory pathways.


Genesis of body surface potential distribution in right bundle branch block.
In order to investigate the specific sites of conduction block in the three types (I, II, III) of right bundle branch block (RBBB) classified by body surface isopotential maps, the simulation of ventricular propagation process and mathematically reconstructed maps were used. Four assumptions were introduced from the results of clinical observations and animal experiments. The maps reconstructed from two of these assumptions, in which the conduction block was placed on the main stem of the right bundle branch, showed two different patterns at late stages of excitation, and these two kinds of map resembled Types I and II in clinical maps, respectively. The maps reconstructed from the other two assumptions, in which the site of the conduction block was located mainly in the Purkinje system of the right ventricular free wall, resembled Type I at the late stage of excitation in one of two assumptions and agreed with Type III through all stages of excitation in other case. Based on the above results, it is speculated that the differences of ranges and degrees of conduction block ascribed to abnormal activation in the Purkinje system of the right ventricular free wall are responsible for the genesis of clinical RBBB map patterns.


Alternative- and focal therapy trends for prostate cancer: a total population analysis of in-patient treatments in Germany from 2006 to 2019.
Focal therapy (FT) offers an alternative approach for prostate cancer (PCa) treatment in selected patients. However, little is known on its actual establishment in health care reality. We defined FT as high-intensity focused ultrasound (HIFU), hyperthermia ablation, cryotherapy, transurethral ultrasound ablation (TULSA) or vascular-targeted photodynamic therapy (VTP) TOOKAD®. We analyzed the nationwide German hospital billing database for a PCa diagnosis in combination with FT. For analyses on the hospital level, we used the reimbursement.INFO tool based on hospitals' quality reports. The study period was 2006 to 2019. We identified 23,677 cases of FT from 2006 to 2019. Considering all PCa cases with surgery, radiotherapy or FT, the share of FT was stable at 4%. The annual caseload of FT increased to a maximum of 2653 cases in 2008 (p < 0.001) and then decreased to 1182 cases in 2014 (p < 0.001). Since 2015, the cases of FT remained on a plateau around 1400 cases per year. The share of HIFU was stable at 92-96% from 2006 to 2017 and decreased thereafter to 75% in 2019 (p = 0.015). In 2019, VTP-TOOKAD® increased to 11.5% and TULSA to 6%. In 2006, 21% (62/299) of urological departments performed FT and 20 departments reached > 20 FT procedures. In 2019, 16% (58/368) of urological departments performed FT and 7 departments reached > 20 FT. In 2019, 25 urological departments offered FT other than HIFU: 5 centers hyperthermia ablation, 11 centers VTP TOOKAD®, 3 centers cryotherapy, 6 centers TULSA. The FT development in Germany followed the Gartner hype cycle. While HIFU treatment is the most commonly performed FT, the share of newer FT modalities such as VTP-TOOKAD® and TULSA is remarkably increasing.


STAT4 targets KISS1 to promote the apoptosis of ovarian granulosa cells.
In mammals, it is known that the estradiol-17β (E2) is mainly synthetized in ovarian granulosa cells (GCs), and the excessive apoptosis of GCs induces the follicular atresia. Many studies have implicated the essential role of KISS1, with the pro-synthetic effect of E2 and the anti-apoptotic effect on GCs, in the mammalian folliculogenesis, and several STAT4 potential binding sites were previously predicted on the promoter of KISS1 in pigs. However, the biological effects of STAT4 on GCs and the molecular regulation between STAT4 and KISS1 remained largely unknown. Using the porcine GCs as the cellular model, the overexpression plasmid, small interfering RNA, 5'-deletion and luciferase assay were applied to investigate the molecular mechanisms for STAT4 regulating the expression of KISS1. In this study, the STAT4 negatively regulated the mRNA and protein levels of KISS1 in porcine GCs, and the mRNA level of STAT4 was observed to significantly decrease from immature to mature follicles, which was inversed with that of KISS1. The relative luciferase activity of KISS1 promoter was significantly increased with deletion of the fourth potential binding site (- 305/- 295), and ChIP further confirmed that the STAT4 bound at - 305/- 295 region of KISS1. Besides, the STAT4 significantly regulated the mRNA levels of PDK1, FOXO3 and TSC2 of PI3K signaling pathway to promote the cell apoptosis and the percentage of cells at G0/G1 phase of cell cycle in GCs. Alternatively, the STAT4 significantly decreased the mRNA levels of CYP17, 3B-HSD, 17B-33 HSD, ESR1, and ESR2, as well as the concentration of E2 in GCs. Furthermore, interfering with the expression of STAT4 was observed to significantly stimulate the pro-synthetic effect of E2 and anti-apoptotic effect of KISS1 in GCs. Collectively, the STAT4 might directly target at - 305/- 295 region of KISS1 to negatively regulate the transcription of KISS1, promote the cell apoptosis via PI3K signaling pathway, suppress the synthesis of E2 through the estrogen signaling pathway in porcine GCs. These proposed works could provide useful insight in further investigations on the molecular functionalities of STAT4 and KISS1 in the folliculogenesis of mammals.


Sensitive and specific quantification of the anticancer agent ZD1839 (Gefitinib) in plasma by on-column focusing capillary liquid chromatography-tandem mass spectrometry.
The development of an on-column focusing gradient capillary LC method coupled to tandem mass spectrometry (quadrupole-linear ion trap) for the quantitative determination of the anticancer agent ZD1839 (Gefitinib, Iressa) in blood plasma is described. Plasma samples (0.2 ml) were extracted with methyl tert-butyl ether. The analytes of interest, ZD1839 and the internal standard [(2)H8]ZD1839 (ZD1839-d8) were eluted on a 50 mm x 1 mm, 5 microm particle size, capillary ODS Hypersil column using an aqueous ammonium acetate gradient at 40 microl/min. Mass spectrometric detection was performed by a Q-Trap tandem mass spectrometer with electrospray positive ionisation, and monitored in the multiple reaction monitoring transitions 447 >128 and 455 >136, respectively. The limit of quantification of ZD18395 was 0.1 ng/ml. The method proved to be robust, allowing quantification of ZD1839 with sufficient precision, accuracy and sensitivity.


Field improvement in a uniaxial centered lens composed of two stacked-volume holographic elements.
Arrangements are described for the recording of volume holograms with two sections that, when stacked together, work as uniaxial centered lenses and allow one to solve the problem of angular selectivity in the imaging of wide objects. The performance of such systems is examined qualitatively, and suggestions aimed at improving these designs are proposed.


Correlations between ADC values and molecular markers of Ki-67 and HIF-1α in hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related deaths. Cell proliferativity and hypoxia have important impact on the response to radiotherapy or chemotherapy. The purpose of this study was to investigate the association of apparent diffusion coefficient (ADC) values and the molecular markers Ki-67 and hypoxia inducible factor-α (HIF-α) in hepatocellular carcinoma (HCC). Forty-seven patients diagnosed with HCC were included in this study. All patients performed diffusion-weighted magnetic resonance imaging (DW-MRI) before any anticancer treatment. The ADC maps were automatically calculated on a Syngo workstation. The Ki-67 and HIF-1α expression were assessed by immunohistochemistry. The Pearson correlation test was used to assess the correlation between ADC values and Ki-67 and HIF-1α expression. Ki-67 staining was clearly identified based on the brown nuclear staining in tumor cells. High Ki-67 expression was correlated with low differentiation (p=0.028). A significant correlation was observed between HIF-1α expression and maximum diameter (p=0.014). The mean ADC value was (0.983±0.21)×10(-3) mm(2)/s. The level of Ki-67 expression was correlated inversely with the ADC values (r=-0.371, p=0.01). There was a significant positive correlation between the ADC values and HIF-1α expression (r=0.389, p=0.007). The ADC values were observed to correlate significantly with the molecular markers Ki-67 and HIF-1α. Our results suggest that the ADC values on DW-MRI may be used as a measure of cell proliferativity and hypoxia in hepatocellular carcinoma.


Validation of an animal model of right ventricular dysfunction and right bundle branch block to create close physiology to postoperative tetralogy of Fallot.
In the past 5 years a few number of studies and case reports have come out focusing on biventricular (BiV) stimulation for treatment of congenital heart disease related ventricular dysfunction. The few available studies include a diverse group of pathophysiological entities ranging from a previously repaired tetralogy of Fallot (TOF) to a functional single ventricle anatomy. Patient's status is too heterogeneous to build important prospective study. To well understand the implication of prolonged electromechanical dyssynchrony we performed a chronic animal model that mimics essential parameters of postoperative TOF. Significant pulmonary regurgitation, mild stenosis, as well as right ventricular outflow tract (RVOT) scars were induced in 15 piglets to mimic repaired TOF. 4 months after hemodynamics and dyssynchrony parameters were compared with a control group and with a population of symptomatic adult with repaired TOF. Comparing the animal model with the animal control group on echocardiography, RV dilatation, RV and LV dysfunction, broad QRS complex and dyssynchrony were observed on the animal model piglets. Moreover, epicardial electrical mapping showed activation consistent with a right bundle branch block. The animal models displayed the same pathophysiological parameters as the post TOF repair patients in terms of QRS duration, pulmonary regurgitation biventricular dysfunction and dyssynchrony. This chronic swine model mimics electromechanical ventricular activation delay, RV and LV dysfunction, as in adult population of repair TOF. It does appear to be a very useful and interesting model to study the implication of dyssynchrony and the interest of resynchronization therapy in TOF failing ventricle.


Management of acute neck pain: A case series describing immediate and short term clinical outcomes following use of the Multifidus Isometric Technique.
Acute facet capsular entrapment results in sudden onset of pain and reduced ability to perform active cervical motions. The Multifidus Isometric Technique (MIT) is a type of manual therapy intervention theorized to target the entrapped facet capsule and pull the entrapped synovial folds from the facet joint resulting in decreased pain and increased function. To describe immediate MIT clinical outcomes for patients with acute neck pain. Consecutive patients (n = 30; 70% female) with sudden onset of neck pain received MIT within 48 hours of symptom onset. Clinical outcome measures included: 1) 11-point Numeric Pain Rating Scale (NPRS); 2) cervical AROM, and 3) the Neck Disability Index (NDI). Paired-sample t-testing was used to assess for within and between-session changes in outcome measure scores. Within-session NPRS improvements were observed during all cervical AROM movements (p < .01), with initial treatment associated with greater improvements on average (M = 2.4 ± 1.6 points) compared to the second treatment (M = 0.6 ± 0.9 points). Similarly, most within-session AROM measures improved during both sessions (p < .01) with greater average improvements observed following the initial treatment session and for cervical rotation to the symptomatic side (M = 26.5 ± 9.6 vs. 8.0 ± 9.7°). Between-session NDI scores improved (M = 15.3 ± 9.8, p < .01) with approximately 60% of patients achieving a minimally clinically important difference of 14 percentage points. The MIT is a potentially beneficial intervention for patients with acute neck pain. Future studies consisting of longer follow-up time points and comparison treatment groups are needed to test MIT effectiveness.


Poland syndrome: Fortuitous discovery of a familial case in Togo.
Poland syndrome is a rare malformative disease. It is characterized by agenesis or hypoplasia of the pectoralis major muscle associated or not with a malformation of the ipsilateral thoracic limb. The authors report the fortuitous discovery of a familial case of Poland syndrome in Togo. He was a 25-year-old young man, with no known pathological history, examined as part of a physical fitness assessment and who presented with thoracic asymmetry. The clinical and radiological explorations made it possible to conclude to a Poland syndrome in its minor form without any other associated malformation. In addition, three other paternal uncles of the patient presented with the same clinical symptomatology. In the absence of a clear indication, the patient received no treatment. Poland syndrome is a rare congenital malformation. The abnormalities encountered are cutaneous-glandular, osteo-cartilaginous and muscular. The etiopathogenesis of the disease is unknown. The diagnosis is clinical and is based on the demonstration of agenesis of the pectoralis major muscle. The disease does not often lead to functional discomfort; therefore the treatment is not systematic and has only aesthetic value. A rare congenital disease, Poland syndrome can occur sporadically or in families. Its treatment depends above all on the psychological repercussion of the disease.


The Structural, Photocatalytic Property Characterization and Enhanced Photocatalytic Activities of Novel Photocatalysts Bi₂GaSbO₇ and Bi₂InSbO₇ during Visible Light Irradiation.
In order to develop original and efficient visible light response photocatalysts for degrading organic pollutants in wastewater, new photocatalysts Bi₂GaSbO₇ and Bi₂InSbO₇ were firstly synthesized by a solid-state reaction method and their chemical, physical and structural properties were characterized. Bi₂GaSbO₇ and Bi₂InSbO₇ were crystallized with a pyrochlore-type structure and the lattice parameter of Bi₂GaSbO₇ or Bi₂InSbO₇ was 10.356497 Å or 10.666031 Å. The band gap of Bi₂GaSbO₇ or Bi₂InSbO₇ was estimated to be 2.59 eV or 2.54 eV. Compared with nitrogen doped TiO₂, Bi₂GaSbO₇ and Bi₂InSbO₇, both showed excellent photocatalytic activities for degrading methylene blue during visible light irradiation due to their narrower band gaps and higher crystallization perfection. Bi₂GaSbO₇ showed higher catalytic activity compared with Bi₂InSbO₇. The photocatalytic degradation of methylene blue followed by the first-order reaction kinetics and the first-order rate constant was 0.01470 min-1, 0.00967 min-1 or 0.00259 min-1 with Bi₂GaSbO₇, Bi₂InSbO₇ or nitrogen doped TiO₂ as a catalyst. The evolution of CO₂ and the removal of total organic carbon were successfully measured and these results indicated continuous mineralization of methylene blue during the photocatalytic process. The possible degradation scheme and pathway of methylene blue was also analyzed. Bi₂GaSbO₇ and Bi₂InSbO₇ photocatalysts both had great potential to purify textile industry wastewater.


The effect of cilostazol on the expression of matrix metalloproteinase-1 and type I procollagen in ultraviolet-irradiated human dermal fibroblasts.
Cilostazol is a selective inhibitor of type III phosphodiesterase that inhibits platelet aggregation. Cilostazol is a useful vasodilator, antithrombotic, and cardiotonic agent. Ultraviolet B (UVB) irradiation increases the production of matrix metalloproteinase-1 (MMP-1) during skin photoaging. The UVB-induced increase of MMP-1 results in connective tissue damage, and the skin becomes wrinkled and aged. Here, we investigated the capacity of cilostazol to inhibit MMP-1 expression in UVB-irradiated human dermal fibroblasts. Cultured human dermal fibroblasts were irradiated with UVB, followed by the addition of cilostazol to the culture medium. Post-treatment with cilostazol attenuated UVB-induced production of MMP-1 and prevented the reduction of type I procollagen. Cilostazol inhibited UVB irradiation-induced phosphorylation of the mitogen-activated protein kinase (MAPK) signaling molecules Jun-N-terminal kinase (JNK) and p38 kinase, as well as activator protein-1 (AP-1) in dermal fibroblasts. Overall, these results demonstrate that cilostazol regulates UVB-induced MMP-1 expression and type I procollagen synthesis by inhibiting MAPK signaling and AP-1 activity. Therefore, we suggest that cilostazol may be useful for the prevention and treatment of skin photodamage caused by UVB-irradiation.


Vitamin D deficiency in gestational diabetes mellitus and the role of the placenta.
The aim of this study was to evaluate the relationships between maternal vitamin D levels and gestational diabetes mellitus (GDM) and differences in the placental production of vitamin D receptor (VDR), CYP24A, and CYP27B1. Forty normal pregnant women and 20 women with GDM were included in this study. Serum levels of 25-hydroxyvitamin D (25[OH]D) were measured with enzyme-linked immunosorbent assay. The expression and production of VDR, CYP27B1, and CYP24A1 in the placenta were evaluated with real time-polymerase chain reaction and Western blot, respectively. We found that 27.5% of normal pregnant women and 85% of women with GDM had vitamin D deficiency, with serum 25(OH)D levels <20 ng/mL. Serum levels of 25(OH)D were lower in women with GDM than normal pregnant women (P < .01). The production of CYP24A1 protein and messenger RNA expression was significantly higher in placental tissue from patients with GDM than in those from normal pregnancies; however, the production of CYP27B1 and VDR protein and messenger RNA expression were not different between 2 the groups. In this study, vitamin D deficiency was associated with GDM. Given that 25(OH)D is hydroxylated by CYP27B1 to the bioactive 1,25(OH)2D form, and CYP24A1 catabolizes both 25(OH)D and 1,25(OH)2D to the inactive metabolites, respectively, our data indicate that the elevated activity of CYP24A1 in the placenta may play a key role in the development of vitamin D deficiency in GDM.


[Thoracic imaging in cystic fibrosis : Radiography].
Patients with cystic fibrosis (CF) require regular follow-up examinations, usually from birth onwards, using imaging techniques. The conventional chest x‑ray examination is the only technique recommended by the guidelines at this age. The examination can be performed at every age and is well standardized. The radiation dose to the patient is small and does not require any sedation. The typical structural changes of the bronchi and the lung parenchyma can be depicted. Typical findings are air-trapping, bronchiectasis, peribronchial cuffing, bronchial wall thickening, mucus plugging, nodular opacities, atelectasis and/or consolidations and hilar lymphadenopathy. Different scoring systems have been developed to allow for a relatively easy but reproducible assessment of the severity of the disease. The most important ones for daily clinical practice are the Chrispin-Norman score, Brasfield score, and the Wisconsin score. All of them show a good correlation with pulmonary function parameters and the clinical picture of the patients. It is recommended to take an annual X‑ray for patients with CF, and in addition in individual cases with clinical deterioration. Computed tomography (CT) can be performed if relevant information for treatment is expected to gained.


Evaluation of regression methods when immunological measurements are constrained by detection limits.
The statistical analysis of immunological data may be complicated because precise quantitative levels cannot always be determined. Values below a given detection limit may not be observed (nondetects), and data with nondetects are called left-censored. Since nondetects cannot be considered as missing at random, a statistician faced with data containing these nondetects must decide how to combine nondetects with detects. Till now, the common practice is to impute each nondetect with a single value such as a half of the detection limit, and to conduct ordinary regression analysis. The first aim of this paper is to give an overview of methods to analyze, and to provide new methods handling censored data other than an (ordinary) linear regression. The second aim is to compare these methods by simulation studies based on real data. We compared six new and existing methods: deletion of nondetects, single substitution, extrapolation by regression on order statistics, multiple imputation using maximum likelihood estimation, tobit regression, and logistic regression. The deletion and extrapolation by regression on order statistics methods gave biased parameter estimates. The single substitution method underestimated variances, and logistic regression suffered loss of power. Based on simulation studies, we found that tobit regression performed well when the proportion of nondetects was less than 30%, and that taken together the multiple imputation method performed best. Based on simulation studies, the newly developed multiple imputation method performed consistently well under different scenarios of various proportion of nondetects, sample sizes and even in the presence of heteroscedastic errors.


Comparison of different versions of simulation studies following OECD 309 using 14C-labelled ionic compounds - Does it make a difference?
Simulation studies following OECD guideline 309 (Aerobic Mineralisation in Surface Water - Simulation Biodegradation Test) are performed to determine the biodegradation of chemicals in surface water. The aim of our study was to perform different versions of this test to identify possible shortcomings. In order to investigate the influence of a chemical charge on the degradation, we used 14C-labelled 4-n-dodecylbenzenesulfonic acid sodium salt (14C-DS-), 4-n-dodecylbenzyltrimethylammonium chloride (14C-DA+) and 4-n-dodecylphenol (14C-DP) Additionally, abiotic degradation of these compounds and the effectiveness of different sterilization methods was assessed. The sampling technique of withdrawing sub-samples provided insufficient recoveries for 14C-DS- and 14C-DP which were mainly below 80% of applied radioactivity (AR). Therefore, the suspended sediment test was repeated using 14C-DS-, 14C-DA+ and 14C-DP and whole flasks were harvested at each sampling time resulting in superior recoveries. For 14C-DS- and 14C-DP, mineralisation was the major route of dissipation with 14CO2 evolution of 75.3% and 69.0% AR, respectively, after 62 days. Mineralisation of 14C-DA+ accounted for only 6.7% AR. Non-extractable residues (NER) of all compounds were similar after 62 days (7.3% - 9.2% AR). Sterile test conditions were best achieved using γ-irradiated sediment and autoclaved water treated with sodium azide. Under sterile conditions, mineralisation of the test compounds as well as NER formation of 14C-DS- and 14C-DP were negligible, whereas small amounts of NER were formed for 14C-DA+ (5.5% AR). We showed that OECD guideline 309 provides too much scope for the experimental setup which affects the outcome of the test and thus, needs further refinement.


Strategies to manage paraplegia risk after endovascular stent repair of descending thoracic aortic aneurysms.
Paraplegia is a recognized complication after endovascular stent repair of descending thoracic aortic aneurysms. A management algorithm employing neurologic assessment, somatosensory evoked potential monitoring, arterial pressure augmentation, and cerebrospinal fluid drainage evolved to decrease the risk of postoperative paraplegia. Patients in thoracic aortic aneurysm stent trials from 1999 to 2004 were analyzed for paraplegic complications. Lower extremity strength was assessed after anesthesia and in the intensive care unit. A loss of lower extremity somatosensory evoked potential or lower extremity strength was treated emergently to maintain a mean arterial pressure 90 mmHg or greater and a cerebrospinal fluid pressure 10 mm Hg or less. Seventy-five patients (male = 49, female = 26, age = 75 +/- 7.4 years) had descending thoracic aortic aneurysms repaired with endovascular stenting. Lumbar cerebrospinal fluid drainage (n = 23) and somatosensory evoked potential monitoring (n = 15) were performed selectively in patients with significant aneurysm extent or with prior abdominal aortic aneurysm repair (n = 17). Spinal cord ischemia occurred in 5 patients (6.6%); two had lower extremity somatosensory evoked potential loss after stent deployment and 4 developed delayed-onset paraplegia. Two had full recovery in response to arterial pressure augmentation alone. Two had full recovery and one had near-complete recovery in response to arterial pressure augmentation and cerebrospinal fluid drainage. Spinal cord ischemia was associated with retroperitoneal bleed (n = 1), prior abdominal aortic aneurysm repair (n = 2), iliac artery injury (n = 1), and atheroembolism (n = 1). Early detection and intervention to augment spinal cord perfusion pressure was effective for decreasing the magnitude of injury or preventing permanent paraplegia from spinal cord ischemia after endovascular stent repair of descending thoracic aortic aneurysm. Routine somatosensory evoked potential monitoring, serial neurologic assessment, arterial pressure augmentation, and cerebrospinal fluid drainage may benefit patients at risk for paraplegia.


Standard clinical computed tomography fails to precisely visualise presence, course and branching points of deep cerebral perforators.
Standard computed tomography (CT) images have earned a well-established position in neuroimaging. Despite that, CT is somehow limited by its resolution, which does not enable to distinctively visualise structures smaller than 300 μm in diameter. Perforating arteries, most of which measure 100-400 μm in diameter, supply important subcortical structures (thalamus, basal ganglia, internal capsule). Consequently, pathologies affecting these vessels (e.g. lacunar strokes) can have a devastating clinical outcome. The aim of our study was to assess standard CT's ability to visualise perforators and compare it with microscopic and micro-CT pictures. We have obtained 6 brainstem and 17 basal ganglia specimens. We infused them with barium sulphate contrast medium administered into either vertebral or internal cerebral artery. After that, the specimens were fixed in formalin and subsequently a series of CT, micro-CT and microscopic examinations were performed. The median number of visualised perforators in brainstem and basal ganglia specimens was 8 and 3, respectively for CT and 18 and 7 for micro-CT (p < 0.05). Standard CT failed to clearly visualise branching points and vessels smaller than 0.25-0.5 mm (1-2 voxels) in diameter. Parallel vessels, like lenticulostriate arteries could not be differentiated in standard CT due to their proximity being smaller that the resolution. Basing on our results, we infer that CT is a poor modality for imaging of the perforators, presenting both quantitative and qualitative flaws in contrast with micro-CT.


Hematological complications with the St. Jude valve and reduced-dose Coumadin.
We examined hematological complications in 415 patients having valve replacement with the St. Jude mechanical prosthesis (212, aortic valve replacement [AVR]; 159, mitral valve replacement [MVR]; and 44, AVR + MVR). There were 164 men and 251 women with a mean age of 59 years (range, 20 to 88 years). Preoperatively 386 patients were in New York Heart Association functional classes III and IV. There were 154 associated procedures (37%), the most common being myocardial revascularization. Overall hospital mortality was 7.5% (31/415), 7% after AVR, 8% after MVR, and 7% after AVR + MVR. All operative survivors were anticoagulated with Coumadin (crystalline warfarin sodium) to maintain the prothrombin time at 1.5 times control. During a mean follow-up of 21 months (range, 6 to 60 months), there were 29 late deaths (7.6%) and 5 patients (1.3%) lost to follow-up. No patient experienced structural valve degeneration. At 48 months, actuarial freedom from thromboembolism was 87% +/- 3% after AVR and 91% +/- 9% after MVR; from anticoagulation-related hemorrhage, 97% +/- 3% after AVR and 91% +/- 3% after MVR; and from hemolysis, 100% after AVR and 98% +/- 2% after MVR. Freedom from all valve-related morbidity at 4 years was 82% +/- 5% after AVR and 75% +/- 10% after MVR. Actuarial survival at 48 months was 80% +/- 4% after AVR and 65% +/- 7% after MVR.


Photo-oxygenation to support nitrification in an algal-bacterial consortium treating artificial wastewater.
Oxygenation by photosynthesis in a mixed culture of algae and nitrifiers was investigated for its potential to support nitrification. An open photo-bioreactor (1L fermentor; 30°C) was inoculated with an enriched culture of nitrifiers obtained from activated sludge and a pure culture of Scenedesmus sp. The reactor was illuminated (60 μmol/m2 s) and operated as a sequencing batch reactor with 50% discharge per cycle and sludge retention time of 15 or 30 days. Full nitrification of synthetic wastewater (50 mgNH4+-N/L) without mechanical aeration was achieved after less than 14 cycles (18.5 h React per 24h cycle) and the biomass steadily increased until a concentration of 1.9 g VSS/L with 29 mg chl-a/L. The maximum observed nitrification rate was 7.7 mgNH4+-N/Lh. A mass balance showed that ammonium removal was primarily by nitrification (81-85%) rather than by ammonium uptake by algae.


Multipurpose dissociation cell for enhanced ETD of intact protein species.
We describe and characterize an improved implementation of ETD on a modified hybrid linear ion trap-Orbitrap instrument. Instead of performing ETD in the mass-analyzing quadrupole linear ion trap (A-QLT), the instrument collision cell was modified to enable ETD. We partitioned the collision cell into a multi-section rf ion storage and transfer device to enable injection and simultaneous separate storage of precursor and reagent ions. Application of a secondary (axial) confinement voltage to the cell end lens electrodes enables charge-sign independent trapping for ion-ion reactions. The approximately 2-fold higher quadrupole field frequency of this cell relative to that of the A-QLT enables higher reagent ion densities and correspondingly faster ETD reactions, and, with the collision cell's longer axial dimensions, larger populations of precursor ions may be reacted. The higher ion capacity of the collision cell permits the accumulation and reaction of multiple full loads of precursor ions from the A-QLT followed by FT Orbitrap m/z analysis of the ETD product ions. This extends the intra-scan dynamic range by increasing the maximum number of product ions in a single MS/MS event. For analyses of large peptide/small protein precursor cations, this reduces or eliminates the need for spectral averaging to achieve acceptable ETD product ion signal-to-noise levels. Using larger ion populations, we demonstrate improvements in protein sequence coverage and aggregate protein identifications in LC-MS/MS analysis of intact protein species as compared to the standard ETD implementation.


Dual effects of the small-conductance Ca2+-activated K+ current on human atrial electrophysiology and Ca2+-driven arrhythmogenesis: an in silico study.
By sensing changes in intracellular Ca2+, small-conductance Ca2+-activated K+ (SK) channels dynamically regulate the dynamics of the cardiac action potential (AP) on a beat-to-beat basis. Given their predominance in atria versus ventricles, SK channels are considered a promising atrial-selective pharmacological target against atrial fibrillation (AF), the most common cardiac arrhythmia. However, the precise contribution of SK current (ISK) to atrial arrhythmogenesis is poorly understood, and may potentially involve different mechanisms that depend on species, heart rates, and degree of AF-induced atrial remodeling. Both reduced and enhanced ISK have been linked to AF. Similarly, both SK channel up- and downregulation have been reported in chronic AF (cAF) versus normal sinus rhythm (nSR) patient samples. Here, we use our multiscale modeling framework to obtain mechanistic insights into the contribution of ISK in human atrial cardiomyocyte electrophysiology. We simulate several protocols to quantify how ISK modulation affects the regulation of AP duration (APD), Ca2+ transient, refractoriness, and occurrence of alternans and delayed afterdepolarizations (DADs). Our simulations show that ISK activation shortens the APD and atrial effective refractory period, limits Ca2+ cycling, and slightly increases the propensity for alternans in both nSR and cAF conditions. We also show that increasing ISK counteracts DAD development by enhancing the repolarization force that opposes the Ca2+-dependent depolarization. Taken together, our results suggest that increasing ISK in human atrial cardiomyocytes could promote reentry while protecting against triggered activity. Depending on the leading arrhythmogenic mechanism, ISK inhibition may thus be a beneficial or detrimental anti-AF strategy.NEW & NOTEWORTHY Using our established framework for human atrial myocyte simulations, we investigated the role of the small-conductance Ca2+-activated K+ current (ISK) in the regulation of cell function and the development of Ca2+-driven arrhythmias. We found that ISK inhibition, a promising atrial-selective pharmacological strategy against atrial fibrillation, counteracts the reentry-promoting abbreviation of atrial refractoriness, but renders human atrial myocytes more vulnerable to delayed afterdepolarizations, thus potentially increasing the propensity for ectopic (triggered) activity.


European postcoital tests: opinions and practice.
To assess differences in opinion and practice with regard to the postcoital test in Europe. Multilingual questionnaire survey among heads of departments of obstetrics and gynaecology with large fertility clinics in 16 European countries. Of 203 heads of departments, each responsible for 882 infertility cases per year (95% CI 657-1107) 145 (71%) responded. INFORMATION SOUGHT: Use of the postcoital test: its timing in relation to cycle and coitus, methodology used for the test, cut-off level of normality and treatments applied for abnormal test results. The postcoital test is used in 92% (and routinely in 68%) of departments. There are large differences in timing of the test in relation to menstrual cycle and coitus, in microscopic magnification used, and in cut-off levels of normality. More than 10 different treatments are applied for abnormal test results. Guidelines of the World Health Organisation are not followed and divergence in practice and opinion is wide enough to question whether infertile couples are better off with than without the test.


A new sulphated nor-sesquiterpene from mangrove Laguncularia racemosa (L.) Gaertn. F.
A new sulphated nor-sesquiterpene, 3alpha-hydroxysulphonyloxy-5alpha,6alpha-epoxy-megastigmen-9-one (1), and a known sulphated lipid, 5'-(hydroxysulphonyloxy) jasmonic acid (2), have been isolated from the twigs and leaves of mangrove Laguncularia racemosa (L.) Gaertn. F. The structure of the new compound was elucidated on the basis of detailed analysis of spectroscopic data and chemical reaction.


G-C3N4 sheet nanoarchitectonics with island-like crystalline/amorphous homojunctions towards efficient H2 and H2O2 evolution.
Photocatalystic evolution of H2O2 from water and oxygen has attracted significant attention because of environmentally friendly. The absorption in visible and hydrophilic feature of graphitic carbon nitride (g-C3N4) make it a good candidate. In this paper, a rapid post-treatment at high temperature was developed to obtain g-C3N4 nanosheets with abundant crystalline/amorphous interfaces to form homojunctions, which optimized uniplanar carrier mobility dynamics. The conversion from bulk to two-dimensional g-C3N4 resulted from the breakage of interplanar hydrogen bonds and interlayer Van der Waals force. The unique morphology not only rendered photocatalyst with larger specific surface area but also inhibited the robust volume recombination of charge carriers. The accelerated charge carriers flow at the interface, interplane and interlayer together ameliorated the separation and transfer of electrons and holes. A new-emerged n→π* transition ameliorated the poor light utilization efficiency. Beyond the increased photocatalytic H2 evolution property (779.2 μmol g-1 h-1), optimized sample displayed a H2O2 evolution activity as high as 4877.1 μM g-1 h-1 under visible light illumination, which was ∼5.8 times of that of bulk g-C3N4. Detailed photocatalytic mechanism investigation manifested that the two-step single-electron oxygen reduction process occupied the dominant status in H2O2 evolution. This work proposed a novel strategy for obtaining g-C3N4 homojunctions as a promising bi-functional metal-free catalyst to be applied in clean energy production field.


The circadian rhythm of LH release can be shifted by injections of a benzodiazepine in female golden hamsters.
Three daily injections of the short-acting benzodiazepine, triazolam, induced pronounced phase shifts in the onset of both the circadian surge in pituitary LH release and the circadian rhythm of locomotor activity in ovariectomized hamsters treated with estrogen. Both the magnitude and the direction of the phase shifts in these two rhythms were similar. These results indicate that a master circadian clock underlying diverse behavioral and endocrine rhythms can be reset by treatment with triazolam.


Mechanochemical endovenous Ablation versus RADiOfrequeNcy Ablation in the treatment of primary great saphenous vein incompetence (MARADONA): study protocol for a randomized controlled trial.
Radiofrequency ablation (RFA) is associated with an excellent outcome in the treatment of great saphenous vein (GSV) incompetence. The use of thermal energy as a treatment source requires the instillation of tumescence anesthesia. Mechanochemical endovenous ablation (MOCA) combines mechanical endothelial damage, using a rotating wire, with the infusion of a liquid sclerosant. Tumescence anesthesia is not required. Preliminary experiences with MOCA showed good results and low post-procedural pain. The MARADONA (Mechanochemical endovenous Ablation versus RADiOfrequeNcy Ablation) trial is a multicenter randomized controlled trial in which 460 patients will be randomly allocated to MOCA or RFA. All patients with primary GSV incompetence who meet the eligibility criteria will be invited to participate in this trial. The primary endpoints are anatomic and clinical success at a one-year follow-up, and post-procedural pain. The secondary endpoints are technical success, complications, operation time, procedural pain, disease-specific quality of life, time taken to return to daily activities and/or work, and cost-efficiency analyses after RFA or MOCA. Both groups will be evaluated on an intention to treat base. The MARADONA trial is designed to show equal results in anatomic and clinical success after one year, comparing MOCA with RFA. In our hypothesis MOCA has an equal anatomic and clinical success compared with RFA, with less post-procedural pain. Clinicaltrials NCT01936168.


Genome-wide survey of repetitive DNA elements in the button mushroom Agaricus bisporus.
Repetitive DNA elements are ubiquitous constituents of eukaryotic genomes. The biological roles of these repetitive elements, supposed to impact genome organization and evolution, are not completely elucidated yet. The availability of whole genome sequence offers the opportunity to draw a picture of the genome-wide distribution of these elements and provide insights into potential mechanisms of genome plasticity. The present study uses in silico approaches to describe tandem repeats and transposable elements distribution in the genome of the button mushroom, Agaricus bisporus. Transposable elements comprised 12.43% of the assembled genome, and 66% of them were found clustered in the centromeric or telomeric regions. Methylation of retrotransposon has been demonstrated. A total of 1996 mini-, 4062 micro-, and 37 satellites motifs were identified. The microsatellites appeared widely and evenly spread over the whole genome sequence, whereas the minisatellites were not randomly distributed. Indeed, minisatellites were found to be associated with transposable elements clusters. Telomeres exhibited a specific sequence with a T(n)AG(n) signature. A comparison between the two available genome sequences of A. bisporus was also performed and sheds light on the genetic divergence between the two varieties. Beyond their role in genome structure, repeats provide a virtually endless source of molecular markers useful for genetic studies in this cultivated species.


[Study on the immunoglobulin class of the antibody anti-e in acute HBsAg+ hepatitis and in asymptomatic carriers of HBsAg (author's transl)].
The AA. have determined the immunoglobulinic appartenence class of HBeAb in patients with acute hepatitis HBsAg+ and in asymptomatic carriers of HBsAg. In these the HBeAb belongs constantly to the IgG class, while in acute hepatitis patients this antibody results present in the IgM and IgG fractions of the sera. Especially, in patients with acute hepatitis the presence of HBeAb was observed only in some sera obtained during the tardive phase of disease.


Construction and evaluation of food-grade vectors for Lactococcus lactis using aspartate aminotransferase and alpha-galactosidase as selectable markers.
We report development of two food-grade cloning vectors for Lactococcus lactis, which utilize either a lactococcal aspartate aminotransferase gene (aspC), or Bifidobacterium longumalpha-galactosidase gene (aglL) as selectable markers. The theta-replicon of lactococcal plasmid, pW563, was combined with aspC and a multiple cloning site. When electroporated into L. lactis JLS400 (AspC-), the resulting vector, pSUW611 (3.9 kbp), restores ability of the mutant to grow in milk thus allowing for selection of the transformants. The vector is stable during 100 generations of nonselective growth (0.2% loss per generation). The second vector, pSUW711 (5.1 kbp), was constructed by exchanging aspC with aglL under the control of usp45 promoter. Lactococcus lactis transformed with pSUW711 produced distinctive colonies within 48-72 h on melibiose-containing plates. Expression of two Lactobacillus helveticus peptidases was attempted using these new vehicles. Introduction of pepN on pSUW611 and pSUW711 into L. lactis led to a sixfold, or 27-fold increase in aminopeptidase activity, respectively. However, no changes in endopeptidase activity were recorded upon transformation with pSUW611 carrying pepO2 under control of three different promoters. Attempts were also made to construct high copy variants of pSUW711. The aspC and aglL can be employed as food-grade genetic markers for L. lactis. The vectors, pSUW611 and pSUW711, were successfully used to express Lact. helveticus PepN in L. lactis. Two novel food-grade vectors were developed which provide simple and convenient selection and maintenance in L. lactis.


Bactericidal antibody against a representative epidemiological meningococcal serogroup B panel confirms that MATS underestimates 4CMenB vaccine strain coverage.
4CMenB (Bexsero), a vaccine developed against invasive meningococcal disease caused by capsular group B strains (MenB), was recently licensed for use by the European Medicines Agency. Assessment of 4CMenB strain coverage in specific epidemiologic settings is of primary importance to predict vaccination impact on the burden of disease. The Meningococcal Antigen Typing System (MATS) was developed to predict 4CMenB strain coverage, using serum bactericidal antibody assay with human complement (hSBA) data from a diverse panel of strains not representative of any specific epidemiology. To experimentally validate the accuracy of MATS-based predictions against strains representative of a specific epidemiologic setting. We used a stratified sampling method to identify a representative sample from all MenB disease isolates collected from England and Wales in 2007-2008, tested the strains in the hSBA assay with pooled sera from infant and adolescent vaccinees, and compared these results with MATS. MATS predictions and hSBA results were significantly associated (P=0.022). MATS predicted coverage of 70% (95% CI, 55-85%) was largely confirmed by 88% killing in the hSBA (95% CI, 72-95%). MATS had 78% accuracy and 96% positive predictive value against hSBA. MATS is a conservative predictor of strain coverage by the 4CMenB vaccine in infants and adolescents.


Cataract-causing variant Q70P damages structural stability of βB1-crystallin and increases its tendency to form insoluble aggregates.
Congenital cataract is the primary cause of childhood blindness worldwide. As the predominant structural protein, βB1-crystallin plays an important role in maintaining lens transparency and cellular homeostasis. Numerous cataract-causing mutations of βB1-crystallin have been identified with unclear pathogenic mechanism. We previously identified the mutation Q70P (Q to P at residue position 70) of βB1-crystallin linked to congenital cataract in a Chinese family. In this work, we investigated the potential molecular mechanism of βB1-Q70P in the congenital cataract at the molecular, protein, and cellular levels. We purified recombinant βB1 wild-type (WT) and Q70P proteins and compared their structural characteristics and biophysical properties by spectroscopic experiments under physiological temperature and environmental stresses (ultraviolet irradiation, heat stress, oxidative stress). Notably, βB1-Q70P significantly changed the structures of βB1-crystallin and exhibited lower solubility at physiological temperature. Meanwhile, βB1-Q70P was prone to aggregation in eukaryotic and prokaryotic cells, and was more sensitive to environmental stresses, along with impaired cellular viability. Furthermore, the molecular dynamics simulation indicated that the mutation Q70P damaged secondary structures and hydrogen bond network of βB1-crystallin, which were essential for the first Greek-key motif. This study delineated the pathological mechanism of βB1-Q70P and provided novel insights into treatment and prevention strategies for cataract-associated βB1 mutations.


Modelling of mycobacterial load reveals bedaquiline's exposure-response relationship in patients with drug-resistant TB.
Bedaquiline has been shown to reduce time to sputum culture conversion (SCC) and increase cure rates in patients with drug-resistant TB, but the influence of drug exposure remains uncharacterized. To investigate whether an exposure-response relationship could be characterized by making better use of the existing information on pharmacokinetics and longitudinal measurements of mycobacterial load. Quantitative culture data in the form of time to positivity (TTP) in mycobacterial growth indicator tubes obtained from a randomized placebo-controlled Phase IIb registration trial were examined using non-linear mixed-effects methodology. The link to individual bedaquiline exposures and other patient characteristics was evaluated. The developed model included three simultaneously fitted components: a longitudinal representation of mycobacterial load in patients, a probabilistic component for bacterial presence in sputum samples, and a time-to-event model for TTP. Data were described adequately, and time to SCC was well predicted. Individual bedaquiline exposure was found to significantly affect the decline in mycobacterial load. Consequently, the proportion of patients without SCC at week 20 is expected to decrease from 25% (95% CI 20%-31%) without bedaquiline to 17% (95% CI 13%-21%), 12% (95% CI 8%-16%) and 7% (95% CI 4%-11%), respectively, with half the median, median and double the median bedaquiline exposure observed in patients with standard dosing. Baseline bacterial load and level of drug resistance were other important predictors. To our knowledge, this is the first successful description of bedaquiline's exposure-response relationship and may be used when considering dose optimization. Characterization of this relationship was possible by integrating quantitative information in existing clinical data using novel models.


Meta-analysis of randomized controlled trials comparing intracoronary and intravenous administration of glycoprotein IIb/IIIa inhibitors in patients with ST-elevation myocardial infarction.
Glycoprotein IIb/IIIa receptor inhibitors (GPIs) have been widely adopted as an adjuvant regimen during primary percutaneous coronary intervention in patients with ST-segment elevation myocardial infarction, but whether intracoronary administration of these potent antiplatelet agents conveys better efficacy and safety over the intravenous route has not been well addressed. A meta-analysis was performed by a systematic search of the published research for randomized controlled trials comparing intracoronary versus intravenous administration of GPIs in patients with ST-segment elevation myocardial infarction. Eight studies involving 686 patients in the intracoronary arm and 660 in the intravenous arm met the inclusion criteria. Postprocedural Thrombolysis In Myocardial Infarction (TIMI) grade 3 flow (odds ratio [OR] 1.46, 95% confidence interval [CI] 1.08 to 1.98, p <0.05) and myocardial reperfusion grade 2 or 3 (OR 1.78, 95% CI 1.29 to 2.46, p <0.001) were markedly more often achieved in patients who received intracoronary boluses of GPIs than those receiving the intravenous strategy. Intracoronary administration resulted in a reduced incidence of mortality (OR 0.44, 95% CI 0.21 to 0.92, p <0.05), target vessel revascularization (OR 0.53, 95% CI 0.29 to 0.99, p <0.05), and the composite end point of major adverse cardiac events (OR 0.48, 95% CI 0.31 to 0.76, p <0.005) at 30-day follow-up. No significant difference was found in terms of major or minor bleeding (OR 1.14, p = 0.71, and OR 0.86, p = 0.47 respectively). In conclusion, intracoronary administration of GPIs yielded favorable outcomes in postprocedural blood flow restoration and 30-day clinical prognosis in patients with ST-segment elevation myocardial infarction. The intracoronary use of GPIs can be recommended as a preferred regimen during primary percutaneous coronary intervention.


Pretreatment evaluation of chronic hepatitis C: risks, benefits, and costs.
Chronic hepatitis C (CHC) infection affects nearly 4 million people in the United States. Treatment with interferon alfa-2b has been limited by its cost and low likelihood of long-term response. To examine the cost-effectiveness of alternative pretreatment management strategies for patients with CHC. Decision and cost-effectiveness analysis using a Markov model to examine prevalence of genotypes, viral load, and histological characteristics in relation to the sustained response rate with treatment. Data were based on a previously published decision model and a MEDLINE literature search for hepatitis C, biopsy, and liver from 1966 to 1996. A hypothetical population of patients with CHC infection and elevated serum alanine aminotransferase level. Combinations of liver biopsy, genotyping, and quantitative viral load determination prior to a single 6-month course of interferon alfa-2b; empirical interferon treatment; and conservative management. Proportion of sustained responders, lifetime costs, life expectancy, and quality-adjusted life expectancy. Strategies involving hepatitis C virus (HCV) RNA testing had marginal cost-effectiveness ratios up to $4400 per discounted quality-adjusted life-year gained but would miss up to 36% of sustained responders. Empirical interferon treatment had a marginal cost-effectiveness ratio of $12400 per discounted quality-adjusted life-year gained and reached all potential sustained responders. Strategies involving liver biopsy were more expensive and would miss 6% of sustained responders and yield slightly lower life expectancies. Routine liver biopsy before treatment with interferon increases the cost of managing patients with CHC without improving health outcomes. Using quantitative HCV RNA testing to guide therapy misses some potential sustained responders. Empirical interferon treatment has a marginal cost-effectiveness ratio within the bounds of other commonly accepted therapies and misses none of the sustained responders.


Alteration of mouse oocyte quality after a subchronic exposure to depleted Uranium.
Gametes and embryo tissues are known to represent a sensitive target to environmental toxicants exposure. Oocyte quality can impact subsequent developmental competence, pregnancy course and even adult health. The major health concern from depleted uranium (DU) is mainly centred on its chemotoxic properties as a heavy metal. Little attention was paid to the impact of uranium on female gamete quality. The aim of this research was to evaluate the effect of DU on mouse oocyte quality after 49 days of subchronic contamination in drinking water and to correlate the observed effects with the amount of DU accumulated in organs. Four different DU concentrations were investigated: 0 (control), 10 (DU10), 20 (DU20) and 40 mg L(-1) (DU40). DU did not influence the intensity of ovulation but affected oocyte quality. The proportion of healthy oocytes was reduced by half (P<0.001) from 20 mg L(-1) compared with control group (0.537; 0.497; 0.282 and 0.239 in control, DU10, DU20 and DU40 groups respectively) whereas no accumulation of DU was recorded in the ovaries whatever the dose tested. Abnormal perivitelline space (P<0.001) or absence of the 1st polar body (P<0.001) was identified as the main characteristic of DU impact. In the context of this study, the NOAEL for oocyte quality was determined at 10 mg L(-1) in drinking water (1.9 mg kg(-1)day(-1)). An increase in the dose of contamination over 20 mg L(-1) did not amplify the proportion of oocytes contracting a specific alteration but conducted to a diversification in oocytes abnormalities. Further investigations are necessary to correlate morphologic assessment of female gamete with its developmental competence.


Esc4p, a new target of Mec1p (ATR), promotes resumption of DNA synthesis after DNA damage.
The DNA damage-responsive protein kinases ATM and ATR phosphorylate SQ/TQ motifs that lie in clusters in most of their in vivo targets. Budding yeast Esc4p contains a cluster of SQ/TQ motifs, suggesting that it might be a target of Mec1p/Tel1p (yeast ATR/ATM). Here it is reported that Esc4p is phosphorylated by Mec1p in response to DNA damage during DNA replication and that cells lacking Esc4p are hypersensitive to DNA damage specifically during S phase. Esc4p is not required for the intra-S-phase checkpoint but is essential for resumption of chromosome replication after DNA damage, and its role in promoting restart appears to be distinct from that of Rad53p. Mutation of Esc4p SQ/TQ motifs phosphorylated by Mec1p or mutation of the BRCT domains of Esc4p also renders cells unable to restart DNA replication after DNA damage and causes hypersensitivity to genotoxins. These results identify Esc4p as an important new S-phase-specific target of Mec1p.


Biochemical characterization of halorhodopsin in native membranes.
Procedures are described for selectively radiolabeling the protein moiety (haloopsin) or the chromophoric prosthestic group (retinal) of the light-driven chloride pump halorhodopsin in intact cells of Halobacterium halobium. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autofluorography, two retinal-binding polypeptides are observed to band near the known molecular weight of the halorhodopsin chromophoric polypeptide (25,000). Synthesis of one of these polypeptides is controlled by retinal and is sufficient for generation of complete halorhodopsin function. The other is constitutively produced by the cells and differs chemically from the haloopsin protein as indicated by differences in their V8 protease digestion patterns. V8 protease cleavage of haloopsin in its native membrane is compared with that of the protein in denaturing and nondenaturing detergents. Protease cleavage sites available in the denatured haloopsin molecule are hidden in its native membrane-integrated conformation and in nondenaturing detergent micelles. Treatment with a variety of proteases indicates susceptibility of a short terminal region of the haloopsin chain in its native conformation.


High crystallinity of tunicate cellulose nanofibers for high-performance engineering films.
Tunicate cellulose nanofibers (CNFs) have received widespread attention as renewable and eco-friendly engineering materials because of their high crystallinity and mechanical stiffness. Here, we report the effects of disintegration process conditions on structure-property relationships of tunicate CNFs. By varying the hydrolysis time, we could establish a correlation between crystallinity of the CNFs with linearity and stiffness, which produces different molecular ordering within their nanostructured films. Despite having identical raw materials, tensile strength and thermal conductivity of the resulting layered films varied widely, ranging from 95.6 to 205 MPa and from 1.08 to 2.37 W/mK respectively. Furthermore, nanolayered CNF films provided highly anisotropic thermal conductivities with an in- and through-plane ratio of 21.5. Our systematic investigations will provide general and practical strategies in tailoring material properties for emerging engineering applications, including flexible paper electronics, heat sink adhesives and biodegradable, implantable devices.


In Silico Modeling of Small Molecule Carboxamides as Inhibitors of SARS-CoV 3CL Protease: An Approach Towards Combating COVID-19.
The quantitative structure-activity relationship (QSAR) approach is most widely used for the prediction of biological activity of potential medicinal compounds. A QSAR model is developed by correlating the information obtained from chemical structures (numerical descriptors/ independent variables) with the experimental response values (the dependent variable). In the current study, we have developed a QSAR model to predict the inhibitory activity of small molecule carboxamides against severe acute respiratory syndrome coronavirus (SARS-- CoV) 3CLpro enzyme. Due to the structural similarity of this enzyme with SARS-CoV-2, the causative organism of the recent pandemic, the former may be used for the development of therapies against coronavirus disease 19 (COVID-19). The final multiple linear regression (MLR) model was based on four two-dimensional descriptors with definite physicochemical meaning. The model was strictly validated using different internal and external quality metrics. The model showed significant statistical quality in terms of determination coefficient (R2=0.748, adjusted R2 or R2 adj = 0.700), cross-validated leave-one-out Q2 (Q2=0.628) and external predicted variance R2 pred = 0.723. The final validated model was used for the prediction of external set compounds as well as to virtually design a new library of small molecules. We have also performed a docking analysis of the most active and least active compounds present in the dataset for comparative analysis and to explain the features obtained from the 2D-QSAR model. The derived model may be useful to predict the inhibitory activity of small molecules within the applicability domain of the model only based on the chemical structure information prior to their synthesis and testing.