The Role of ONECUT1 Variants in Monogenic and Type 2 Diabetes Mellitus.
ONECUT1 (also known as HNF6) is a transcription factor involved in pancreatic development and β-cell function. Recently, biallelic variants in ONECUT1 were reported as a cause of neonatal diabetes mellitus (NDM) in two subjects, and missense monoallelic variants were associated with type 2 diabetes and possibly maturity-onset diabetes of the young (MODY). Here we examine the role of ONECUT1 variants in NDM, MODY, and type 2 diabetes in large international cohorts of subjects with monogenic diabetes and >400,000 subjects from UK Biobank. We identified a biallelic frameshift ONECUT1 variant as the cause of NDM in one individual. However, we found no enrichment of missense or null ONECUT1 variants among 484 individuals clinically suspected of MODY, in whom all known genes had been excluded. Finally, using a rare variant burden test in the UK Biobank European cohort, we identified a significant association between heterozygous ONECUT1 null variants and type 2 diabetes (P = 0.006) but did not find an association between missense variants and type 2 diabetes. Our results confirm biallelic ONECUT1 variants as a cause of NDM and highlight monoallelic null variants as a risk factor for type 2 diabetes. These findings confirm the critical role of ONECUT1 in human β-cell function.


An acceptability and safety study of the Duet cervical barrier and gel delivery system in Zimbabwe.
Adherence problems with coitally dependent, female-initiated HIV prevention methods have contributed to several trials' failure to establish efficacy. Continuous use of a cervical barrier with once-daily cleaning and immediate reinsertion may simplify use for women and improve adherence. We assessed the acceptability and safety of precoital and continuous use of the Duet, a cervical barrier and gel delivery system, in Zimbabwean women. Using a two-arm crossover design with a parallel observation group, we randomized 103 women in a 2:2:1 ratio: (1) to use the Duet continuously for 14 days, followed by a minimum of seven days of washout and then 14 days of precoital use; (2) to use the same Duet regimens in reverse order; or (3) for observation only. Women were aged 18 to 40 years; half were recruited from a pool of previous diaphragm study participants and the other half from the general community. Acceptability and adherence were assessed through an interviewer-administered questionnaire at each of two follow-up visits. Safety was monitored through pelvic speculum exams and report of adverse events. The proportion of women who reported consistent Duet use during sex was virtually identical during continuous and precoital regimens (88.6% vs. 88.9%). Partner refusal was the most common reason cited for non-use during sex in both use regimens. Not having the device handy was the most common reason cited for non-daily use (in the continuous regimen). Most women were "very comfortable" using it continuously (86.3%) and inserting it precoitally (92.8%). The most favoured Duet attribute was that it did not interfere with "natural" sex (55%). The least favoured Duet attribute was the concern that it might come out during sex (71.3%). No serious adverse events were reported during the study; 57 participants reported 90 adverse events classified as mild or moderate. There were no statistically significant differences in: (1) the proportion of women reporting adverse events; (2) the severity of events among those using the Duet and observational controls; or (3) event severity reported during each regimen use period. In this study, the Duet was found to be acceptable and safe when inserted precoitally or used continuously for 14 days. Assignment to use of the Duet continuously did not increase adherence to the Duet during sex. Future HIV prevention trials should evaluate use of the Duet (precoitally and continuously) with promising microbicide candidates.


[Challenges in the implementation of HPV vaccination for cancer prevention].
In France, every year, HPVs cause approximately 6,300 cancers -cervix, anus, oropharynx, penis, vulva, vagina, oral cavity, larynx- representing 2% of all incident cancers, 35,000 precancerous le-sions of the cervix, and at least 50,000 condylomas in the under- 30s. All 3,000 new cases of cervical cancers are attributable to HPV infections. Cervical cancer prevention is based on the combination of two complementary strategies: vaccination of 11 to 14-years-old girls against HPV and organized screening of cervical cancer for women between 25 and 65 years old. In France, the vaccination coverage rate -VCR- against HPV for 2 vaccine doses is less than 25% while it reaches more than 80% in the United Kingdom, Portugal, Australia, etc. This low level of VCR can be explained by the mistrust in these vaccines especially regarding their effectiveness and safety. However, more than 10 years after their commercialization, HPV vaccines have already demonstrated, in many countries, their effectiveness against HPV infections, genital warts and precancerous cervical lesions. In addition, no association between vaccines and autoimmune diseases has been demonstrated, either by national or international surveillance authorities, or by scientific publications, after the commercialization of 200 million doses worldwide. As the International Papillomavirus Society has declared that the combination of a high VCR against HPV and a high participation in cervical cancer screening, combined with appropriate treatment, can lead to the elimination of cervical cancer as a public health problem, it is important to better communicate the scientific findings in order to better understand vaccination's benefits.


Xenopus laevis FGF16 activates the expression of genes coding for the transcription factors Sp5 and Sp5l.
Fibroblast growth factors (FGFs) comprise a family of signalling molecules with essential roles in early embryonic development across animal species. The role of FGFs in mesoderm formation and patterning in Xenopus has been particularly well studied. However, little is known about FGF16 in Xenopus. Using in situ hybridisation, we uncover the expression pattern of FGF16 during early Xenopus laevis development, which has not been previously described. We show that the zygotic expression of FGF16 is activated in the mesoderm of the early gastrula as a ring around the blastopore, with its first accumulation at the dorsal side of the embryo. Later, FGF16 expression is found in the otic vesicle, the branchial arches and the anterior pituitary, as well as in the chordal neural hinge region of the tailbud. In addition, we show that FGF16 can activate the MAPK pathway and expression of sp5 and sp5l. Like FGF16, sp5 is expressed in the otic vesicle and the branchial arches, with all three of these genes being expressed in the tailbud. These data provide evidence that FGF16 is present in the early mesoderm and can activate the expression of developmentally important transcription factors.


Interplay among nocturnal activity, melatonin, corticosterone and performance in the invasive cane toad (Rhinella marinus).
Most animals conduct daily activities exclusively either during the day or at night. Here, hormones such as melatonin and corticosterone, greatly influence the synchronization or regulation of physiological and behavioral cycles needed for daily activity. How then do species that exhibit more flexible daily activity patterns, responses to ecological, environmental or life-history processes, regulate daily hormone profiles important to daily performance? This study examined the consequences of (1) nocturnal activity on diel profiles of melatonin and corticosterone and (2) the effects of experimentally increased acute melatonin levels on physiological and metabolic performance in the cane toad (Rhinella marinus). Unlike inactive captive toads that had a distinct nocturnal melatonin profile, nocturnally active toads sampled under field and captive conditions, exhibited decreased nocturnal melatonin profiles with no evidence for any phase shift. Nocturnal corticosterone levels were significantly higher in field active toads than captive toads. In toads with experimentally increased melatonin levels, plasma lactate and glucose responses following recovery post exercise were significantly different from control toads. However, exogenously increased melatonin did not affect resting metabolism in toads. These results suggest that toads could adjust daily hormone profiles to match nocturnal activity requirements, thereby avoiding performance costs induced by high nocturnal melatonin levels. The ability of toads to exhibit plasticity in daily hormone cycles, could have broad implications for how they and other animals utilize behavioral flexibility to optimize daily activities in response to natural and increasingly human mediated environmental variation.


Differential expression of progestin receptor isoforms in the hypothalamus, pituitary, and endometrium of rhesus macaques.
The progestin receptor exists in at least two isoforms: a long form (PR-B) and a short form (PR-A), which can be separated and detected with Western blot analysis. It has been suggested from in vitro transfection experiments that differential expression of the two isoforms may provide one mechanism for tissue specific actions of progesterone (P). However, more information from in vivo experimentation is needed. It has been reported that P down-regulates the expression of PR in the endometrium and pituitary of E primed macaques. However, PR protein and PR messenger RNA expression in the hypothalamus is maintained with P treatment of E-primed macaques. Thus, there is tissue-specific regulation of PR by its cognate ligand in the nonhuman primate. To gain insight into the tissue-specific regulation of PR by P, we questioned whether differential expression of the isoforms of PR exists in the endometrium, pituitary, and hypothalamus of rhesus monkeys. The expression of PR-A and PR-B was examined after E (28-30 days) and E + P (14 days E + 14 days E + P) treatment in the primate endometrium, pituitary, and hypothalamus. After E or E + P treatment, the levels of PR-A were 5 times higher than PR-B in the endometrium. PR-A was 1.6-fold higher than PR-B in the pituitary. In the hypothalamus, the ratio of A to B ranged from less than 1 (B exceeds A) to unity (A and B equimolar). There was no difference in the ratio of A to B between E-treated and E + P-treated groups in any tissue examined. These observations (a) provide further support of the hypothesis that differential expression of the isoforms of PR may subserve the tissue specific actions of P and (b) also suggest that P does not differentially affect the expression of the isoforms of its cognate receptor in the endometrium, pituitary, or hypothalamus.


Structural features of incremental line-like striations in mandibular condylar cartilage of c-src-deficient mice.
Mandibular condylar cartilage is sensitive to masticatory force, while mice lacking the c-src gene (c-src-deficient mice) have osteopetrosis and tooth eruption failure. The purpose of this study was to investigate the morphology of the mandibular condyle in these mice, which were maintained with a soft-food diet for 240 days after birth. The condylar head in the c-src-deficient mice showed slight deformity in shape before weaning, but showed remarkable undergrowth after weaning. No significant morphological or histological differences were detected between the mandibular condyle in wild-type mice fed soft food and those fed hard food, indicating that osteopetrosis, as well as abnormal masticatory force, influences the morphology of the mouse mandibular condyle, and that malocclusion rather than dietary consistency may have greater influence. After 70 days, incremental line-like striations consisting of cartilaginous and non-cartilaginous layers were detected in the mandibular condyle of the c-src-deficient mice, but not in the tibial growth plate. Immunostaining of aggrecan, collagen types II and X, and osteopontin was detected in the cartilaginous layers, but not in the non-cartilaginous layers showing collagen type I immunostaining. Chondrocyte lacunae were not eroded in the cartilaginous layers, and complete circumferential mineralisation around the lacunae and impaired osteoclast (chondroclast) function can account for this phenomenon. However, repeated cessation of chondrocyte differentiation may be required to completely explain the formation of the striations. These results indicate that the mandibular condyle in the c-src-deficient mice has unique structural features, adding to its deformity.


MicroRNA-29c mediates initiation of gastric carcinogenesis by directly targeting ITGB1.
Gastric cancer (GC) remains difficult to cure due to heterogeneity in a clinical challenge and the molecular mechanisms underlying this disease are complex and not completely understood. Accumulating evidence suggests that microRNAs (miRNAs) play an important role in GC, but the role of specific miRNAs involved in this disease remains elusive. We performed next generation sequencing (NGS)-based whole-transcriptome profiling to discover GC-specific miRNAs, followed by functional validation of results. NGS-based miRNA profiles were generated in matched pairs of GCs and adjacent normal mucosa (NM). Quantitative RT-PCR validation of miR-29c expression was performed in 274 gastric tissues, which included two cohorts of matched GC and NM specimens. Functional validation of miR-29c and its gene targets was undertaken in cell lines, as well as K19-C2mE and K19-Wnt1/C2mE transgenic mice. NGS analysis revealed four GC-specific miRNAs. Among these, miR-29c expression was significantly decreased in GC versus NM tissues (p<0.001). Ectopic expression of miR-29c mimics in GC cell lines resulted in reduced proliferation, adhesion, invasion and migration. High miR-29c expression suppressed xenograft tumour growth in nude mice. Direct interaction between miR-29c and its newly discovered target, ITGB1, was identified in cell lines and transgenic mice. MiR-29c expression demonstrated a stepwise decrease in wild type hyperplasia-dysplasia cascade in transgenic mice models of GC. MiR-29c acts as a tumour suppressor in GC by directly targeting ITGB1. Loss of miR-29c expression is an early event in the initiation of gastric carcinogenesis and may serve as a diagnostic and therapeutic biomarker for patients with GC.


Refining the phenotype of borderline personality disorder: Diagnostic criteria and beyond.
Borderline personality disorder (BPD) is a heterogeneous disorder, and previous analyses have parsed its phenotype in terms of subtypes or underlying traits. We refined the BPD construct by testing a range of latent variable models to ascertain whether BPD is composed of traits, latent classes, or both. We also tested whether subtypes of BPD could be distinguished by anger, aggressiveness, antisocial behavior, and mis-trustfulness, additional putative indicators drawn from Kernberg's (1967, 1975) theory of BPD. In a mixed clinical and nonclinical sample (N = 362), a factor mixture model consisting of two latent classes (symptomatic and asymptomatic) and a single severity dimension fit the BPD criteria, as defined by the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV), data better than latent class or factor analytic approaches. In the second analytic phase, finite mixture modeling of the symptomatic latent class (n = 100) revealed four BPD subtypes: angry/aggressive, angry/mistrustful, poor identity/low anger, and prototypical. Our results support a hybrid categorical-dimensional model of the BPD DSM-IV criteria. The BPD subtypes emerging from this model have important implications for treatment and etiological research.


Inhaled Loxapine for Agitation in Intoxicated Patients: A Case Series.
Episodes of agitation are frequent in intoxicated patients who have a substance use disorder, a psychiatric disorder or both (dual diagnosis). For managing the agitation, it is necessary to act promptly in a safe environment and addressing any underlying etiology. Inhaled loxapine improves symptoms of agitation in adults with psychiatric disorders (eg, schizophrenia) within 10 minutes of administration. Recently, some reports have documented the usefulness of loxapine in dual diagnoses patients with agitation. However, the efficacy of loxapine in intoxicated patients has not been deeply addressed. This report describes a case series of 12 patients (with addiction or dual disorder) who received inhaled loxapine for symptoms of psychomotor agitation during intoxication with different substances (eg, alcohol, cannabis, or cocaine) at 1 center in Spain. Data from 12 patients were reviewed, 5 patients were attended at the emergency room, 4 at the addiction and dual diagnosis unit, and 3 were treated during hospitalization for detoxification. All patients were under effects of substances. They had substance use disorder (including cannabis, cocaine, alcohol, hypnotics, and hallucinogens), and almost all (90%) presented 1 or more psychiatric disorders. One dose of inhaled loxapine was effective in 9 patients (75%), and in 3 patients, a second dose was required. Only mild dizziness was reported in 1 patient after the second dose. The acute agitation was effectively and quickly managed with inhaled loxapine in all intoxicated patients and enabled the appropriate clinical evaluation of the agitated state and the patient's management.


Investigations into the Potential Role of Metabolites on the Anti-Leukemic Activity of Imatinib, Nilotinib and Midostaurin.
The efficacy and side-effects of drugs do not just reflect the biochemical and pharmacodynamic properties of the parent compound, but often comprise of cooperative effects between the properties of the parent and active metabolites. Metabolites of imatinib, nilotinib and midostaurin have been synthesised and evaluated in assays to compare their properties as protein kinase inhibitors with the parent drugs. The N-desmethyl-metabolite of imatinib is substantially less active than imatinib as a BCR-ABL1 kinase inhibitor, thus providing an explanation as to why patients producing high levels of this metabolite show a relatively low response rate in chronic myeloid leukaemia (CML) treatment. The hydroxymethylphenyl and N-oxide metabolites of imatinib and nilotinib are only weakly active as BCR-ABL1 inhibitors and are unlikely to play a role in the efficacy of either drug in CML. The 3-(R)-HO-metabolite of midostaurin shows appreciable accumulation following chronic drug administration and, in addition to mutant forms of FLT3, potently inhibits the PDPK1 and VEGFR2 kinases (IC50 values <100 nM), suggesting that it might contribute to drug efficacy in acute myeloid leukaemia patients. The case studies discussed here provide further examples of how the synthesis and characterisation of metabolites can make important contributions to understanding the clinical efficacy of drugs.


Comparative drug systemic exposure and clinical efficacy against resistant nematodes in lambs treated with different albendazole formulations.
A pharmaco-parasitological assessment of four different albendazole (ABZ) formulations was carried out in lambs infected with multiple resistant gastrointestinal (GI) nematodes. The comparative drug systemic exposure profiles (ABZ sulphoxide plasma concentrations) and anthelmintic efficacies (clinical endpoint measured through the faecal nematode eggs reduction counts) were determined for a reference formulation (RF) and three different test (T1, T2, T3) generic ABZ preparations. Fifty (50) Corriedale lambs naturally infected with multiple resistant GI nematodes were allocated into five experimental groups (n = 10). Animals in each group received treatment with either the RF, one of the test ABZ formulations (5 mg/kg by the intraruminal route) or were kept as untreated control. Blood samples were collected over 48 h post-treatment. ABZ parent drug was not recovered in the bloodstream. The ABZ sulphoxide (ABZSO) and sulphone (ABZSO(2) ) metabolites were measured in plasma by ultraviolet high-performance liquid chromatography over 36-48 h post-treatment. A faecal nematode egg count reduction test (FECRT) was performed at day 10th post-treatment to lambs from all treated and untreated groups, which indicated the predominance of nematodes with high level of resistance to ABZ. Both ABZSO C(max) and AUC(0-LOQ) values obtained for the RF (pioneer product) were significantly higher (P < 0.05) than those obtained for the T1 and T3 preparations. Based on the currently available bioequivalence criteria, the test (generic) ABZ formulations under evaluation could not be considered equivalent to the RF regarding the rate (C(max) ) and extent (AUC(0-LOD) ) of drug absorption (indirectly estimated through the ABZSO metabolite). A large variation in nematode egg counts did not permit to obtain statistically significant differences among formulations. However, a favourable trend in the efficacy against the most resistant nematodes was observed for the formulations with the highest ABZSO systemic exposure.


Permeability of the ellagitannin geraniin and its metabolites in a human colon adenocarcinoma Caco-2 cell culture model.
Ellagitannins, found abundantly in berries, pomegranates, walnuts and almonds, have been increasingly investigated for their health benefits. Geraniin (GE), an ellagitannin, found predominantly in herbal plants, as well has been shown to exhibit a number of biological activities. Like many hydrolysable tannins, geraniin is water-soluble and readily undergoes hydrolysis in the presence of hot water, weak acids and weak bases to yield several metabolites including corilagin (CO), ellagic acid (EA) and gallic acid (GA). There are numerous studies on the pharmacological effectiveness of GE, CO and GA. However, the intestinal permeability of GE and CO has never been investigated before. Caco-2 cell transport assay was utilized to evaluate the in vitro permeability of GE and its metabolites. GE, CO and EA were found to have no apparent permeability (Papp) while GA displayed a Papp value of 31.3 ± 1.1 × 10-6 cm s-1. Mass balance studies showed a loss of geraniin and its metabolites during transport. Chemical stability studies in the transport buffers revealed that GE and CO were hydrolyzed in the HBSS buffers. Experiments using lysed cells revealed that GE and its metabolites were metabolized during transport. Absorption and desorption studies confirmed the accumulation of EA inside the cells. The above results indicate that the compounds have poor oral absorption. To consider these compounds or their natural extracts as oral nutraceutical candidates, formulation strategies are mandatory.


DESMAN: a new tool for de novo extraction of strains from metagenomes.
We introduce DESMAN for De novo Extraction of Strains from Metagenomes. Large multi-sample metagenomes are being generated but strain variation results in fragmentary co-assemblies. Current algorithms can bin contigs into metagenome-assembled genomes but are unable to resolve strain-level variation. DESMAN identifies variants in core genes and uses co-occurrence across samples to link variants into haplotypes and abundance profiles. These are then searched for against non-core genes to determine the accessory genome of each strain. We validated DESMAN on a complex 50-species 210-genome 96-sample synthetic mock data set and then applied it to the Tara Oceans microbiome.


Thermal conductivity of the porcine heart tissue.
The porcine heart was used as a model for studying the thermal changes in myocardium at cooling and re-warming during open heart surgery. A section of the heart septum was excised and tissue was cut into two similar square slices. The same shape of the tissue, cut from the surface from the upper lateral wall of the left ventricle, covered with epicardium and fat, was taken for another measurement. A thin (<0.5 mm) square thermal source of the same length of the side as the tissue samples was put between the two slices of tissue. This set was placed in the middle of two identical copper cylinders (2r=50 mm, height=55 mm) used to keep the outer side of the specimen at controlled room temperature. Thermal conductivity of the heart tissue was determined at controlled thermal power, and known difference of the temperature at the edge of the tissue and at the middle of the heater, when steady state was reached. Thermal conductivity calculated from the temperature difference and the geometry of heater and samples was 0.75 W/m.K for septal heart tissue, and 0.60 W/m.K for the lateral wall ventricle tissue with epicardium and fat.


RACE1, a Japanese Blumeria graminis f. sp. hordei isolate, is capable of overcoming partially mlo-mediated penetration resistance in barley in an allele-specific manner.
Loss-of-function mutation of the MILDEW RESISTANCE LOCUS O (Mlo) gene confers durable and broad-spectrum resistance to powdery mildew fungi in various plants, including barley. In combination with the intracellular nucleotide-binding domain and leucine-rich repeat receptor (NLR) genes, which confer the race-specific resistance, the mlo alleles have long been used in barley breeding as genetic resources that confer robust non-race-specific resistance. However, a Japanese Blumeria graminis f. sp. hordei isolate, RACE1, has been reported to have the potential to overcome partially the mlo-mediated penetration resistance, although this is yet uncertain because the putative effects of NLR genes in the tested accessions have not been ruled out. In this study, we examined the reproducibility of the earlier report and found that the infectious ability of RACE1, which partially overcomes the mlo-mediated resistance, is only exerted in the absence of NLR genes recognizing RACE1. Furthermore, using the transient-induced gene silencing technique, we demonstrated that RACE1 can partially overcome the resistance in the host cells with suppressed MLO expression but not in plants possessing the null mutant allele mlo-5.


Adenosine deaminase, a key enzyme in DNA precursors control, is a new p73 target.
The discovery of the p73 and p63 genes, homologous to p53 tumor suppressor has uncovered a family of transcription factors and widened the scenario of cell cycle control and apoptosis. We have identified a putative p53-responsive element in the human adenosine deaminase (ADA) gene, an important enzyme involved in nucleotide metabolism, the deficit of which causes the inhibition of DNA synthesis and repair. Here, we demonstrate that the ectopic expression of p73 isoforms leads to the ADA gene upregulation, showing for the first time a correlation between p73 and ADA. We found that p73 promotes ADA gene expression following a dNTP unbalance generated by ADA enzyme deficiency and 2'deoxyadenosine accumulation. The abrogation of p73 transcriptional activity by the specific dominant-negative p73DD abolishes ADA induction. By contrast, the ADA gene does not appear to be a functional p53 target in the physiological conditions we tested. On the whole, our results contribute to the emerging picture that p73 could play a different role from p53 in normal growth and development by inducing alternative target genes, which are not shared by p53.


Nickel enzyme maturation in Helicobacter hepaticus: roles of accessory proteins in hydrogenase and urease activities.
Helicobacter hepaticus, a causative agent of chronic hepatitis and hepatocellular carcinoma in mice, possesses a hydrogenase and a urease, both of which are nickel-containing enzymes. Analysis of the genome sequence of H. hepaticus revealed a full set of accessory genes which are required for the nickel maturation of each enzyme in other micro-organisms. Erythromycin-resistant mutants were constructed in four of these genes, hypA, hypB, ureE and ureG. Controls for polar effect were provided for hypA or hypB mutants by disrupting each gene located immediately downstream, i.e. hp0809 or hypC, respectively. Urease and hydrogenase activities were determined for each strain with or without supplemented nickel in the medium. As expected, the ureE and the ureG mutants had negligible urease activity, but they retained normal levels of hydrogenase activity. Urease levels could not be increased by the addition of nickel to the medium. The H. hepaticus hypA and hypB strains were deficient in both urease and hydrogenase activities, suggesting that both gene products act in a similar fashion as their counterparts in H. pylori. However, in contrast with the analogous mutants of H. pylori, the addition of nickel into the growth medium failed to restore either urease or hydrogenase enzyme levels in the H. hepaticus hypA or hypB mutants, indicating a probably unique role for these genes in the mouse liver pathogen.


Waveform feature extraction based on tauberian approximation.
A technique is presented for feature extraction of a waveform y based on its Tauberian approximation, that is, on the approximation of y by a linear combination of appropriately delayed versions of a single basis function x, i.e., y(t) = ¿M i = 1 aix(t - ¿i), where the coefficients ai and the delays ¿i are adjustable parameters. Considerations in the choice or design of the basis function x are given. The parameters ai and ¿i, i=1, . . . , M, are retrieved by application of a suitably adapted version of Prony's method to the Fourier transform of the above approximation of y. A subset of the parameters ai and ¿i, i = 1, . . . , M, is used to construct the feature vector, the value of which can be used in a classification algorithm. Application of this technique to the classification of wide bandwidth radar return signatures is presented. Computer simulations proved successful and are also discussed.


Jittery, a Mutator distant relative with a paradoxical mobile behavior: excision without reinsertion.
The unstable mutation bz-m039 arose in a maize (Zea mays) stock that originated from a plant infected with barley stripe mosaic virus. The instability of the mutation is caused by a 3.9-kb mobile element that has been named Jittery (Jit). Jit has terminal inverted repeats (TIRs) of 181 bp, causes a 9-bp direct duplication of the target site, and appears to excise autonomously. It is predicted to encode a single 709-amino acid protein, JITA, which is distantly related to the MURA transposase protein of the Mutator system but is more closely related to the MURA protein of Mutator-like elements (MULEs) from Arabidopsis thaliana and rice (Oryza sativa). Like MULEs, Jit resembles Mutator in the length of the element's TIRs, the size of the target site duplication, and in the makeup of its transposase but differs from the autonomous element Mutator-Don Robertson in that it encodes a single protein. Jit also differs from Mutator elements in the high frequency with which it excises to produce germinal revertants and in its copy number in the maize genome: Jit-like TIRs are present at low copy number in all maize lines and teosinte accessions examined, and JITA sequences occur in only a few maize inbreds. However, Jit cannot be considered a bona fide transposon in its present host line because it does not leave footprints upon excision and does not reinsert in the genome. These unusual mobile element properties are discussed in light of the structure and gene organization of Jit and related elements.


[The effect of straw meal on the crude protein and amino acid metabolism and digestibility of crude nutrients in broiler hen breeds. 1. Problems, experimental review and N-excretion in the urine].
The metabolization of the straw N and the influence of the straw on N excretion in urine were studied in 2 experiments with colostomized broiler hens and with 15N labelled wheat straw as well as 15N labelled wheat. In experiment 1 the test animals divided up into 4 groups received 0 g, 20 g, 30 g and 40 g straw meal per animal and day in addition to 120 g mixed feed. The daily 15N-excess (15N') intake from the straw was 18.4 mg, 27.5 mg and 36.7 mg. The amount of 15N' daily consumed with the labelled wheat in experiment 2 was 119.7 mg. 40 g straw meal resulted in a significantly increased amount of urine (p less than 0.05). The amounts of urine N and uric acid N were only increased as a tendency. On average the productive N decreased as a consequence of the straw meal supplement from 1070 mg/animal and day (control) to 764 mg/animal and day after 40 g straw meal supplement. The productive 15N' of the labelled wheat was not influenced by the straw meal supplement. The productive 15N' of the straw increased from 3.8 mg/animal and day (20 g straw) to 13.4 mg/animal and day (40 g straw). In contrast to 15N wheat, straw as a 15N source resulted in a lower labelling of uric acid N in comparison with urine N. It can be assumed that the changed metabolization of the straw N is influenced by microbial processes in the intestines.


Development of an ultrasensitive immunoassay for detecting tartrazine.
We have developed an ultrasensitive indirect competitive enzyme-linked immunosorbent assay for the determination of tartrazine. Two carboxylated analogues of tartrazine with different spacer lengths, and one derivative from commercial tartrazine after a little chemical modification, were synthesized as haptens in order to produce antibodies specific to tartrazine. The effect of sulfonic acid groups on the hapten structure of tartrazine was also studied carefully for the first time. A most specific monoclonal antibody against tartrazine was created and exhibited an IC50 value of 0.105 ng/mL and a limit of detection of 0.014 ng/mL, with no cross-reactivity to other structurally-related pigments. The established immunoassay was applied to the determination of tartrazine in fortified samples of orange juice and in real positive samples of carbonated beverages.


The syp enhancer sequence plays a key role in transcriptional activation by the σ54-dependent response regulator SypG and in biofilm formation and host colonization by Vibrio fischeri.
Biofilm formation by Vibrio fischeri is a complex process that requires multiple regulators. One such regulator, the NtrC-like response regulator SypG, controls biofilm formation and host colonization by V. fischeri via its impact on transcription of the symbiosis polysaccharide (syp) locus. SypG is predicted to activate syp transcription by binding to the syp enhancer (SE), a conserved sequence located upstream of four syp promoters. In this study, we performed an in-depth analysis of the sequences necessary for SypG to promote syp transcription and biofilm formation. We found that the SE sequence is necessary for SypG-mediated syp transcription, identified individual bases necessary for efficient activation, and determined that SypG is able to bind to syp promoter regions. We also identified SE sequences outside the syp locus and established that SypG recognizes these sequences as well. Finally, deletion of the SE sequence upstream of sypA led to defects in both biofilm formation and host colonization that could be restored by reintroducing the SE sequence into its native location in the chromosome. This work thus fills in critical gaps in knowledge of the Syp regulatory circuit by demonstrating a role for the SE sequence in SypG-dependent control of biofilm formation and host colonization and by identifying new putative regulon members. It may also provide useful insights into other bacteria, such as Vibrio vulnificus and Vibrio parahaemolyticus, that have syp-like loci and conserved SE sequences.


Ronidazole in high concentrations in drinking water for treatment and prevention of diarrhea in swine dysentery.
Ronidazole administered in the drinking water at concentrations of 0.012, 0.006, and 0.003% was effective for the treatment of swine dysentery. All groups of medicated swine had more survivors, fewer days of hemorrhagic and nonhemorrhagic diarrhea, greater feed consumption, greater weight gain, and more favorable feed efficiency than did nonmedicated swine. The only adverse clinical sign and lesion observed in the medicated swine comprised the drug-delayed-augmented swine dysentery which affected 3 swine in a group given 0.003% ronidazole. Several of the swine given the 0.012% concentration developed nonhemorrhagic diarrhea while on medication, probably because of the high concentration of the drug. Diarrhea did not recur in swine after the withdrawal of medication in both of 2 groups given the 0.012% concentration, in 3 of 4 groups given the 0.006% concentration, or in 1 of 4 groups given the 0.003% concentration. One or more swine were susceptible to reexposure to swine dysentery in both groups given the 0.012% concentration, in 1 group given the 0.006% concentration and in 1 group given the 0.003% concentration. Large spirochetes were observed in fecal smears from all exposed swine which developed either hemorrhagic or nonhemorrhagic diarrhea.


Headache, drugs and sleep.
Headache and sleep mechanisms share multiple levels of physiological interaction. Pharmacological treatment of headache syndromes may be associated with a broad range of sleep disturbances, either as a direct result of the pharmacology of the drug used, or by unmasking physiological alterations in sleep propensity seen as part of the headache symptom complex. This review summarises known sleep and circadian effects of various drugs commonly used in the management of headache disorders, with particular attention paid to abnormal sleep function emerging as a result of treatment. Literature searches were performed using MEDLINE, PubMed, and the Cochrane database using search terms and strings relating to generic drug names of commonly used compounds in the treatment of headache and their effect on sleep in humans with review of additional pre-clinical evidence where theoretically appropriate. Medications used to treat headache disorders may have a considerable impact on sleep physiology. However, greater attention is needed to characterise the direction of the changes of these effects on sleep, particularly to avoid exacerbating detrimental sleep complaints, but also to potentially capitalise on homeostatically useful properties of sleep which may reduce the individual burden of headache disorders on patients.


Cl-Amidine Prevents Histone 3 Citrullination and Neutrophil Extracellular Trap Formation, and Improves Survival in a Murine Sepsis Model.
Sepsis refers to the presence of a serious infection that correlates with systemic and uncontrolled immune activation. Posttranslational histone modification plays an important role in chromatin decondensation, which is regulated by citrullination. Citrullinated histone H3 (H3cit) has been identified as a component of neutrophil extracellular traps (NETs), which are released into the extracellular space as part of the neutrophil response to infection. The conversion of arginine to citrulline residues on histones is catalyzed by peptidylarginine deiminase 4 (PAD4). This study's goals were to characterize the presence of PAD4-catalyzed H3cit and NET formation during the onset of sepsis and elucidate the effects on the immune response when this mechanism of action is blocked. Adult C57BL/6 male mice were treated with Cl-amidine, an inhibitor of PAD4, 1 h prior to sepsis induced by cecal ligation and puncture (CLP). Twenty-four hours after CLP, cytokine levels, H3cit protein expression, neutrophil counts, and NET production were evaluated in the peritoneal cavity. Survival studies were also performed. Here we demonstrate that Cl-amidine treatment prior to CLP improves overall survival in sepsis and the abrogation of PAD4 has minimal effects on the proinflammatory immune response to sepsis, while it has no effect on overall neutrophil migration to the peritoneum.


The Effect of Fire Smoke Exposure on Firefighters' Lung Function: A Meta-Analysis.
Firefighters are exposed to a range of harmful substances during firefighting. Exposure to fire smoke has been associated with a decrease in their lung function. However, the cause-effect relationship between those two factors is not yet demonstrated. This meta-analysis aimed to evaluate the potential associations between firefighters' occupational exposure and their lung function deterioration. Studies were identified from PubMed, Web of Science, Scopus and Science Direct databases (August 1990-March 2021). The studies were included when reporting the lung function values of Forced Expiratory Volume in 1 s (FEV1) or Forced Vital Capacity (FVC). The meta-analyses were performed using the generic inverse variance in R software with a random-effects model. Subgroup analysis was used to determine if the lung function was influenced by a potential study effect or by the participants' characteristics. A total of 5562 participants from 24 studies were included. No significant difference was found between firefighters' predicted FEV1 from wildland, 97.64% (95% CI: 91.45-103.82%; I2 = 99%), and urban fires, 99.71% (95% CI: 96.75-102.67%; I2 = 98%). Similar results were found for the predicted FVC. Nevertheless, the mean values of firefighters' predicted lung function varied significantly among studies, suggesting many confounders, such as trials' design, statistical methods, methodologies applied, firefighters' daily exposure and career length, hindering an appropriate comparison between the studies.


Ion mobility mass spectrometry uncovers regioselectivity in the carboxylate-assisted C-H activation of palladium N-heterocyclic carbene complexes.
Carboxylate-assisted Pd-catalyzed C-H bond activation constitutes a mild and versatile synthetic tool to efficiently and selectively cleave inert C-H bonds. Herein, we demonstrate a simple method to experimentally evaluate both reactivity and selectivity in such systems using mass spectrometry (MS) methods. The N-heterocyclic carbene (NHC) cations [(NHC)PdX]+, bearing as X- ligand bases commonly used to promote the C-H activation (carboxylates and bicarbonate), are generated in the gas-phase by ESI-MS. Their C-H bond activation at the N-bound groups of the NHC is then studied using Collision Induced Dissociation (CID) experiments. Ion Mobility Spectrometry (IM)-MS is exploited to identify a number of regioisomers associated with the distinctive site selective C-H activations. It is demonstrated that such C-H activation concomitant with acetic acid release occurs from a mixture of activated [(NHC-H)Pd(CH3CO2H)]+ and non-activated [(NHC)Pd(CH3CO2)]+ complexes. The identity of the X-type ligands (X = Cl-, carboxylates and bicarbonate) has a significant impact on the regioisomer branching ratio upon CID conditions. IM-MS in conjunction with a DFT mechanistic study is presented for the acetate-assisted C-H activation of the [(NHC)Pd(CH3CO2)]+ cation featuring butyl and aryl as N-donor groups.


Zebrafish as a Model System for the Study of Severe CaV2.1 (α1A) Channelopathies.
The P/Q-type CaV2.1 channel regulates neurotransmitter release at neuromuscular junctions (NMJ) and many central synapses. CACNA1A encodes the pore-containing α1A subunit of CaV2.1 channels. In humans, de novo CACNA1A mutations result in a wide spectrum of neurological, neuromuscular, and movement disorders, such as familial hemiplegic migraine type 1 (FHM1), episodic ataxia type 2 (EA2), as well as a more recently discovered class of more severe disorders, which are characterized by ataxia, hypotonia, cerebellar atrophy, and cognitive/developmental delay. Heterologous expression of CaV2.1 channels has allowed for an understanding of the consequences of CACNA1A missense mutations on channel function. In contrast, a mechanistic understanding of how specific CACNA1A mutations lead in vivo to the resultant phenotypes is lacking. In this review, we present the zebrafish as a model to both study in vivo mechanisms of CACNA1A mutations that result in synaptic and behavioral defects and to screen for effective drug therapies to combat these and other CaV2.1 channelopathies.


Exploring contextual factors impacting the implementation of and engagement with a digital platform supporting psychosis recovery: A brief report.
Individuals with schizophrenia often demonstrate poor engagement in treatment and challenges with illness self-management. App4independence (A4i) is a digital health platform that was developed with the purpose of addressing the aforementioned challenges. While digital interventions can support patient care, there is a paucity of research on implementing such interventions in clinical settings. To describe the contextual factors that impacted the implementation of and engagement with A4i across three different clinical implementation sites, a descriptive approach, guided by implementation science frameworks, was employed to understand how people, culture, process, and technology impacted the implementation of A4i. Descriptive statistics were used to present user engagement data across each site implementation. Additionally, the lessons learned from each implementation were described narratively. Overall, 53 patients were onboarded to A4i in Context 1, 8 in Context 2, and 65 within Context 3, with retention rates over 90 days of 100%, 100%, and 96%, respectively. The adoption, engagement, and sustained use of the A4i platform varied across each implementation site and were affected by implementation strategies within the sociotechnical domains of people, culture, process, and technology. Despite differences in implementation processes, engagement with A4i remained consistently high. Customized educational materials, digital navigators, and technical support served as facilitators in the adoption of A4i.


The adsorptive removal of disinfection by-product precursors in a high-SUVA water using iron oxide-coated pumice and volcanic slag particles.
The main objective of this work was to study the effectiveness of iron oxide-coated pumice and volcanic slag particles in removing disinfection by-product (DBP) precursors from a raw drinking water source with high specific UV absorbance (SUVA(254)) value. Iron oxide coating of particles significantly increased dissolved organic carbon (DOC) uptakes and decreased DBP formation after chlorination compared to uncoated particles. pH values close to neutral levels during adsorption and chlorination provided DOC, trihalomethane and haloacetic acid reductions around 60-75% employing 6 g/L coated particle dosage. Higher degree of DOC and DBP reductions (>85%) were obtained with increasing particle dose. The uptake of bromide by iron oxide surfaces was negligible and increasing bromide concentrations (up to 550 μg/L) did not negatively impact the DOC uptake. However, due to competition between natural organic matter (NOM) and bicarbonate for the iron oxide surfaces, increasing bicarbonate alkalinity levels reduced DOC uptakes. Overall, the results indicated that the iron oxide-coated pumice/slag particles are effective adsorbents to remove NOM and control DBP formation in waters with relatively high DOC and SUVA(254) levels. However, they may not be effective for waters with alkalinity levels above 250 mg CaCO(3)/L.


Enrichment of probiotic ice cream with different dietary fibers: Structural characteristics and culture viability.
This study evaluated the effect of 5 dietary fibers (apple, orange, oat, bamboo, and wheat) on the physicochemical, rheological, and textural characteristics; sensory properties; and culture viability of probiotic ice cream stored at -18°C for 180 d. The presence of orange and apple fibers increased the titratable acidity, decreased the lightness (color) value of the ice creams, and enhanced the red and yellow coloration. Compared with the control sample, the consistency indices and apparent viscosities of the experimental samples increased with the addition of all dietary fibers except oat fiber. The highest viscosity was obtained in the sample fortified with apple fiber, whereas the ice cream containing orange fiber showed the highest hardness after d 60 of storage. The addition of orange and apple fibers significantly increased melting resistance; however, panelists did not generally like these samples in terms of taste-flavor. All ice creams had viable counts of Lactobacillus acidophilus of ≥7 log cfu/g during storage except the samples with orange and bamboo fiber. Bifidobacterium lactis counts were also found to be >6 log cfu/g in those samples until d 150 of storage.


Influence of morphology of colloidal nanoparticle gels on ion transport and rheology.
We develop a simple model to probe the ion transport and mechanical properties of low volume fraction colloidal nanoparticle gels. Specifically, we study the influence of the morphology of gels on ion diffusion and the corresponding roles of affinity to and enhanced ion transport along nanoparticle surfaces. We employ kinetic Monte Carlo simulations to simulate ion transport in the colloidal gels, and we perform nonequilibrium molecular dynamics to study their viscoelastic behavior. Our results indicate that in the presence of enhanced diffusion pathways for ions along the particle surface, morphology has a significant influence on the diffusivity of ions. We demonstrate that some gel morphologies can exhibit simultaneously enhanced ion transport and mechanical properties, thus illustrating a strategy to decouple ion transport and mechanical strength in electrolytes.


[Comparative determination of progesterone in the blood plasma by radioimmunological and chromato-mass-fragmentographic methods].
A comparative determination of progesterone in the blood plasma was performed using a radioimmunoassay (RIA) and 2 variants of a chromato-mass-fragmentographic (CMF) method. A deuterated internal standard, capillary gas-liquid chromatography and 3-enol-3-tret-BDMS and 3.20-bis-enol-bis-BDMS progesterone ethers as chromatographied derivatives were employed. Both derivatives made it possible to perform CMF determination using simplified variants of the method. The variants of the CMF method were employed as references for RIA of progesterone. Comparison of the results of the CMF and RIA methods indicated RIA significance (r = +0.99) in using antiserum to the conjugate 3-(O-carboxy methyl)-oxime of progesterone with bovine serum albumin injected to rabbits.


[The prevention of renal failure has to improve].
For some years there has been evidence at population level that prevention of end-stage kidney disease is feasible. However, the number of people receiving renal replacement therapy is still rising as a consequence of population ageing. With a greater focus on prevention through better screening and more active treatment, the number of people with end-stage kidney disease can be reduced. The authors argue that screening for chronic kidney damage should be improved, not only in people with diabetes but also in those with hypertension or a family history of cardiovascular disease, those eligible for a Prevention Consultation, and those from lower socio-economic classes. There is also a need for better treatment of people diagnosed with chronic kidney damage.


Lipid concentration effects on the amyloidogenic apoC-II(60-70) peptide: a computational study.
Molecular dynamics simulations were implemented to investigate the effects of phospholipid concentration on the conformation and dynamics of the amyloidogenic peptide apoC-II(60-70). The results showed a progressive reduction in the solvent accessible surface area of apoC-II(60-70) with increasing lipid concentration, accompanied by increased lipid-peptide interactions. Favorable peptide interaction sites with lipids were found to be the aromatic residues, Tyr63 and Phe67. The high stability of lipid-peptide contacts resulted in reduced conformational flexibility of the peptide. A significant change in the secondary structure of apoC-II(60-70) peptide was observed with increasing lipid concentration. At lower concentrations (1-3 lipids per peptide), the peptide adopted extended beta-strand conformations, caused by contacts with the lipids, which reduced the intramolecular interactions within the peptide. In contrast, a higher lipid concentration (4-6 lipids per peptide) had a restraining effect on the peptide's flexibility by trapping it in a particular conformation. Such behavior can be suggested as inhibiting fibril formation, because of the lipid-induced inability of the peptide to adopt fibril competent conformations. This finding complements our recent ThT fluorescence results, which revealed that the 4:1 lipid to peptide ratio is sufficient to cause fibril inhibition in apoC-II(60-70).


An Absorption Mitigation Technique for Received Signal Strength-Based Target Localization in Underwater Wireless Sensor Networks.
Localization is an indispensable technology for underwater wireless sensor networks (UWSNs). In what concerns UWSNs, the accurate location information is not only the requirement of the marine field applications but also the basis of the other corresponding research, for instance, network routing and topology control. Recently, an astonishing surge of interest has been drawn in the received signal strength (RSS)-based scheme due to cost-effectiveness and synchronization-free compared with others. However, unlike the terrestrial wireless sensor networks (WSNs), the acoustic signal may suffer the absorption loss in the underwater environment besides the path loss, which degrades the localization accuracy and limits the capability of the RSS-based technology in UWSNs. In this context, a robust localization method with an absorption mitigation technique (AMT) is developed. First, an RSS-based analytically tractable measurement model is conducted, where the maximum likelihood estimator (MLE) is derived. Nevertheless, it is quite challenging to solve the problem using MLE under a non-convex expression. Therefore, by exploiting certain approximations, the considered localization problem is converted into an optimization expression with a maximum absorption loss involved. A min-max strategy is then presented, with which the problem is turned to minimize the worst situation of the absorption loss. After a simple manipulation, the problem is further investigated as a generalized trust region sub-problem (GTRS) framework. Although the GTRS is a non-convex scheme, the solution can be obtained through an iteration method by introducing a multiplier. In addition, the closed-form expression of the Cramer-Rao lower bound (CRLB) of the analytically tractable measurement model is derived. Numerical simulations demonstrate the effectiveness of the proposed method compared with the state-of-the-art approaches in different scenarios.


Vector analysis as a fast and easy method to compare gene expression responses between different experimental backgrounds.
Gene expression studies increasingly compare expression responses between different experimental backgrounds (genetic, physiological, or phylogenetic). By focusing on dynamic responses rather than a direct comparison of static expression levels, this type of study allows a finer dissection of primary and secondary regulatory effects in the various backgrounds. Usually, results of such experiments are presented in the form of Venn diagrams, which are intuitive and visually appealing, but lack a statistical foundation. Here we introduce Vector Analysis (VA) as a simple, yet principled, approach to comparing expression responses in different experimental backgrounds. VA enables the automatic assignment of genes to response prototypes and provides statistical significance estimates to eliminate spurious response patterns. The application of VA to a real dataset, comparing nutrient starvation responses in wild type and mutant Arabidopsis plants, reveals that consistent patterns of expression behavior are present in the data and are reliably detected by the algorithm. Vector analysis is a flexible, easy-to-use technique to compare gene expression patterns in different experimental backgrounds. It compares favorably with the classical Venn diagram approach and can be implemented manually using spreadsheets, such as Excel, or automatically by using the supplied software.


Epigenetic switches in clag3 genes mediate blasticidin S resistance in malaria parasites.
Malaria parasites induce changes in the permeability of the infected erythrocyte membrane to numerous solutes, including toxic compounds. In Plasmodium falciparum, this is mainly mediated by PSAC, a broad-selectivity channel that requires the product of parasite clag3 genes for its activity. The two paralogous clag3 genes, clag3.1 and clag3.2, can be silenced by epigenetic mechanisms and show mutually exclusive expression. Here we show that resistance to the antibiotic blasticidin S (BSD) is associated with switches in the expression of these genes that result in altered solute uptake. Low concentrations of the drug selected parasites that switched from clag3.2 to clag3.1 expression, implying that expression of one or the other clag3 gene confers different transport efficiency to PSAC for some solutes. Selection with higher BSD concentrations resulted in simultaneous silencing of both clag3 genes, which severely compromises PSAC formation as demonstrated by blocked uptake of other PSAC substrates. Changes in the expression of clag3 genes were not accompanied by large genetic rearrangements or mutations at the clag3 loci or elsewhere in the genome. These results demonstrate that malaria parasites can become resistant to toxic compounds such as drugs by epigenetic switches in the expression of genes necessary for the formation of solute channels.


[Effect of various cations on the secretory and acid-forming function of the stomach].
The data obtained suggest that the sodium ions (0.5-2.0 g) reduce the gastric juice secretion and its acidity whereas magnesium and calcium ions increase the latter. Potassium ions exert practically no effect on the acidity. The suppressing effect of the sodium ions seems to be connected with an increase excretion of sodium chloride with urine as well as with increased diuresis in general. Saluretics suppress the secretion and acidity of the gastric juice on account of increased diuresis and subsequent excretion of large amount of chlorides as well as by means of serotonin release from the Ec cells of the gastro-intestinal tract, the action of serotonin being actualized on the level of the gastric glands' cells, parietal those in particular.


The Natural History of Symptomatic Fractures in Children and Adolescents with Osteogenesis Imperfecta Type 1: A Cohort Study from Western Australia.
The fracture experience of children and adolescents with osteogenesis imperfecta (OI) type 1 is not well described in the literature. We present data on symptomatic long bones and axial skeleton fractures of all patients aged 0 to 18 years with OI type 1 seen at a specialized bone clinic in Western Australia in the period 2008 to 2020 using a retrospective chart review method. The cohort consisted of 44 patients (21 males, 23 females). Median (interquartile range [IQR]) age was 11.3 (6.2 to 17) years, giving a total of 520 patient-years in the study during which 197 fractures were experienced. The mean fracture rate was 379 fractures per 1000 patient-years (95% confidence interval [CI]: 310 to 440); however, the experience for fractures varied from ≤1 fracture in 23% (n = 10) to two to 20 in 77% (n = 34) of the cohort. Twenty-one patients (48.5%) received bisphosphonates during the study period. In logistic regression, age, but not sex or family history of OI, was a significant predictor of fracture risk. The highest total fracture rate was observed in the age group 0 to <3 years at 469 fractures/1000 patient-years, which declined to 140 fractures/1000 patient-years in the age group 15 to 18 years. The lower limbs were the site of 49.7% of all fractures. The highest rate for lower limb fracture was in the age group 0 to <3 years at 331 fractures/1000 patient-years, decreasing to 0 fractures/1000 patient-years in the age group 15 to 18 years. Upper limb fracture rates increased from 100 fractures/1000 patient-years in the 0 to <3 years age group to 307 fractures/1000 patient-years in the 9 to <12 years age group and then declining to 70 fractures/1000 years in the 15 to 18 years age group. In pediatric patients with OI type 1, fracture risk is highest in early life, especially in the lower limbs. Multidisciplinary care of children with OI should have a particular focus on strategies to prevent these fractures. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.


2-chlorodeoxyadenosine (cladribine) in the treatment of hairy cell leukemia.
Hairy cell leukemia is an uncommon chronic B cell lymphoproliferative disorder in Taiwan and only sporadic cases have been noted. This disease is often found in middle-aged men with the typical manifestations of pancytopenia and splenomegaly. While tartrate-resistant acid phosphatase (TRAP) stain of the neoplastic cells in peripheral blood smear is positive in 90% cases, bone marrow examination is still necessary for diagnosis. For treatment, splenectomy and interferon have been the standard strategies. Recently, newly-developed purine analogues, 2-chlorodeoxyadenosine (cladribine) and 2'-deoxycorfomycin (pentostatin), were found to achieve long lasting complete remission. Here we reported a 48-year-old men who was found to have splenomegaly and bicytopenia incidentally at routine physical checkup. Although the TRAP staining of peripheral blood showed negative result, hairy cell leukemia was finally diagnosed by bone marrow biopsy. The following splenectomy showed a typical picture of hairy cell leukemia on pathology, but cytopenia recovered only partially after surgery. One course of chemotherapy with cladribine (0.1 mg/kg/day continuous intravenous infusion for 7 days) was given after recovery from major surgery. The white cell and platelet counts normalized after treatment and he carried a stable condition thereafter. Since the disease is rare and purine analogues are newly developed, there has been no report on using 2-clorodeoxy-adenosine against hairy cell leukemia in our country. Here we suggest that newly developed purine analogues such as cladribine are the first choice of treatment in cases of hairy cell leukemia.


Assemblage structure and spatial diversity patterns of kelp forest-associated fishes in Southern Patagonia.
Knowledge of the ecology of the fish fauna associated with kelp (primarily Macrocystis pyrifera) forests in Southern Patagonia is scarce, especially in how abiotic and biotic variables influence their structure, diversity, and distribution. This information is important for the management and conservation of this unique ecosystem, which has minimal anthropogenic impacts at present. We analyzed data from 122 quantitative underwater transects conducted within kelp forests at 61 stations from Chile's southern Patagonian fjords to the Cape Horn and Diego Ramirez archipelagos and the southern tip of Argentina, including the Mitre Peninsula and Isla de los Estados. In total, 25 fish species belonging to 13 families were observed. Multivariate analysis indicated that there are significant differences in fish assemblage structure among locations and wave exposures, which was driven primarily by Patagonotothen sima and Paranotothenia magellanica, which occurred on exposed and semi-exposed stations. P. cornucola was mainly distributed across sheltered stations of the Kawésqar National Park. Temperature, salinity, depth, and kelp density influenced fish assemblage structure, with the highest diversity in areas with the lowest temperature and greater depth at Isla de los Estados. In contrast, species richness, diversity, abundance, and biomass were all lower in areas with high density of the understory kelp Lessonia spp., which might be driven by the absence of P. tessellata, P. squamiceps and P. cornucola, the most important species in terms of occurrence, abundance, and biomass. Our study provides the first broad-scale description of the fish assemblages associated with kelp forests along the southern cone of South America based on non-invasive visual transects, improving our knowledge of the distribution of fish assemblages across several environmental conditions in this vast and little-studied area.


RETRACTED: Hydroxychloroquine in the Treatment of COVID-19: A Multicenter Randomized Controlled Study.
RETRACTED ARTICLE: The COVID-19 pandemic is showing an exponential growth, mandating an urgent need to develop an effective treatment. Indeed, to date, a well-established therapy is still lacking. We aimed to evaluate the safety and efficacy of hydroxychloroquine (HCQ) added to standard care in patients with COVID-19. This was a multicenter, randomized controlled trial conducted at three major university hospitals in Egypt. One hundred ninety-four patients with confirmed diagnosis of COVID-19 were included in the study after signing informed consent. They were equally randomized into two arms: 97 patients administrated HCQ plus standard care (HCQ group) and 97 patients administered only standard care as a control arm (control group). The primary endpoints were recovery within 28 days, need for mechanical ventilation, or death. The two groups were matched for age and gender. There was no significant difference between them regarding any of the baseline characteristics or laboratory parameters. Four patients (4.1%) in the HCQ group and 5 (5.2%) patients in the control group needed mechanical ventilation (P = 0.75). The overall mortality did not differ between the two groups, as six patients (6.2%) died in the HCQ group and 5 (5.2%) died in the control group (P = 0.77). Univariate logistic regression analysis showed that HCQ treatment was not significantly associated with decreased mortality in COVID-19 patients. So, adding HCQ to standard care did not add significant benefit, did not decrease the need for ventilation, and did not reduce mortality rates in COVID-19 patients.


Carboxylate- and Phosphonate-Modified Polyethylenimine: Toward the Design of Actinide Decorporation Agents.
Plutonium (Pu) is an anthropogenic element involved in the nuclear industry cycle. Located at the bottom of the periodic table within the actinide family, it is a chemical toxic but also a radiological toxic, regardless of isotopy. After nearly 80 years of Pu industrialization, it has become clear that inhalation and wounds represent the two main ways a person may become contaminated after an accident. In order to reduce the deleterious health effects of Pu, it is crucial to limit chronic exposure by removing it or preventing its incorporation into the body. Diethylenetriaminepentaacetic acid (DTPA) has emerged as the gold standard for Pu decorporation, although it suffers from very short retention time in serum. Other molecules like the hydroxypyridonate family with high chemical affinity have also been considered. We have been considering alternative polymeric chelates and, in particular, polyethylenimine (PEI) analogues of DTPA (the carbonate or phosphonate version), which may present a real breakthrough in Pu decorporation not only because of their higher loading capacity but also because of their indirect vectorization properties correlated with a specific biodistribution into the lungs, bone, kidney, or liver. In the first part of this Forum Article, new data on the structural characterization of the complexation of PuIV with polyethylenimine methylphosphonate (PEI-MP) were obtained using the combination of extended X-ray absorption fine structure spectroscopy and ab initio molecular dynamics (AIMD) calculations. The use of thorium (Th) as a Pu chemical surrogate is also discussed because its unique oxidation state is IV+ in solution. In the second part of the paper, we put this new set of data on PEI-MP-Pu into perspective with use of the PEI platform to complex ThIV and PuIV. Uptake curves of ThIV witth polyethylenimine methylcarboxylate (PEI-MC) are compared with those of PEI-MP and DTPA, and the AIMD data are discussed.


Kinetics of CD4+ T cell repopulation of lymphoid tissues after treatment of HIV-1 infection.
Potent combinations of antiretroviral drugs diminish the turnover of CD4+ T lymphocytes productively infected with HIV-1 and reduce the large pool of virions deposited in lymphoid tissue (LT). To determine to what extent suppression of viral replication and reduction in viral antigens in LT might lead correspondingly to repopulation of the immune system, we characterized CD4+ T lymphocyte populations in LT in which we previously had quantitated viral load and turnover of infected cells before and after treatment. We directly measured by quantitative image analysis changes in total CD4+ T cell counts, the CD45RA+ subset, and fractions of proliferating or apoptotic CD4+ T cells. Compared with normal controls, we documented decreased numbers of CD4+ T cells and increased proliferation and apoptosis. After treatment, proliferation returned to normal levels, and total CD4+ T and CD45RA+ cells increased. We discuss the effects of HIV-1 on this subset based on the concept that renewal mechanisms in the adult are operating at full capacity before infection and cannot meet the additional demand imposed by the loss of productively infected cells. The slow increases in the CD45RA+ CD4+ T cells are consistent with the optimistic conclusions that (i) renewal mechanisms have not been damaged irreparably even at relatively advanced stages of infection and (ii) CD4+ T cell populations can be partially restored by control of active replication without eradication of HIV-1.


A global assessment of climate-water quality relationships in large rivers: an elasticity perspective.
To uncover climate-water quality relationships in large rivers on a global scale, the present study investigates the climate elasticity of river water quality (CEWQ) using long-term monthly records observed at 14 large rivers. Temperature and precipitation elasticities of 12 water quality parameters, highlighted by N- and P-nutrients, are assessed. General observations on elasticity values show the usefulness of this approach to describe the magnitude of stream water quality responses to climate change, which improves that of simple statistical correlation. Sensitivity type, intensity and variability rank of CEWQ are reported and specific characteristics and mechanism of elasticity of nutrient parameters are also revealed. Among them, the performance of ammonia, total phosphorus-air temperature models, and nitrite, orthophosphorus-precipitation models are the best. Spatial and temporal assessment shows that precipitation elasticity is more variable in space than temperature elasticity and that seasonal variation is more evident for precipitation elasticity than for temperature elasticity. Moreover, both anthropogenic activities and environmental factors are found to impact CEWQ for select variables. The major relationships that can be inferred include: (1) human population has a strong linear correlation with temperature elasticity of turbidity and total phosphorus; and (2) latitude has a strong linear correlation with precipitation elasticity of turbidity and N nutrients. As this work improves our understanding of the relation between climate factors and surface water quality, it is potentially helpful for investigating the effect of climate change on water quality in large rivers, such as on the long-term change of nutrient concentrations.


Serum gamma-glutamyl transferase: a novel biomarker for coronary artery disease.
Atherosclerosis is a chronic inflammatory process, in which oxidative stress is the key event. Gamma-glutamyl transferase (GGT) is a cellular production of oxidants. We aimed to elucidate the relationship of serum GGT levels and coronary artery disease (CAD) in a Chinese population. A total of 513 adult subjects who had undergone coronary angiography were enrolled in the study. Clinical characteristics, coronary angiography, and serum samples were collected from all the patients and analyzed for the serum GGT, blood lipids, and cardiovascular risk factors. Subjects with CAD had significantly increased activity of serum GGT (p=0.003). Serum GGT levels exhibited positive correlations with alcohol intake (β=0.177, p<0.001), coronary complexity (β=0.068, p<0.001), and triacylglycerol (β=0.058, p<0.001). High-density lipoprotein cholesterol levels (β=0.157, p=0.008) and age (β=0.004, p=0.002) were negatively correlated with serum GGT in the CAD group. The coronary complexity presented a negative correlation with Ig-apolipoprotein AI (β=-2.517, p=0.001) and positive correlations with smoking (β=0.640, p<0.001), Ig-GGT (β=0.613, p=0.004), Ig high sensitivity-C reactive protein (β=0.320, p<0.001), and hypertension (β=0.286, p<0.026). The study showed a positive correlation between serum GGT and CAD in a Chinese population. Serum GGT levels may be a potential biomarker for CAD.


[Synaptic transmission in the frog spinal cord exposed to intensive microwave radiation].
Microwave (6.45 GHz, CW, SAR 0.1-2 W/g) effects on mono- and polysynaptic transmission, temporal and spatial EPSP summation were investigated in isolated frog spinal cord. No changes were observed if the preparation temperature remained constant during irradiation. VHF and conventional heating produced the same effects. The data obtained confirm the thermal mechanism of VHF influences on synaptic transmission processes.


Dietary habits, physical activity and obesity in Hong Kong residents.
Recent data from the Hong Kong Population Health Survey 2003/04 commissioned by the Department of Health estimate that 17.8% of the population aged 15 and above are overweight (BMI 23.0-24.9 kg/m(2)) and 21.1% are obese (BMI > or = 25.0 kg/m(2)). Similar to many industrialized societies, it is generally believed that chronic positive energy balance as a consequence of excess energy intake (energy dense processed food typically high in fat, protein and refined carbohydrates with a low fiber content) together with a sedentary life is the main culprit. Against this background, a territory-wide 'email@example.com' campaign (http://www.eatsmart.gov.hk) organized by the Department of Health, with support from academic institutes and voluntary organizations, has been commissioned since November 2005. So far several important guidelines have been published including the 'Nutritional Guidelines on Lunch Box/Snacks for Primary School Students'. We certainly welcome a long term vision and a direction from policy makers relating to a population and public health approach to obesity. The prevention of obesity requires a review of dietary and physical activity habits and then measures to change our habits in a practical and reliable way.


Apparent diffusion coefficient and its standard deviation from diffusion-weighted imaging in preoperative predicting liver invasion by T3-staged resectable gallbladder carcinoma.
To evaluate apparent diffusion coefficient (ADC) and its standard deviation (SDADC) in preoperative predicting liver invasion by T3-staged gallbladder carcinoma (GBC). Forty-one consecutive patients with T3-staged resectable GBC were included and divided into two sets with (n=27) and without (n=14) liver invasion. All patients underwent DWI at b-values of 0, 20, 50, 80, 100, 200, 400, 600, 800, and 1,000 s/mm2 with a 3 T magnetic resonance imaging scanner before surgery. ADC and SDADC of tumour-adjacent and tumour-distant liver tissues were measured on DWI, and were compared by Mann-Whitney U-tests. If there was a significant difference in any derived parameter, the area under the receiver operating characteristic curve (AUC) was used to assess performance of this parameter to predict liver invasion. DWI could differentiate between patients with and without liver invasion when b = 0, 1,000 s/mm2 (AUCs of ADC and SDADC were 0.697 and 0.714, respectively). In patients with liver invasion, mean ADC and SDADC of tumour-adjacent liver tissue were lower than of tumour-distant liver tissue when b = 0, 800 s/mm2, and = 0, 1,000 s/mm2 (all p-values <0.05). To differentiate tumour-adjacent from tumour-distant liver tissues in patients with liver invasion, AUCs of ADC were 0.687 (b = 0, 800 s/mm2) and 0.680 (b = 0, 1,000 s/mm2), and AUCs of SDADC were 0.673 (b = 0, 800 s/mm2) and 0.731 (b = 0, 1,000 s/mm2). DWI could have potential value in preoperative predicting liver invasion by T3-staged GBC.


Dysregulated TGF-β signaling alters bone microstructure in a mouse model of Loeys-Dietz syndrome.
Loeys-Dietz syndrome (LDS) is a connective tissue disorder characterized by vascular and skeletal abnormalities resembling Marfan syndrome, including a predisposition for pathologic fracture. LDS is caused by heterozygous mutations in the genes encoding transforming growth factor-β (TGF-β) type 1 and type 2 receptors. In this study, we characterized the skeletal phenotype of mice carrying a mutation in the TGF-β type 2 receptor associated with severe LDS in humans. Cortical bone in LDS mice showed significantly reduced tissue area, bone area, and cortical thickness with increased eccentricity. However, no significant differences in trabecular bone volume were observed. Dynamic histomorphometry performed in calcein-labeled mice showed decreased mineral apposition rates in cortical and trabecular bone with normal numbers of osteoblasts and osteoclasts. Mechanical testing of femurs by three-point bending revealed reduced femoral strength and fracture resistance. In vitro, osteoblasts from LDS mice demonstrated increased mineralization with enhanced expression of osteoblast differentiation markers compared with control cells. These changes were associated with impaired TGF-β1-induced Smad2 and Erk1/2 phosphorylation and upregulated TGF-β1 ligand mRNA expression, compatible with G357W as a loss-of-function mutation in the TGF-β type 2 receptor. Paradoxically, phosphorylated Smad2/3 in cortical osteocytes measured by immunohistochemistry was increased relative to controls, possibly suggesting the cross-activation of TGF-β-related receptors. The skeletal phenotype observed in the LDS mouse closely resembles the principal structural features of bone in humans with LDS and establishes this mouse as a valid in vivo model for further investigation of TGF-β receptor signaling in bone.


Assessing local capacity for health intervention.
Because of their location within the practice realm, participatory, community-based public health coalitions offer many lessons about implementing and sustaining local interventions. This paper presents a case study of capacity assessment at the local level. Capacity evaluation methods are presented, with emphasis on the theoretical framework used to guide the evaluation. The capacity evaluation framework presented herein was theoretically based and designed to generate practical information to facilitate the adoption of a locally tailored youth obesity prevention program, VERB Summer Scorecard (VSS). Using multiple methods, four aspects of community capacity were assessed, including community, knowledge and skills, resources, and power. Within each category, factors that facilitated or impeded program implementation were distinguished. The evaluation protocol was designed to generate information increasing community capacity to sustain a community-based obesity prevention program. Capacity tables were used as a program-planning tool and as a system for sharing implementation and sustainability requirements with potential adopters. This case study also explores how to use capacity assessment results to empower coalitions to serve as catalysts for development of local programs in other communities.


The antiobesity and metabolic activities of LY79771 in obese and normal mice.
Without affecting the appetite, Compound LY79771 reduced the body weight of obese Avy/a mice, prevented young Avy/a mice from becoming obese and ob/ob mice from becoming overtly obese. This weight-controlling effect of LY79771 was reversible upon withdrawal of treatment. The major portion of the weight change induced by the compound was due to a reduction in triacylglycerol content of the body. Hyperinsulinemia and hyperglycemia of the obese mice were also improved. In spite of its potent lipolytic activity, only a 20 to 25 per cent excess of fatty acid was found in the liver after dosing. No accumulation of fatty acid was observed in the carcass. A single dose of LY79771 also stimulated metabolism as indicated by the increase of CO2 expiration. The transient rise of body temperature that occurred after each injection in a long-term treatment regimen suggested that the compound was thermogenic. LY79771 had no effect on the weight of normal mice.


Multimodal regulation of E2F1 gene expression by progestins.
An analysis of mRNA expression in T47D breast cancer cells treated with the synthetic progestin R5020 revealed a subset of progesterone receptor (PR) target genes that are enriched for E2F binding sites. Following up on this observation, we determined that PR-B acts in both direct and indirect manners to positively upregulate E2F1 expression in T47D cells. The direct effects of PR on E2F1 expression were confirmed by chromatin immunoprecipitation (ChIP) analysis, which indicated that the agonist-bound receptor was recruited to several enhancer elements proximal to the E2F1 transcript. However, we also noted that cycloheximide partially inhibits R5020 induction of E2F1 expression, indicating that the ligand-dependent actions of PR on this gene may involve additional indirect regulatory pathways. In support of this hypothesis, we demonstrated that treatment with R5020 significantly increases both hyperphosphorylation of Rb and recruitment of E2F1 to its own promoter, thus activating a positive feedback loop that further amplifies its transcription. Furthermore, we established that PR-mediated induction of Krüppel-like factor 15 (KLF15), which can bind to GC-rich DNA within the E2F1 promoter, is required for maximal induction of E2F1 expression by progestins. Taken together, these results suggest a new paradigm for multimodal regulation of target gene expression by PR.


Expression cassettes for formaldehyde and fluoroacetate resistance, two dominant markers in Saccharomyces cerevisiae.
We employed two genes in constructing yeast expression cassettes for dominant, selectable markers. The Saccharomyces cerevisiae gene SFA1, encoding formaldehyde dehydrogenase, was placed under the control of the GPD1 promoter and CYC1 terminator. The Moraxella sp. strain B gene dehH1, encoding fluoroacetate dehalogenase, was placed under the control of both the GPD1 and CYC1 promoters. With these constructs it was possible to select directly for yeast strains resistant to either formaldehyde or fluoroacetate. Both selective agents are completely metabolized and inexpensive, making them very useful in the pursuit of yeast gene functions and for industrial applications. An additional advantage of the formaldehyde dehydrogenase marker is that it is an S. cerevisiae gene, thus allowing 'all yeast' constructs.


Sensory transduction in Escherichia coli: role of a protein methylation reaction in sensory adaptation.
The behavioral response of Escherichia coli to the addition of a stimulatory compound is transient; thus the organism undergoes sensory adaptation. When the compound is removed, E. coli undergoes the inverse process, called deadaptation, and very rapidly regains its sensitivity to the stimulus. In this communication we demonstrate that the previously reported methylation of several cytoplasmic membrane proteins is correlated with, and very likely controls, the state of adaptation of the cell. In the absence of an added stimulus these proteins are methylated to a basal level. When the bacteria are stimulated by the addition of an attractant, the extent of methylation increases over a period of several minutes to a new level, which is maintained as long as the attractant is present. The magnitude of the increase in methylation is a function of the size of the stimulus and is directly proportional to the duration of the behavioral response. Upon removal of the attractant the level of methylation very rapidly falls to the basal value. Previously we have shown that adaptation requires methionine, but maintenance of the adapted state and de-adaptation do not [Springer, M. S., Goy, M. F. & Adler, J. (1975) Proc. Natl. Acad. Sci. USA 74, 183-187]; here we demonstrate that methylation requires methionine but maintenance of an attractant-induced level of methylation and the demethylation that occurs following removal of the attractant do not. These results strongly indicate a role for protein methylation in sensory adaptation.


Noncovalently Linked Metallacarboranes on Functionalized Magnetic Nanoparticles as Highly Efficient, Robust, and Reusable Photocatalysts in Aqueous Medium.
A successful homogeneous photoredox catalyst has been fruitfully heterogenized on magnetic nanoparticles (MNPs) coated with a silica layer, keeping intact its homogeneous catalytic properties but gaining others due to the easy magnetic separation and recyclability. The amine-terminated magnetic silica nanoparticles linked noncovalently to H[3,3'-Co(1,2-C2B9H11)2]- (H[1]), termed MSNPs-NH2@H[1], are highly stable and do not produce any leakage of the photoredox catalyst H[1] in water. The magnetite MNPs were coated with SiO2 to provide colloidal stability and silanol groups to be tethered to amine-containing units. These were the MSNPs-NH2 on which was anchored, in water, the cobaltabis(dicarbollide) complex H[1] to obtain MSNPs-NH2@H[1]. Both MSNPs-NH2 and MSNPs-NH2@H[1] were evaluated to study the morphology, characterization, and colloidal stability of the MNPs produced. The heterogeneous MSNP-NH2@H[1] system was studied for the photooxidation of alcohols, such as 1-phenylethanol, 1-hexanol, 1,6-hexanediol, or cyclohexanol among others, using catalyst loads of 0.1 and 0.01 mol %. Surfactants were introduced to prevent the aggregation of MNPs, and cetyl trimethyl ammonium chloride was chosen as a surfactant. This provided adequate stability, without hampering quick magnetic separation. The results proved that the catalysis could be speeded up if aggregation was prevented. The recyclability of the catalytic system was demonstrated by performing 12 runs of the MSNPs-NH2@H[1] system, each one without loss of selectivity and yield. The cobaltabis(dicarbollide) catalyst supported on silica-coated magnetite nanoparticles has proven to be a robust, efficient, and easily reusable system for the photooxidation of alcohols in water, resulting in a green and sustainable heterogeneous catalytic system.


Reflecting on our good intentions: A critical discourse analysis of women's health and empowerment discourses in sexual and gender-based violence policies relevant to southern Africa.
Decades of 'feminist' sexual- and gender-based violence (SGBV) policies have produced limited change in southern African SGBV rates. Recent critiques highlight ongoing colonial legacies in such policymaking, arguing that these legacies limit the potential for liberatory change. Further, reflecting on such discourses can highlight reasons behind global public health intervention failure. To promote reflexivity among public health actors who create, reproduce, and implement SGBV policies, this paper presents a critical discourse analysis of how women's empowerment is constructed in foundational global and national health and development policies bearing on SGBV in Zambia. The analysis identifies neoliberal feminist discourses of empowerment: (i) the protection of women, which perpetuates a saviour complex; (ii) the promotion of equality to men, which excludes those deemed unworthy; (iii) the eradication of harmful cultural norms, which challenge the preservation of African values; and (iv) (neoliberal) empowerment through women's attained employment and capital, which empowers women within unequal economic relations rather than liberating women from those relations. The author critiques such neoliberal empowerment discourses for failing to structurally transform the conditions for women's liberation. This paper offers a first step to the dismantling of colonial structures in SGBV policies by unpacking and promoting reflexivity about such discourses.


Establishment of immortalized primary cell from the critically endangered Bonin flying fox (Pteropus pselaphon).
The Bonin flying fox (Pteropus pselaphon) is one of the most critically endangered species of animals. The number of this species is estimated to be around 150; being classified at the top rank in the list by International Union of Animal Conservation. Our group previously showed that expression of CDK4, CYCLIN D1, and telomerase reverse transcriptase (TERT) efficiently induce immortalization of human, bovine, swine, monkey, and buffalo-derived cells. In this manuscript, we successfully established the primary cells from Bonin flying fox. We introduced CDK4, CYCLIN D1, and TERT into the primary cells. The established cells showed efficient expression of introduced genes at the protein level. Furthermore, the established cells were free from senescence, indicating it reached to immortalization. Moreover, we showed that interspecies somatic cell nuclear transfer of Bonin flying fox derived cell into bovine embryo allowed the development of the embryo to 8 cell stages. Our established cell has the potential to contribute to species conservation.


The change of the COVID-19 pandemic on medical education in Korea: a national survey of medical schools.
This study aims to investigate how medical schools in Korea managed their academic affairs and student support in response to the coronavirus disease 2019 (COVID-19) pandemic and their plans for the post-COVID-19 era. An online survey was conducted, and a link to a questionnaire was distributed to all the administrators of the 40 medical schools in Korea. The final analysis data involved responses from 33 medical schools and 1,342 students. Frequency analysis, cross-tabulation analysis, Fisher's exact test, and one-way analysis of variance were applied for statistical analysis. Regarding instruction methods, most medical schools in Korea transitioned from in-person learning to video-on-demand learning (51.5%) and real-time online learning (42.4%). Among the school leaders, 36.4% planned to continue offering online classes combined with in-person classes beyond the end of the pandemic. Among the students, the online class concentration and participation score was 3.0 points or lower, but the class understanding score was 3.6 points, above a moderate level. Students cited the shorter times needed to attend school and being able to take classes repeatedly as advantages of online classes, and over one-third of medical schools intended to continue with a hybrid of in-person and online learning even after the COVID-19 pandemic restrictions end.


Research Resource: The androgen receptor modulates expression of genes with critical roles in muscle development and function.
Androgen signaling through the androgen receptor (AR), a ligand-dependent transcription factor within the steroid receptor superfamily, plays an important role in the development and maintenance of many tissues. In muscle, androgens act as anabolic agents that increase both muscle mass and strength; however, a key unanswered question is the mechanism through which AR-mediated gene expression leads to these effects. To gain further insight into the mechanism of AR action in muscle, we identified AR-binding sites in primary human muscle cells using ChIP-on-Chip (chromatin immunoprecipitation coupled with tiling microarray detection of genomic fragments). Through this analysis, we identified 32,518 potential AR-binding sites throughout the genome that were enriched upon androgen treatment. Sequence analysis of these regions indicated that approximately 90% possess a consensus androgen response element or half-site. Among the identified AR-binding sites are genes known to be directly regulated by AR, confirming the validity of our methodology. Additionally, we identified a number of novel AR targets, including genes and micro-RNAs implicated in muscle differentiation and function, suggesting a direct role for AR-mediated transcription in muscle development. Intriguingly, binding sequences for the Mef2 family of transcription factors were enriched in the AR-bound regions, and we show that several Mef2c-dependent genes are direct targets of AR, suggesting a functional interaction between Mef2c and AR in skeletal muscle. Our results provide new insights into the mechanisms by which androgens promote muscle growth and validate AR as a potential therapeutic target for sarcopenia, muscle wasting, and other androgen-related muscle disorders.


Functional signal- and paradigm-dependent linear relationships between synaptic activity and hemodynamic responses in rat somatosensory cortex.
Linear relationships between synaptic activity and hemodynamic responses are critically dependent on functional signal etiology and paradigm. To investigate these relationships, we simultaneously measured local field potentials (FPs) and optical intrinsic signals in rat somatosensory cortex while delivering a small number of electrical pulses to the hindpaw with varied stimulus intensity, number, and interstimulus interval. We used 570 and 610 nm optical signals to estimate cerebral blood volume (CBV) and oxygenation, respectively. The spatiotemporal evolution patterns and trial-by-trial correlation analyses revealed that CBV-related optical signals have higher fidelity to summed evoked FPs (SigmaFPs) than oxygenation-derived signals. CBV-related signals even correlated with minute SigmaFP fluctuations within trials of the same stimulus condition. Furthermore, hemodynamic signals (CBV and late oxygenation signals) increased linearly with SigmaFP while varying stimulus number, but they exhibited a threshold and steeper gradient while varying stimulus intensity, suggesting insufficiency of the homogeneity property of linear systems and the importance of spatiotemporal coherence of neuronal population activity in hemodynamic response formation. These stimulus paradigm-dependent linear and nonlinear relationships demonstrate that simple subtraction-based analyses of hemodynamic signals produced by complex stimulus paradigms may not reflect a difference in SigmaFPs between paradigms. Functional signal- and paradigm-dependent linearity have potentially profound implications for the interpretation of perfusion-based functional signals.


Ofloxacin levels after intravitreal injection. Effects of trauma and inflammation.
This study was carried out to get an insight into the ofloxacin elimination after intravitreal injection in rabbits. We also studied the effects of trauma and inflammation on the vitreous ofloxacin levels after intravitreal injection of ofloxacin. A penetrating eye injury in the right eye was inflicted on 24 rabbits and another 12 animals were used as control. A standardized intraocular inflammation was induced by intravitreal injection of a suspension of Staphylococcus aureus in half of the traumatized eyes. Ofloxacin (200 microg/0.1 ml) was injected into the midvitreous cavity of both traumatized and control right eyes, and samples were obtained at 2, 8, 24 and 48 h after injection. Drug concentrations were measured using high-pressure liquid chromatography analysis. Vitreous levels of ofloxacin were above the MIC(90) at 2 and 8 h in all groups for most of the common microorganisms causing endophthalmitis and also at 24 h in traumatized-infected eyes. At the second hour, the mean vitreous concentrations of ofloxacin both in traumatized and traumatized-infected eyes were lower than that in the control eyes (p < 0.05). At 8 h, the mean vitreous concentrations of ofloxacin in the traumatized and in the traumatized-infected eyes were higher than that in the control eyes (p < 0.05). At 24 h, the mean ofloxacin concentration was higher in the traumatized-infected eyes than that in control (p < 0.01) and traumatized eyes (p < 0.05), and also higher in the traumatized eyes than that in the control eyes (p < 0.05). The mean ofloxacin concentrations in the traumatized and traumatized-infected eyes were significantly higher (p < 0.01) than those in the controls at 48 h. The elimination half-life of ofloxacin in the control eyes was 5.65 h and trauma and inflammation prolonged the half-life to 9.47 and 9. 72 h, respectively. Clearance of ofloxacin is fast and appears to be reduced by trauma and inflammation. Therapeutic drug levels in traumatized-infected eyes were maintained up to 24 h. This may be an important pharmacokinetic advantage in treating endophthalmitis unless the dose used has local toxicity and allows a longer dose interval when the dose is repeated.


SYR2, a gene necessary for syringomycin growth inhibition of Saccharomyces cerevisiae.
The Pseudomonas syringae cyclic lipodepsipeptide syringomycin inhibits the growth of Saccharomyces cerevisiae. A novel yeast gene, SYR2, was found to complement two syringomycin-resistant S. cerevisiae mutants. SYR2 was cloned, sequenced, and shown to encode a 349 amino acid protein located in the endoplasmic reticulum. SYR2 was identical to SUR2, which is involved in survival during nutritional starvation. Gene disruption or overexpression of SYR2 did not affect cell viability or ergosterol levels, but did influence cellular phospholipid levels. The findings suggest that phospholipids are important for the growth inhibitory action of syringomycin.