Topography of histone H3-H4 interaction with the Hat1-Hat2 acetyltransferase complex.
Chaperones influence histone conformation and intermolecular interaction in multiprotein complexes, and the structures obtained with full-length histones often provide more accurate and comprehensive views. Here, our structure of the Hat1-Hat2 acetyltransferase complex bound to Asf1-H3-H4 shows that the core domains of H3 and H4 are involved in binding Hat1 and Hat2, and the N-terminal tail of H3 makes extensive interaction with Hat2. These findings expand the knowledge about histone-protein interaction and implicate a function of Hat2/RbAp46/48, which is a versatile histone chaperone found in many chromatin-associated complexes, in the passing of histones between chaperones.


Glutathione and its dependent enzymes' modulatory responses to toxic metals and metalloids in fish--a review.
Toxic metals and metalloid are being rapidly added from multiple pathways to aquatic ecosystem and causing severe threats to inhabiting fauna including fish. Being common in all the type of aquatic ecosystems such as freshwater, marine and brackish water fish are the first to get prone to toxic metals and metalloids. In addition to a number of physiological/biochemical alterations, toxic metals and metalloids cause enhanced generation of varied reactive oxygen species (ROS) ultimately leading to a situation called oxidative stress. However, as an important component of antioxidant defence system in fish, the tripeptide glutathione (GSH) directly or indirectly regulates the scavenging of ROS and their reaction products. Additionally, several other GSH-associated enzymes such as GSH reductase (GR, EC 184.108.40.206), GSH peroxidase (EC 220.127.116.11), and GSH sulfotransferase (glutathione-S-transferase (GST), EC 22.214.171.124) cumulatively protect fish against ROS and their reaction products accrued anomalies under toxic metals and metalloids stress conditions. The current review highlights recent research findings on the modulation of GSH, its redox couple (reduced glutathione/oxidised glutathione), and other GSH-related enzymes (GR, glutathione peroxidase, GST) involved in the detoxification of harmful ROS and their reaction products in toxic metals and metalloids-exposed fish.


A cross-sectional study on the knowledge, attitude, and practice of pregnant women regarding medication use and restriction during pregnancy.
The use of medication during pregnancy is a complex issue that requires careful consideration to avoid potential harm to the developing fetus. Despite the existence of guidelines and restrictions on medication use during pregnancy, pregnant women in Nigeria often have limited access to information regarding drug use and may rely on their own judgment or the advice of non-professionals when making decisions about medication use. To assess Nigerian pregnant women's knowledge, attitude, and practice toward the use of drugs and restriction/contraindications in pregnancy. A descriptive cross-sectional study was conducted among pregnant women who visited the Federal Medical Center, Umuahia, from January to February 2023. A self-administered questionnaire was answered by conveniently sampled participants. Data were collected about their sociodemographic characteristics, knowledge, attitude, and practice. Descriptive statistics and inferential statistics utilizing the Pearson's chi-Square test were used for data analysis. The level of significance was set at p < 0.05. One hundred and fifty-two pregnant women completed the questionnaire (60.8% response rate) and were mostly married (n = 148; 95.0%). Majority were between the age of 25 to 35 years (n = 107; 70.4%), had a university/bachelor's degree (n = 94; 61.8%), and were business women (n = 85; 56.0%). About 35.5% of the respondents had good knowledge of medication restrictions in pregnancy. The majority of the respondents had a positive attitude (n = 98; 64.5%) and good practice (n = 139; 91.4%). The findings established that a substantial proportion of the women had a positive attitude and good practice of medication use and restrictions in pregnancy. However, the level of knowledge was surprisingly poor among the study group. There is a need to educate pregnant women on safe medication use and avoidance during pregnancy.


Chickpea chlorotic stunt virus: a threat to cool-season food legumes.
Chickpea chlorotic stunt virus (CpCSV, genus Polerovirus, family Solemoviridae), first reported in Ethiopia in 2006, causes an economically important yellowing and stunting disease in legume crops such as chickpea, faba bean, field pea, and lentil in most production areas of North Africa and Central and West Asia. Disease epidemics have been reported in Ethiopia, Syria, and Tunisia. The virus is transmitted persistently by aphids of the species Aphis craccivora and Acyrthosiphon pisum and naturally infects several legume and non-legume hosts. CpCSV exists as at least two geographic strain groups that differ in their genome sequence and serological and biological properties. In addition, a genetically divergent isolate proposed to be a member of a distinct polerovirus species has been reported from pea and faba bean in China. The ssRNA genome of the Ethiopian isolate has 5900 nucleotides, is encapsidated in isometric particles of ~ 28 nm diameter, and is suggested to have evolved by recombination of cucurbit aphid-borne yellows virus- and soybean dwarf virus-like parents. Moreover, a number of newly reported poleroviruses are suggested to have evolved by recombination between CpCSV and other parental poleroviruses. Identification of sources of resistance and further knowledge on disease epidemiology, including specific strains, vectors, and alternate hosts in different growing areas, are required for devising effective disease management strategies. Modern biotechnology tools such as next-generation sequencing, molecular markers, and agroinoculation-based resistance screening techniques can expedite future research and management efforts. This review addresses various aspects of CpCSV, including its properties, ecology, the disease it causes, management options, and future research perspectives.


Maximizing diversity within and among teams in a large-scale project.
This work aims to improve an earlier methodology for assigning personnel to diverse three-member teams. Notably, the original algorithm focused only on diversity within teams, to ensure that conflicting interests are represented in each team. While this indeed created diverse teams, in many cases different teams featured the same combination of conflicting interests. The client for the original project, a government agency, asked for a methodology that produces more combinations. Hence, the current study presents an approach for boosting diversity among teams. That is, the new method maximizes the differences among teams, not only differences within teams. This was achieved by limiting the number of times each combination appears, while maintaining maximal diversity within teams as well. The upgraded algorithm is scalable and fast converging.•We suggest an integer linear programming algorithm for creating thousands of diverse three-member teams that represent the conflicting interests of different groups.•The algorithm imposes penalty costs on potential assignments based on their deviations from the project's requirements and sets upper bound constraints on the frequency of different assignments.•The algorithm is efficient, scalable, and converges to maximal diversity within seconds.


Emotion regulation, perfectionism, and eating disorder symptoms in adolescents: the mediating role of cognitive eating patterns.
Poor emotion regulation, along with elevated perfectionism, is recognised as a risk factor for the development of eating disorder (ED) symptoms. Therefore, the aim of our study was to examine the relationship between emotion regulation difficulties and perfectionism with ED symptoms, while controlling for emotional eating, uncontrolled eating, and cognitive restraint as mediators. In total, 482 adolescents (246 girls and 236 boys; M = 15.00, SD = 0.31) participated in this study. Based on the participants' body height and weight measured by the medical team during their regular preventive health examinations, the body mass index (BMI) was calculated. Participants also answered the Difficulties in Emotion Regulation Scale, Multidimensional Perfectionism Scale, Three-Factor Eating Questionnaire, and Eating Disorder Examination Questionnaire. According to the results obtained, adolescent girls in our sample reported more difficulties in emotion regulation, more uncontrolled and emotional eating, and more ED symptoms. A significant direct effect was found for difficulties in emotion regulation and ED symptoms but not for perfectionism. In addition, only emotional eating and cognitive restraint (not uncontrolled eating) mediated the relationship between difficulties in emotion regulation and perfectionism and ED symptoms. The results suggest that difficulties in emotion regulation may be more important than perfectionism in explaining ED symptoms in a sample of healthy adolescents. When adolescents experience problems in emotion regulation in combination with emotional or uncontrolled eating, they might be at a higher risk of experiencing concerns about weight and a variety of other ED symptoms, and this should be considered when planning preventive interventions for adolescents.


Assembly of motor proteins, PomA and PomB, in the Na+-driven stator of the flagellar motor.
PomA and PomB are transmembrane proteins that form the stator complex in the sodium-driven flagellar motor of Vibrio alginolyticus and are believed to surround the rotor part of the flagellar motor. We constructed and observed green fluorescent protein (GFP) fusions of the stator proteins PomA and PomB in living cells to clarify how stator proteins are assembled and installed into the flagellar motor. We were able to demonstrate that GFP-PomA and GFP-PomB localized to a cell pole dependent on the presence of the polar flagellum. Localization of the GFP-fused stator proteins required their partner subunit, PomA or PomB, and the C-terminal domain of PomB, which has a peptidoglycan-binding motif. Each of the GFP-fused stator proteins was co-isolated with its partner subunit from detergent-solubilized membrane. From these lines of evidence, we have demonstrated that the stator proteins are incorporated into the flagellar motor as a PomA/PomB complex and are fixed to the cell wall via the C-terminal domain of PomB.


Detection and isolation of H5N1 influenza virus from large volumes of natural water.
Various species of aquatic or wetlands birds can be the natural reservoir of avian influenza A viruses of all hemagglutinin (HA) subtypes. Shedding of the virus into water leads to transmission between waterfowl and is a major threat for epidemics in poultry and pandemics in humans. Concentrations of the influenza virus in natural water reservoirs are often too low to be detected by most methods. The procedure was designed to detect and isolate low concentrations of the influenza virus in large volumes of water without the need for costly installations and reagents. The virus was adsorbed onto formalin-fixed erythrocytes and subsequently isolated in chicken embryos. Sensitivity of the method was determined using a reverse-genetic H5N1 virus. A concentration as low as 0.03 of the 50% egg infection dose per milliliter (EID50/ml) of the initial volume of water was effectively detected. The probability of detection was approximately 13%, which is comparable to that of detecting the influenza virus M-gene by PCR amplification. The method can be used by field workers, ecologists, ornithologists, and researchers who need a simple method to isolate H5N1 influenza virus from natural reservoirs. The detection and isolation of virus in embryonated chicken eggs may help epidemiologic, genetic, and vaccine studies.


Tramadol utilization among patients with higher risk of adverse drug events: A claims analysis of commercially insured and Medicare Advantage members.
To assess prescribing of tramadol among patients with contraindications and higher risks of adverse events in a large population of commercially insured and Medicare Advantage members. We performed a cross-sectional analysis evaluating tramadol utilization in patients with higher risk of adverse outcomes. This study utilized 2016-2017 data from the Optum Clinformatics Data Mart. Patients with at least one tramadol prescription without a cancer or sickle cell diagnosis during the study period. We first determined if tramadol was prescribed among patients with contraindications or risk factors for adverse outcomes. We then determined if patient demographic or clinical factors were associated with the use of tramadol in these higher-risk scenarios using multivariable logistic regression models. Among patients with at least one prescription for tramadol, 19.66 percent (99 percent CI: 19.57-19.75) concurrently received an interacting cytochrome P450 isoenzyme medication, 19.24 percent (99 percent CI: 19.15-19.33) concurrently received a serotonergic medication, and 7.93 percent (99 percent CI: 7.88-8.00) concurrently received a benzodiazepine. Additionally, 1.59 percent (99 percent CI: 1.56-1.61) of patients who received tramadol also had a seizure disorder, while 0.55 percent (99 percent CI: 0.53-0.56) of patients were under the age of 18. Overall, nearly one in three patients (31.17 percent) received tramadol in the presence of at least one of these risks (99 percent CI: 31.06-31.27). Almost one in three patients prescribed tramadol had a clinically significant drug interaction or contraindication for use, suggesting that prescribers often disregard these concerns. Real-world studies are needed to better understand the likelihood of harms associated with the use of tramadol in these contexts.


1H-NMR study of the effect of synthetic polymers on the fluidity, transition temperature and fusion of dipalmitoyl phosphatidylcholine small vesicles.
The interaction of water-soluble polymers with dipalmitoyl phosphatidylcholine small vesicles and the effect on vesicle fusion were studied by means of 1H-NMR spectrometry. The motion of dipalmitoyl phosphatidylcholine molecules decreased on interaction with the polymers and was detected as a change in the signal intensity. The interaction behavior of polymers is very sensitive to the chemical structure of the applied polymers. Poly(styrene sulfonic acid) and poly(ethylene glycol) decreased the motion of the choline methyl group, predominantly through coulombic and hydrophobic interaction forces, respectively. For example, in the case of the poly(styrene sulfonic acid)-containing system, the signal intensity of the choline methyl group was decreased about 15% while those of the hydrophobic methylene and terminal methyl groups were scarcely decreased by the addition of polymer to a final concentration of 4.0 x 10(-2) unit mol/l. These polymers are considered to interact with the surface of the vesicle membrane. On the other hand, poly(L-glutamic acid) and poly(N-vinyl-2-pyrrolidone) decreased the signal intensities of not only the choline methyl group, but also those of the hydrophobic methylene and terminal methyl groups. This result suggests that part of these polymers might be incorporated into the hydrophobic region of the vesicle membrane. Addition of the non-ionic polymers inhibited vesicle fusion considerably. This effect was explained by the stabilization of dipalmitoyl phosphatidylcholine vesicles by complexation with these polymers.


Solid-phase preparation of hydantoins through a new cyclization/cleavage step.
An efficient process for the solid-phase synthesis of hydantoins has been developed. The amino acid starting material is anchored to the resin from its carboxylic acid end through formation of a very stable amide bond. After introduction of different functional groups, the cleavage/cyclization step can be performed in acidic or basic conditions.


Allergy to various components of topical preparations in stasis dermatitis and leg ulcer.
Patients suffering from stasis dermatitis and leg ulcers (192) were found to give an allergic reaction in epicutaneous tests to one or more components of topical preparations in 69.2% of cases. The most common reactions were obtained with neomycin 34.0% and its component framycetin 31.2%, wool alcohols 22.9%, balsam of Peru 21.4%, chlorquinaldon 16.7%, bacitracin 13.1%, chinoform 10.9%, dequalin 9.9%, formaldehyde 4.2% and parabens 3.6%. The incidence of allergic reactions reflects local use of preparations containing various allergens as well as their capacity to produce alllergy. Allergy may in some cases interfere with effective topical therapy. Debrisan dextran polymer beads were found to be suitable for treatment of ulcers of multiallergic patients without risk of contact allergy.


Improving patient selection for immuno-oncology phase 1 trials: External validation of six prognostic scores in a French Cancer Center.
We compared the performance of six prognostic scores (Royal Marsden Hospital, MDACC: MD Anderson Clinical Center and MDACC + NLR: neutrophil-to-lymphocyte ratio, MD Anderson - immune checkpoint inhibitors (MDA-ICI), GRIm: Gustave Roussy Immune Score and LIPI: Lung Immune Prognostic Index) in predicting overall survival (OS) in phase I trial patients treated with immune checkpoint inhibitors (ICI). Medical records of patients with advanced solid tumors enrolled in ICI phase I trials between 2015 and 2018 at Institut Universitaire du Cancer de Toulouse-Oncopole were reviewed. The performance of prognostic scores on OS was compared using different criteria. A total of 259 patients were included. Median age was 63 years (range: 18-83). Main primary cancers were melanoma (19%), head and neck (16%), lung (13%) and bladder (10%). With a median follow-up of 15 months (95% confidence interval [CI] = [11.6;17.5]), median OS was 12.5 months (95% CI = [10.3;16.0]). All scores were associated with OS. The MDACC, LIPI and GRIm scores performed better than the others. Concordance of risk group assignment between the scoring systems was poor. According to our results, the MDACC, GRIm and LIPI scores better suited to ICI phase I settings. Adequate scoring would allow better patient selection in early ICI trials, especially during the critical period of dose escalation, and in proof-of-concept expansion cohorts.


Short- and Long-Term Evaluation of General Practitioners' Competences After a Training in Melanoma Diagnosis: Refresher Training Sessions May Be Needed.
General practitioners (GPs) are first-line clinicians in melanoma diagnosis. It is, therefore, important to ensure that they maintain their melanoma diagnostic accuracy over time. The objective of this study was to assess the short- and long-term competences of GPs after a training session in naked-eye melanoma diagnosis. An interventional prospective study was conducted whereby, over a 6-month period, GPs attended a 1-h melanoma diagnostic training session. To assess their acquired competences, GPs were asked to fill in a questionnaire on basic melanoma knowledge and to evaluate 10 clinical images of pigmented skin lesions prior to training, immediately after and 1 year later. In total, 89 GPs completed the questionnaire prior and immediately after training. As expected, the number of GPs who appropriately managed [Formula: see text] 50% of the melanoma cases increased after training (P < 0.001). One year after training, only 27 (30%) of the 89 GPs completed the questionnaire. This number of participants was too low to obtain significant figures but the GPs' mean overall score of appropriately managed clinical cases was much lower than in the immediate post-test. In conclusion, although this short training improved the GPs' diagnostic accuracy and management of melanoma in the short-term, participating GPs do not seem to have maintained these competences in the long-term. Further studies are needed to assess whether refresher training sessions are able to sustain acquired diagnostic and management skills.


Induction of delayed hypersensitivity to turkey herpesvirus in fowls.
Delayed hypersensitivity, as determined by the wattle skin reaction, to turkey herpesvirus developed in chickens sensitised with the infected cell antigen. A significant increase in skin thickness occurred in the sensitised wattles as compared with the controls.


Collagen XV is necessary for modeling of the extracellular matrix and its deficiency predisposes to cardiomyopathy.
The extracellular matrix (ECM) is a major determinant of the structural integrity and functional properties of the myocardium in common pathological conditions, and changes in vasculature contribute to cardiac dysfunction. Collagen (Col) XV is preferentially expressed in the ECM of cardiac muscle and microvessels. We aimed to characterize the ECM, cardiovascular function and responses to elevated cardiovascular load in mice lacking Col XV (Col15a1(-/-)) to define its functional role in the vasculature and in age- and hypertension-associated myocardial remodeling. Cardiac structure and vasculature were analyzed by light and electron microscopy. Cardiac function, intraarterial blood pressure, microhemodynamics, and gene expression profiles were studied using echocardiography, telemetry, intravital microscopy, and PCR, respectively. Experimental hypertension was induced with angiotensin II or with a nitric oxide synthesis inhibitor. Under basal conditions, lack of Col XV resulted in increased permeability and impaired microvascular hemodynamics, distinct early-onset and age-dependent defects in heart structure and function, a poorly organized fibrillar collagen matrix with marked interstitial deposition of nonfibrillar protein aggregates, increased tissue stiffness, and irregularly organized cardiomyocytes. In response to experimental hypertension, Col15a1 gene expression was increased in the left ventricle of wild-type mice, and mRNA expression of natriuretic peptides (ANP and BNP) and ECM modeling were abnormal in Col15a1(-/-) mice. Col XV is necessary for ECM organization in the heart, and for the structure and functions of microvessels. Col XV deficiency leads to a complex cardiac phenotype and predisposes the subject to pathological responses under cardiac stress.


Microbial community of predatory bugs of the genus Macrolophus (Hemiptera: Miridae).
The predatory mirids of the genus Macrolophus are key natural enemies of various economically important agricultural pests. Both M. caliginosus and M. pygmaeus are commercially available for the augmentative biological control of arthropod pests in European greenhouses. The latter species is known to be infected with Wolbachia -inducing cytoplasmic incompatibility in its host- but the presence of other endosymbionts has not been demonstrated. In the present study, the microbial diversity was examined in various populations of M. caliginosus and M. pygmaeus by 16S rRNA sequencing and denaturing gradient gel electrophoresis. Besides Wolbachia, a co-infection of 2 Rickettsia species was detected in all M. pygmaeus populations. Based on a concatenated alignment of the 16S rRNA gene, the gltA gene and the coxA gene, the first is phylogenetically related to Rickettsia bellii, whereas the other is closely related to Rickettsia limoniae. All M. caliginosus populations were infected with the same Wolbachia and limoniae-like Rickettsia strain as M. pygmaeus, but did not harbour the bellii-like Rickettsia strain. Interestingly, individuals with a single infection were not found. A PCR assay on the ovaries of M. pygmaeus and M. caliginosus indicated that all endosymbionts are vertically transmitted. The presence of Wolbachia and Rickettsia in oocytes was confirmed by a fluorescence in situ hybridisation. A bio-assay comparing an infected and an uninfected M. pygmaeus population suggested that the endosymbionts had minor effects on nymphal development of their insect host and did not influence its fecundity. Two species of the palaearctic mirid genus Macrolophus are infected with multiple endosymbionts, including Wolbachia and Rickettsia. Independent of the origin, all tested populations of both M. pygmaeus and M. caliginosus were infected with three and two endosymbionts, respectively. There was no indication that infection with endosymbiotic bacteria had a fitness cost in terms of development and fecundity of the predators.


Inducible cell-specific mouse models for paired epigenetic and transcriptomic studies of microglia and astroglia.
Epigenetic regulation of gene expression occurs in a cell type-specific manner. Current cell-type specific neuroepigenetic studies rely on cell sorting methods that can alter cell phenotype and introduce potential confounds. Here we demonstrate and validate a Nuclear Tagging and Translating Ribosome Affinity Purification (NuTRAP) approach for temporally controlled labeling and isolation of ribosomes and nuclei, and thus RNA and DNA, from specific central nervous system cell types. Analysis of gene expression and DNA modifications in astrocytes or microglia from the same animal demonstrates differential usage of DNA methylation and hydroxymethylation in CpG and non-CpG contexts that corresponds to cell type-specific gene expression. Application of this approach in LPS treated mice uncovers microglia-specific transcriptome and epigenome changes in inflammatory pathways that cannot be detected with tissue-level analysis. The NuTRAP model and the validation approaches presented can be applied to any brain cell type for which a cell type-specific cre is available.


Upright CT of the knee: the effect of weight-bearing on joint alignment.
To prospectively compare patellofemoral and femorotibial alignment in supine non-weight-bearing computed tomography (NWBCT) and upright weight-bearing CT (WBCT) and assess the differences in joint alignment. NWBCT and WBCT images of the knee were obtained in 26 patients (mean age, 57.0 ± 15.9 years; range, 21-81) using multiple detector CT for NWBCT and cone-beam extremity CT for WBCT. Two musculoskeletal radiologists independently quantified joint alignment by measuring femorotibial rotation, tibial tuberosity-trochlear groove distance (TTTG), lateral patellar tilt angle, lateral patellar shift, and medial and lateral femorotibial joint space widths. Significant differences between NWBCT and WBCT were sought using Wilcoxon signed-rank test (P-value < 0.05). Significant differences were found for femorotibial rotation (the NWBCT mean changed from 2.7° ± 5.1 (reader 1)/2.6° ± 5.6 (reader 2) external rotation to WBCT 0.4° ± 7.7/0.2° ± 7.5 internal rotation; P = 0.009/P = 0.004), TTTG (decrease from NWBCT (13.8 mm ± 5.1/13.9 mm ± 3.9) to WBCT (10.5 mm ± 5.0/10.9 mm ± 5.2; P = 0.008/P = 0.002), lateral patellar tilt angle (decrease from NWBCT (15.6° ± 6.7/16.9° ± 7.4) to WBCT (12.5° ± 7.7/15.0° ± 6.2; P = 0.011/P = 0.188). The medial femorotibial joint space decreased from NWBCT (3.9 mm ± 1.4/4.5 mm ± 1.3) to WBCT (2.9 mm ± 2.2/3.5 mm ± 2.2; P = 0.003/P = 0.004). Inter-reader agreement ranged from 0.52-0.97. Knee joint alignment changes significantly in the upright weight-bearing position using CT when compared to supine non-weight-bearing CT. • Cone-beam extremity CT offers upright weight-bearing examinations of the lower extremities. • Knee alignment changes significantly in an upright position compared to supine position. • Tibial tuberosity-trochlear groove distance (TTTG) is less pronounced in a weight-bearing position. • The weight-bearing position leads to a decrease of the lateral patellar tilt angle.


Clinical presentation of prostate cancer in black South Africans.
Compared with White Americans, Black American men are at a significant increased risk of presenting with prostate cancer (PCa) and associated mortality, suggesting a link to African-ancestry. However, PCa status within Africa is largely unknown. We address the clinical presentation of PCa within Black South African men. Over 1,000 participants with or without PCa have enrolled in the Southern African Prostate Cancer Study (SAPCS). Using genome-wide profiling we establish a unique within Africa population substructure. Adjusting for age, clinical variables were assessed, compared against Black Americans and between rural and urban localities while addressing potential socio-demographic confounders. We report a significant difference in the distribution of prostate specific antigen (PSA) levels skewed towards higher PSA levels in the PCa cases (83.0% present with a PSA ≥ 20 µg/L; median PSA = 98.8 µg/L) relative to men with no detectable PCa (18.5% present with a PSA ≥ 20 µg/L; median PSA = 9.1 µg/L). Compared with Black Americans, Black South Africans presented with significantly more aggressive disease defined by Gleason score >7 (17% and 36%, respectively) and PSA ≥ 20 µg/L (17.2% and 83.2%, respectively). We report exasperated disease aggression defined by Gleason score >7 (P = 0.0042) and poorly differentiated tumor grade (P < 0.0001) within rural versus urban localities. Black South African men present with higher PSA levels and histopathological tumor grade compared with Black Americans, which is further escalated in men from rural localities. Our data suggests that lack of PSA testing may be contributing to an aggressive PCa disease phenotype within South African men.


Estrogen-induced transcription factor EGR1 regulates c-Kit transcription in the mouse uterus to maintain uterine receptivity for embryo implantation.
Early growth response 1 (Egr1) is a key transcription factor that mediates the action of estrogen (E2) to establish uterine receptivity for embryo implantation. However, few direct target genes of EGR1 have been identified in the uterus. Here, we demonstrated that E2 induced EGR1-regulated transcription of c-Kit, which plays a crucial role in cell fate decisions. Spatiotemporal expression of c-Kit followed that of EGR1 in uteri of ovariectomized mice at various time points after E2 treatment. E2 activated ERK1/2 and p38 to induce EGR1, which then activated c-Kit expression in the uterus. EGR1 transfection produced rapid and transient induction of c-KIT in a time- and dose-dependent manner. Furthermore, luciferase assays to measure c-Kit promoter activity confirmed that a functional EGR1 binding site(s) (EBS) was located within -1 kb of the c-Kit promoter. Site-directed mutagenesis and chromatin immunoprecipitation-PCR for three putative EBS within -1 kb demonstrated that the EBS at -818/-805 was critical for EGR1-dependent c-Kit transcription. c-Kit expression was significantly increased in the uterus on day 4 and administration of Masitinib, a c-Kit inhibitor, effectively interfered with embryo implantation. Collectively, our results showed that estrogen induces transcription factor EGR1 to regulate c-Kit transcription for uterine receptivity for embryo implantation in the mouse uterus.


Whole body bone mineral content is similar at discharge from the hospital in premature infants receiving fortified breast milk or preterm formula.
Prematurely born infants, especially those with very low birth weight (<1500 g) are at risk for metabolic bone disease. The influence of the type of oral feeding regimen and of other potential determinants of whole bone mineral content in prematurely born infants, when they approached full gestation, were evaluated. Previous studies have mainly examined effects at the level of regional bone. 34 infants (21 males and 13 females), all born between 25.4 and 33.7 weeks of gestation, were studied before discharge. Whole body bone mineral content measurements were made just before hospital discharge using a commercial densitometer (Hologic QDR 4500, Hologic Inc, Waltham, MA) at a median age of 40 days (range, 10 to 115 days) after birth. Expressed as a percentage of whole body mass, bone mass ranged between 0.86% and 1.99%, was similar between girls and boys and correlated positively with birth weight SD (r=0.42; P<0.05) and body weight SD (r=0.35; P<0.05). No difference in bone mass percentage was found between the different types of oral feedings (fortified human milk and preterm formula) or medications studied (corticoids and diuretics). Whereas prenatal and postnatal weight gain determines the degree of bone mineralization of premature infants, it appears that the type of oral feeding does not affect differently the postnatal bone mineralization of premature infants, when assessed at the moment of discharge.


Genetic predictors of acute toxicities related to radiation therapy following lumpectomy for breast cancer: a case-series study.
The cytotoxic effects of radiation therapy are mediated primarily through increased formation of hydroxyl radicals and reactive oxygen species, which can damage cells, proteins and DNA; the glutathione S-transferases (GSTs) function to protect against oxidative stress. We hypothesized that polymorphisms encoding reduced or absent activity in the GSTs might result in greater risk for radiation-associated toxicity. Women receiving therapy in radiation units in Germany following lumpectomy for breast cancer (1998-2001) provided a blood sample and completed an epidemiological questionnaire (n = 446). Genotypes were determined using Sequonom MALDI-TOF (GSTA1, GSTP1) and Masscode (GSTM1, GSTT1). Biologically effective radiotherapy dose (BED) was calculated, accounting for differences in fractionation and overall treatment time. Side effects considered were grade 2c and above, as classified using the modified Common Toxicity Criteria. Predictors of toxicity were modelled using Cox regression models in relation to BED, with adjustment for treating clinic, photon field, beam energy and boost method, and potential confounding variables. Low activity GSTP1 genotypes were associated with a greater than twofold increase in risk for acute skin toxicities (adjusted hazard ratio 2.28, 95% confidence interval 1.04-4.99). No associations were noted for the other GST genotypes. These data indicate that GSTP1 plays an important role in protecting normal cells from damage associated with radiation therapy. Studies examining the effects of GSTP1 polymorphisms on toxicity, recurrence and survival will further inform individualized therapeutics based on genotypes.


Monoclonal antibodies that cross-react with the E1 glycoprotein of different alphavirus serogroups: characterization including passive protection in vivo.
A panel of four monoclonal antibodies (mAb) were produced that cross-react with representatives of two different togavirus serogroups, namely sindbis (SIN) and Semliki Forest (SF) viruses, by ELISA and ADCMC assays. Three of these mAb, IgG2a and IgG2b isotypes, passively protected C3H/Hej mice against 10 and 100 LD50 of SF challenge and one, IgM, did not protect against either challenge dose, or even at 1 LD50. All these mAb were cross-reactive with the E1 glycoprotein of the viruses by immunoblotting in which three different patterns of reactivity were evident, suggesting that three epitopes were involved. The patterns depended upon whether the mAb recognized E1 extracted from purified virions or infected cells and whether SDS-PAGE and immunoblotting were done in the presence or absence of beta-mercaptoethanol. One mAb (IgM) reacted with nonreduced or reduced E1 from either virions or cells suggesting recognition of a linear epitope. The other three mAb reacted with nonreduced but not reduced E1 from virions suggesting that recognition depends upon conformational epitopes. These three mAb reacted also with nonreduced E1 extracted from SF-infected cells whereas only one reacted with nonreduced E1 extracted from SIN-infected cells.


Human immunodeficiency virus-related gastrointestinal pathology: a southern Africa perspective with review of the literature (part 2: neoplasms and noninfectious disorders).
Human immunodeficiency virus (HIV) infection is rife in sub-Saharan Africa and in southern Africa in particular. Despite the increasing availability of antiretroviral therapy in this region, HIV-associated neoplasms remain common and frequently involve the gastrointestinal tract, which may also demonstrate other noninfectious, HIV-related pathology. To review the histopathologic findings and distinguishing features of neoplastic and noninfectious, HIV-associated gastrointestinal disorders in southern Africa and relate those findings to the documented international literature. The available literature on this topic was reviewed and supplemented with personal experience in a private histopathology practice in South Africa. In southern Africa, a diverse range of HIV-related neoplasms and noninfectious gastrointestinal disorders is seen, but published data for the region are scarce. The gastrointestinal disorders include drug-associated pathology, gastrointestinal manifestations of the immune reconstitution inflammatory syndrome, idiopathic chronic esophageal ulceration, and the controversial entity of HIV enteropathy.


Selective inhibition of natural killer activity by the monoclonal antibodies OKT10 and VEP10 at the single cell level.
The monoclonal antibodies, VEP10 and OKT10, which have been shown to recognize determinants on human natural killer (NK) cells, inhibit large granular lymphocyte (LGL) NK activity against K562, MOLT4, and CEM tumor target cells in the single cell conjugate agarose assay. Inhibition of NK activity by monoclonal antibodies was expressed independently of effector-target cell binding, as inhibitory activity could be demonstrated when the monoclonal antibodies VEP10 and OKT10 were added to preformed conjugates or to the LGLs and targets prior to the binding event. In addition, this inhibition was exerted on the effector cell and not the target cell since VEP10 and OKT10 did not react with determinants on K562 target cells. Furthermore, the 4F2 monoclonal antibody, which reacted with determinants on the LGL and all of the targets used, effected no inhibition of NK activity. Inhibition of killing by OKT10 and VEP10 was specific to endogenous NK activity since the same antibodies did not inhibit antibody-dependent cellular cytotoxicity (ADCC), mixed lymphocyte-generated NK, or cytotoxic T lymphocyte (CTL) activities.


2D Ultrasound image processing in identifying responses of urogenital structures to pelvic floor muscle activity.
We present the analysis of sequences of voluntarily and reflexly generated pelvic floor muscle (PFM) contractions on the urethra, vagina, and rectum. A succession of observations were taken of perineal ultrasound at a frequency of 3.5 MHz imaging using a curved linear array probe in the sagittal plane and the videos captured and stored. An edge extraction algorithm was used to outline the coordinates of the symphysis pubis, urethra, and rectum interfaces on a frame-by-frame basis for sequences of 10-20 s. During each PFM contraction, the trajectory of the boundary of each structure was evaluated, colour coded, and overlaid to characterize the sequential history of the ensuing movement. The resulting image analysis was focused to reveal the anatomical displacement of the urogenital structures, which enables the evaluation of their biomechanical parameters in terms of displacement, velocity, and acceleration at any point in time. On the basis of these observations, the biomechanical mechanisms of pelvic floor muscle responses to voluntary and reflex contractions can be identified. It is concluded that a considerable amount of new, potentially useful clinical information can be revealed from video recordings of perineal ultrasound using the image analysis approach proposed.


Changes of the complement system and rheological indicators after therapy with rheohemapheresis.
In the last 10 years, many studies have been published on the role of the complement system in microcirculation disorders. However, as for the changes of complement components after rheohemapheresis, there is still a lack of detailed data in the literature. Complement changes may play an important role in pathogenesis of some microcirculation disorders, such as age-related macular degeneration and acute hearing loss. The objective of this study was to investigate the effect of rheohemapheresis on the basic complement pathways. 32 patients were treated with rheohemapheresis, including 16 patients (10 men and 6 women) for age-related macular degeneration (AMD), mean age 69.7 ± 6.06 years (range 62-87 years) and 16 patients (11 men and 5 women) aged 56.4 ± 11.5 (range 34-73 years) for acute hearing loss. Rheohemapheresis led to a significant drop of all three complement-activation pathways in both groups of patients. Moreover, complement factor H was also reduced. The observed reduction in all three basic complement activation pathways after rheohemapheresis could be clinically important. The search continues both to find substances which influence complement systems and to develop more effective new drugs that require less frequent administration and that provide improved intraocular therapy for AMD patients.


Light and temperature signal crosstalk in plant development.
Light and temperature are two of the most important environmental stimuli regulating plant development. Recent advances have suggested considerable interaction between these signalling pathways at the molecular level. Studies of both flowering and germination have shown the phytochrome family of plant photoreceptors to display altered functional hierarchies at different growth temperatures. The existence of common signalling components in both light and temperature sensing has additionally been proposed. More recently, light quality signals have been shown to regulate plant-freezing tolerance in an ambient temperature-dependent manner. Together, these data suggest that complex crosstalk between light-signalling and temperature-signalling pathways is fundamental to the growth and development of plants in natural environments.


Tyrosine-phosphorylated Hic-5 inhibits epidermal growth factor-induced lamellipodia formation.
The focal adhesion protein Hic-5, a homologue to paxillin, has been shown to be tyrosine-phosphorylated in fibroblasts in response to stimuli such as osmotic stress, serum, LPA and endothelin. However, the function of this modification to Hic-5 is unclear. Herein, we show that Hic-5 is tyrosine-phosphorylated on residues 38 and 60 following epidermal growth factor (EGF) treatment of COS-7 cells, coincident with an increase in peripheral actin reorganization. To explore the role of Hic-5 phosphorylation in this process, we introduced wild-type (WT) and mutant Hic-5 constructs into COS-7 cells and determined that EGF-induced lamellipodia formation was suppressed by WT Hic-5. This effect required localization to focal adhesions as well as phosphorylation of Hic-5 as overexpression of both a non-targeting and a non-phosphorylatable Hic-5 failed to inhibit peripheral actin reorganization. Interestingly, overexpression of non-phosphorylatable Y31/118F or WT paxillin did not affect lamellipodia formation, indicating that this effect is specific to Hic-5. The EGF-induced lamellipodia were Rac-dependent and overexpressed WT Hic-5, but not non-phosphorylatable Hic-5 inhibited Rac activation. Our data suggest that Hic-5 tyrosine phosphorylation functions to regulate signaling associated with lamellipodia formation, a process fundamental to cell motility.


Carotid atherosclerosis in young-aged IDDM associated with diabetic retinopathy and diastolic blood pressure.
To determine whether young patients with IDDM already have atherosclerosis and what factors would relate to atherosclerosis, we examined the intimal-medial thickness (IMT) of the common carotid artery by ultrasonography. Subjects were 29 young patients with IDDM (aged 17-39 years, duration 4-31 years) without manifest macroangiopathy and 13 healthy controls of comparable age (22-29 years). The carotid artery IMT of young patients with IDDM were significantly higher than those of controls (0.60 +/- 0.09 vs. 0.46 +/- 0.02 mm, P < 0.0001). The levels of IMT significantly correlated to diastolic blood pressure (r = 0.45, P < 0.02), and were higher in those with proliferative retinopathy than those without retinopathy (0.66 +/- 0.09 vs. 0.55 +/- 0.08 mm, P < 0.02). The levels of IMT showed no significant correlation to the attained age, duration of IDDM, HbA1c, systolic blood pressure, and cholesterol level. These findings suggest the usefulness of this examination for the early detection of diabetic macroangiopathy, and point to a close relationship between microangiopathy and macroangiopathy in IDDM.


Tuberculous otomastoiditis complicated by thrombosis of the sigmoid sinus in an infant.
Otomastoiditis is a rare and exceptional complication of tuberculosis; its pathogenesis has been debated since its recognition as a distinct condition. The evolution and prognosis in general are modified after the end of antituberculosis treatment. In our observation, we report the case of an infant with tuberculous otomastoiditis complicated by sigmoid sinus thrombosis.


Bacterial Skin Infections.
Skin and soft tissue infections account for 0.5% of outpatient visits to primary care. Skin and soft tissue infections can usually be managed in an outpatient setting. However, there are certain circumstances as discussed in this article that require more urgent care or inpatient management. Primary care providers should be able to diagnose, manage, and provide appropriate follow-up care for these frequently seen skin infections. This article provides family physicians with a comprehensive review of the assessment and management of common bacterial skin infections.


Evaluation of vestibular system with vHIT in industrial workers with noise-induced hearing loss.
Industrial workers are frequently exposed to noise and the noise can cause hearing loss. The relationship between vestibular abnormalities and long-term exposure to occupational noise has been reported. The aim of this study was to use vHIT (video head impulse test) to evaluate the semicircular canal's function in industrial workers with noise-induced hearing loss. In the present study, we included 36 male patients (aged 28-55 years, mean 44.4 ± 6.1 years) with hearing loss due to noise exposure from working in the steel and metal industry for at least 4 years, and a control group of 30 healthy men (aged 34-50 years, mean 41.1 ± 4.5 years) without hearing loss. The gain of the vestibulo-ocular reflex for all 6 semicircular canals was examined by vHIT in 66 patients. Canal deficit was detected in 20 (55.5%) of 36 patients in the noise exposure hearing loss group and was detected in 2 (6.6%) of 30 participants in the control group. There was significant loss of capacity for VOR gain in patients with noise exposure hearing loss. In the present study we found that exposure to noise can cause vestibular dysfunction. And the vHIT is easily applicable, quick and practical and can individually evaluate all semicircular channels. Therefore, to evaluate vestibular function with vHIT is quick and practical.


Improved Cytotoxicity and Induced Apoptosis in HeLa Cells by Co-loading Vitamin E Succinate and Curcumin in Nano-MIL-88B-NH2.
One of the strategies for improved therapeutic effects in cancer therapy is combination chemotherapy. In this study, a flexible nano-MOF (Fe-MIL-88B-NH2 ) was synthesized in a sonochemical process, then co-loaded with α-tocopheryl succinate (TOS) and curcumin (CCM). The anticancer activity of co-loaded Fe-MIL-88B-NH2 (Fe-MIL-88B-NH2 /TOS@CCM) against the HeLa cells was compared with that of the single-loaded counterpart (Fe-MIL-88B-NH2 @CCM). MTT analysis indicates improved cytotoxicity of Fe-MIL-88B-NH2 /TOS@CCM. The data from the cell apoptosis assay indicated more apoptosis in the case of the co-loaded nano-MOF. This study indicates the positive effect of the presence of TOS on enhancing the anticancer effect of Fe-MIL-88B-NH2 @CCM to prepare a more efficient drug delivery nanosystem.


Comparison of a dot immunobinding assay and the serum agglutination test for detecting serological responses in vaccinated and unvaccinated chickens following challenge with Salmonella gallinarum.
A dot immunobinding assay (DIA) was compared with the standard tube agglutination test (SAT) for detecting antibodies in sera from vaccinated and unvaccinated chickens challenged with Salmonella gallinarum var duisburg . The virulence plasmid-cured S. gallinarum vaccine was given at 7 and 21 days, and the challenge at 35 days. The DIA was very sensitive and detected antibodies to high titres in challenged birds. It also detected high titres in vaccinated, unchallenged birds which failed to react by SAT.


Optimisation of itraconazole therapy using target drug concentrations.
Itraconazole is a new triazole compound with a broad spectrum of activity against a number of fungal pathogens, including Aspergillus species. The drug is being used increasingly as prophylaxis in patients with immunodepression. Itraconazole is highly lipophilic and only ionised at low pH. The absolute availability of capsules in healthy volunteers under fasting conditions is about 55% and is increased after a meal. Itraconazole is 99.8% bound to human plasma proteins and its apparent volume of distribution is about 11 L/kg. The drug is extensively metabolised by the liver. Among the metabolites, hydroxy-itraconazole is of particular interest because its antifungal activity measured in vitro is similar to that of the parent drug and its plasma concentration is 2 to 3 times higher than that of itraconazole. Mean total itraconazole blood clearance determined in healthy volunteers following a single intravenous infusion was 39.6 L/h. After a single oral dose, the terminal elimination half-life of itraconazole is about 24 hours. The drug exhibits a dose-dependent pharmacokinetic behaviour. Renal failure does not affect the pharmacokinetic properties of itraconazole; however, little is known about the effects of hepatic insufficiency. In immunocompromised patients the absorption of itraconazole is affected by gastrointestinal disorders caused by diseases and cytotoxic chemotherapy. The pharmacokinetics of itraconazole may be significantly altered when the drug is coadministered with certain other agents. Itraconazole is a potent inhibitor of cytochrome P450 (CYP) 3A4 and, thus, can also considerably change the pharmacokinetics of other drugs. Such changes may have clinically relevant consequences. Itraconazole appears to be well tolerated. Gastrointestinal disturbances and dizziness are the most frequently reported adverse effects. Clinical studies in patients with haemotological malignancies suggest that plasma concentrations [measured by high performance liquid chromatography (HPLC)] > or = 250 micrograms/L itraconazole, or 750 to 1000 micrograms/L for itraconazole plus hydroxy-itraconazole, are required for effective prophylactic antifungal activity. It seems that a curative effect may be enhanced by ensuring that itraconazole plasma concentrations exceed 500 micrograms/L. The marked intra- and inter-patient variability in the pharmacokinetics of the drug, and the fact that it is impossible to predict steady-state plasma concentrations from the initial dosage are major factors obscuring any clear relationship between dose and plasma concentrations and clinical efficacy. Thus, in patients with life-threatening fungal infections treated with itraconazole drug, plasma concentrations should be regularly monitored to ensure sufficient drug exposure for antifungal activity.


Laparoscopic Transanal Total Mesorectal Excision (taTME) for Rectal Cancer.
Proper treatment of adenocarcinoma of the rectum demands a systematic, multidisciplinary approach where surgery remains the cornerstone of treatment. An evolving shift toward minimally invasive surgical approaches for rectal cancer continues to be hampered by the challenges of reliable pelvic exposure and adequate instrumentation for rectal dissection, distal rectal division, and low pelvic anastomosis. The laparoscopic transanal total mesorectal excision surgery has been developed as a novel alternative that provides solutions to many of the limitations of conventional open, laparoscopic and robotic proctectomy. This manuscript will describe the procedure in detail and attempt to define its role as the optimal surgical approach for rectal resection.


Surgeon Learning Curve for the Sinus Tarsi Approach to Intra-articular Calcaneus Fractures - Improvement with Experience.
To analyze the relationship between surgeon experience with the sinus tarsi approach (STA) and outcomes in the treatment of displaced intra-articular calcaneus fractures (DIACF). Single level 1 trauma center. Retrospective.Patients/Participants: 103 consecutive DIACF (OTA/AO 82C; Sanders II-IV) treated operatively using STA from 2015 to 2021. Open management using the STA performed by two fellowship-trained orthopaedic traumatologists. Quality of anatomic reduction based on postoperative CT scans and standard radiographs. Sixty-six patients met inclusion criteria. Patients were primarily men (75.8%) with mean age 41 years (range 20-71 years), including 14 smokers (21.2%), 9 diabetics (13.6%), and 10 open fractures (15.2%). Sanders III fractures were most common (68.2% vs 28.5% and 6.1% Sanders II/IV respectively). Reduction quality was predominantly Good (59.1%, n=39) or Excellent (25.8%, n=17). Complications included wound necrosis (1), superficial infection (1), deep infection (1), and symptomatic posttraumatic arthritis requiring arthrodesis (3). There was a 29.3% reduction in likelihood of surgical complication with each year in surgeon experience with the STA and an 8.9% reduction per case (p<0.001). The likelihood of achieving a Good or Excellent reduction was 1.8 and 2.3 times greater than achieving a Fair reduction, respectively, for each year increase in surgeon experience with the STA (p=0.012 and 0.007, respectively). For each successive case, there was a 1.2 times greater likelihood of achieving a Good reduction (p=0.03). Surgeon experience plays a critical role in outcomes. We found that outcomes (reduction, complications) improve with each cumulative case and year of experience with the STA to treat DIACF. Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.


High-frequency sonography of the volar digital nerves of the hand.
The purpose of this study is to describe our technique for high-frequency sonography of the volar digital nerves and to define the course, caliber, and morphology of normal volar digital nerves. The volar digital nerves of 10 volunteers were imaged in the long axis using a linear ultrasound transducer. Height, weight, glove size, and hand dominance were recorded. Each radial and ulnar nerve was divided into four segments. Segment thickness and depth were measured. A mixed model analysis for repeated measures was utilized (α = 0.05). The average nerve thickness (NT) was 1.1 mm (± 0.01 SE), and the average nerve depth (ND) was 2.8 mm (± 0.04 SE). Neither NT nor ND demonstrated any significant correlation with height, age, weight, body mass index, or glove size. Nerves ranged in thickness from 0.7 to 1.5 mm and in depth from 0.7 to 6.8 mm. High-frequency sonography permits high-resolution imaging of the volar digital nerves.


Structure-dependent activities of hydroxylated polybrominated diphenyl ethers on human estrogen receptor.
Polybrominated diphenyl ethers (PBDEs) have been shown to affect the estrogen receptor (ER) signaling pathway, and one of the proposed disruption mechanisms is direct binding of hydroxylated PBDE (OH-PBDE) to ER. In this paper, the binding affinity of 22 OH-PBDEs with different degrees of bromination to ER was assessed quantitatively using a surface plasmon resonance biosensor technique. Seven OH-PBDEs were found to bind directly with ER with KD ranging from 1.46x10(-7) M to 7.90x10(-6) M, and the affinity is in the order of 6-OH-BDE-047>/=4'-OH-BDE-049>4'-OH-BDE-017>6'-OH-BDE-099>/=5'-OH-BDE-099>2'-OH-BDE-007>3'-OH-BDE-028. In MVLN luciferase gene reporter assays, 10 low-brominated OH-PBDEs induced luciferase activity alone, but are 10(5) to 10(7) fold less potent than E2. Their estrogenic activity is in the order of 4'-OH-BDE-049>4'-OH-BDE-017>2'-OH-BDE-007>3'-OH-BDE-028>3-OH-BDE-047>/=3'-OH-BDE-007. The good correlation between estrogenic activity and ER binding affinity of the low-brominated OH-PBDEs strongly suggest that these compounds induce ER transcriptional activity by binding directly with ER. The other 12 high-brominated OH-PBDEs inhibited luciferase activity of E2 to various degrees, demonstrating their antagonistic activity. Molecular docking analysis of the ER/OH-PBDE complexes revealed two distinctive binding modes between low- and high-brominated OH-PBDEs which provided rationale for the difference in their ER activity.


'Naturalization' of textile disperse dyes through glycoconjugation: the case of a bis(2-hydroxyethyl) group containing azo dye.
A family of five strictly related glycoconjugated azo dyes (GADs), characterized by the presence of the same chromophore and a variable number (1-4) of deprotected hexose units, has been prepared by employing succinate bridges for connecting the azo dye and the sugar portions. The modulation of the hydrophilic portion determines the appreciable changes in the water solubility of GADs. In all the cases, however, hydrophobic fibres (polyester) were homogeneously dyed with GADs at temperatures lower than that used for original azo dyes, at atmospheric pressure, and avoiding the use of surfactants. Furthermore, GADs show an interesting multipurpose character leading to dyeing well also the natural fibres as, for instance, wool. The presence of a variable number of hexose units in the different GADs determines some changes in the colour intensity of dyed fabrics, but in all the cases an appreciable rubbing and water fastness were maintained.


Cleistocalyx nervosum var. paniala berry fruit protects neurotoxicity against endoplasmic reticulum stress-induced apoptosis.
Oxidative and endoplasmic reticulum (ER) stresses cause neuronal damage leading to neurodegenerative disorders. Cleistocalyx nervosum var. paniala (CNP) berry fruit has been shown to possess powerful antioxidant properties. Here, we investigated the neuroprotective effect of CNP extract against glutamate-mediated oxidative/ER stress-induced cell death in mouse hippocampal neuronal HT22 cells. CNP extract was clarified for its radical scavenging activities, total phenolic and anthocyanin contents. The key anthocyanin cyanidin-3-glucoside was used as a marker to standardize the extract used in the study. We found that pretreated cells with CNP extract (0.05-1 μg/ml) prevented neuronal cell death in response to 5 mM glutamate evaluated by cell viability MTT, LDH and apoptosis/necrosis Annexin V/propidium iodide co-staining assays. For mechanistic approach, glutamate-induced cell death through reactive oxygen species (ROS)-mediated ER stress pathways, indicating the increase of ROS and ER stress signature molecules including calpain, caspases-12 and C/EBP homologous proteins (CHOP). CNP extract inhibited ROS production. Moreover, the extract also suppressed the specific-ER stress apoptotic proteins level in glutamate-induced cells by upregulating the gene expression of cellular antioxidant enzymes (SODs, CAT, GPx and GSTs). Taken together, our results provide information about and the molecular mechanism of CNP extract as a promising neuroprotectant and antioxidant.


Decadal decline of dominant copepod species in the North Sea is associated with ocean warming: Importance of marine heatwaves.
Time-series are crucial to understand the status of zooplankton communities and to anticipate changes that might affect the entire food web. Long-term time series allow us to understand impacts of multiple environmental and anthropogenic stressors, such as chemical pollution and ocean warming, on the marine ecosystems. Here, a recent time series (2018-2022) of abundance data of four dominant calanoid and one harpacticoid copepod species from the Belgian Part of the North Sea was combined with previously collected (2009-2010, 2015-2016) datasets for the same study area. The time series reveals a significant decrease (up to two orders of magnitude) in calanoid copepod abundance (Temora longicornis, Acartia clausi, Centropages spp., Calanus helgolandicus), while this was not the case for the harpacticoid Euterpina acutifrons. We applied generalized additive models to quantify the relative contribution of temperature, nutrients, salinity, primary production, turbidity and pollution (anthropogenic chemicals, i.e., polychlorinated biphenyls and polycyclic aromatic hydrocarbons) to the population dynamics of these species. Temperature, turbidity and chlorophyll a concentrations were the only variables consistently showing a relative high contribution in all models predicting the abundances of the selected species. The observed heat waves which occurred during the summer periods of the investigated years coincided with population collapses (versus population densities in non-heatwave years) and are considered the most likely cause for the observed copepod abundance decreases. Moreover, the recorded water temperatures during these heatwaves correspond to the physiological thermal limit of some of the studied species. As far as we know, this is the first study to observe ocean warming and marine heat waves having such a dramatic impact (population collapse) on the dominant zooplankton species in shallow coastal areas.


Assessment of cyclic changes in the diameter of the aortic annulus using speckle-tracking trans-esophageal echocardiography.
It is uncertain whether dynamic variation in the diameter of the aortic annulus occurs during the cardiac cycle in humans. The purpose of this study was to analyze cyclic changes of the aortic annulus using speckle-tracking trans-esophageal echocardiography. The subjects were 40 patients with aortic stenosis and 40 controls. Absolute and relative changes in the diameter of the aortic annulus and the times at which the maximum and minimum diameters occurred during the cardiac cycle were determined using speckle-tracking trans-esophageal echocardiography. The maximum and minimum diameters were 22.9 ± 2.7 and 20.0 ± 2.9 mm, respectively, in controls. The change in diameter of the aortic annulus was 2.9 ± 0.7 mm, and the relative change was 12.9 ± 3.5%. The maximum aortic annulus diameter was reached at the onset of aortic valve opening, and the minimum diameter occurred in the rapid filling phase. The change in diameter of the aortic annulus was significantly smaller (2.2 ± 0.6 mm vs. 2.9 ± 0.7 mm, p < 0.0001), and the time to reach the maximum diameter was significantly longer (98.5 ± 17.5 ms vs. 83.4 ± 18.2 ms, p = 0.0004), in the aortic stenosis group than in the control group. The study found that dynamic changes of the aortic annulus occur in the cardiac cycle and can be measured using speckle-tracking trans-esophageal echocardiography. We also found that aortic stenosis has an effect on the extent and timing of these changes. This suggests that accurate assessment of aortic annulus diameter requires consideration of the timing of the cardiac cycle.


Combined anti-fibrotic and anti-inflammatory properties of JAK-inhibitors on macrophages in vitro and in vivo: Perspectives for scleroderma-associated interstitial lung disease.
Janus kinase (JAK) inhibitors (also termed Jakinibs) constitute a family of small drugs that target various isoforms of JAKs (JAK1, JAK2, JAK3 and/or tyrosine kinase 2 (Tyk2)). They exert anti-inflammatory properties linked, in part, to the modulation of the activation state of pro-inflammatory M1 macrophages. The exact impact of JAK inhibitors on a wider spectrum of activation states of macrophages is however still to be determined, especially in the context of disorders involving concomitant activation of pro-inflammatory M1 macrophages and profibrotic M2 macrophages. This is especially the case in autoimmune pulmonary fibrosis like scleroderma-associated interstitial lung disease (ILD), in which M1 and M2 macrophages play a key pathogenic role. In this study, we directly compared the anti-inflammatory and anti-fibrotic effects of three JAK inhibitors (ruxolitinib (JAK2/1 inhibitor); tofacitinib (JAK3/2 inhibitor) and itacitinib (JAK1 inhibitor)) on five different activation states of primary human monocyte-derived macrophages (MDM). These three JAK inhibitors exert anti-inflammatory properties towards macrophages, as demonstrated by the down-expression of key polarization markers (CD86, MHCII, TLR4) and the limited secretion of key pro-inflammatory cytokines (CXCL10, IL-6 and TNFα) in M1 macrophages activated by IFNγ and LPS or by IFNγ alone. We also highlighted that these JAK inhibitors can limit M2a activation of macrophages induced by IL-4 and IL-13, as notably demonstrated by the down-regulation of the M2a associated surface marker CD206 and of the secretion of CCL18. Moreover, these JAK inhibitors reduced the expression of markers such as CXCL13, MARCO and SOCS3 in alternatively activated macrophages induced by IL-10 and dexamethasone (M2c + dex) or IL-10 alone (M2c MDM). For all polarization states, Jakinibs with inhibitory properties over JAK2 had the highest effects, at both 1 μM or 0.1 μM. Based on these in vitro results, we also explored the effects of JAK2/1 inhibition by ruxolitinib in vivo, on mouse macrophages in a model of HOCl-induced ILD, that mimics scleroderma-associated ILD. In this model, we showed that ruxolitinib significantly prevented the upregulation of pro-inflammatory M1 markers (TNFα, CXCL10, NOS2) and pro-fibrotic M2 markers (Arg1 and Chi3L3). These results were associated with an improvement of skin and pulmonary involvement. Overall, our results suggest that the combined anti-inflammatory and anti-fibrotic properties of JAK2/1 inhibitors could be relevant to target lung macrophages in autoimmune and inflammatory pulmonary disorders that have no efficient disease modifying drugs to date.


Successful reduction of a high defibrillation threshold by a combined implantation of a subcutaneous array and azygos vein lead.
A 72-year-old man with nonischemic cardiomyopathy was referred because his implantable cardioverter defibrillator had failed to terminate spontaneous ventricular fibrillation (VF). Defibrillation threshold (DFT) testing confirmed that 830-V shocks failed to defibrillate VF despite optimization of the biphasic waveform and reversal of shock polarity. The placement of a new right ventricular lead and the addition of a subcutaneous array failed to defibrillate VF at 830 V. The combination of a subcutaneous array and azygos vein coil successfully defibrillated VF. The mechanism for successful DFT reduction was likely greater current supplied to the posterior basal left ventricle by the azygos vein lead.


Raman spectroscopy combined with deep learning for rapid detection of melanoma at the single cell level.
Melanoma is an aggressive and metastatic skin cancer caused by genetic mutations in melanocytes, and its incidence is increasing year by year. Understanding the gene mutation information of melanoma cases is very important for its precise treatment. The current diagnostic methods for melanoma include radiological, pharmacological, histological, cytological and molecular techniques, but the gold standard for diagnosis is still pathological biopsy, which is time consuming and destructive. Raman spectroscopy is a rapid, sensitive and nondestructive detection method. In this study, a total of 20,000 Surface-enhanced Raman scattering (SERS) spectra of melanocytes and melanoma cells were collected using a positively charged gold nanoparticles planar solid SERS substrate, and a classification network system based on convolutional neural networks (CNN) was constructed to achieve the classification of melanocytes and melanoma cells, wild-type and mutant melanoma cells and their drug resistance. Among them, the classification accuracy of melanocytes and melanoma cells was over 98%. Raman spectral differences between melanocytes and melanoma cells were analyzed and compared, and the response of cells to antitumor drugs were also evaluated. The results showed that Raman spectroscopy provided a basis for the medication of melanoma, and SERS spectra combined with CNN classification model realized classification of melanoma, which is of great significance for rapid diagnosis and identification of melanoma.


A Rapid Advice Guideline for the Prevention of Novel Coronavirus Through Nutritional Intervention.
An unexpected and sudden outbreak of a novel infection known as a coronavirus (COVID-19) has imposed important problems to global well-being and economy. Based upon current researches, this virus is spreading from one human to another through respiratory droplets, i.e. cough and sneeze. Till now, there has not been any specific treatment found for this virus. Hence, there is a critical need to discover alternative techniques to cope with the current scenario. This review conducted an online search for prevention of coronavirus infection with the help of nutritional interventions. It has been observed that the effect of the virus is mostly on the individual with low immunity, individual affected with diseases like diabetes, and individual using any immune-suppressed drug or having past history of major surgeries or severe medical conditions. Therefore, consuming foods which boost immunity helps in preventing respiratory-related disorder or suppressing diseases-related problems, which could be helpful in controlling the spread of this virus. In conclusion, it has been suggested that before the beginning of generalised treatments and interventions in each infected patient, nutritional status should be evaluated, as it can help in creating a specific nutrition intervention for the infected individual.


Facile and reversible double dearomatization of pyridines in non-phosphine MnI complexes with N,S-donor pyridinophane ligand.
Single and double dearomatization of pyridine rings was observed in MnI complexes with an N2S2 pyridinophane ligand via deprotonation of one or two CH2 arms, respectively. In contrast to other N,S-donor pincer-like systems, the dearomatized (N2S2)Mn species were found to be stable, with the dearomatization being reversible.


Evaluating the behavior and principle of deep eutectic solvent on ephedrine-type alkaloid extraction from Ephedrae Herba.
In this study, deep eutectic solvent (DES), as a new green solvent, was used to extract bioactive alkaloids from Ephedrae Herba using supersonic extraction. In a variety of tested hydrophilic and hydrophobic DESs, DES composed of choline chloride and xylitol was proved to be the most efficient solvent. Factors affecting extraction efficiency, including the mole ratio of hydrogen bond acceptor/hydrogen bond donor, water contention, and solid/liquid ratio, were optimized individually. Under optimal conditions, the yield of ephedrine (EP) and pseudoephedrine obtained using this new method was 14.24 and 4.32 mg/g, respectively, which was higher than that using the traditional solvent (acidified water and methanol). Furthermore, the extraction mechanism of DES and EP was investigated using molecular dynamics simulation study. Structural properties such as radial distribution functions and average number of hydrogen bonds were then computed. The results showed that hydrogen bonds and van der Waals forces are important driving forces of extraction; in addition, the hydrogen bonds between the Cl atom of choline chloride and N atom of EP played a dominant part in the extraction process. Based on the extraction principle, the extraction method using choline chloride as extraction solvent was also discussed.


Automatic Sleep Stage Scoring Using Time-Frequency Analysis and Stacked Sparse Autoencoders.
We developed a machine learning methodology for automatic sleep stage scoring. Our time-frequency analysis-based feature extraction is fine-tuned to capture sleep stage-specific signal features as described in the American Academy of Sleep Medicine manual that the human experts follow. We used ensemble learning with an ensemble of stacked sparse autoencoders for classifying the sleep stages. We used class-balanced random sampling across sleep stages for each model in the ensemble to avoid skewed performance in favor of the most represented sleep stages, and addressed the problem of misclassification errors due to class imbalance while significantly improving worst-stage classification. We used an openly available dataset from 20 healthy young adults for evaluation. We used a single channel of EEG from this dataset, which makes our method a suitable candidate for longitudinal monitoring using wearable EEG in real-world settings. Our method has both high overall accuracy (78%, range 75-80%), and high mean [Formula: see text]-score (84%, range 82-86%) and mean accuracy across individual sleep stages (86%, range 84-88%) over all subjects. The performance of our method appears to be uncorrelated with the sleep efficiency and percentage of transitional epochs in each recording.


4-Phenylbutyric Acid Reveals Good Beneficial Effects on Vital Organ Function via Anti-Endoplasmic Reticulum Stress in Septic Rats.
Sepsis and septic shock are the common complications in ICUs. Vital organ function disorder contributes a critical role in high mortality after severe sepsis or septic shock, in which endoplasmic reticulum stress plays an important role. Whether anti-endoplasmic reticulum stress with 4-phenylbutyric acid is beneficial to sepsis and the underlying mechanisms are not known. Laboratory investigation. State Key Laboratory of Trauma, Burns and Combined Injury. Sprague-Dawley rats. Using cecal ligation and puncture-induced septic shock rats, lipopolysaccharide-treated vascular smooth muscle cells, and cardiomyocytes, effects of 4-phenylbutyric acid on vital organ function and the relationship with endoplasmic reticulum stress and endoplasmic reticulum stress-mediated inflammation, apoptosis, and oxidative stress were observed. Conventional treatment, including fluid resuscitation, vasopressin, and antibiotic, only slightly improved the hemodynamic variable, such as mean arterial blood pressure and cardiac output, and slightly improved the vital organ function and the animal survival of septic shock rats. Supplementation of 4-phenylbutyric acid (5 mg/kg; anti-endoplasmic reticulum stress), especially administered at early stage, significantly improved the hemodynamic variables, vital organ function, such as liver, renal, and intestinal barrier function, and animal survival in septic shock rats. 4-Phenylbutyric acid application inhibited the endoplasmic reticulum stress and endoplasmic reticulum stress-related proteins, such as CCAAT/enhancer-binding protein homologous protein in vital organs, such as heart and superior mesenteric artery after severe sepsis. Further studies showed that 4-phenylbutyric acid inhibited endoplasmic reticulum stress-mediated cytokine release, apoptosis, and oxidative stress via inhibition of nuclear factor-κB, caspase-3 and caspase-9, and increasing glutathione peroxidase and superoxide dismutase expression, respectively. Anti-endoplasmic reticulum stress with 4-phenylbutyric acid is beneficial to septic shock. This beneficial effect of 4-phenylbutyric acid is closely related to the inhibition of endoplasmic reticulum stress-mediated oxidative stress, apoptosis, and cytokine release. This finding provides a potential therapeutic measure for clinical critical conditions, such as severe sepsis.


Extended TAQing system for large-scale plant genome reorganization.
We previously developed a large-scale genome restructuring technology called the TAQing system. It can induce genomic rearrangements by introducing transient and conditional formation of DNA double-strand breaks (DSBs) via heat activation of a restriction enzyme TaqI, which can cleave DNA at 5'-TCGA-3' sequences in the genome at higher temperatures (37-42°C). Such heat treatment sometimes confers lethal damage in certain plant species and TaqI cannot induce rearrangements in AT-rich regions. To overcome such problems we developed an extended TAQing (Ex-TAQing) system, which enables the use of a wider range of restriction enzymes active at standard plant-growing temperatures. We established the Ex-TAQing system using MseI that can efficiently cleave DNA at room temperature (at temperatures ranging from 22 to 25°C) and the 5'-TTAA-3' sequence which is highly abundant in the Arabidopsis genome. A synthetic intron-spanning MseI gene, which was placed downstream of a heat-shock-inducible promoter, was conditionally expressed upon milder heat treatment (33°C) to generate DSBs in Arabidopsis chromosomes. Genome resequencing revealed various types of genomic rearrangements, including copy number variations, translocation and direct end-joining at MseI cleavage sites. The Ex-TAQing system could induce large-scale rearrangements in diploids more frequently (17.4%, n = 23) than the standard TAQing system. The application of this system to tetraploids generated several strains with chromosomal rearrangements associated with beneficial phenotypes, such as high salinity stress tolerance and hypersensitivity to abscisic acid. We have developed the Ex-TAQing system, allowing more diverse patterns of genomic rearrangements, by employing various types of endonucleases and have opened a way to expand the capacity for artificial genome reorganization.


Myocardial injury in patients with SARS-CoV-2 pneumonia: Pivotal role of inflammation in COVID-19.
Infection by SARS-CoV-2 may result in a systemic disease and a proportion of patients ranging 15%-44% experienced cardiac injury (CI) diagnosed by abnormal troponin levels. The aim of the present study was to analyse the clinical characteristics of a large series of hospitalized patients for COVID-19 in order to identify predisposing and/or protective factors of CI and the outcome. This is an observational, retrospective study on patients hospitalized in two Italian centres (San Raffaele Hospital and Cremona Hospital) for COVID-19 and at least one high-sensitivity cardiac troponin (hs-cTnt) measurement during hospitalization. CI was defined if at least one hs-cTnt value was above the 99th percentile. The primary end-point was the occurrence of CI during hospitalization. We included 750 patients (median age 67, IQR 56-77 years; 69% males), of whom 46.9% had history of hypertension, 14.7% of chronic coronary disease and 22.3% of chronic kidney disease (CKD). Abnormal troponin levels (median troponin 74, IQR 34-147 ng/l) were detected in 390 patients (52%) during the hospitalization. At multivariable analysis age, CKD, cancer, C-reactive protein (CRP) levels were independently associated with CI. Independent predictors of very high troponin levels were chronic kidney disease and CRP levels. Patients with CI showed higher rate of all-cause mortality (40.0% vs. 9.1%, p = 0.001) compared to those without CI. This large, multicentre Italian study confirmed the high prevalence of CI and its prognostic role in hospitalized patients with COVID-19, highlighting the leading role of systemic inflammation for the occurrence of CI.


Fascia iliaca compartment block for analgesia in total hip replacement: A randomized controlled study protocol.
Pain management after the total joint arthroplasty is still challenging, but worthy of attention, because good pain management can improve the outcomes of patient. It is still controversial whether fascia iliaca compartment block (FICB) can effectively decrease the opioid consumption and pain after total hip replacement (THR) owing to the number of published investigations is small. The purpose of this present study is to assess the efficacy and safety of FICB for postoperative analgesia after THR. This is a single center, placebo-controlled randomized trial which is performed in accordance with the SPIRIT Checklist for randomized studies. It was authorized via the Chifeng Municipal Hospital institutional review committee (H2020-19-8). 100 patients undergoing THR will be included in this study. Patients are randomly divided into 2 groups: FICB group or Non-FICB group, FICB with 5mgmL of epinephrine and 40 mL of ropivacaine 0.2%. Primary outcomes are pain score at different time point. Visual analog scale is used to assess the pain (10: the maximum possible pain and 0: absent pain). The secondary outcomes are the postoperative complications, length of hospital stay and total consumption of opioid. All the needed analyses are implemented through utilizing SPSS for Windows Version 15.0. Figure 1 will show the primary and secondary outcomes. This trial can provide an evidence for the use of FICB for analgesia after THR.


Analysis of the B10.D2-H-2dml mutant with monoclonal antibodies.
We compared H-2D products of B10.D2 and the mutant B10.D2-H-2dml spleen cells using two monoclonal antibodies designated D3.179 and S11.7. D3.179 reacts with public specificities and appears to define more than one product as shown by cytotoxicity and binding studies. Strong reactivity was obtained in both binding and cytotoxicity to B10.D2 and marginal cytotoxicity and low binding were observed with B10.D2-H-2dml cells. The second monoclonal antibody S11.7, in contrast, reacts only with H-2Dd bearing strains and gave strong binding to B10.D2-H-2dml and weaker binding to B10.D2. Taken together, the observations indicate that the mutant not only expresses H-2D encoded antigens but bears quantitatively greater amounts of some specificities than the parent. A product of approximately 46,000 daltons was immunoprecipitated from B10.D2 cells by D3.179. Very small amounts of this product were seen in B10.D2-H-2dml immunoprecipitates.


The roles of endothelin and its receptors in cigarette smoke-associated pulmonary hypertension with chronic lung disease.
Chronic exposure to cigarette smoke is the major risk factor for the development of pulmonary hypertension (PH) with chronic lung disease (i.e. PH group III). The pathogenesis of smoke-associated PH group III in chronic obstructive pulmonary disease (COPD) involves cigarette smoke exposure-induced damage to lung tissue and dysfunction of pulmonary system with increased synthesis and release of endothelin-1 (ET-1), hypoxia, inflammation, pulmonary vascular remodeling. Many studies have demonstrated that cigarette smoke exposure induces activation of mitogen-activated protein kinase (MAPK) signal pathway that leads to up-regulation of ET-1 and its receptors with the receptor-mediated enhanced contraction, proliferation of pulmonary vascular smooth muscle cells, pulmonary vascular remodeling, elevated pulmonary arterial pressure and finally PH group III. This mini-review article aims to summarize the current state of understanding on the roles of cigarette smoke-induced up-regulation of ET-1 and its receptors in the development of PH group III. Understanding the underlying molecular mechanisms that cigarette smoke exposure leads to PH group III may provide a novel strategy for the treatment.


Persistent atrial standstill developed in a patient with rheumatic heart disease: electrophysiological and histological study.
A case with persistent atrial standstill is presented which developed from atrial fibrillation associated with rheumatic combined valvular heart disease. In addition to clinical and electrocardiographic findings, electrophysiological and histological studies by using microelectrode technique and electron microscopy, respectively, were carried out on specimens from the right atrial appendage resected at mitral valve surgery. Scattered but severe loss of myocardial cells in the atria and deterioration of the action potential of these poorly sustained myocardial cells were confirmed. The suggestive evidence is also shown that the deteriorated action potential observed by micro-electrode technique in this case may be composed of calcium current.


Cell-free detection and characterization of a novel nerve growth factor-activated protein kinase in PC12 cells.
We have developed a cell-free assay to detect and characterize nerve growth factor (NGF)-activated protein kinase activity. Cultured PC12 cells were briefly exposed to NGF, and extracts of these were assayed for phosphorylating activity using exogenously added tyrosine hydroxylase as substrate. Tyrosine hydroxylase was employed since it is an endogenous substrate of NGF-regulated kinase activity and is activated by phosphorylation. In the cell-free assay, extracts prepared from NGF-treated cells yielded a 2-3-fold greater incorporation of phosphate into tyrosine hydroxylase as compared with extracts of control, NGF-untreated cells. Activation did not occur, however, if NGF was added directly to cell extracts. The NGF-stimulated phosphorylating activity appeared to be due to regulation of a protein kinase rather than of a phosphoprotein phosphatase. Characterization of the kinase (designated as kinase N) showed that it is soluble, is detectably activated within 1-3 min after cells are exposed to NGF and maximally activated by 10 min, is half-maximally activated with 0.5 nM NGF and maximally activated with 1 nM NGF, is detectable in the presence of either Mg2+ or Mn2+ but does not require Ca2+, does not require nonmacromolecular cofactors, can use histone H1 as a substrate, and exhibits a 2-fold increase in apparent Vmax in response to NGF but does not undergo a significant change in apparent Km for either ATP or GTP. A number of characteristics of kinase N were assessed including susceptibility to inhibitors, substrate specificity, cofactor requirements, ATP dependence, and lack of down-regulation by prolonged expose to a phorbol ester. These studies indicated that it lacks tyrosine kinase activity and is distinct from a variety of well-characterized protein kinases including cAMP-dependent protein kinase, protein kinase C (Ca2+/phospholipid-dependent enzyme), Ca2+/calmodulin-dependent kinase, and casein kinase II. Preliminary purification data show that the kinase has a basic pI and that it has an apparent Mr of 22,000-25,000. The only amino acid in tyrosine hydroxylase found to be phosphorylated by the semipurified kinase is serine.


Specificity of homework compliance effects on treatment outcome in CBT: evidence from a controlled trial on panic disorder and agoraphobia.
Although homework assignments are an integral component of cognitive-behavioral therapy (CBT) and relate to positive therapy outcomes, it is unclear whether specific homework types and their completion have specific effects on outcome. Data from N = 292 patients (75% female, mean age 36 years) with panic disorder and agoraphobia and treated with standardized CBT were analyzed with homework compliance quality and quantity for different types of homework serving as predictors for different outcome variables. Quality ratings of homework completion were stronger outcome predictors than quantitative compliance ratings. Exposure homework was a better outcome predictor than homework relating to psychoeducation and self-monitoring. Different aspects of homework compliance and specific homework types might differentially relate to CBT outcome.


[Cloning and immunogenicity of conservative region of adhesin gene of Helicobacter pylori].
To construct a candidate strain of Helicobacter pylori (Hp) that expresses the proteins of the conservative region of 4 adhesins (BabA2, AlpA, AlpB, and HopZ) and study its immunogenicity. The DNA of Hp was extracted. Primers were designed according to the C-terminal structural gene sequence (called CB) of AlpA. The CB gene was amplified by PCR and inserted into the prokaryotic expression vector pET-22b (+) and expressed in BL21 (DE3) strain of Escherichia coli. The product of expression, CB, was purified by affinity chromatography and identified by Western blot analysis. ELISA assay was used to measure the CB-specific antibody in the specimens of serum of 55 Hp infected patients. Rapid urease test (RUT) was used on biopsy specimens collected by gastroscopy as parallel control. A recombinant plasmid pET-22b (+)/CB was constructed with the conservative region of the 4 adhesins. DNA sequencing showed one open reading frame of 588 bp encoding a polypeptide of 195 amino acids. The recombinant CB (rCB) protein, with a molecular weight of 22.5KD, amounted to 29% of the total bacterial protein. The purity of purified rCB was 96%. Western blot analysis showed that the rCB protein could be specifically recognized by the serum from Hp infected patients. The kappa coefficient was 0.76 for evaluation by ELISA and RUT results. CB has the potential to be used as a vaccine against Hp infection.


Dietary fat influences electric membrane properties of neurons in cell culture.
1. SJL/J mice were maintained on semipurified diets which differed in the ratio of polyunsaturated/saturated fatty acid content (P/S). Exposure was from conception and was maintained for periods ranging from 6 to 34 weeks. 2. Neural cell cultures were prepared from dorsal root ganglia (DRG). After 6 and 20 days of culture, neuronal electric membrane properties were determined quantitatively by intracellular recording. 3. A number of significant differences were observed for the two dietary conditions. DRG from mice on the low-P/s diet had an increase in the rate of fall of both phases of repolarization which, in conjunction with the reduced action potential overshoot, led to a reduced action potential duration. This shift to shorter-duration action potentials was accompanied by a shift to more monophasic falling phases. The low-P/S neurons also exhibited a decreased afterhyperpolarization, decreased specific membrane resistance, and decreased membrane electrical time constant compared to high-P/S neurons. 4. It was concluded that the P/S ratio in the diet can have a significant effect on the electric properties of neurons. The high-P/S neurons tended to have action potentials with biphasic repolarizations and longer durations. In contrast, the low-P/S neurons tended to have action potentials with monophasic repolarizations and shorter durations. Moreover, the known ionic dependence of these two types of action potentials suggested that the low-P/S diet resulted in action potentials with a more exclusive Na dependence, while the high-P/S diet resulted in action potentials with both Na and Ca dependence.


Membrane-based torque magnetometer: Enhanced sensitivity by optical readout of the membrane displacement.
The design and realization of a torque magnetometer is reported that reads the deflection of a membrane by optical interferometry. The compact instrument allows for low-temperature measurements of tiny crystals less than a microgram with a significant improvement in sensitivity, signal-to-noise ratio as well as data acquisition time compared with conventional magnetometry and offers an enormous potential for further improvements and future applications in different fields. Magnetic measurements on single-molecule magnets demonstrate the applicability of the membrane-based torque magnetometer.


Prepubertal sons of substance abusers: influences of parental and familial substance abuse on behavioral disposition, IQ, and school achievement.
In order to better understand the transgeneration liability for a substance abuse disorder, we investigated the impact of parental and familial substance abuse disorders on prepubertal boys. Specifically, the influence of each parent's substance abuse history and the effects of significant family aggregation of substance abuse disorders were tested as predictors of the child's behavioral disposition, IQ, and school achievement scores, while controlling for socioeconomic status (SES). Sons of substance abusing fathers were found to have higher externalizing and internalizing problem-behavior scores, lower IQ scores, and lower school achievement scores. Internalizing and externalizing problem-behavior scores were most strongly associated with bilineal parental substance abuse, whereas SES and paternal substance abuse were most strongly associated with IQ and school performance scores. The results are compatible with the hypothesis that although paternal substance abuse has an adverse impact on the son's functioning, bilineal parental substance abuse is associated with the greatest behavioral deviancy among prepubertal males and is associated with a greater liability for substance abuse.


Generation of interconnected vesicles in a liposomal cell model.
We introduce an experimental method based upon a glass micropipette microinjection technique for generating a multitude of interconnected vesicles (IVs) in the interior of a single giant unilamellar phospholipid vesicle (GUV) serving as a cell model system. The GUV membrane, consisting of a mixture of soybean polar lipid extract and anionic phosphatidylserine, is adhered to a multilamellar lipid vesicle that functions as a lipid reservoir. Continuous IV formation was achieved by bringing a micropipette in direct contact with the outer GUV surface and subjecting it to a localized stream of a Ca2+ solution from the micropipette tip. IVs are rapidly and sequentially generated and inserted into the GUV interior and encapsulate portions of the micropipette fluid content. The IVs remain connected to the GUV membrane and are interlinked by short lipid nanotubes and resemble beads on a string. The vesicle chain-growth from the GUV membrane is maintained for as long as there is the supply of membrane material and Ca2+ solution, and the size of the individual IVs is controlled by the diameter of the micropipette tip. We also demonstrate that the IVs can be co-loaded with high concentrations of neurotransmitter and protein molecules and displaying a steep calcium ion concentration gradient across the membrane. These characteristics are analogous to native secretory vesicles and could, therefore, serve as a model system for studying secretory mechanisms in biological systems.


Parenteral nutrition solution retrieved by lumbar puncture following left saphenous vein catheterization.
Two extremely preterm infants had silastic lines inserted via the left great saphenous vein for intravenous feeding. Clinical deterioration approximately 10 days later led to lumbar puncture as part of a sepsis screen. Parenteral nutrition solution was recovered in the cerebrospinal fluid and heralded potentially life-threatening catheter malposition. The cases illustrate another percutaneous central line-associated morbidity, which is avoidable by careful verification of line position. All previously reported cases of this unusual complication are reviewed and we describe its likely mechanism. We postulate that the phenomenon is a complication peculiar to percutaneous catheterization of the left lower limb.


Genomic features of Mycoplasma bovis subtypes currently circulating in France.
Mycoplasma (M.) bovis is a major etiological agent of bovine respiratory disease, which is the most economically costly disease of cattle worldwide. Cattle disease surveillance on M. bovis is increasingly using gene-based techniques, such as multilocus sequence typing (MLST), or genome-based techniques such as core genome MLST that both require only partial genomic data. However, accurate up-to-date surveillance also demands complete, circular genomes that can be used as reference to track the evolution of the different lineages. Yet, in France, two of the main subtypes currently circulating still have no representing genome in public databases. Here, to address this gap, we provide and compare three new complete M. bovis genomes obtained from recent clinical isolates that represent major subtypes circulating in France and Europe. Genomes were obtained using a hybrid assembly strategy (Illumina and Nanopore) with fine-tuning of settings and inputs used in the Unicycler assembly pipeline, such as size selection of reads and quality trimming of the FASTQ files. The main characteristics and synteny of the genomes were compared. The three genomes mainly differed by their content in terms of mobile genetic elements, i.e. integrative conjugative elements (ICE) and insertion sequences (IS), a feature that impacts their structure. For instance, strain L15527, representing subtype3 (st3), harbours an exceptionally high number of ICEs, which results in a bigger-sized genome than all those previously described and could be associated with the propensity of st3 to gain and fix mutations through chromosomal transfer mechanisms. In contrast, strain F9160, of st1, is very close to the PG45 type strain isolated in 1961 in the USA, and harbours a huge number of IS. These features may be associated with an evolution towards a host-restricted state or in a "closed" host or environment reservoir until a recent re-emergence. Whole-genome comparison of the three French M. bovis subtypes provides valuable resources for future studies combining epidemiology, phylogenetic data, and phylodynamic methods.


Testing the relationships between energy consumption, CO2 emissions, and economic growth in 24 African countries: a panel ARDL approach.
This study complements existing literature by examining the nexus between energy consumption (EC), CO2 emissions (CE), and economic growth (GDP; gross domestic product) in 24 African countries using a panel autoregressive distributed lag (ARDL) approach. The following findings are established. First, there is a long-run relationship between EC, CE, and GDP. Second, a long-term effect from CE to GDP and EC is apparent, with reciprocal paths. Third, the error correction mechanisms are consistently stable. However, in cases of disequilibrium, only EC can be significantly adjusted to its long-run relationship. Fourth, there is a long-run causality running from GDP and CE to EC. Fifth, we find causality running from either CE or both CE and EC to GDP, and inverse causal paths are observable. Causality from EC to GDP is not strong, which supports the conservative hypothesis. Sixth, the causal direction from EC to GDP remains unobservable in the short term. By contrast, the opposite path is observable. There are also no short-run causalities from GDP, or EC, or EC, and GDP to EC. Policy implications are discussed.