Investigation of the instability of plasmids directing the expression of Met-prochymosin in Escherichia coli.
The causes of the instability of a multicopy plasmid, pCT70, which directs the expression of calf prochymosin in Escherichia coli, were investigated. Plasmid pAT153 and its derivative, pCT54, were stable for more than 90 generations in continuous culture with glucose limitation. The multicopy plasmid pCT66, which expressed very low levels of prochymosin due to poor translational efficiency, and low copy number plasmids which efficiently expressed the prochymosin gene, were also stable. These results indicated that high level translation of the recombinant gene was the cause of the instability of pCT70. The maximum specific growth rate of E. coli(pCT70) was reduced by 30% compared with E. coli(pCT66). To fulfil the requirements of a production system, a dual origin plasmid with controllable copy number was developed. Both this plasmid (pMG165) and a derivative which contained the prochymosin gene (pMG168) were stable when maintained at low copy number. When the copy number of plasmid pMG168 was increased by putting replication under the control of the lambda PR promoter and the cI857 temperature sensitive repressor, expression of prochymosin was achieved. This strategy enables large-scale production of prochymosin without the need for antibiotic selection or other methods of preventing plasmid loss.


Methodological framework for functional characterization of plant microRNAs.
Since the beginning of this century, microRNAs (miRNAs), which are tiny RNA molecules, have become one of the major research topics on gene expression regulation in both animals and plants. The major task of miRNA study is to elucidate how the miRNAs are expressed in vivo, how they exert regulatory effects on their targets, and how they can be qualitatively or quantitatively cloned. For these purposes, the methodology of miRNA study has been developed and significantly improved in recent years. The focus here is on a number of powerful methods for plant miRNA research including bioinformatics tools and experimental approaches being used for upstream or downstream analysis of miRNAs or miRNA cloning. Some discrepancies exist in the miRNA research methodology between plants and animals, for example, 5' modified RACE (Rapid Amplification of cDNA Ends) can be used for cleavage target validation only in plants. However, numerous common methods are shared by these two miRNA research areas. Thus, this review will enhance our understanding of miRNA research methodology in organisms.


Burton's Agammaglobulinemia and COVID-19.
During the SARS-CoV-2 global pandemic, many patients who have co-morbid conditions are considered high risk for morbidity and mortality; however, those who are immunodeficient are at higher risk of becoming seriously ill. In this article, we present a 26-year old male with a history of X-linked agammaglobulinemia who presented to the hospital with fever and chills after exposure to a SARS-CoV-2 positive individual. The patient had a prolonged course in the hospital, but his symptoms improved quickly after receiving convalescent plasma. This case highlights the clinical course of a patient with severe immunoglobulin deficiency and a possible treatment approach for patients with concomitant agammaglobulinemia and COVID-19.


Mutations identified in engineered Escherichia coli with a reduced genome.
Characterizing genes that regulate cell growth and survival in model organisms is important for understanding higher organisms. Construction of strains harboring large deletions in the genome can provide insights into the genetic basis of cell growth compared with only studying wild-type strains. We have constructed a series of genome-reduced strains with deletions spanning approximately 38.9% of the E. coli chromosome. Strains were constructed by combining large deletions in chromosomal regions encoding nonessential gene groups. We also isolated strains Δ33b and Δ37c, whose growth was partially restored by adaptive laboratory evolution (ALE). Genome sequencing of nine strains, including those selected following ALE, identified the presence of several Single Nucleotide Variants (SNVs), insertions, deletions, and inversions. In addition to multiple SNVs, two insertions were identified in ALE strain Δ33b. The first was an insertion at the promoter region of pntA, which increased cognate gene expression. The second was an insertion sequence (IS) present in sibE, encoding the antitoxin in a toxin-antitoxin system, which decreased expression of sibE. 5 strains of Δ37c independently isolated following ALE harboring multiple SNVs and genetic rearrangements. Interestingly, a SNV was identified in the promoter region of hcaT in all five strains, which increased hcaT expression and, we predict, rescued the attenuated Δ37b growth. Experiments using defined deletion mutants suggested that hcaT encodes a 3-phenylpropionate transporter protein and is involved in survival during stationary phase under oxidative stress. This study is the first to document accumulation of mutations during construction of genome-reduced strains. Furthermore, isolation and analysis of strains derived from ALE in which the growth defect mediated by large chromosomal deletions was rescued identified novel genes involved in cell survival.


Regulation of urea production by glucose infusion in vivo.
We have investigated the acute in vivo regulation of urea production in normal postabsorptive volunteers by administering a primed constant infusion of 15N2-urea to measure urea production during the constant intravenous infusion of equivalent molar quantities of exogenous nitrogen, given as alanine or glutamine, either with or without a simultaneous infusion of glucose at 4 mg.kg-.min-1. These responses were compared with the response to the infusion of glucose alone. Both amino acid infusions elicited significant (P less than 0.05) and identical (26%) increases in urea production over 4 h. When the glucose infusion was added to the amino acid infusions, urea production remained constant, despite the comparable increases in plasma total nonessential amino nitrogen, as were observed with the amino acid infusions alone. Glucose infused alone elicited a significant (P less than 0.05) reduction (18%) in urea production but no corresponding change in plasma total amino nitrogen. We conclude that 1) infused glucose or its hormonal response suppresses urea production by blunting the normal hepatic ureagenic response to a fixed nitrogen load, 2) this suppressive effect is not mediated via a reduction in substrate (nitrogen) supply, and 3) the inhibition of hepatic gluconeogenesis from amino acids represents one component of this suppressive effect, and direct suppression of urea cycle activity probably represents another component.


The Effect of COVID-19 Pandemic on Eye-Related Emergency Department Visits: A Comparison of 2-Year Results.
The aim of this study was to examine the clinical and demographic impact of the COVID-19 pandemic on emergency department admissions to a tertiary eye care hospital. Records of admissions to the ophthalmology emergency department during the pre-pandemic period (Group 1) and pandemic period (Group 2) were retrospectively reviewed to cover the period between March 15, 2019, and March 15, 2021. Application numbers, demographic characteristics of patients were recorded. The cases were grouped by age, sex and diagnoses, and the findings were compared within and between the groups. A total of 161, 941 patients (Group 1: 103, 178 and Group 2: 58, 763) were admitted to the emergency department of our hospital within 2 years. All admission diagnoses were significantly lower in the pandemic period than in the pre-pandemic period (p=0.001). However, the rate of sight-threatening cases or cases requiring urgent treatment (retinal diseases, uveitis, etc.) was significantly higher in the pandemic period than in the pre-pandemic period (p=0.001). The most common admission diagnoses were ocular infectious/inflammatory diseases while the foreign body on the ocular surface/ocular trauma group was second. Admissions for allergic and infectious diseases were significantly less during the pandemic period (p=0.001 and p=0.002, respectively). In both periods, the number of admissions of male patients was significantly higher than that of females. The most frequent applicant age group was the 17-40 years age group. This observational study reveals the changes caused by the COVID-19 pandemic in ocular emergency admissions. The data from our study may be helpful in planning healthcare delivery during and after the pandemic period.


Starvation kinetics.
Reactions that take place in shock waves are slow compared to the energy present in the translational degrees of freedom. One explanation for the slow burning in a detonation is that successive surface layers of the solid or liquid must react sequentially. Another mechanism, which can also account for the slowness of reactions in shock waves in gases, is the basis for starvation kinetics: the bond that is breaking must draw its activation energy from a vibrational reservoir in disequilibrium with the translational degrees of freedom of the reacting molecule.


Viroid-induced RNA silencing of GFP-viroid fusion transgenes does not induce extensive spreading of methylation or transitive silencing.
Viroid infection is associated with the production of short interfering RNAs (siRNAs), a hallmark of post-transcriptional gene silencing (PTGS). However, viroid RNAs autonomously replicating in the nucleus have not been shown to trigger the degradation of homologous RNA in the cytoplasm. To investigate the potential of viroids for the induction of gene silencing, non-infectious fragments of potato spindle tuber viroid (PSTVd) cDNA were transcriptionally fused to the 3' end of the green fluorescent protein (GFP)-coding region. Introduction of such constructs into tobacco plants resulted in stable transgene expression. Upon PSTVd infection, transgene expression was suppressed and partial de novo methylation of the transgene was observed. PSTVd-specific siRNA was detected but none was found corresponding to the gfp gene. Methylation was restricted almost entirely to the PSTVd-specific part of the transgene. Neither a gfp transgene construct lacking viroid-specific elements was silenced nor was de novo methylation detected, when it was introduced into the genetic background of the PSTVd-infected plant lines containing silenced GFP:PSTVd transgenes. The absence of gfp-specific siRNAs and of significant methylation within the gfp-coding region demonstrated that neither silencing nor DNA methylation spread from the initiator region into adjacent 5' regions.


Left ventricular twist was decreased in isolated left bundle branch block with preserved ejection fraction.
Left ventricular (LV) rotation and twist play an important role in LV contraction and relaxation. Left bundle branch block (LBBB) deteriorates both diastolic and systolic functions. We evaluated the LV twist in patients with LBBB and preserved ejection fraction (EF) (>50%) to determine twist as a potential marker for subtle myocardial dysfunction. This observational cross-sectional study included 34 LBBB patients with preserved EF who were free from ischemic and valvular disease (Group 1) and 36 healthy controls (Group 2). All patients underwent 2-D Doppler and 2-D speckle tracking echocardiography. LV apical, basal rotation, and twist were evaluated in both groups and compared accordingly. In addition, subjects were dichotomized considering the median twist value of the study population. Binary logistic regression analysis was performed to determine the independent variables associated with inframedian twist. Baseline clinical characteristics were similar in LBBB patients and controls. Mean apical rotation (2.5°±1.9° vs. 4.4°±2.9°; p=0.002), basal rotation (-2.9°±2.3° vs. -4.1°±2.7°; p=0.05), and twist (5.4°±3° vs. 8.6°±3.3°; p<0.001) were decreased in group 1. Parameters related to intra- and interventricular mechanical dyssynchrony, such as longitudinal left ventricular dyssynchrony index (LVdys) and preejection interval of LV, interventricular mechanical delay (IVMD), and left posterior wall contractions (SPMWD) were significantly higher in the LBBB group. The median twist value of the studied population was 6.65°. Binary logistic regression analysis showed that only presence of LBBB was independently associated with inframedian twist (OR=6.250; 95% CI: 2.215-17.632; p<0.001). The LBBB might have induced the reduction of LV twist by diminishing the LV rotation before inducing a prominent effect on the left ventricular ejection fraction (LVEF). Therefore, twist may be considered as a marker for subtle LV dysfunction in LBBB with substantially normal EF.


Retrospective study of pediatric facial cellulitis of odontogenic origin.
The aims of this retrospective study were to investigate the clinical characteristics of pediatric facial cellulitis of odontogenic origin and to compare the clinical differences between upper and lower facial infections and between genders. Fifty-six children (28 boys and 28 girls) diagnosed with facial cellulitis of odontogenic origin during the year 2003 were selected for study. The average age was 5.72 +/- 2.70 years. Clinical investigations included sex, location of cellulitis, peak occurrence month, symptoms of infection, preference of first visit, hospitalization status, timing of surgical or dental interventions, need for incision and drainage, source of infection, and management of the primary odontogenic origin. Children were divided into upper face and lower face groups depending on the location of the infection. The chi and Student t tests were computed to assess the differences in the various study variables between upper and lower face infections and gender. Upper and lower face infections were reported in 57.1 and 42.9% of children, respectively. The source of infection was mostly from the deciduous posterior teeth. The clinical symptoms included a frequent toothache before swelling, and less commonly fever and trismus. Hospitalization was needed in 53.6% of the patients, and the mean length of hospitalization was 5.03 +/- 3.80 days. Surgical intervention such as incision and drainage was needed in only 25% of the cases. The majority of the patients had either surgical or dental interventions after subsidence of the swelling. The peak occurrence month was February. The source of infection in the upper face infections was significantly different from that of lower face infections (P < 0.01). Other study variables were not statistically significant for upper and lower face infections (P > 0.05). The differences in upper and lower face infections and differences in gender were not clinically significant. Surgical or dental interventions can be delayed through the proper use of antibiotics. With correct diagnosis, antibiotic treatment and appropriate timing for surgical or dental interventions, rapid resolution of the infection is expected.


Cell growth of the mouse SDHC mutant cells was suppressed by apoptosis throughout mitochondrial pathway.
SDHC E69 cells, which overproduce superoxide anions in their mitochondria, were previously established that had a mutation in the SDHC gene of complex II of the respiratory chain. We now demonstrate that tumors formed by NIH 3T3 and SDHC E69 cells showed significant histological differences. Cytoplasmic cytochrome c release from mitochondria was significantly elevated in SDHC E69 cells and was likely caused by superoxide anion overproduction from mitochondria. In addition, the p53 and Ras signal transduction pathways were activated by oxidative stress and may play a key role in the supernumerary apoptosis in SDHC E69 cells. Our results suggest that the development and growth characteristics of hereditary paragangliomas, which are defective in the same complex of electron transport as mouse SDHC E69 cells, may be caused by apoptosis induction by mitochondrial oxidative stress.


[The effect of chemical noxae on the health status of a group of workers in the chemical industry of Săvineşti].
The paper presents the study on the effect of the industrial xenobiotics against some biochemical indicators, in view to detect, precociously, the probable significant changes. There are presented the medium, minimum, maximum and median value for the following parameters: total proteins, haemoglobin and glucose for 112 workers exposed or not exposed to specific noxae--benzen, phenol, cyclohexanone. The analysis of the results shows decreased values for some workers, even at the inferior limit, in rapport with the normal values, for the parameters studied. These persons have been directed for complex investigations, in view to determine the causes and to establish the adequate treatment.


More than energy cost: multiple benefits of the long Achilles tendon in human walking and running.
Elastic strain energy that is stored and released from long, distal tendons such as the Achilles during locomotion allows for muscle power amplification as well as for reduction of the locomotor energy cost: as distal tendons perform mechanical work during recoil, plantar flexor muscle fibres can work over smaller length ranges, at slower shortening speeds, and at lower activation levels. Scant evidence exists that long distal tendons evolved in humans (or were retained from our more distant Hominoidea ancestors) primarily to allow high muscle-tendon power outputs, and indeed we remain relatively powerless compared to many other species. Instead, the majority of evidence suggests that such tendons evolved to reduce total locomotor energy cost. However, numerous additional, often unrecognised, advantages of long tendons may speculatively be of greater evolutionary advantage, including the reduced limb inertia afforded by shorter and lighter muscles (reducing proximal muscle force requirement), reduced energy dissipation during the foot-ground collisions, capacity to store and reuse the muscle work done to dampen the vibrations triggered by foot-ground collisions, reduced muscle heat production (and thus core temperature), and attenuation of work-induced muscle damage. Cumulatively, these effects should reduce both neuromotor fatigue and sense of locomotor effort, allowing humans to choose to move at faster speeds for longer. As these benefits are greater at faster locomotor speeds, they are consistent with the hypothesis that running gaits used by our ancestors may have exerted substantial evolutionary pressure on Achilles tendon length. The long Achilles tendon may therefore be a singular adaptation that provided numerous physiological, biomechanical, and psychological benefits and thus influenced behaviour across multiple tasks, both including and additional to locomotion. While energy cost may be a variable of interest in locomotor studies, future research should consider the broader range of factors influencing our movement capacity, including our decision to move over given distances at specific speeds, in order to understand more fully the effects of Achilles tendon function as well as changes in this function in response to physical activity, inactivity, disuse and disease, on movement performance.


Inactivity of Peptidase ClpP Causes Primary Accumulation of Mitochondrial Disaggregase ClpX with Its Interacting Nucleoid Proteins, and of mtDNA.
Biallelic pathogenic variants in CLPP, encoding mitochondrial matrix peptidase ClpP, cause a rare autosomal recessive condition, Perrault syndrome type 3 (PRLTS3). It is characterized by primary ovarian insufficiency and early sensorineural hearing loss, often associated with progressive neurological deficits. Mouse models showed that accumulations of (i) its main protein interactor, the substrate-selecting AAA+ ATPase ClpX, (ii) mitoribosomes, and (iii) mtDNA nucleoids are the main cellular consequences of ClpP absence. However, the sequence of these events and their validity in human remain unclear. Here, we studied global proteome profiles to define ClpP substrates among mitochondrial ClpX interactors, which accumulated consistently in ClpP-null mouse embryonal fibroblasts and brains. Validation work included novel ClpP-mutant patient fibroblast proteomics. ClpX co-accumulated in mitochondria with the nucleoid component POLDIP2, the mitochondrial poly(A) mRNA granule element LRPPRC, and tRNA processing factor GFM1 (in mouse, also GRSF1). Only in mouse did accumulated ClpX, GFM1, and GRSF1 appear in nuclear fractions. Mitoribosomal accumulation was minor. Consistent accumulations in murine and human fibroblasts also affected multimerizing factors not known as ClpX interactors, namely, OAT, ASS1, ACADVL, STOM, PRDX3, PC, MUT, ALDH2, PMPCB, UQCRC2, and ACADSB, but the impact on downstream metabolites was marginal. Our data demonstrate the primary impact of ClpXP on the assembly of proteins with nucleic acids and show nucleoid enlargement in human as a key consequence.


Deoxyribonucleic acid reassociation in the classification of the 'rhodochrous' complex and allied taxa.
The degree of binding was determined between DNA preparations from gordonae and rhodochrous strains and uracil-labelled DNA from five reference strains, Nocardia asteroides NK20, N. pellegrino PII, Gordona bronchialis TI, G. terrae T5 and rhodochrous strain R90. Most of the rhodochrous and pellegrino strains fell into one of two genetically homogeneous taxa. The nucleotide sequence homology data also suggested that G. bronchialis, G. rubra, and more equivocally G. terrae, formed distinct species. However, the values for DNA relatedness between these species, and between then and the rhodochrous homology groups, were comparatively low. Only a small degree of nucleotide sequence homology was found between the N. asteroides reference system and the rest of the taxa studied. The nucleotide composition of the DNA preparations from 29 of the 30 test strains fell between 63 and 69 mol% guanine plus cytosine.


HIV among men who have sex with men in the Caribbean: reaching the left behind.
To present the epidemiology, social and cultural factors driving the HIV epidemic among men who have sex with men (MSM) in the Caribbean region and to highlight the regional and national responses, and what remains to be addressed to close the gaps in order to ending AIDS by 2030. A literature review was performed in the following databases: PubMed and Scopus. Articles published in the past 10 years were selected. The outcomes of interest were sociocultural risk factors, description of regional and national efforts and potential challenges and barriers to effective control of the epidemic among MSM. This report concentrates exclusively on publications related to MSM living in the Caribbean countries. 11 peer-reviewed studies, 9 grey literature reports and programme frameworks were thematically analysed. The prevalence of HIV among MSM is high and the rates also do vary among Caribbean countries. Several factors influence the epidemic among MSM in the Caribbean but stigma and discrimination underlie the social vulnerability and play a central role in driving the HIV epidemic. To end the AIDS epidemic by 2030, MSM can no longer be kept unchecked in the era of the Sustainable Development Goals with the motto 'Leave no one behind'.


[Low intensity laser radiation in complex therapy of patients with vascular obliterating atherosclerosis of low extremities].
It is shown that laser therapy used in a multiple-modality treatment of patients with obliterating atherosclerosis of vessels of the interior limbs presenting with stage I-III ischemia permits achieving a substantial clinical effect manifested subjectively by fewer compliants or disappearance thereof in a proportion of patients, which fact is corraborated by objective findings such as increase in peripheral, volumetric blood flow and lower degree, in some patients, of ischemia of the extremities, improvement in processes of microcirculation and hemocoagulation. The use of laser radiation and pneumocompression combined in treatment of patients with chronic arterial insufficiency of the lower extremities of atherosclerotic genesis has been shown to have a more marked and appreciable effect. The studies made broaden our possibilities of conservative non-medicamentous treatment of obliterating atherosclerosis of vessels of the lower extremities.


High-speed collagen fiber modeling and orientation quantification for optical coherence tomography imaging.
Quantifying collagen fiber architecture has clinical and scientific relevance across a variety of tissue types and adds functionality to otherwise largely qualitative imaging modalities. Optical coherence tomography (OCT) is uniquely suited for this task due to its ability to capture the collagen microstructure over larger fields of view than traditional microscopy. Existing image processing techniques for quantifying fiber architecture, while accurate and effective, are very slow for processing large datasets and tend to lack structural specificity. We describe here a computationally efficient method for quantifying and visualizing collagen fiber organization. The algorithm is demonstrated on swine atria, bovine anterior cruciate ligament, and human cervical tissue samples. Additionally, we show an improved performance for images with crimped fiber textures and low signal to noise when compared to similar methods.


[Renal involvement in convalescents after icterohemorrhagic leptospirosis].
Icterohemorrhagic spirochetosis convalescents develop slowly regressing renal dysfunctions most frequent of which are chronic renal failure, pyelonephritis and tubulointersticial nephritis, arterial hypertension. Renal disorders may be due to immunopathological reactions followed by activation of bacterial microflora. Damage from commissures at the sites of prior hemorrhage is also possible. In bovine leptospirosis renal damage is not so severe but it tends to progression. So, renal affection in leptospirosis is a specific pathology observed in any clinical form of the disease and demands surveillance in the regions of local focuses.


Adult physical activity levels and associated factors in rural communities of Minas Gerais State, Brazil.
To estimate the physical activity level and its association with sociodemographic factors in adults living in rural areas. Cross-sectional population study including 567 adults in two rural communities from the Jequitinhonha Valley, Southeastern Brazil, during the years of 2008 and 2009. Physical activity levels were assessed with the adapted long version of the International Physical Activity Questionnaire. A cut-off point of 150 minutes per week was used in the analyses for the domains: occupational, household, leisure and commuting. The sociodemographic factors studied were sex, skin color, age, marital status, education and self-reported health. Bivariate analysis (chi-square test, p< 0.05) and multiple logistic regression analysis were performed. The prevalence of subjects practicing 150 min/wk or more of work related physical activity was 82.9% (95% CI: 77.8;88.0) of those currently working. The equivalent proportions for the other domains were: household 63.5% (95% CI: 59.6; 67.4); leisure time 10.1% (95% CI: 7.6;12.6) and commuting 32.0% (95% CI: 28.2;35.8%). Men were more active than women in leisure time, commuting and occupational domains, while women were more active in the household domain. Leisure time physical activity was more prevalent in younger subjects, those with higher levels of education and among those of black or mixed skin color. Commuting physical activity was more frequent among younger women and among men and women in excellent/good health. Men with higher level of schooling were less active in the commuting domain. The prevalence of physically active adults in this rural area was high, but the levels of leisure time physical activity were low and followed patterns similar to those observed in urban areas, in relation to age, sex and educational status.


In vitro binding of N-acetoxy-N-2-acetylaminofluorene to DNA in chromatin.
The binding of N-acetoxy-N-2-acetylaminofluorene to DNA in native and partially dehistonized chicken erythrocyte chromatin was studied. The amounts of carcinogen bound to DNA were measured, after removal of proteins with phenol, by using the absorption ratio A305/A260 or by counting the radioactivity of 14C-labeled carcinogen. Measurements of uncovered zones of DNA in chromatin were made by comparison of results obtained with free DNA and with chromatin at increasing ratios of carcinogen/nucleotide. The proportion of DNA accessible to the carcinogen was found to be 15% in native chicken erythrocyte chromatin and about 22% in native calf thymus chromatin. The amount of accessible DNA increases to 55% in chicken erythrocyte chromatin depleted of histones H1 and H5. Formaldehyde unwinding performed on DNA extracted from chromatin after modification showed an increasing number of defects in the double helix with the amount of DNA-fixed carcinogen. At high ratios of carcinogen/nucleotide, the recoveries of DNA (by phenol method) and of histones (by acidic extraction) decreased with increasing ratios. This suggests a covalent linkage between proteins and DNA.


Overlapping roles of the cytoplasmic and mitochondrial redox regulatory systems in the yeast Saccharomyces cerevisiae.
Thioredoxins are small, highly conserved oxidoreductases which are required to maintain the redox homeostasis of the cell. Saccharomyces cerevisiae contains a cytoplasmic thioredoxin system (TRX1, TRX2, and TRR1) as well as a complete mitochondrial thioredoxin system, comprising a thioredoxin (TRX3) and a thioredoxin reductase (TRR2). In the present study we have analyzed the functional overlap between the two systems. By constructing mutant strains with deletions of both the mitochondrial and cytoplasmic systems (trr1 trr2 and trx1 trx2 trx3), we show that cells can survive in the absence of both systems. Analysis of the redox state of the cytoplasmic thioredoxins reveals that they are maintained independently of the mitochondrial system. Similarly, analysis of the redox state of Trx3 reveals that it is maintained in the reduced form in wild-type cells and in mutants lacking components of the cytoplasmic thioredoxin system (trx1 trx2 or trr1). Surprisingly, the redox state of Trx3 is also unaffected by the loss of the mitochondrial thioredoxin reductase (trr2) and is largely maintained in the reduced form unless cells are exposed to an oxidative stress. Since glutathione reductase (Glr1) has been shown to colocalize to the cytoplasm and mitochondria, we examined whether loss of GLR1 influences the redox state of Trx3. During normal growth conditions, deletion of TRR2 and GLR1 was found to result in partial oxidation of Trx3, indicating that both Trr2 and Glr1 are required to maintain the redox state of Trx3. The oxidation of Trx3 in this double mutant is even more pronounced during oxidative stress or respiratory growth conditions. Taken together, these data indicate that Glr1 and Trr2 have an overlapping function in the mitochondria.


AR DriveSim: An Immersive Driving Simulator for Augmented Reality Head-Up Display Research.
Optical see-through automotive head-up displays (HUDs) are a form of augmented reality (AR) that is quickly gaining penetration into the consumer market. Despite increasing adoption, demand, and competition among manufacturers to deliver higher quality HUDs with increased fields of view, little work has been done to understand how best to design and assess AR HUD user interfaces, and how to quantify their effects on driver behavior, performance, and ultimately safety. This paper reports on a novel, low-cost, immersive driving simulator created using a myriad of custom hardware and software technologies specifically to examine basic and applied research questions related to AR HUDs usage when driving. We describe our experiences developing simulator hardware and software and detail a user study that examines driver performance, visual attention, and preferences using two AR navigation interfaces. Results suggest that conformal AR graphics may not be inherently better than other HUD interfaces. We include lessons learned from our simulator development experiences, results of the user study and conclude with limitations and future work.


Exercise in a healthy heart program: a cohort study.
To assess the effects of exercise on resting heart rate (RHR), weight, lipid profile, and blood pressure. We hypothesized that the participants who increased their physical activity would show improvement in their cardiovascular risk factors compared to those who did not. Retrospective chart review over the mean duration of 4.9 years of follow-up. Healthy Heart Program Prevention Clinic at St. Paul's Hospital, Vancouver, British Columbia, Canada. We reviewed 300 charts of patients randomly selected from those who attended the Prevention Clinic between 1984 and 2009. 248 (82.7%) patients were referred for primary prevention and 52 (17.3%) for secondary prevention. Weight, RHR, lipid profile, and blood pressure were recorded at the initial and last visit. During a mean of 4.9 years of follow-up, 55% of participants improved their exercise. The mean decrease in the RHR for these patients (group 1) was 5.9 beats per minute (bpm) versus the mean increase of 0.3 bpm for the "no change" group (group 2) (P < 0.01). The mean net weight increase in group 1 was 0.06 kg/year versus 0.25 kg/year in group 2. Because of medications, all patients had a significant improvement in their lipid profiles. Furthermore, there was a statistically significant greater reduction in Framingham Risk Score (FRS) in group 1 versus group 2 (11.8% versus 15.1%, P < 0.01). Participation in the program significantly reduces modifiable risk factors for cardiovascular disease. Improved exercise regimen results in lower RHR and greater reduction in FRS. However, even in a Prevention Program, despite strong advocacy of the importance of exercise, a significant percentage of participants does not improve their exercise habits.


Kinetics and selectivity of mechanism-based inhibition of guinea pig hepatic and pulmonary cytochrome P450 by N-benzyl-1-aminobenzotriazole and N-alpha-methylbenzyl-1-aminobenzotriazole.
The time dependence for mechanism-based inactivation of cytochrome P450 (P450)-dependent 7-pentoxyresorufin O-depentylation (PROD), 7-ethoxyresorufin O-deethylation (EROD), and 7-methoxyresorufin O-demethylation (MROD) activities by N-benzyl-1-aminobenzotriazole (BBT) and N-alpha-methylbenzyl-1-aminobenzotriazole (alpha MB) was investigated in hepatic and pulmonary microsomes from phenobarbital-treated guinea pigs. In the presence of NADPH, both compounds inhibited P450-dependent catalytic activity in a time- and concentration-dependent manner. Inactivation of hepatic PROD activity was more rapid (t1/2 = 13.2 vs. 155 min) for 0.1 microM alpha MB when compared with equimolar BBT. On the other hand, hepatic EROD inactivation was more rapid (t1/2 = 8.1 vs. 11 min) with 0.1 microM BBT, compared with equimolar alpha MB. Inactivation of pulmonary PROD activity was the most rapid and potent, with an apparent half-life for inactivation of t1/2 = 0.94 and 32.2 min for 0.025 microM alpha MB and BBT, respectively. Incubation of hepatic microsomes for 45 min in the presence of NADPH and 10 microM BBT or alpha MB resulted in > 90% inhibition of PROD, EROD, and MROD activities. After washing by repeated sedimentation and resuspension, inhibition of PROD (78%; 93% for BBT and alpha MB, respectively), EROD (80% and 50%), and MROD (15% and 3%) activities was reversed to varying degrees. We conclude that BBT and alpha MB are rapidly metabolized to products that inhibit individual P450 isozymes by both mechanism-based (P4502B and P4501A1) and reversible (P4501A2) mechanisms. Of the two inhibitors, alpha MB is relatively more potent and selective for guinea pig lung P4502B isozyme(s).


A layer-by-layer ZnO nanoparticle-PbS quantum dot self-assembly platform for ultrafast interfacial electron injection.
Absorbent layers of semiconductor quantum dots (QDs) are now used as material platforms for low-cost, high-performance solar cells. The semiconductor metal oxide nanoparticles as an acceptor layer have become an integral part of the next generation solar cell. To achieve sufficient electron transfer and subsequently high conversion efficiency in these solar cells, however, energy-level alignment and interfacial contact between the donor and the acceptor units are needed. Here, the layer-by-layer (LbL) technique is used to assemble ZnO nanoparticles (NPs), providing adequate PbS QD uptake to achieve greater interfacial contact compared with traditional sputtering methods. Electron injection at the PbS QD and ZnO NP interface is investigated using broadband transient absorption spectroscopy with 120 femtosecond temporal resolution. The results indicate that electron injection from photoexcited PbS QDs to ZnO NPs occurs on a time scale of a few hundred femtoseconds. This observation is supported by the interfacial electronic-energy alignment between the donor and acceptor moieties. Finally, due to the combination of large interfacial contact and ultrafast electron injection, this proposed platform of assembled thin films holds promise for a variety of solar cell architectures and other settings that principally rely on interfacial contact, such as photocatalysis.


Sulfadiazine modified PDMS as a model material with the potential for the mitigation of posterior capsule opacification (PCO).
Cataract surgery, while the most common surgical procedure performed, leads to posterior capsule opacification in approximately 30% of cases. Transforming growth factor beta 2 (TGF-β2) and matrix metalloproteinases (MMPs) have been shown to play important roles in the cellular processes leading to posterior capsule opacification. Delivery of inhibitors to MMPs may have the potential to inhibit the initial cascade of events that lead to PCO. However, delivery of these molecules via tethering has proven difficult. In this work, sulfadiazine was tethered to polydimethylsiloxane (PDMS) via a polyethylene glycol (PEG) spacer as a potential MMPI mimic. Surface characterization using a variety of methods demonstrated successful modification with the antibiotic. The surfaces were examined with lens epithelial cells to determine their effect on these cellular processes, including cell transdifferentiation and production of extracellular matrix components. The presence of TGF-β2 in the cell culture media was found to stimulate the production of ECM components such as collagen, fibronectin, and laminin, as well as alpha smooth muscle actin (α-SMA), and the migration marker Rho by HLE-B3 and FHL124 cells. In all cases, these effects were decreased but not completely eradicated by the presence of sulfadiazine on the PDMS surfaces. While the level of inhibition necessary for inhibition of PCO in vivo is unknown, these results suggest that IOL surface modification with sulfadiazine has the potential to reduce cellular changes associated with PCO. Furthermore, the results demonstrate for the first time that changes consistent with inhibition of fibrosis may be elicited by surfaces modified with sulfadiazine.


Effects of Physical Distancing to Control COVID-19 on Public Health, the Economy, and the Environment.
Physical distancing measures are important tools to control disease spread, especially in the absence of treatments and vaccines. While distancing measures can safeguard public health, they also can profoundly impact the economy and may have important indirect effects on the environment. The extent to which physical distancing measures should be applied therefore depends on the trade-offs between their health benefits and their economic costs. We develop an epidemiological-economic model to examine the optimal duration and intensity of physical distancing measures aimed to control the spread of COVID-19. In an application to the United States, our model considers the trade-off between the lives saved by physical distancing-both directly from stemming the spread of the virus and indirectly from reductions in air pollution during the period of physical distancing-and the short- and long-run economic costs that ensue from such measures. We examine the effect of air pollution co-benefits on the optimal physical distancing policy and conduct sensitivity analyses to gauge the influence of several key parameters and uncertain model assumptions. Using recent estimates of the association between airborne particulate matter and the virulence of COVID-19, we find that accounting for air pollution co-benefits can significantly increase the intensity and duration of the optimal physical distancing policy. To conclude, we broaden our discussion to consider the possibility of durable changes in peoples' behavior that could alter local markets, the global economy, and our relationship to nature for years to come.


Epidemiology of cutaneous tuberculosis in Japan: a retrospective study from 1906 to 2002.
Interest in tuberculosis has recently been revived, especially in association with the onset of acquired immunodeficiency syndrome (AIDS). In Japan, the incidence of tuberculosis has declined since the 1950s, but has shown a tendency to increase in the past 10 years. To determine the patterns of prevalence of cutaneous tuberculosis in Japan, data for cutaneous tuberculosis cases recorded between 1906 and 2002 were obtained from the dermatologic clinic of Kyushu University Hospital. The population of tuberculosis-infected patients was stratified into groups encompassing 10-year periods. Each patient group was then classified as having true tuberculosis or tuberculid, as well as being classified by gender. The total number of patients with cutaneous tuberculosis analyzed was 1324. The incidence of cutaneous tuberculosis, both true tuberculosis and tuberculid, decreased between 1906 and 1935, increased between 1936 and 1955, and decreased again from 1955. The incidence of true tuberculosis was higher than that of tuberculid between 1906 and 1925. The incidence rates of true tuberculosis and tuberculid were equal from 1926 to 1945, and since 1946 the incidence of tuberculid has been higher than that of true tuberculosis. The incidence of tuberculid decreased between 1986 and 1995, but showed a dramatic increase between 1996 and 2000. The frequency of cutaneous tuberculosis was highest in the 10-49 year age group before 1980, but since 1981 has been highest in groups over 40 years of age. The incidence of cutaneous tuberculosis in Japan is decreasing and shows a shift towards older generations. However, there is an increase in the incidence of tuberculid, especially in elderly female patients.


Sociodemographic disparities and potential biases in persistent pain estimates: Findings from 5 waves of the Irish Longitudinal Study on Ageing (TILDA).
Pain is a prevalent, debilitating condition among older adults. Much evidence on this topic comes from cohort studies, which may be affected by attrition and measurement bias. Little is known about the impact of these biases on pain estimates for European older adults. Additionally, there is a lack of longitudinal research on pain and sociodemographic disparities in Irish older adults. We analysed data from 8171 participants (aged ≥50 at baseline) across five waves of the Irish Longitudinal Study on Ageing. Longitudinal pain severity and sociodemographic disparities in pain were explored visually and using a latent growth curve model. Using multivariate logistic regression, we examined bias due to attrition at later waves associated with reported pain at Wave 1. Measurement biases due to reporting heterogeneity were assessed by investigating associations between sociodemographic factors and pain-related disability for given pain levels. Wave 1 severe pain was associated with increased odds of attrition due to death by Wave 5 (AOR: 1.63, 95% CI: 1.20, 2.19). Not having private health insurance was associated with increased odds of pain-related disability at Wave 1, controlling for pain severity (AOR: 1.37, 95% CI: 1.15, 1.64). These results suggested mortality bias and reporting heterogeneity measurement bias, respectively. Sex, education level, and private health insurance status disparities in pain were observed longitudinally. Mortality bias and reporting heterogeneity measurement bias must be accounted for to improve older adult pain estimates. There is a need for policymakers to address sociodemographic disparities in older adult pain levels. This study highlights a need to address bias in the estimation of pain in observational studies of older adults. Understanding the sources and extent of these biases is important so that health practices and policies to address pain disparities can be guided by accurate estimates. Women, those with lower educational attainment, and those without private health insurance were found to have the highest pain burden longitudinally, suggesting a need for targeted interventions for these groups in Ireland and internationally.


Hexagonal Close-packed Iron Hydride behind the Conventional Phase Diagram.
Hexagonal close-packed iron hydride, hcp FeHx, is absent from the conventional phase diagram of the Fe-H system, although hcp metallic Fe exists stably over extensive temperature (T) and pressure (P) conditions, including those corresponding to the Earth's inner core. In situ X-ray and neutron diffraction measurements at temperatures ranging from 298 to 1073 K and H pressures ranging from 4 to 7 GPa revealed that the hcp hydride was formed for FeHx compositions when x < 0.6. Hydrogen atoms occupied the octahedral interstitial sites of the host metal lattice both partially and randomly. The hcp hydride exhibited a H-induced volume expansion of 2.48(5) Å3/H-atom, which was larger than that of the face-centered cubic (fcc) hydride. The hcp hydride showed an increase in x with T, whereas the fcc hydride showed a corresponding decrease. The present study provides guidance for further investigations of the Fe-H system over an extensive x-T-P region.


Clinical syndromes associated with ovarian neoplasms: a comprehensive review.
Functional ovarian neoplasms have unique clinical manifestations related to hormone overproduction and may give rise to a broad spectrum of clinical syndromes. Sex cord-stromal tumors, the most common functional ovarian neoplasms, are associated with either hyperestrogenism (as in granulosa cell tumor and thecoma) or hyperandrogenism (as in Sertoli-Leydig cell tumor and Leydig cell tumor). Other, less common ovarian neoplasms that may have endocrine or nonendocrine syndromic manifestations include germ cell tumors associated with the excessive production of human chorionic gonadotropin (eg, choriocarcinoma, dysgerminoma), monodermal teratomas (eg, carcinoid tumor, struma ovarii) associated with carcinoid syndrome and hyperthyroidism, and primary epithelial ovarian cancers associated with paraneoplastic syndromes. The application of diagnostic algorithms based on patient demographic information, clinical manifestations, laboratory findings, and cross-sectional imaging features may help identify ovarian neoplasms in complex clinical settings.


A brief review on Group A Streptococcus pathogenesis and vaccine development.
Streptococcus pyogenes, also known as Group A Streptococcus (GAS), is a Gram-positive human-exclusive pathogen, responsible for more than 500 000 deaths annually worldwide. Upon infection, GAS commonly triggers mild symptoms such as pharyngitis, pyoderma and fever. However, recurrent infections or prolonged exposure to GAS might lead to life-threatening conditions. Necrotizing fasciitis, streptococcal toxic shock syndrome and post-immune mediated diseases, such as poststreptococcal glomerulonephritis, acute rheumatic fever and rheumatic heart disease, contribute to very high mortality rates in non-industrialized countries. Though an initial reduction in GAS infections was observed in high-income countries, global outbreaks of GAS, causing rheumatic fever and acute poststreptococcal glomerulonephritis, have been reported over the last decade. At the same time, our understanding of GAS pathogenesis and transmission has vastly increased, with detailed insight into the various stages of infection, beginning with adhesion, colonization and evasion of the host immune system. Despite deeper knowledge of the impact of GAS on the human body, the development of a successful vaccine for prophylaxis of GAS remains outstanding. In this review, we discuss the challenges involved in identifying a universal GAS vaccine and describe several potential vaccine candidates that we believe warrant pursuit.


Shifts in dietary carbohydrate-lipid exposure regulate expression of the non-alcoholic fatty liver disease-associated gene PNPLA3/adiponutrin in mouse liver and HepG2 human liver cells.
Patatin-like phospholipase domain containing 3 (PNPLA3, adiponutrin) has been identified as a modifier of lipid metabolism. To better understand the physiological role of PNPLA3/adiponutrin, we have investigated its regulation in intact mice and human hepatocytes under various nutritional/metabolic conditions. PNPLA3 gene expression was determined by real-time PCR in liver of C57BL/6 mice after dietary treatments and in HepG2 cells exposed to various nutritional/metabolic stimuli. Intracellular lipid content was determined in HepG2 cells after siRNA-mediated knockdown of PNPLA3. In vivo, mice fed a high-carbohydrate (HC) liquid diet had elevated hepatic lipid content, and PNPLA3 mRNA and protein expression, compared to chow-fed mice. Elevated expression was completely abrogated by addition of unsaturated lipid emulsion to the HC diet. By contrast, in mice with high-fat diet-induced steatosis, Pnpla3 expression did not differ compared to low-fat fed mice. In HepG2 cells, Pnpla3 expression was reversibly suppressed by glucose depletion and increased by glucose refeeding, but unchanged by addition of insulin and glucagon. Several unsaturated fatty acids each significantly decreased Pnpla3 mRNA, similar to lipid emulsion in vivo. However, Pnpla3 knockdown in HepG2 cells did not alter total lipid content in high glucose- or oleic acid-treated cells. Our results provide evidence that PNPLA3 expression is an early signal/signature of carbohydrate-induced lipogenesis, but its expression is not associated with steatosis per se. Under lipogenic conditions due to high-carbohydrate feeding, certain unsaturated fatty acids can effectively suppress both lipogenesis and PNPLA3 expression, both in vivo and in a hepatocyte cell line.


Zwitterionic Conductive Hydrogel-Based Nerve Guidance Conduit Promotes Peripheral Nerve Regeneration in Rats.
In recent years, conductive biomaterials have been widely used to enhance peripheral nerve regeneration. However, most biomaterials use electronic conductors to increase the conductivity of materials. As information carriers, electronic conductors always transmit discontinuous electrical signals, while biological systems essentially transmit continuous signals through ions. Herein, an ion-based conductive hydrogel was fabricated by simple copolymerization of the zwitterionic monomer sulfobetin methacrylate and hydroxyethyl methacrylate. Benefiting from the excellent mechanical stability, suitable electrical conductivity, and good cytocompatibility of the zwitterionic hydrogel, the Schwann cells cultured on the hydrogel could grow and proliferate better, and dorsal root ganglian had an increased neurite length. The zwitterionic hydrogel-based nerve guidance conduits were then implanted into a 10 mm sciatic nerve defect model in rats. Morphological analysis and electrophysiological data showed that the grafts achieved a regeneration effect close to that of the autologous nerve. Overall, our developed zwitterionic hydrogel facilitates efficient and efficacious peripheral nerve regeneration by mimicking the electrical and mechanical properties of the extracellular matrix and creating a suitable regeneration microenvironment, providing a new material reserve for the repair of peripheral nerve injury.


Highly efficient blue- and white-organic light emitting diodes base on triple-emitting layer.
We have demonstrated highly efficient blue phosphorescent organic light-emitting diodes (PHOLEDs) using iridium (III) bis[(4,6-di-fluoropheny)-pyridinato-N,C2] picolinate (Flrpic) doped in three kinds of host materials, such as 9-(4-(triphenylsilyl)phenyl)-9H-carbazole (SPC), N,N'-dicarbazolyl-3,5-benzene (mCP), and 2,2',2"-(1,3,5-benzenetriyl)tris-[1-phenyl-1H-benzimidazole] (TPBi) as triple-emitting layer (T-EML). The properties of device with T-EML using the stepwise structure was found to be superior to the other blue PHOLEDs and exhibited a maximum luminous efficiency of 23.02 cd/A, a maximum external quantum efficiency of 11.09%, and a maximum power efficiency of 14.89 lm/W, respectively. An optimal blue device has improving charge balance and triplet excitons confinement within emitting layers (EMLs) each. Additionally, we also fabricated white PHOLED using a phosphorescent red dopant, bis(2-phenylquinolinato)-acetylacetonate iridium III (Ir(pq)2acac) doped in mCP and TPBi between blue EMLs. The properties of white PHOLED showed a maximum luminous efficiency and a maximum external quantum efficiency of 33.03 cd/A and 16.95%, respectively. It also showed the white emission with CIEx,y coordinates of (x = 0.36, y = 0.39) at 10 V.


MODEL-FREE FORWARD SCREENING VIA CUMULATIVE DIVERGENCE.
Feature screening plays an important role in the analysis of ultrahigh dimensional data. Due to complicated model structure and high noise level, existing screening methods often suffer from model misspecification and the presence of outliers. To address these issues, we introduce a new metric named cumulative divergence (CD), and develop a CD-based forward screening procedure. This forward screening method is model-free and resistant to the presence of outliers in the response. It also incorporates the joint effects among covariates into the screening process. With a data-driven threshold, the new method can automatically determine the number of features that should be retained after screening. These merits make the CD-based screening very appealing in practice. Under certain regularity conditions, we show that the proposed method possesses sure screening property. The performance of our proposal is illustrated through simulations and a real data example.


Expression of intercellular adhesion molecule-1 in immune response of inner ear.
To understand the role of intercellular adhesion molecule-1 (ICAM-1) in immune response of the inner ear, inner ear immune response was induced in rats by inoculation of keyhole limpet hemocyanine (KLH) into the scala tympani of the animals who had been systemically sensitized. The expression of ICAM-1 in the inner ear was immunohistochemically examined. ICAM-1 was found in the epithelium of the spiral modiolar vein (SMV) with its collecting venules (CVs) as early as 6 h after challenge. Expression of ICAM-1 was observed on the epithelium of the endolymphatic sac (ES) and perisaccular region at 12 h. The intensity of ICAM-1 staining reached its peak within 24-48 h in these sites of the inner ear. By day 28, most specimens were devoid of appreciable staining for ICAM-1. Our study demonstrates that adhesion molecules play an important role in extravasation of inflammatory cells from the systemic circulation in the process of inner ear immune response. It also shows that cytokines that control expression of adhesion molecules may be released by cells outside ES, besides those cells in the ES.


Investigation of gender role behaviors in boys with hypospadias: comparative study with unaffected boys and girls.
The purpose of the study was (1) to investigate gender role behaviors of boys with hypospadias compared with groups of unaffected boys and girls using parental reports and direct observations; and (2) to directly observe effects of socialization (mothers' presence) on children's gender role behaviors. Ages of 19 children with hypospadias ranged from 3 to 7 years, and each of them were matched to controls of unaffected boys and girls by age. All the children participated with their mothers. Children's gender role behaviors and their mothers' behaviors were evaluated using an observation coding system. Mothers also completed questionnaires regarding their children's gender role behaviors. Results indicated no atypical gender role behavior for the boys with hypospadias and no direct effects of socialization on their gender role behaviors. However, differences were found in negative communicative behaviors between boys with hypospadias and unaffected boys, suggesting a possible role of socialization.


Effect of confounding variables on hemodynamic response function estimation using averaging and deconvolution analysis: An event-related NIRS study.
Slow and rapid event-related designs are used in fMRI and functional near-infrared spectroscopy (fNIRS) experiments to temporally characterize the brain hemodynamic response to discrete events. Conventional averaging (CA) and the deconvolution method (DM) are the two techniques commonly used to estimate the Hemodynamic Response Function (HRF) profile in event-related designs. In this study, we conducted a series of simulations using synthetic and real NIRS data to examine the effect of the main confounding factors, including event sequence timing parameters, different types of noise, signal-to-noise ratio (SNR), temporal autocorrelation and temporal filtering on the performance of these techniques in slow and rapid event-related designs. We also compared systematic errors in the estimates of the fitted HRF amplitude, latency and duration for both techniques. We further compared the performance of deconvolution methods based on Finite Impulse Response (FIR) basis functions and gamma basis sets. Our results demonstrate that DM was much less sensitive to confounding factors than CA. Event timing was the main parameter largely affecting the accuracy of CA. In slow event-related designs, deconvolution methods provided similar results to those obtained by CA. In rapid event-related designs, our results showed that DM outperformed CA for all SNR, especially above -5 dB regardless of the event sequence timing and the dynamics of background NIRS activity. Our results also show that periodic low-frequency systemic hemodynamic fluctuations as well as phase-locked noise can markedly obscure hemodynamic evoked responses. Temporal autocorrelation also affected the performance of both techniques by inducing distortions in the time profile of the estimated hemodynamic response with inflated t-statistics, especially at low SNRs. We also found that high-pass temporal filtering could substantially affect the performance of both techniques by removing the low-frequency components of HRF profiles. Our results emphasize the importance of characterization of event timing, background noise and SNR when estimating HRF profiles using CA and DM in event-related designs.


Circadian rhythms in the vegetative state.
To evaluate whether vegetative state patients maintain circadian rhythms. An observational study of five single cases. Five chronic vegetative state patients underwent clinical and neurological evaluations and 2-week continuous temperature measurements. The two patients with traumatic brain injury showed well-formed circadian temperature rhythms and had more reflexive behaviours and relatively low cortical and sub-cortical atrophy, whereas the three patients from anoxic-hypoxic origin demonstrated no cycles or rhythmic behaviour. The presence of periods of wakefulness does not imply preserved sleep-wake cycling capacity, nor preserved circadian rhythms and it should not be taken as a distinguishing feature for the definition of the vegetative state.


Periconceptional undernutrition in sheep accelerates maturation of the fetal hypothalamic-pituitary-adrenal axis in late gestation.
We investigated the effects of moderate maternal periconceptional undernutrition from 60 d before to 30 d after mating on fetal hypothalamic-pituitary-adrenal axis function in late gestation. Ewes were sampled regularly during the period of undernutrition for circulating hormone levels. Vascular catheters were inserted into ewes and their singleton fetuses at 112 d gestation (term, 145 d), and fetal ACTH(1-24) and metyrapone challenge tests were performed at 127 and 128 d. Postmortems were performed at 132 d. Fetuses of undernourished ewes (UN, n = 12) had elevated baseline cortisol concentrations (P < 0.05), compared with fetuses of ad libitum-fed ewes (n = 10). There were no differences between groups in fetal responses to ACTH challenge, but only UN fetuses demonstrated ACTH and 11-deoxycortisol responses to metyrapone (P < 0.05). UN fetuses had increased mRNA levels for proopiomelanocortin and prohormone convertase-1, but not -2, in the pars intermedia of the pituitary gland (P < 0.05). Glucocorticoid receptor mRNA levels were not different between groups in pituitary or hypothalamus. Maternal cortisol and ACTH levels during undernutrition were profoundly suppressed (P < 0.001), rather than elevated, in UN ewes. Furthermore, the normal pregnancy rise in maternal serum progesterone concentrations was delayed in undernourished mothers. These data demonstrate that events around the time of conception have profound effects on fetal hypothalamic-pituitary-adrenal development in late gestation and that factors other than fetal exposure to excess glucocorticoids may be important.


Overexpression of a gene encoding a cytochrome P450, CYP78A9, induces large and seedless fruit in arabidopsis.
An activation tagging screen in which the cauliflower mosaic virus 35S enhancer was inserted randomly into an Arabidopsis genome homozygous for the floral homeotic mutation apetala2-1 (ap2-1) resulted in a line (28-5) with extraordinarily wide, heart-shaped ovaries. The ovary of the 28-5 ap2-1 mutant shows an oval shape because of increased numbers of enlarged cells. When the ap2-1 mutation is crossed out of the genetic background, more elongated rather than wider fruits are obtained. Normally, Arabidopsis fruits will develop to a normal size only when the ovules are present and fertilized. In the 28-5 single mutant, the siliques keep growing despite failure of fertilization and can reach nearly normal size. When wild-type pollen was used to pollinate the mutant pistil, the pollinated 28-5 silique became >10% longer and 40% wider than a wild-type silique, although producing very few seeds. The enhancer insertion in line 28-5 acts by hyperactivating a cytochrome P450 gene, CYP78A9. The pistil of 28-5 ap2-1 mutant flowers shows a structure similar to that of Capsella bursa-pastoris, a distant mustard relative of Arabidopsis, suggesting that the processes regulated by the CYP78A9-encoded protein may be involved in evolutionary control of carpel shape.


Optimum synthesis for time scale generation.
A time scale can be regarded as a synthesis of readings from precise clocks. Usually such synthesis is based on the principle of weighted averaging, which balances the contribution of each clock according to its noise level. It is well known that there are five different noise processes in precise clocks. Therefore, a good synthesis should balance each of those noise levels. Most existing algorithms control only one or two noise types. If an algorithm can control all five noise types simultaneously, we consider it to be optimum. The key point of constructing an optimum algorithm is the separation of all five noise types. In this paper, an optimum algorithm is presented using the half-integrating/half-differentiating model by which the five noise types are separated correctly. Performances of the new algorithm are demonstrated with simulated and real data.


Antibiotic biosynthesis following horizontal gene transfer: new milestone for novel natural product discovery?
Bacteria obtain a significant proportion of their genetic diversity via acquisition of DNA from distantly related organisms, a phenomenon known as horizontal gene transfer. The focus of horizontal gene transfer investigations has been primarily on the impact of this phenomenon on the ecological and/or pathogenic characteristics of bacterial species, with very little effort devoted to investigating horizontal gene transfer as a means of drug discovery. Here, we describe a novel approach to harness the power of horizontal gene transfer to produce novel chemotherapeutic molecules, a process that is easily scalable. We describe the state of the art in this field and discuss the current limiting factors associated with this phenomenon. Utilising a horizontal gene transfer method, we have identified and characterised a novel antimicrobial compound. Production of this antibiotic, termed rhodostreptomycin, is associated with the transfer of DNA from a species of Streptomyces to Rhodococcus by an as yet identified mechanism. We believe that horizontal gene transfer may represent the future of natural product discovery and engineering.


Phosphoenolpyruvate carboxylase kinase is a novel protein kinase regulated at the level of expression.
Phosphorylation of phosphoenolpyruvate carboxylase plays a key role in the control of plant metabolism. Phosphoenolpyruvate carboxylase kinase is a Ca2+-independent enzyme that is activated by a process involving protein synthesis in response to a range of signals in different plant tissues. The component whose synthesis is required for activation has not previously been identified, nor has the kinase been characterised at a molecular level. We report the cloning of phosphoenolpyruvate carboxylase kinase from the Crassulacean Acid Metabolism plant Kalanchoë fedtschenkoi and the C3 plant Arabidopsis thaliana. Surprisingly, phosphoenolpyruvate carboxylase kinase is a member of the Ca2+/calmodulin-regulated group of protein kinases. However, it lacks the auto-inhibitory region and EF hands of plant Ca2+-dependent protein kinases, explaining its Ca2+-independence. Its sequence is novel in that it comprises only a protein kinase catalytic domain with no regulatory regions; it appears to be the smallest known protein kinase. In K. fedtschenkoi, the abundance of phosphoenolpyruvate carboxylase kinase transcripts increases during leaf development. The transcript level in mature leaves is very low during the photoperiod, reaches a peak in the middle of the dark period and correlates with kinase activity. It exhibits a circadian oscillation in constant conditions. Protein kinases are typically regulated by second messengers, phosphorylation or protein/protein interactions. Phosphoenolpyruvate carboxylase kinase is an exception to this general rule, being controlled only at the level of expression. In K. fedtschenkoi, its expression is controlled both developmentally and by a circadian oscillator.


[Selective heterologous communication among human lung epithelial cells, fibroblasts, and lung cancer PG cells].
To further understand the roles of gap junctional intercellular communication in carcinogenesis and progression of human lung cancer. The heterologous communication was characterized among human lung epithelial cells HLEC, human lung fibroblasts HLF, human lung giant carcinoma PG cells and its Cx43 transfectants PG/C4 by using a preloading assay. Cx43 immunofluorescent staining and Northern blot were performed to examine Cx43 gene expression. Although both human lung fibroblasts HLF and epithelial cells HLEC expressed Cx43 and had very strong homologous communication, HLEC cells were unable to communicate with HLF cells. The human lung carcinoma cell line PG was defective of both homologous communication, and heterologous communication with its epithelial origin HLEC. Transfection of Cx43 into PG cells, which rescued PG cells' homologous communication and enhanced heterologous communication with HLF cells, could restore heterologous coupling with HLEC cells. Selective heterologous communication exists among different kinds of human lung cells, when human lung carcinoma cells lost coupling with its epithelial origin, Cx43 gene expression is not enough to establish heterologous communication between them.


Concomitant Arthroscopy With Labral Reconstruction and Periacetabular Osteotomy for Hip Dysplasia.
In the setting of true hip dysplasia, the high prevalence of intra-articular pathology may lead to recurrent symptoms and failure after periacetabular osteotomy (PAO). Femoral neck osteochondroplasty, microfracture, removal of loose bodies, and labral repair are examples of procedures that are performed with concomitant arthroscopy. When damage to the labrum is too severe to repair, reconstruction instead of extensive debridement before PAO can be more effective in restoring the labral seal to maintain joint lubrication and chondral protection. This Technical Note describes a method for concomitant hip arthroscopy with circumferential labral reconstruction with allograft and PAO.


A thermodynamic characterization of the interaction of 8-anilino-1-naphthalenesulfonic acid with native globular proteins: the effect of the ligand dimerization in the analysis of the binding isotherms.
8-Anilino-1-naphthalenesulfonic acid (ANS) is a popular fluorescence probe, broadly used for the analysis of proteins, but the nature of its interaction with proteins and the high increase in the fluorescence intensity that takes place upon such process are still unclear. In the last few years, isothermal titration calorimetry has been used to characterize the nature of the interaction of this dye with proteins. The analysis of the binding isotherms of these studies has not considered the dimerization equilibrium of ANS, which is pH dependent, and it can result in serious errors in the data analysis. In the present work we have developed a suitable data analysis by which this process is taken into account. To study the binding of the dye to proteins at different pH values, we have used the Abl-SH3 domain. Our results suggest that at pH 3 and 5, where the dimerization of the ANS is important, electrostatic interactions are significant for the binding of ANS to the Abl-SH3 domain. However, at pH 7, ANS behaves mostly as monomer and the interaction with the protein is mainly hydrophobic. The pH dependent behavior of the ANS binding to proteins can be explained in terms of ionization states of both, the protein and the ANS.


A Botanical Composition Mitigates Cartilage Degradations and Pain Sensitivity in Osteoarthritis Disease Model.
Osteoarthritis (OA) is a degenerative joint disease characterized by a progressive articular cartilage degradation manifested with significant functional impairment in consort with signs and symptoms of inflammation, stiffness, and loss of mobility. Current OA management is inadequate due to the lack of nominal therapies proven to be effective in hampering disease progression where symptomatic therapy focused intervention masks the primary etiology leading to irreversible structural damage. In this study, we describe the effect of UP1306, a composition containing a proprietary blend of two standardized extracts from the heartwood of Acacia catechu and the root bark of Morus alba, in maintaining joint structural integrity and alleviating OA associated symptoms in monosodium-iodoacetate (MIA)-induced rat OA disease model. Data from pain sensitivity, histopathology, and glycosaminoglycan (GAG) level were analyzed. Diclofenac at 10 mg/kg was used as a reference compound. Ex vivo proteoglycan protection model demonstrated 31.5%, 50.0%, and 54.8% inhibitions of proteoglycan degradations from UP1306 at concentrations of 50, 100, and 200 μg/mL, respectively. The merit of combining two bioflavonoid standardized extracts from A. catechu and M. alba was demonstrated in their Ex vivo synergistic proteoglycan protection activity. In the MIA in vivo OA model, administered orally at 500 mg/kg, UP1306 resulted in reductions of 17.5%, 29.0%, 34.4%, 33.5%, and 40.9% through week 1-5 in pain sensitivity, statistically significant improvements in articular cartilage matrix integrity, and minimal subchondral bone damage. Therefore, UP1306 could potentially be considered as an alternative remedy from natural sources for the management of OA and/or its associated symptoms.


Handedness and its genetic influences are associated with structural asymmetries of the cerebral cortex in 31,864 individuals.
Roughly 10% of the human population is left-handed, and this rate is increased in some brain-related disorders. The neuroanatomical correlates of hand preference have remained equivocal. We resampled structural brain image data from 28,802 right-handers and 3,062 left-handers (UK Biobank population dataset) to a symmetrical surface template, and mapped asymmetries for each of 8,681 vertices across the cerebral cortex in each individual. Left-handers compared to right-handers showed average differences of surface area asymmetry within the fusiform cortex, the anterior insula, the anterior middle cingulate cortex, and the precentral cortex. Meta-analyzed functional imaging data implicated these regions in executive functions and language. Polygenic disposition to left-handedness was associated with two of these regional asymmetries, and 18 loci previously linked with left-handedness by genome-wide screening showed associations with one or more of these asymmetries. Implicated genes included six encoding microtubule-related proteins: TUBB, TUBA1B, TUBB3, TUBB4A, MAP2, and NME7-mutations in the latter can cause left to right reversal of the visceral organs. There were also two cortical regions where average thickness asymmetry was altered in left-handedness: on the postcentral gyrus and the inferior occipital cortex, functionally annotated with hand sensorimotor and visual roles. These cortical thickness asymmetries were not heritable. Heritable surface area asymmetries of language-related regions may link the etiologies of hand preference and language, whereas nonheritable asymmetries of sensorimotor cortex may manifest as consequences of hand preference.


VAR2CSA Serology to Detect Plasmodium falciparum Transmission Patterns in Pregnancy.
Pregnant women constitute a promising sentinel group for continuous monitoring of malaria transmission. To identify antibody signatures of recent Plasmodium falciparum exposure during pregnancy, we dissected IgG responses against VAR2CSA, the parasite antigen that mediates placental sequestration. We used a multiplex peptide-based suspension array in 2,354 samples from pregnant women from Mozambique, Benin, Kenya, Gabon, Tanzania, and Spain. Two VAR2CSA peptides of limited polymorphism were immunogenic and targeted by IgG responses readily boosted during infection and with estimated half-lives of <2 years. Seroprevalence against these peptides reflected declines and rebounds of transmission in southern Mozambique during 2004-2012, reduced exposure associated with use of preventive measures during pregnancy, and local clusters of transmission that were missed by detection of P. falciparum infections. These data suggest that VAR2CSA serology can provide a useful adjunct for the fine-scale estimation of the malaria burden among pregnant women over time and space.


Spatiotemporal land use regression models of fine, ultrafine, and black carbon particulate matter in New Delhi, India.
Air pollution in New Delhi, India, is a significant environmental and health concern. To assess determinants of variability in air pollutant concentrations, we develop land use regression (LUR) models for fine particulate matter (PM2.5), black carbon (BC), and ultrafine particle number concentrations (UFPN). We used 136 h (39 sites), 112 h (26 sites), 147 h (39 sites) of PM2.5, BC, and UFPN data respectively, to develop separate morning (0800-1200) and afternoon (1200-1800) models. Continuous measurements of PM2.5 and BC were also made at a single fixed rooftop site located in a high-income residential neighborhood. No continuous measurements of UFPN were available. In addition to spatial variables, measurements from the fixed continuous monitoring site were used as independent variables in the PM2.5 and BC models. The median concentrations (and interquartile range) of PM2.5, BC, and UFPN at LUR sites were 133 (96-232) μg m(-3), 11 (6-21) μg m(-3), and 40 (27-72) × 10(3) cm(-3) respectively. In addition (a) for PM2.5 and BC, the temporal variability was higher than the spatial variability; (b) the magnitude and spatial variability in pollutant concentrations was higher during morning than during afternoon hours. Further, model R(2) values were higher for morning (for PM2.5, BC, and UFPN, respectively: 0.85, 0.86, and 0.28) than for afternoon models (0.73, 0.69, and 0.23); (c) the PM2.5 and BC concentrations measured at LUR sites all over the city were strongly correlated with measured concentrations at a fixed rooftop site; (d) spatial patterns were similar for PM2.5 and BC but different for UFPN; (e) population density and road variables were statistically significant predictors of pollutant concentrations; and (f) available geographic predictors explained a much lower proportion of variability in measured PM2.5, BC, and UFPN than observed in other LUR studies, indicating the importance of temporal variability and suggesting the existence of uncharacterized sources.


Dysfunction in the βII spectrin-dependent cytoskeleton underlies human arrhythmia.
The cardiac cytoskeleton plays key roles in maintaining myocyte structural integrity in health and disease. In fact, human mutations in cardiac cytoskeletal elements are tightly linked to cardiac pathologies, including myopathies, aortopathies, and dystrophies. Conversely, the link between cytoskeletal protein dysfunction and cardiac electric activity is not well understood and often overlooked in the cardiac arrhythmia field. Here, we uncover a new mechanism for the regulation of cardiac membrane excitability. We report that βII spectrin, an actin-associated molecule, is essential for the posttranslational targeting and localization of critical membrane proteins in heart. βII spectrin recruits ankyrin-B to the cardiac dyad, and a novel human mutation in the ankyrin-B gene disrupts the ankyrin-B/βII spectrin interaction, leading to severe human arrhythmia phenotypes. Mice lacking cardiac βII spectrin display lethal arrhythmias, aberrant electric and calcium handling phenotypes, and abnormal expression/localization of cardiac membrane proteins. Mechanistically, βII spectrin regulates the localization of cytoskeletal and plasma membrane/sarcoplasmic reticulum protein complexes, including the Na/Ca exchanger, ryanodine receptor 2, ankyrin-B, actin, and αII spectrin. Finally, we observe accelerated heart failure phenotypes in βII spectrin-deficient mice. Our findings identify βII spectrin as critical for normal myocyte electric activity, link this molecule to human disease, and provide new insight into the mechanisms underlying cardiac myocyte biology.


Comparative analysis of the efficacy and functionality of abdominoplasty versus minimally invasive techniques in the surgical treatment of diastasis rectus abdominis in postpartum women.
This review aims to describe the surgical options available for treatment of diastasis recti in postpartum women, as well as compare the effectiveness of these treatment options. Historically, diastasis recti has been repaired through open procedures, such as abdominoplasty. More recently, studies have explored other methods for the treatment of diastasis recti, including various minimally invasive surgical options. Twelve studies ranging from 2015 to 2022 were included in this analysis. Studies were identified using PubMed, EMBASE, and Cochrane Library. Studies that met the inclusion criteria were analyzed descriptively. Statistical comparison of surgical outcomes between studies was performed using Fisher's Exact Test in SPSS. Minimally invasive approaches were categorized as laparoscopic preaponeurotic approaches, robotic approaches, and enhanced-view/extended totally extraperitoneal (eTEP) approaches. These techniques were compared to two open approaches: abdominoplasty and miniabdominoplasty. There were no significant differences in the rate of seromas, surgical site infections/complications, or hematomas between abdominoplasty and minimally invasive surgical techniques (p > 0.05). Among the minimally invasive techniques, no significant differences in readmission rates were reported (p > 0.05). Additionally, no significant differences in recurrence rates were seen following minimally invasive or abdominoplasty repairs, except for the increased recurrence rates seen with the r-TARRD robotic technique (p < 0.05). Although current data on minimally invasive approaches is limited, our review reveals that both open and minimally invasive approaches are viable options for diastasis recti repair in postpartum women. Identifying the optimal approach for diastasis recti repair should rely on the patient's desired treatment outcome. If the patient indicates a desire for the removal of excess abdominal subcutaneous tissue, abdominoplasty may be a better surgical approach. Alternatively, if the patient puts a greater emphasis on shorter recovery time and smaller surgical incisions/scars, minimally invasive approaches may be a better surgical option.


Competitive fitness and homologous recombination of SARS-CoV-2 variants of concern.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants continue to emerge and cocirculate in humans and wild animals. The factors driving the emergence and replacement of novel variants and recombinants remain incompletely understood. Herein, we comprehensively characterized the competitive fitness of SARS-CoV-2 wild type (WT) and three variants of concern (VOCs), Alpha, Beta and Delta, by coinfection and serial passaging assays in different susceptible cells. Deep sequencing analyses revealed cell-specific competitive fitness: the Beta variant showed enhanced replication fitness during serial passage in Caco-2 cells, whereas the WT and Alpha variant showed elevated fitness in Vero E6 cells. Interestingly, a high level of neutralizing antibody sped up competition and completely reshaped the fitness advantages of different variants. More importantly, single clone purification identified a significant proportion of homologous recombinants that emerged during the passage history, and immune pressure reduced the frequency of recombination. Interestingly, a recombination hot region located between nucleotide sites 22,995 and 28,866 of the viral genomes could be identified in most of the detected recombinants. Our study not only profiled the variable competitive fitness of SARS-CoV-2 under different conditions, but also provided direct experimental evidence of homologous recombination between SARS-CoV-2 viruses, as well as a model for investigating SARS-CoV-2 recombination.


Evaluation of nuclear factor κB and chemokine receptor CXCR4 co-expression in patients with prostate cancer in the Radiation Therapy Oncology Group (RTOG) 8610.
To determine the frequency of nuclear factor κB (NFκB) and the chemokine receptor CXCR4 co-expression in prostate cancer specimens from men with locally advanced disease. Paraffin-embedded samples from patients enrolled on the Radiation Therapy Oncology Group (RTOG) 8610 trial underwent immunohistochemical staining for NFκB and CXCR4. The amount of NFκB and CXCR4 was scored by a 'blinded' pathologist for the percentage of cells stained (0-100%) and staining intensity (0-3 +). Cox proportional hazard models were used for overall survival and disease-free survival to examine if NFκB and/or CXCR4 expression were associated with patient outcomes with and without adjustment for covariates. Available material and successful staining allowed NFκB and CXCR4 status to be determined for 55 and 63 patients, respectively. Both NFκB and CXCR4 status were available for 51 patients. Of these, 53% were 2/3 + for cytoplasmic NFκB staining and 56% were 2/3 + for CXCR4. In all, 18 of the 51 patients were 2/3 + for both NFκB and CXCR4 (P = 0.129). Ten of 11 patients with 3 + NFκB had 2/3 + CXCR4 (P= 0.004). In this small study, neither NFκB nor CXCR4 were associated with prostate cancer outcomes. High NFκB expression is associated with CXCR4 expression and they are co-expressed in about one third of patients with clinically localized prostate cancer. Larger studies to accurately determine the frequency of co-expression and prognostic utility of NFκB and CXCR4 alone and in combination are warranted.


Population genomic response to geographic gradients by widespread and endemic fishes of the Arabian Peninsula.
Genetic structure within marine species may be driven by local adaptation to their environment, or alternatively by historical processes, such as geographic isolation. The gulfs and seas bordering the Arabian Peninsula offer an ideal setting to examine connectivity patterns in coral reef fishes with respect to environmental gradients and vicariance. The Red Sea is characterized by a unique marine fauna, historical periods of desiccation and isolation, as well as environmental gradients in salinity, temperature, and primary productivity that vary both by latitude and by season. The adjacent Arabian Sea is characterized by a sharper environmental gradient, ranging from extensive coral cover and warm temperatures in the southwest, to sparse coral cover, cooler temperatures, and seasonal upwelling in the northeast. Reef fish, however, are not confined to these seas, with some Red Sea fishes extending varying distances into the northern Arabian Sea, while their pelagic larvae are presumably capable of much greater dispersal. These species must therefore cope with a diversity of conditions that invoke the possibility of steep clines in natural selection. Here, we test for genetic structure in two widespread reef fish species (a butterflyfish and surgeonfish) and eight range-restricted butterflyfishes across the Red Sea and Arabian Sea using genome-wide single nucleotide polymorphisms. We performed multiple matrix regression with randomization analyses on genetic distances for all species, as well as reconstructed scenarios for population subdivision in the species with signatures of isolation. We found that (a) widespread species displayed more genetic subdivision than regional endemics and (b) this genetic structure was not correlated with contemporary environmental parameters but instead may reflect historical events. We propose that the endemic species may be adapted to a diversity of local conditions, but the widespread species are instead subject to ecological filtering where different combinations of genotypes persist under divergent ecological regimes.


Specific down-regulation of parathyroid hormone (PTH) receptors and responses to PTH by tumour necrosis factor alpha and retinoic acid in UMR 106-06 osteoblast-like osteosarcoma cells.
Parathyroid hormone (PTH) and PTH-related protein (PTHrP) act via PTH receptors in bone to stimulate bone resorption. Bone resorption is also stimulated by certain cytokines, which are produced in bone and bone marrow. The effects of such cytokines on the PTH-receptor system were studied in the osteoblast-like osteosarcoma cell line UMR 106-06. 125I-labelled PTHrP-(1-84)-peptide bound specifically to the cells, and PTHrP-(1-34) and -(1-84) competed with equimolar affinity for binding to UMR 106-06 cells. The specific binding of 125I-PTHrP-(1-84) could be completely blocked by PTH. Therefore 125I-PTHrP-(1-84) bound to a classical receptor in UMR 106-06 cells. Preincubation for 3 days with either tumour necrosis factor alpha (TNF alpha) or retinoic acid (RA) both decreased the specific binding of 125I-PTHrP-(1-84) to about 40% of control levels. These effects were specific for PTH binding, since there was little effect on 125I-salmon-calcitonin binding. Both TNF alpha and RA required 24 h exposure to cells to produce a measurable effect. The decrease in 125I-PTHrP-(1-84) binding was due to a reduced number of binding sites, with little apparent change in affinity. Half-maximal effects were seen with 1 ng of TNF alpha/ml, whereas 1 microM-RA was needed to observe the loss of PTH receptors. Combinations of RA and TNF alpha produced a greater effect than that of either agonist alone. The loss of PTH receptors was accompanied by a specific loss of PTH-stimulated cyclic AMP production. Preincubation with TNF alpha increased the basal plasminogen activator (PA) activity in the cells and decreased the amplitude of the response of PA activity to PTH compared with control cells. Furthermore TNF alpha decreased sensitivity to PTH (50% stimulation of PA activity with 0.1 nM-PTH in control cells versus 50% stimulation with 0.3 nM-PTH in TNF alpha-treated cells). In contrast, TNF alpha pretreatment increased the amplitude of the response of PA activity to calcitonin, whereas sensitivity to calcitonin was not altered. These data are consistent with a specific down-regulation of PTH receptors in osteoblast-like UMR 106-06 cells after exposure to TNF alpha or RA. The loss of PTH receptors is accompanied by a decreased responsiveness to PTH, as measured with the PA system in these cells. A loss of PTH receptors could modulate PTH responses in osteoblasts, either in the local control of bone formation and resorption, or in pathological conditions such as humoral hypercalcaemia of malignancy.


STAT3 Activation Impairs the Stability of Th9 Cells.
Th9 cells regulate multiple immune responses, including immunity to pathogens and tumors, allergic inflammation, and autoimmunity. Despite ongoing research into Th9 development and function, little is known about the stability of the Th9 phenotype. In this study, we demonstrate that IL-9 production is progressively lost in Th9 cultures during several rounds of differentiation. The loss of IL-9 is not due to an outgrowth of cells that do not secrete IL-9, as purified IL-9 secretors demonstrate the same loss of IL-9 in subsequent rounds of differentiation. The loss of IL-9 production correlates with increases in phospho-STAT3 levels within the cell, as well as the production of IL-10. STAT3-deficient Th9 cells have increased IL-9 production that is maintained for longer in culture than IL-9 in control cultures. IL-10 is responsible for STAT3 activation during the first round of differentiation, and it contributes to instability in subsequent rounds of culture. Taken together, our results indicate that environmental cues dictate the instability of the Th9 phenotype, and they suggest approaches to enhance Th9 activity in beneficial immune responses.


B and T cells in a cat with thymic lymphosarcoma.
The mixed antiglobulin reaction and the formation of nonimmune rosettes with guinea-pig red blood cells (RBC) distinguished feline B and T cells, respectively. In a cat with thymic lymphosarcoma, the cells reacting in these tests formed separate, nonoverlapping populations. The malignant cells were large lymphoblasts replacing the normal thymus and infiltrating local lymph nodes, where they localized only in the paracortical, i.e., thymus-dependent areas. Cells from the nodes could therefore be identified as malignant or normal by their size. The mixed antiglobulin reaction showed that the malignant cells did not carry the surface Ig characteristic of B cells, whereas these malignant cells formed nonimmune rosettes with guinea-pig RBC. Among lymph node cells, most surviving normal and small lymphocytes, from outside the thymus-dependent areas, reacted as B cells. The morphologic evidence therefore corroborated the test results, which indicated that the formation of rosettes with guinea-pig RBC seems a reliable means for the demonstration of T cells in the cat.


Analyzing Cross-country Pandemic Connectedness During COVID-19 Using a Spatial-Temporal Database: Network Analysis.
Communicable diseases including COVID-19 pose a major threat to public health worldwide. To curb the spread of communicable diseases effectively, timely surveillance and prediction of the risk of pandemics are essential. The aim of this study is to analyze free and publicly available data to construct useful travel data records for network statistics other than common descriptive statistics. This study describes analytical findings of time-series plots and spatial-temporal maps to illustrate or visualize pandemic connectedness. We analyzed data retrieved from the web-based Collaborative Arrangement for the Prevention and Management of Public Health Events in Civil Aviation dashboard, which contains up-to-date and comprehensive meta-information on civil flights from 193 national governments in accordance with the airport, country, city, latitude, and the longitude of flight origin and the destination. We used the database to visualize pandemic connectedness through the workflow of travel data collection, network construction, data aggregation, travel statistics calculation, and visualization with time-series plots and spatial-temporal maps. We observed similar patterns in the time-series plots of worldwide daily flights from January to early-March of 2019 and 2020. A sharp reduction in the number of daily flights recorded in mid-March 2020 was likely related to large-scale air travel restrictions owing to the COVID-19 pandemic. The levels of connectedness between places are strong indicators of the risk of a pandemic. Since the initial reports of COVID-19 cases worldwide, a high network density and reciprocity in early-March 2020 served as early signals of the COVID-19 pandemic and were associated with the rapid increase in COVID-19 cases in mid-March 2020. The spatial-temporal map of connectedness in Europe on March 13, 2020, shows the highest level of connectedness among European countries, which reflected severe outbreaks of COVID-19 in late March and early April of 2020. As a quality control measure, we used the aggregated numbers of international flights from April to October 2020 to compare the number of international flights officially reported by the International Civil Aviation Organization with the data collected from the Collaborative Arrangement for the Prevention and Management of Public Health Events in Civil Aviation dashboard, and we observed high consistency between the 2 data sets. The flexible design of the database provides users access to network connectedness at different periods, places, and spatial levels through various network statistics calculation methods in accordance with their needs. The analysis can facilitate early recognition of the risk of a current communicable disease pandemic and newly emerging communicable diseases in the future.


[Interference of hemolysis on haptoglobin determined by kinetic immunonephelometry and comparison between phenotypes].
The aim of the study was to check if a slight and non visible hemolysis to naked eye such as that induced by blood coagulation could interfere in the immunonephelometric measurement of haptoglobin, and if this interference was dependent on the protein phenotype. Results confirmed that blood coagulation induced a non visible hemolysis whose intensity markedly varied from one specimen to another. Under our conditions (kinetic measurement with a Beckman Coulter immunonephelometer), we observed with the sera a negative interference linked to the hemolysis induced by blood coagulation when compared to corresponding plasmas obtained with lithium heparinate (p < 0,005). It was checked also that this anticoagulant did not induce a positive interference. Hemolysis interference was not found phenotype dependent. These results lead us to recommend to perform haptoglobin measurements on heparinised plasmas.


Comparative in vitro activities of amoxicillin-clavulanic acid, cefuroxime, cephalexin, and cephalothin against trimethoprim-resistant Escherichia coli isolated from stools of children attending day-care centers.
A high prevalence of fecal colonization with trimethoprim-resistant Escherichia coli was found in diapered children attending day-care centers in Houston, Tex. In the present study, 100 isolates of E. coli resistant to multiple antibiotics, including trimethoprim (100%), sulfisoxazole (100%), streptomycin (94%), and ampicillin (87%), were obtained over a 5-month period from stool samples of diapered children attending four day-care centers and tested for their susceptibilities to amoxicillin-clavulanic acid, cefuroxime, cephalexin, and cephalothin. The MICs for 50 and 90% of strains tested were 16 and 32 micrograms/ml, respectively, for amoxicillin-clavulanic acid, 4 and 16 micrograms/ml, respectively, for cefuroxime, 4 and 64 micrograms/ml, respectively, for cephalexin, and 32 and greater than 64 micrograms/ml, respectively, for cephalothin. Although all three oral beta-lactams tested were generally active at concentrations likely to be achieved in urine, cefuroxime and cephalexin were more potent and are thus more likely to be inhibitory at the concentrations needed for systemic infections.


Vitamin C Attenuates Chronic Chlorpyrifos-induced Alteration of Neurobehavioral Parameters in Wistar Rats.
Oxidative stress is one of the molecular mechanisms in chlorpyrifos toxicity. The present study was designed to evaluate the attenuating effect of vitamin C on chlorpyrifos-induced alteration of neurobehavioral performance and the role of muscle acetylchloinesterase (AChE), glycogen and lipoperoxidation in the accomplishment of this task. Male rats were randomly assigned into 4 groups with the following regimens: soya oil (S/oil), vitamin C (VC), chlorpyrifos (CPF) and vitamin C+CPF (VC+CPF). The regimens were administered by gavage once daily for a period of 17 weeks. Neurobehavioral parameters measuring efficiency of locomotion, motor strength, righting reflex and excitability were evaluated at day 0 (pretreatment value), weeks 8 and 16. The rats were sacrificed at week 17 and evaluated for muscle glycogen and malonaldehyde (MDA) concentrations and AChE activity. The result showed that deficits in locomotion efficiency, motor strength, righting reflex and excitability score induced by chronic CPF were mitigated but not completely abolished by vitamin C. The reduced muscle AChE activity and concentrations of glycogen and MDA evoked by chronic CPF were ameliorated by vitamin C. The study therefore showed that improvement in muscle AChE activity, glycogen concentration and reduced lipoperoxidation by vitamin C may be partly responsible for the mitigation of the chronic CPF-induced sensorimotor performance.