Intestinal hemodynamics in dogs with chronic one-kidney, one-wrapped hypertension.
In pentobarbital-anesthetized dogs with chronic one-kidney, one-wrapped perinephritic hypertension, the status of blood flow and resistance differs among vascular beds. There is normal blood flow with elevated resistance in the limb and elevated blood flow with normal resistance in the ileum. In the present study we investigated the mechanism of elevated flow and normal resistance in the ileum by measuring blood flow, pressure, O2 consumption, reactive hyperemia, pressure-flow relationships, resistance at maximal vasodilation, and nonentrapment (shunting) of 9-microns microspheres in vascularly isolated, blood-perfused ilea of 11 24-h fasted dogs with chronic (greater than 4 wk duration) one-kidney perinephritic hypertension [1KH; PA = 163 +/- 8 (SD) mmHg] and 15 fasted normotensive one-kidney, sham-wrapped dogs (1KC; PA = 108 +/- 7 mmHg). In 1KH ileal blood flow was elevated by 35% (P less than 0.001), but ileal vascular resistance was unchanged. Pressure-flow relationships were similar in hypertensive and normotensive ilea, with little evidence for significant autoregulation in either. We also noted no significant differences in ileal O2 consumption, reactive hyperemia, or resistance at maximal vasodilation. However, we found nonentrapment of microspheres to be a positive function of perfusion pressure (P less than 0.02) and to be increased by 40% during ileal perfusion at levels of perfusion pressure within the hypertensive range (150-200 mmHg). Thus the increased ileal blood flow in 1KH may reflect the effects of a poorly autoregulating tissue, as well as "shunt" flow through pressure-activated channels.(ABSTRACT TRUNCATED AT 250 WORDS)


Estimating toxicity-related biological pathway altering doses for high-throughput chemical risk assessment.
We describe a framework for estimating the human dose at which a chemical significantly alters a biological pathway in vivo, making use of in vitro assay data and an in vitro-derived pharmacokinetic model, coupled with estimates of population variability and uncertainty. The quantity we calculate, the biological pathway altering dose (BPAD), is analogous to current risk assessment metrics in that it combines dose-response data with analysis of uncertainty and population variability to arrive at conservative exposure limits. The analogy is closest when perturbation of a pathway is a key event in the mode of action (MOA) leading to a specified adverse outcome. Because BPADs are derived from relatively inexpensive, high-throughput screening (HTS) in vitro data, this approach can be applied to high-throughput risk assessments (HTRA) for thousands of data-poor environmental chemicals. We envisage the first step of HTRA to be an assessment of in vitro concentration-response relationships across biologically important pathways to derive biological pathway altering concentrations (BPAC). Pharmacokinetic (PK) modeling is then used to estimate the in vivo doses required to achieve the BPACs in the blood at steady state. Uncertainty and variability are incorporated in both the BPAC and the PK parameters and then combined to yield a probability distribution for the dose required to perturb the critical pathway. We finally define the BPADL as the lower confidence bound of this pathway-altering dose. This perspective outlines a framework for using HTRA to estimate BPAD values; provides examples of the use of this approach, including a comparison of BPAD values with published dose-response data from in vivo studies; and discusses challenges and alternative formulations.


Aspergillus lentulus sp. nov., a new sibling species of A. fumigatus.
In a prior study, we identified seven clinical isolates of an Aspergillus sp. that were slow to sporulate in multiple media and demonstrated decreased in vitro susceptibilities to multiple antifungals, including amphotericin B, itraconazole, voriconazole, and caspofungin. These isolates were initially considered to be variants of Aspergillus fumigatus because of differences in mitochondrial cytochrome b sequences and unique randomly amplified polymorphic DNA PCR patterns (S. A. Balajee, M. Weaver, A. Imhof, J. Gribskov, and K. A. Marr, Antimicrob. Agents Chemother. 48: 1197-1203, 2004). The present study was performed to clarify the taxonomic status of these organisms by phylogenetic analyses based on multilocus sequence typing of five genes (the beta-tubulin gene, the rodlet A gene, the salt-responsive gene, the mitochondrial cytochrome b gene, and the internal transcribed spacer regions). Results revealed that four of the seven variant isolates clustered together in a clade very distant from A. fumigatus and distinct from other members of the A. fumigatus group. This new clade, consisting of four members, was monophyletic with strong bootstrap support when the protein-encoding regions were analyzed, indicating a new species status under the phylogenetic species concept. Phenotype studies revealed that the variant isolate has smaller conidial heads with diminutive vesicles compared to A. fumigatus and is not able to survive at 48 degrees C. Our findings suggest the presence of a previously unrecognized, potentially drug-resistant Aspergillus species that we designate A. lentulus.


OSA and atherosclerosis.
Untreated obstructive sleep apnea (OSA) is increasingly recognized as a risk factor contributing to cardiovascular morbidity and mortality. Research in recent decades has uncovered many components of the complex pathological events leading to the atherosclerotic vascular diseases in OSA, which involve heightened oxidative stress as a result of intermittent hypoxia, vascular inflammation, activation of platelet and coagulation cascades, endothelial dysfunction and ultimately the formation of atherosclerotic plagues. The close association of OSA and conventional cardiovascular risk factors including hypertension, diabetes mellitus, dyslipidemia and obesity adds to the adverse cardiovascular sequelae. Further studies are required to clarify further on the pathophysiological processes, and the effect size of OSA therapy, and other potential preventive strategies.


Eurythermic and stenothermic growth of cultured fish cells and their thermosensitivity.
RBCF-1 cells derived from the caudal fin of the goldfish (Carassius auratus) have been established at 37 degrees C, and could grow at a wide range of temperatures (20-37 degrees C). To examine the mechanism(s) of this eurythermic property, we isolated clones from RBCF-1 cells with different optimal growth temperatures. R37 clone grew continuously at 37 degrees C but could not grow at 27 degrees C. R27 clone grew continuously at 27 degrees C but could not grow at 37 degrees C. Thus R37 and R27 cells may be referred to as stenothermic. The R3727 clone, which was derived from R37 cells selected at 27 degrees C, and the cell hybrids of R37 and R27 cells, could proliferate for a long time and form colonies at 27-37 degrees C, like progenitor RBCF-1 cells, so they may be referred to as eurythermic. The shape of colonies of R3727 cells changed with incubation temperature; they resembled those of R37 cells at a higher temperature (37 degrees C), while at a lower temperature, (27 degrees C), they were like those of R27 cells. The lethal effects of heat treatment were compared among RBCF-1 clones. R27 cells were most sensitive, and R37 cells were most resistant. The heat treatment could induce the four major heat-shock proteins (hsp30, hsp42, hsp70 and hsp90). Marked induction of hsp70 and hsp30 was observed at 33 degrees C for R27, at 37 degrees C for R3727 and at 40 degrees C for R37. With R3727 cells, induction of hsps and thermosensitivity were highly dependent on the incubation temperature before heat treatment.


Strain-specific SCAR markers for the detection of Trichoderma harzianum AS12-2, a biological control agent against Rhizoctonia solani, the causal agent of rice sheath blight.
In order to identify a specific marker for T. harzianum AS12-2, a strain capable of controlling rice sheath blight caused by Rhizoctonia solani, UP-PCR was performed using five universal primers (UP) both separately and in pairwise combinations. The application of two UP primers resulted in the amplification of unique fragments from the genomic DNA of T. harzianum AS12-2, clearly distinguishing it from other Trichoderma strains. The unique fragments had no significant sequence homology with any other known sequence available in databases. Based on the sequences of the unique fragments, 14 oligonucleotide primers were designed. Two primer sets amplified a fragment of expected size from the DNA of strain T. harzianum AS12-2 but not from any other examined strains belonging to T. harzianum, to other Trichoderma species assayed, or to other common fungi present in paddy fields of Mazandaran province, Iran. In conclusion, SCAR (sequence characterized amplified regions) markers were successfully identified and rapid, reliable tools were provided for the detection of an effective biocontrol Trichoderma strain, which can facilitate studies of its population dynamics and establishment after release into the natural environment.


Redox-Mediated pH Swing Systems for Electrochemical Carbon Capture.
ConspectusThe rising levels of atmospheric CO2 and their resulting impacts on the climate have necessitated the urgent development of effective carbon capture technologies. Electrochemical carbon capture systems have emerged as a potential alternative to conventional thermal systems based on amine solutions due to their modularity, energy efficiency, and lower environmental impact. Among these, aqueous electrochemical pH swing systems that capitalize on the pH dependence of dissolved inorganic carbon (CO2/HCO3-/CO32-) speciation to capture and release CO2 are of particular interest as they can be flexible in system design and in the range of electrochemical potentials used as well as being environmentally benign. In this Account, we present our recent findings in pH swing-based electrochemical carbon capture using redox-active materials, paving the way toward a sustainable solution for mitigating CO2 emissions.In the first section, we discuss the utilization of molecular redox-active organic materials in electrochemical carbon capture by the pH swing method. This electrochemical system configuration involves homogeneous aqueous electrolytes containing molecular redox-active compounds combined with inert carbon-based electrodes. We first present the development of redox-active amine and oxygen-insensitive neutral red (NR)-based systems. Notably, the discovery of 1-aminopyridinium (1-AP) as an electrochemically reversible compound enables efficient pH swing, leading to an impressive electron utilization of 1.25 mol of CO2 per mole of electrons. Additionally, we explore an oxygen-insensitive neutral red/leuconeutral red (NR/NRH2) redox system, which demonstrates potential applicability to direct air capture (DAC) systems with ambient air as a feed gas.The second section focuses on the utilization of inorganic nanomaterials for redox-active electrodes for pH swing-based electrochemical carbon capture. In this system configuration, we employ redox-active electrodes for inducing reversible pH swings in aqueous electrolytes without interrupting other ionic species, except protons. Specifically, we explore the effectiveness of manganese oxide (MnO2) electrodes for achieving selective CO2 removal from simulated flue gas. We then demonstrate a bismuth/silver (Bi/BiOCl, Ag/AgCl) nanoparticle electrode system as a sodium-insensitive pH swing system for extracting dissolved inorganic carbon (DIC) from simulated seawater with high electrochemical energy efficiency.Overall, these advances in pH swing-based electrochemical carbon capture offer promising preliminary solutions for combating climate change by capturing CO2 from dilute sources such as flue gas and ambient air as well as enabling direct carbon removal from ocean water. While these systems have demonstrated impressive energy efficiency and environmental benefits using redox-active materials, they represent only the beginning of our research journey. Further development and optimization are currently underway as we strive to unlock their full potential for large-scale implementation, paving the way toward a sustainable and carbon-neutral future.


Management of advanced metacarpal aneurysmal cysts: Surgical technique.
The hand is a rare site for aneurysmal bone cysts (ABCs). Depending on their degree of invasiveness, treatment of ABCs ranges from medical therapy to surgical curettage and bone filling. In the most advanced cases, bone resection and reconstruction, with or without adjuvant or neoadjuvant treatment, are indicated. We describe a technique involving segmental bone resection and structural iliac graft reconstruction with carpo-metacarpal plate arthrodesis, for the management of advanced ABC involving the base of the metacarpals. The radio-clinical results of two patients operated on using this technique are presented, at 18 and 36 months. At the last follow-up, the results showed a QDASH of 15 and 10, and a PRWE of 9 and 11. Satisfaction was 10/10 in both cases. Grasp strength was 25 kg and 42 kg versus 28 kg and 40 kg on the opposite side. Pinch strength was 6 kg and 11 kg versus 7 kg and 10 kg. Metacarpophalangeal flexion was 80° and 90°. Extension was complete in both cases. Radiographs showed good graft fusion, with no lysis or signs of ABC recurrence at the last follow-up. Segmental bone resection and iliac autograft reconstruction with carpo-metacarpal plate arthrodesis represents a therapeutic option in the management of advanced ABCs of the metacarpal base.


[Children's oral hygiene: healthy mouth, families supervise?].
Dental caries results from a complex interaction between the host and environmental factors and it is an important public health issue. To determine the prevalence of dental caries in a school population of six and 12 years old children from Leiria; to establish a relationship between dental caries and related known risk factors; parental dental health care, parental control of their children oral hygiene and to compare our results with the National Dental Care Study of 1999. Descriptive and statistical analysis based on a questionnaire and dental examination. In our sample of 248 children, 43% were six years old and 57% 12 years old; 52% were female and 72% reside in a suburban area. The prevalence of dental caries was 42% (48% for the six year and 33% for the 12 years old groups. Dental caries were more prevalent in males (p = 0, 01) and 25% of these had three or more caries. Analysis of the questionnaire revealed that 87% of children brushed their teeth daily, findings that were unrelated to gender, age or residential area. Of this group, 68% brushed their teeth twice or more daily (p = 0,008). This routine was commenced in 32% of children prior to the age of three and these had less dental caries (p = 0,022). With regard to the children's dietary habits, we found them to be similar in both groups. The children who did not sweeten their milk (23 and 24% of the six and 12 years group, respectively) had fewer dental caries (p = 0,031). Dental treatment was required in 53% of 12 year old group and in 41% of six year old group. Dental caries was also less prevalent in children who also had taken fluoride (p = 0,045). We found a significant statistical relationship between parental and their children's dietary habits (p = 0,000). Parents who brushed their teeth twice daily had children with similar oral hygiene habits and had fewer caries (p = 0,002). Parents who oversee their children's dental care (52%) include the group of parents and children that brush their teeth at least twice daily (p = 0,003). Dental caries was more prevalent in the six year old group with a male predominance. Children who brushed their teeth twice a day had less caries and the 12 year old group had more visits to the dentist. To ensure adequate oral hygiene habits it is important that parents supervise their children' habits.


Phylogenetic position and taxonomy of Kusaghiporia usambarensis gen. et sp. nov. (Polyporales).
A large polyporoid mushroom from the West Usambara Mountains in North-eastern Tanzania produces dark brown, up to 60-cm large fruiting bodies that at maturity may weigh more than 10 kg. It has a high rate of mycelial growth and regeneration and was found growing on both dry and green leaves of shrubs; attached to the base of living trees, and it was also observed to degrade dead snakes and insects accidentally coming into contact with it. Phylogenetic analyses based on individual and concatenated data sets of nrLSU, nrSSU and the RPB2 and TEF1 genes showed it, together with Laetiporus, Phaeolus, Pycnoporellus and Wolfiporia, to form a monophyletic group in Polyporales. Based on morphological features and molecular data, it is described as Kusaghiporia usambarensis.


Frequent silencing of a putative tumor suppressor gene melatonin receptor 1 A (MTNR1A) in oral squamous-cell carcinoma.
Array-based comparative genomic hybridization (array-CGH) has good potential for the high-throughput identification of genetic aberrations in cell genomes. In the course of a program to screen a panel of 21 oral squamous-cell carcinoma (OSCC) cell lines for genome-wide copy-number aberrations by array-CGH using our in-house bacterial artificial chromosome arrays, we identified a frequent homozygous deletion at 4q35 loci with approximately 1 Mb in extent. Among the seven genes located within this region, the expression of the melatonin receptor 1 A (MTNR1A) messenger RNA (mRNA) was not detected or decreased in 35 out of the 39 (89%) OSCC cell lines, but was detected in immortalized normal oral epithelial cell line, and was restored in gene-silenced OSCC cells without its homozygous loss after treatment with 5-aza-2'-deoxycytidine. The hypermethylation of the CpG (cytosine and guanine separated by phosphate) island in the promoter region of MTNR1A was inversely correlated with its expression in OSCC lines without a homozygous deletion. Methylation of this CpG island was also observed in primary OSCC tissues. In an immunohistochemical analysis of 50 primary OSCC tumors, the absence of immunoreactive MTNR1A was significantly associated with tumor size and a shorter overall survival in patients with OSCC tumors, and seems to be an independent prognosticator in a multivariate analysis. Exogenous restoration of MTNR1A expression inhibited the growth of OSCC cells lacking its expression. Together with the known tumor-suppressive function of melatonin and MTNR1A in various tumors, our results indicate MTNR1A to be the most likely target for epigenetic silencing at 4q35 and to play a pivotal role during oral carcinogenesis.


Identification of protein-interacting nucleotides in a RNA sequence using composition profile of tri-nucleotides.
The RNA-protein interactions play a diverse role in the cells, thus identification of RNA-protein interface is essential for the biologist to understand their function. In the past, several methods have been developed for predicting RNA interacting residues in proteins, but limited efforts have been made for the identification of protein-interacting nucleotides in RNAs. In order to discriminate protein-interacting and non-interacting nucleotides, we used various classifiers (NaiveBayes, NaiveBayesMultinomial, BayesNet, ComplementNaiveBayes, MultilayerPerceptron, J48, SMO, RandomForest, SMO and SVM(light)) for prediction model development using various features and achieved highest 83.92% sensitivity, 84.82 specificity, 84.62% accuracy and 0.62 Matthew's correlation coefficient by SVM(light) based models. We observed that certain tri-nucleotides like ACA, ACC, AGA, CAC, CCA, GAG, UGA, and UUU preferred in protein-interaction. All the models have been developed using a non-redundant dataset and are evaluated using five-fold cross validation technique. A web-server called RNApin has been developed for the scientific community (http://crdd.osdd.net/raghava/rnapin/).


RedRibbon: A new rank-rank hypergeometric overlap for gene and transcript expression signatures.
High-throughput omics technologies have generated a wealth of large protein, gene, and transcript datasets that have exacerbated the need for new methods to analyse and compare big datasets. Rank-rank hypergeometric overlap is an important threshold-free method to combine and visualize two ranked lists of P-values or fold-changes, usually from differential gene expression analyses. Here, we introduce a new rank-rank hypergeometric overlap-based method aimed at gene level and alternative splicing analyses at transcript or exon level, hitherto unreachable as transcript numbers are an order of magnitude larger than gene numbers. We tested the tool on synthetic and real datasets at gene and transcript levels to detect correlation and anticorrelation patterns and found it to be fast and accurate, even on very large datasets thanks to an evolutionary algorithm-based minimal P-value search. The tool comes with a ready-to-use permutation scheme allowing the computation of adjusted P-values at low time cost. The package compatibility mode is a drop-in replacement to previous packages. RedRibbon holds the promise to accurately extricate detailed information from large comparative analyses.


Prevalence of self-reported early glaucoma eye drop bottle exhaustion and associated risk factors: a patient survey.
One barrier to patient adherence with chronic topical glaucoma treatment is an inadequate amount of medication available between prescription refills. We examined the self-reported prevalence of early exhaustion of glaucoma eye drops prior to a scheduled refill, and associated risk factors. This cross-sectional survey was performed at a University-based clinical practice. Glaucoma patients at the University of Washington who were experienced with eye drop application and were on a steady regimen of self-administered glaucoma drops in both eyes took a survey at the time of clinic examination. The main outcome measure was self-reported early eye drop bottle exhaustion. 236 patients were eligible and chose to participate. In general, patients included were relatively healthy (mean 2.3 comorbid medical conditions). Sixty patients (25.4%) reported any problem with early exhaustion of eye drop bottles, and this was associated with visual acuity ≤ 20/70 in the better eye (P = .049). Twelve patients (5.1%) reported that they "often" (5-7 times per year), "usually" (8-11 times per year) or "always" ran out of eye drops prior to a scheduled refill. Patients affected by this higher level (≥ 5 times yearly) of eye drop bottle exhaustion were more likely to have poor visual acuity in their worse eye ≤ 20/70 (P = .015) and had significantly lower worse-eye logMAR (P = .043). Self-reported early glaucoma bottle exhaustion regularly affected 5% of patients in our population and 25% reported early exhaustion at least once; the main risk factor was poor vision in at least one eye. These results may not be generalizable to a broad patient population, or to those inexperienced with eye drop self-administration. However, this pilot study compels further evaluation and consideration of early eye drop bottle exhaustion in glaucoma patients.


Focusing on basic data and a model of healthcare security for rare diseases: The Multidisciplinary Expert Seminar on Healthcare Security for Rare Diseases in China was held in Beijing.
On August 10, 2019, the Multidisciplinary Expert Seminar on Healthcare Security for Rare Diseases in China was held in Beijing. The seminar was organized by the Shanghai Foundation for Rare Disease and Shanghai Health Development Research Center and advised by the China Alliance of Rare Diseases. Participants in this seminar included government officials, experts in clinical medicine, pharmacy, epidemiology, health economics, and law as well as representatives from rare disease patient organizations. The participating experts cited three key elements of healthcare security, including its concept, data, and mechanism, to solve the problem of health care security for patients with rare diseases at the national level. Collection of basic data and creation of a model of healthcare security for rare diseases were discussed. Data collection should be actively promoted. Creation of a special zone to ensure medical care for patients with rare diseases should be considered. Healthcare security should be classified, which means that basic medical insurance provides better care for rare diseases that respond to treatment, and channels should be established for rare diseases that respond poorly to treatment.


Caesarean delivery rates, determinants and indications in Makassed Hospital, Jerusalem 1993 and 2002.
This study investigated the rising rate of caesarean section (CS) deliveries between 1993 and 2002 (9.4% to 14.4%) and associated factors, including indications for CS and sociodemographic and clinical characteristics based on the register of a major Palestinian teaching hospital. Instrumental deliveries declined from 12.6% to 4.4%. Fetal distress decreased as an indication for CS, while previous CS and breech presentations contributed to the increase. Decision-making for CS needs to frame the benefits and risks of the intervention within the context of women's entire reproductive life-cycle and existing standards of care, avoiding unnecessary and costly CS deliveries to reduce iatrogenic complications and conserve resources.


In vitro study of the NS2-3 protease of hepatitis C virus.
Processing at the C terminus of the NS2 protein of hepatitis C virus (HCV) is mediated by a virus-encoded protease which spans most of the NS2 protein and part of the NS3 polypeptide. In vitro cotranslational cleavage at the 2-3 junction is stimulated by the presence of microsomal membranes and ultimately results in the membrane insertion of the NS2 polypeptide. To characterize the biochemical properties of this viral protease, we have established an in vitro assay whereby the NS2-3 protease of HCV BK can be activated posttranslationally by the addition of detergents. The cleavage proficiency of several deletion and single point mutants was the same as that observed with microsomal membranes, indicating that the overall sequence requirements for proper cleavage at this site are maintained even under these artificial conditions. The processing efficiency of the NS2-3 protease varied according to the type of detergent used and its concentration. Also, the incubation temperature affected the cleavage at the 2-3 junction. The autoproteolytic activity of the NS2-3 protease could be inhibited by alkylating agents such as iodoacetamide and N-ethylmaleimide. Metal chelators such as EDTA and phenanthroline also inhibited the viral enzyme. The EDTA inhibition of NS2-3 cleavage could be reversed, at least in part, by the addition of ZnCl2 and CdCl2. Among the common protease inhibitors tested, tosyl phenylalanyl chloromethyl ketone and soybean trypsin inhibitor inactivated the NS2-3 protease. By means of gel filtration analysis, it was observed that the redox state of the reaction mixture greatly influenced the processing efficiency at the 2-3 site and that factors present in the rabbit reticulocyte lysate, wheat germ extract, and HeLa cell extract were required for efficient processing at this site. Thus, the in vitro assay should allow further characterization of the biochemical properties of the NS2-3 protease of HCV and the identification of host components that contribute to the efficient processing at the 2-3 junction.


[Detection of a system of microsomal monooxygenases in the rodent malaria parasite Plasmodium berghei].
A microsomal fraction from the cells of the malaria parasite of rodent Plasmodium berghei was obtained. The spectral properties of microsomal preparations suggest that P. berghei microsomes contain cytochromes b5 and P-420. Electrophoretic separation of microsomal proteins revealed the presence of proteins whose molecular mass corresponds to NADPH-cytochrome c reductase, cytochrome P-450 and epoxide hydratase. The activities of NADPH-cytochrome c reductase and benzpyrene hydroxylase were determined. The spectral parameters, electrophoretic data and enzymatic activities of microsomal proteins indicate that P. berghei cells contain a cytochrome P-450 monooxygenase system. The interrelationship between the activity of the microsomal monooxygenase system and the resistance of P. berghei cells to the antimalaria preparation chloroquine is discussed.


Lanthanoids Goes Healing: Lanthanoidic Metallopolymers and Their Scratch Closure Behavior.
Metallopolymers represent an interesting combination of inorganic metal complexes and polymers resulting in a variety of outstanding properties and applications. One field of interest are stimuli-responsive materials and, in particular, self-healing polymers. These systems could be achieved by the incorporation of terpyridine-lanthanoid complexes of Eu (III), Tb (III), and Dy (III) in the side chains of well-defined copolymers, which were prepared applying the reversible addition fragmentation chain-transfer (RAFT)-polymerization technique. The metal complexes crosslink the polymer chains in order to form reversible supramolecular networks. These dynamics enable the self-healing behavior. The information on composition, reversibility, and stability of the complexes was obtained by isothermal titration calorimetry (ITC). Moreover, self-healing experiments were performed by using 3D-microscopy and indentation.


Eliminating first trimester markers: will replacing PAPP-A and βhCG miss women at risk for small for gestational age?
Placental analytes are traditionally used for aneuploidy screening, although may be replaced by cell-free fetal DNA. Abnormal analytes also identify women at risk for small for gestational age (SGA). We sought to quantify the proportion of women at risk for SGA by low pregnancy-associated plasma protein-A (PAPP-A) or βhCG who would not otherwise be identified by maternal risk factors. We studied first-trimester PAPPA-A and βhCG from 658 euploid singleton pregnancies from a prospective longitudinal cohort. Analytes were standardized for gestational age in multiples of the median (MoM). SGA was defined as birthweight z-score ≤-1.28. Maternal risk factors included chronic hypertension, pre-gestational diabetes and age ≥40. Mean GA was 38.8 ± 1.9 weeks; 6.8% had a SGA infant. Low PAPP-A and βhCG were identified in 48 (7.4%) and 9 (1.4%) of pregnancies, respectively, of whom 18.9% were SGA (OR 3.0, 95% CI 1.4-6.3). 88% did not have risk factors for SGA. Among women with no risk factors, low PAPP-A was a significant predictor of SGA (OR 3.3, 95% CI 1.5-7.4). Most women with abnormal analytes did not have risk factors for SGA. Eliminating PAPP-A and βhCG may present missed opportunities to identify women at risk for SGA.


Middle cerebral artery Doppler flow velocity waveforms.
Doppler flow velocity-time waveforms of the middle cerebral arteries at their origin from the internal carotid arteries were obtained serially on 14 patients to determine the alterations of these waveforms with gestational age. Satisfactory recordings were obtained from all patients. The middle cerebral artery waveforms demonstrated a typically biphasic pattern with continuing forward flow in diastole. The peak systolic to end-diastolic ratio (A/B ratio) showed a progressive and significant decline from 6.89 +/- 1.48 at around 25 weeks to 4.23 +/- 0.67 at term. Elevation of diastolic velocities and diminished A/B ratio in the middle cerebral arteries were noted in four fetuses, in whom severe antepartum compromise was diagnosed. The ratio also appeared to decrease with worsening of fetal condition. Because the fetus compensates for hypoxia by increasing blood flow to the brain, Doppler middle cerebral artery waveforms may permit evaluation of fetal compromise and hypoxia.


Identification of the ectonucleotidases expressed in mouse, rat, and human Langerhans islets: potential role of NTPDase3 in insulin secretion.
Extracellular nucleotides and adenosine regulate endocrine pancreatic functions such as insulin secretion by Langerhans islet β-cells via the activation of specific P2 and P1 receptors. Membrane-bound ectonucleotidases regulate the local concentration of these ligands and consequently control the activation of their receptors. The objective of this study was to identify and localize the major ectonucleotidases, namely NTPDases and ecto-5'-nucleotidase, present in the endocrine pancreas. In addition, the potential implication of ecto-ATPase activity on insulin secretion was investigated in the rat β-cell line INS-1 (832/13). The localization of ectonucleotidase activity and protein was carried out in situ by enzyme histochemistry and immunolocalization in mouse, rat, and human pancreas sections. NTPDase1 was localized in all blood vessels and acini, and NTPDase2 was localized in capillaries of Langerhans islets and in peripheral conjunctive tissue, whereas NTPDase3 was detected in all Langerhans islet cell types. Interestingly, among the mammalian species tested, ecto-5'-nucleotidase was present only in rat Langerhans islet cells, where it was coexpressed with NTPDase3. Notably, the inhibition of NTPDase3 activity by BG0136 and NF279 facilitated insulin release from INS-1 (832/13) cells under conditions of low glycemia, probably by affecting P2 receptor activation. NTPDase3 activity also regulated the inhibitory effect of exogenous ATP in the presence of a high glucose concentration most likely by controlling adenosine production. In conclusion, all pancreatic endocrine cells express NTPDase3 that was shown to modulate insulin secretion in rat INS-1 (832/13) β-cells. Ecto-5'-nucleotidase is expressed in rat Langerhans islet cells but absent in human and mouse endocrine cells.


PRL-3 mediates the protein maturation of ULBP2 by regulating the tyrosine phosphorylation of HSP60.
Many malignant cells release the NKG2D ligand ULBP2 from their cell surface to evade immunosurveillance by NK cells and CD8 T cells. Although the shedding mechanism remains unclear, various inhibitors of matrix metalloproteinases have been shown to efficiently block the release of soluble ULBP2. The clinical use of these inhibitors, however, is limited because of adverse side effects. Using high-throughput screening technique, we identified a specific inhibitor of phosphatase of regenerating liver 3 (PRL-3) that could reduce the level of soluble ULBP2 in the culture supernatant of various cancer cell lines. Inhibition or gene knockdown of PRL-3 did not reduce ULBP2 shedding, but rather suppressed posttranslational maturation of ULBP2, resulting in intracellular retention of immature ULBP2. We then found that ULBP2 was constitutively associated with heat shock protein HSP60. Complete maturation of ULBP2 required tyrosine phosphorylation of HSP60 which was mediated by PRL-3.


An evolutionary perspective on complex neuropsychiatric disease.
The forces of evolution-mutation, selection, migration, and genetic drift-shape the genetic architecture of human traits, including the genetic architecture of complex neuropsychiatric illnesses. Studying these illnesses in populations that are diverse in genetic ancestry, historical demography, and cultural history can reveal how evolutionary forces have guided adaptation over time and place. A fundamental truth of shared human biology is that an allele responsible for a disease in anyone, anywhere, reveals a gene critical to the normal biology underlying that condition in everyone, everywhere. Understanding the genetic causes of neuropsychiatric disease in the widest possible range of human populations thus yields the greatest possible range of insight into genes critical to human brain development. In this perspective, we explore some of the relationships between genes, adaptation, and history that can be illuminated by an evolutionary perspective on studies of complex neuropsychiatric disease in diverse populations.


The Effect of Intravitreal Azithromycin on the Albino Newborn Rabbit Retina.
To evaluate the effect of intravitreal azithromycin on the retina in a newborn rabbit model. Twelve, two-week old New Zealand albino rabbits were divided into two groups (six in each). The right eyes of six rabbits received 0.75 mg (0.05 mL) azithromycin and the right eyes of the remaining six rabbits 1.5 mg (0.1 mL) azithromycin intravitreally. Left eyes were served as the control and received the same volume of saline. All eyes were enucleated at the third postinjection week. Retinal histology was examined by light microscopy. Apoptosis of the retinal cells was further evaluated by immunohistochemical staining for caspase-3 and in situ terminal deoxynucleotidyl transferase-mediated biotin-deoxyuridine triphosphate nick-end labeling (TUNEL) of DNA fragments. Light microscopy demonstrated no retinal abnormalities in all eyes. However, retinal nuclear DNA fragmentation was evident in both study groups (33.6% with 1.5 mg and 21.4% with 0.75 mg azithromycin) with the TUNEL method. TUNEL staining ratio was statistically higher only in the second group treated with 1.5 mg azithromycin when compared to the control group (p=0.01 Mann Whitney U test). The ratio of caspase-3 positive cells in the two study groups was 21.5% and 20.2%, respectively. Caspase-3 staining ratio was statistically higher in both study groups when compared to the control eyes (p=0.00, p=0.00 respectively). The difference of TUNEL staining ratio between the two study groups was statistically significant (p=0.028), but there were no statistically significant differences in the two study groups by caspase-3 staining (p=0.247). In newborn rabbits, intravitreal azithromycin injection resulted in an apoptotic activity in the photoreceptor, bipolar and ganglion cells. Immunohistochemical analysis suggested that doses of 0.75 mg and 1.5 mg azithromycin, administered intravitreally might be toxic to the newborn rabbit retina.


Intermittent dosing of the farnesyl transferase inhibitor tipifarnib (R115777) in advanced malignant solid tumors: a phase I California Cancer Consortium Trial.
Tipifarnib (R115777) inhibits farnesylation of key proteins that modulate signaling pathways implicated in cell growth and proliferation, including members of the Ras and Rho families. It has broad-spectrum antiproliferative activity in vitro and in vivo. Clinical trials employing a continuous administration schedule have demonstrated dose-limiting neurotoxicity and myelosuppression. Preclinical studies have shown that intermittent oral administration can suppress tumor growth comparable to continuous administration. We conducted a National Cancer Institute-sponsored phase I trial to determine the feasibility of an intermittent dosing schedule of R115777 given orally twice daily on weeks 1 and 3 of a 28-day cycle in patients with malignant solid tumors. Starting dose was 300 mg twice daily (b.i.d.) with escalation by 300 mg b.i.d. increments over six dose levels to a maximum of 1800 mg b.i.d. Dose-limiting toxicity (DLT) was defined as any grade 3 or 4 non-hematologic toxicity, grade 4 thrombocytopenia, grade 4 neutropenia (ANC) with fever (38.3 degrees C or above) or a documented infection. Twenty-one patients with advanced solid tumors, all of whom had prior systemic therapy, were accrued. Grade 3 fatigue was dose limiting for two of three patients at the 900 mg b.i.d. dose level. Although no responses were seen, four of six patients with stable disease remained on study for at least a year (16, 17, 13 and 12 months) before developing progressive disease. Three of these prolonged stable disease patients had non-small cell lung cancer. We conclude that intermittent dosing of R115777 is feasible and tolerable. The recommended phase II dose is 600 mg orally b.i.d. on alternate weeks.


"It Haunts Me": Impact of COVID-19 Deaths on Frontline Clinicians In Acute Care Settings-A Qualitative Study.
The COVID-19 pandemic created unprecedented conditions for clinicians providing end-of-life care in acute care settings, yet almost 2 years since the start of the pandemic, little is known about its impact on clinicians. To qualitatively explore how clinicians experienced their role as they cared for dying patients with COVID-19 during the pandemic and the impact of these experiences on their lives as health care professionals. In-depth, hour-long interviews were conducted by phone in the spring of 2022 at a single time point with 23 physicians and nurses in critical care and emergency department settings from 2 health systems in California. The goal was to elicit perspectives on experiences with end-of-life care and the long-term impact of these experiences. Clinicians were asked to reflect on the different stages of the pandemic, and interviews were analyzed using a thematic analysis. The interviews highlighted 2 primary themes around provision of end-of-life care during the COVID-19 pandemic. First, clinicians described challenges around their clinical duties of providing care to dying patients, which included decision-making dilemmas and professionalism challenges. Second, clinicians described the emotional aspects around end-of-life care situations they experienced, including communication with family members, "being patients' family members" at the end of life, and various degrees of meaning-making and remembrance of these experiences. The findings highlight the sustained and cumulative emotional challenges and burden clinicians are still shouldering more than 2 years after the start of the pandemic.


Genomics and proteomics: Applications in autoimmune diseases.
Tremendous progress has been made over the past decade in the development and refinement of genomic and proteomic technologies for the identification of novel drug targets and molecular signatures associated with clinically important disease states, disease subsets, or differential responses to therapies. The rapid progress in high-throughput technologies has been preceded and paralleled by the elucidation of cytokine networks, followed by the stepwise clinical development of pathway-specific biological therapies that revolutionized the treatment of autoimmune diseases. Together, these advances provide opportunities for a long-anticipated personalized medicine approach to the treatment of autoimmune disease. The ever-increasing numbers of novel, innovative therapies will need to be harnessed wisely to achieve optimal long-term outcomes in as many patients as possible while complying with the demands of health authorities and health care providers for evidence-based, economically sound prescription of these expensive drugs. Genomic and proteomic profiling of patients with autoimmune diseases holds great promise in two major clinical areas: (1) rapid identification of new targets for the development of innovative therapies and (2) identification of patients who will experience optimal benefit and minimal risk from a specific (targeted) therapy. In this review, we attempt to capture important recent developments in the application of genomic and proteomic technologies to translational research by discussing informative examples covering a diversity of autoimmune diseases.


Hippocampal cGMP-dependent protein kinase I supports an age- and protein synthesis-dependent component of long-term potentiation but is not essential for spatial reference and contextual memory.
cGMP-dependent protein kinase I (cGKI) is expressed in the hippocampus, but its role in hippocampal long-term potentiation (LTP) is controversial. In addition, whether cGKI is involved in spatial learning has not been investigated. To address these issues, we generated mice with a hippocampus-specific deletion of cGKI (cGKIhko mice). Unlike conventional cGKI knock-out mice, cGKIhko mice lack gastrointestinal and cardiovascular phenotypes and have a normal life expectancy, which enables us to analyze hippocampal synaptic plasticity and learning in young and adult animals. Hippocampal LTP after repetitive episodes of theta burst stimulation was impaired in adult (12-14 weeks of age) but not in juvenile (3-4 weeks of age) cGKIhko mice. The difference in LTP between adult control and cGKIhko mice was abolished by the protein synthesis inhibitor anisomycin, suggesting that the impairment of LTP in adult cGKIhko mice reflects a defect in late-phase LTP. Despite their deficit in LTP, adult cGKIhko mutants showed normal performance in a discriminatory water maze and had intact contextual fear conditioning. These results suggest that hippocampal cGKI supports an age- and protein synthesis-dependent form of hippocampal LTP, whereas it is dispensable for hippocampus-dependent spatial reference and contextual memory.


Interferon-induced transcriptional and post-transcriptional modulation of factor H and C4 binding-protein synthesis in human monocytes.
Cultured human monocytes synthesize factor H (H) and C4 binding protein (C4-bp), as assessed by measuring their presence in culture fluids, and demonstrating the presence of their corresponding mRNAs in total monocyte RNA by Northern blot analysis and by nuclear run-on experiments. H mRNA (4.3 kb and 1.8 kb) could only be detected when cycloheximide (2.5 micrograms/ml) was present in monocyte culture fluid. Recombinant interferon-gamma (IFN-gamma) and lymphoblastoid interferon-alpha (IFN-alpha) produce dose-related increases in the synthesis of H and C4-bp and in the abundance of C4-bp mRNA (2.5 kb). Increased abundance of H mRNA was also seen when cycloheximide (2.5 micrograms/ml) was present in the cultures. Both cytokines increased the transcription rates of the H and C4-bp genes. These changes in transcription were rapid, with significant increases being observed within 30 min of exposure. Following the removal of the cytokines from the cultures the transcription rates of both genes returned to control levels within 4 h. The effects of combining IFN-alpha and IFN-gamma on H and C4-bp transcription rates, mRNA abundances and protein secretion rates were not quantitatively additive. The half-life of H mRNA in monocytes was 15 min, whereas that of C4-bp mRNA was 2 h 45 min. Neither half-life was altered by IFN-gamma.


Subconjunctival antibiotics in the treatment of endophthalmitis managed without vitrectomy.
To determine if the treatment outcomes for endophthalmitis are influenced by subconjunctival antibiotics. A retrospective, nonrandomized consecutive series of patients with clinically diagnosed bacterial endophthalmitis confirmed with positive cultures who presented between December 1, 1995, and February 28, 2002, was studied. Patients with cataract surgery, glaucoma filtering blebs, or trauma who presented with visual acuity of hand motions or better were included. All patients received intravitreal and topical antibiotics. Management by pars plana vitrectomy or vitreous tap and use or nonuse of subconjunctival antibiotics were at the discretion of the treating physician. There were 59 patients identified; 54 met the follow-up criteria. These patients were divided into two groups based on whether subconjunctival antibiotics were used (group ABX; n = 21) or not used (group noABX; n = 33). The median pretreatment visual acuity was hand motions in both groups. The median age in both groups was 74 years. Etiology, duration of symptoms, vitreous culture organisms, percentage of cases with wound complications such as leaks or vitreous incarceration, and intraocular lens type were similar in the two study groups. Intravitreal and topical antibiotics and corticosteroids used were not significantly different in the two groups, except that topical ceftazidime was used less frequently in group ABX than in group noABX (43% vs. 82%, respectively; P = 0.007). The median follow-up was 13 months in both groups (range: 3-87 months for group ABX and 3-63 months for group noABX). Final visual acuity in groups ABX and noABX was at least 20/50 (33% vs. 39%, respectively), 20/60 to 5/200 (29% vs. 39%, respectively), 4/200 to better than hand motions (0 vs. 3%, respectively), or hand motions or worse (38% vs. 18%, respectively). These differences were not significant (P = 0.37). Reinjection rates (14% vs. 15%, respectively) were also similar in groups ABX and noABX. The additional procedures rate was significantly higher in group ABX than in group noABX (P = 0.024), with cumulative rates of 33% and 3%, respectively, at the 12-month follow-up. These data suggest that subconjunctival antibiotics may not be necessary to treat infectious endophthalmitis managed with otherwise standard tap and injection techniques and topical antibiotics.


Transcatheter aortic-valve replacement for inoperable severe aortic stenosis.
Transcatheter aortic-valve replacement (TAVR) is the recommended therapy for patients with severe aortic stenosis who are not suitable candidates for surgery. The outcomes beyond 1 year in such patients are not known. We randomly assigned patients to transfemoral TAVR or to standard therapy (which often included balloon aortic valvuloplasty). Data on 2-year outcomes were analyzed. A total of 358 patients underwent randomization at 21 centers. The rates of death at 2 years were 43.3% in the TAVR group and 68.0% in the standard-therapy group (P<0.001), and the corresponding rates of cardiac death were 31.0% and 62.4% (P<0.001). The survival advantage associated with TAVR that was seen at 1 year remained significant among patients who survived beyond the first year (hazard ratio, 0.58; 95% confidence interval [CI], 0.36 to 0.92; P=0.02 with the use of the log-rank test). The rate of stroke was higher after TAVR than with standard therapy (13.8% vs. 5.5%, P=0.01), owing, in the first 30 days, to the occurrence of more ischemic events in the TAVR group (6.7% vs. 1.7%, P=0.02) and, beyond 30 days, to the occurrence of more hemorrhagic strokes in the TAVR group (2.2% vs. 0.6%, P=0.16). At 2 years, the rate of rehospitalization was 35.0% in the TAVR group and 72.5% in the standard-therapy group (P<0.001). TAVR, as compared with standard therapy, was also associated with improved functional status (P<0.001). The data suggest that the mortality benefit after TAVR may be limited to patients who do not have extensive coexisting conditions. Echocardiographic analysis showed a sustained increase in aortic-valve area and a decrease in aortic-valve gradient, with no worsening of paravalvular aortic regurgitation. Among appropriately selected patients with severe aortic stenosis who were not suitable candidates for surgery, TAVR reduced the rates of death and hospitalization, with a decrease in symptoms and an improvement in valve hemodynamics that were sustained at 2 years of follow-up. The presence of extensive coexisting conditions may attenuate the survival benefit of TAVR. (Funded by Edwards Lifesciences; ClinicalTrials.gov number, NCT00530894.).


Effect of therapy switch on time to second-line antiretroviral treatment failure in HIV-infected patients.
Switch from first line antiretroviral therapy (ART) to second-line ART is common in clinical practice. However, there is limited knowledge of to which extent different reason for therapy switch are associated with differences in long-term consequences and sustainability of the second line ART. Data from 869 patients with 14601 clinical visits between 1999-2014 were derived from the national cohort database. Reason for therapy switch and viral load (VL) levels at first-line ART failure were compared with regard to outcome of second line ART. Using the Laplace regression model we analyzed the median, 10th, 20th, 30th and 40th percentile of time to viral failure (VF). Most patients (n = 495; 57.0%) switched from first-line to second-line ART without VF. Patients switching due to detectable VL with (n = 124; 14.2%) or without drug resistance mutations (DRM) (n = 250; 28.8%) experienced VF to their second line regimen sooner (median time, years: 3.43 (95% CI 2.90-3.96) and 3.20 (95% 2.65-3.75), respectively) compared with those who switched without VF (4.53 years). Furthermore level of VL at first-line ART failure had a significant impact on failure of second-line ART starting after 2.5 years of second-line ART. In the context of life-long therapy, a median time on second line ART of 4.53 years for these patients is short. To prolong time on second-line ART, further studies are needed on the reasons for therapy changes. Additionally patients with a high VL at first-line VF should be more frequently monitored the period after the therapy switch.


Tissue expansion in reconstruction.
Tissue expansion offers a new alternative in plastic surgery. Its applications are numerous for reconstruction of the scalp, head and neck, trunk, breast, and extremity. Over the past year I have used tissue expansion for reconstruction in 13 patients, with multiple expanders in six patients. Complications included two exposed expanders requiring early removal, and two mechanical malfunctions requiring replacement. Expansion of previously irradiated tissue has been unsatisfactory.


Dynamic range and sensitivity adaptation in a silicon spiking neuron.
We here propose an adaptive procedure that enables a spiking neuron, whether artificial or biological, to make optimal use of its dynamic range and gain.We discuss an analog electronic circuit implementation of this algorithm using a biologically realistic artificial "silicon" neuron. The adaptation procedure adapts the neuron's firing threshold and the sensitivity (or gain) of its current-frequency relationship to match the dc offset (or mean) and the dynamic range (or variance) of the time-varying somatic input current. The neuron extracts the minimum and maximum levels of the reconstructed somatic current signals from the cell's own spike trains. These are used to regulate the somatic leak conductance in order to shift the somatic current-frequency relation and to adjust a calcium-activated potassium conductance to change the dynamic range of the cell's somatic current-frequency relationship. We report experimental data from a test neuron--built using analog subthreshold CMOS VLSI technology--that shows the expected behavior.


Synthesis, Structural Investigations, DNA/BSA Interactions, Molecular Docking Studies, and Anticancer Activity of a New 1,4-Disubstituted 1,2,3-Triazole Derivative.
We report herein a new 1,2,3-triazole derivative, namely, 4-((1-(3,4-dichlorophenyl)-1H-1,2,3-triazol-4-yl)methoxy)-2-hydroxybenzaldehyde, which was synthesized by copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC). The structure of the compound was analyzed using Fourier transform infrared spectroscopy (FTIR), 1H NMR, 13C NMR, UV-vis, and elemental analyses. Moreover, X-ray crystallography studies demonstrated that the compound adapted a monoclinic crystal system with the P21/c space group. The dominant interactions formed in the crystal packing were found to be hydrogen bonding and van der Waals interactions according to Hirshfeld surface (HS) analysis. The volume of the crystal voids and the percentage of free spaces in the unit cell were calculated as 152.10 Å3 and 9.80%, respectively. The evaluation of energy frameworks showed that stabilization of the compound was dominated by dispersion energy contributions. Both in vitro and in silico investigations on the DNA/bovine serum albumin (BSA) binding activity of the compound showed that the CT-DNA binding activity of the compound was mediated via intercalation and BSA binding activity was mediated via both polar and hydrophobic interactions. The anticancer activity of the compound was also tested by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay using human cell lines including MDA-MB-231, LNCaP, Caco-2, and HEK-293. The compound exhibited more cytotoxic activity than cisplatin and etoposide on Caco-2 cancer cell lines with an IC50 value of 16.63 ± 0.27 μM after 48 h. Annexin V suggests the induction of cell death by apoptosis. Compound 3 significantly increased the loss of mitochondrial membrane potential (MMP) levels in Caco-2 cells, and the reactive oxygen species (ROS) assay proved that compound 3 could induce apoptosis by ROS generation.


Thermal fluctuations in the lattice Boltzmann method for nonideal fluids.
We introduce thermal fluctuations in the lattice Boltzmann method for nonideal fluids. A fluctuation-dissipation theorem is derived within the Langevin framework and applied to a specific lattice Boltzmann model that approximates the linearized fluctuating Navier-Stokes equations for fluids based on square-gradient free-energy functionals. The obtained thermal noise is shown to ensure equilibration of all degrees of freedom in a simulation to high accuracy. Furthermore, we demonstrate that satisfactory results for most practical applications of fluctuating hydrodynamics can already be achieved using thermal noise derived in the long-wavelength limit.


Low vitamin d and cardiovascular risk factors in males and females from a sunny, rich country.
Low serum vitamin (vit) D levels are common even in sunny countries. We assessed the prevalence and relationship of low vit D with cardiovascular risk factors in Qatar. Data were collected retrospectively from January 2008 and November 2009. In patients who had low vi t D (< 30 ng/ml ) , demographic and clinical profiles were analyzed and compared in males and females. The overall mean level of vit D among 547 patients was 14.4±11 ng/mL. Among the low vitamin D group, 56% were females (mean age 48±12) and 44% males (mean age 49.6±13). Severely low vit D levels (<10 ng/mL) were found in 231 (46%) patients with mean age of 46±12 years. Compared with females, males with low vitamin D were more likely to have diabetes mellitus (38 vs 22%, p=0.001), dyslipidemia (41 vs 29%, p=0.007), myocardial infarction (5.5 vs 1.5%, p=0.001) and angiographically documented coronary artery disease (CAD) (53 vs 17%, p=0.001). Multivariate logistic regression analysis showed that in the presence of low vit D, age and hypertension were independent predictors of CAD (OR 1.07;95% CI: 1.02-1.11) and OR 8.0; 95% CI: 1.67-39.82), respectively. Our study supports the widespread prevalence of low vit D in sunny regions. Low vit D is associated with 3 times increase in the rate of MI among males. Hypertension increases the risk of CAD 8 times in the presence of low vit D regardless of gender.


Novel sandwich immunoassay detects a shrimp AHPND-causing binary PirABVp toxin produced by Vibrio parahaemolyticus.
The binary PirA/PirB toxin expressed by Vibrio parahaemolyticus (PirABVp) is a virulent complex that causes acute hepatopancreatic necrosis disease (AHPND) in shrimps, affecting the global shrimp farming industry. AHPND is currently diagnosed by detecting pirA and pirB genes by PCR; however, several V. parahaemolyticus strains do not produce the two toxins as proteins. Thus, an immunoassay using antibodies may be the most effective tool for detecting toxin molecules. In this study, we report a sandwich ELISA-based immunoassay for the detection of PirABVp. We utilized a single-chain variable fragment (scFv) antibody library to select scFvs against the PirA or PirB subunits. Phage display panning rounds were conducted to screen and identify scFv antibodies directed against each recombinant toxin subunit. Selected scFvs were converted into IgGs to develop a sandwich immunoassay to detect recombinant and bacterial PirABVp. Antibodies produced as IgG forms showed sub-nanomolar to nanomolar affinities (KD), and a pair of anti-PirA antibody as a capture and anti-PirB antibody as a detector showed a limit of detection of 201.7 ng/mL for recombinant PirABVp. The developed immunoassay detected PirABVp in the protein lysates of AHPND-causing V. parahaemolyticus (VpAHPND) and showed a significant detectability in moribund or dead shrimp infected with a VpAHPND virulent strain compared to that in non-infected shrimp. These results indicate that the developed immunoassay is a reliable method for diagnosing AHPND by detecting PirABVp at the protein level and could be further utilized to accurately determine the virulence of extant or newly identified VpAHPND in the global shrimp culture industry.


Field induced changes in the ring/chain equilibrium of hydrogen bonded structures: 5-methyl-3-heptanol.
Using non-linear dielectric techniques, we have measured the dynamics of 5-methyl-3-heptanol at a temperature at which the Kirkwood correlation factor gK indicates the coexistence of ring- and chain-like hydrogen-bonded structures. Steady state permittivity spectra recorded in the presence of a high dc bias electric field (17 MV/m) reveal that both the amplitude and the time constant are increased by about 10% relative to the low field limit. This change is attributed to the field driven conversion from ring-like to the more polar chain-like structures, and a direct observation of its time dependence shows that the ring/chain structural transition occurs on a time scale that closely matches that of the dielectric Debye peak. This lends strong support to the picture that places fluctuations of the end-to-end vector of hydrogen bonded structures at the origin of the Debye process, equivalent to fluctuations of the net dipole moment or gK. Recognizing that changes in the ring/chain equilibrium constant also impact the spectral separation between Debye and α-process may explain the difference in their temperature dependence whenever gK is sensitive to temperature, i.e., when the structural motifs of hydrogen bonding change considerably.


Numerical and Experimental Study of Mechanisms Involved in Boiling Histotripsy.
The aim of boiling histotripsy is to mechanically fractionate tissue as an alternative to thermal ablation for therapeutic applications. In general, the shape of a lesion produced by boiling histotripsy is tadpole like, consisting of a head and a tail. Although many studies have demonstrated the efficacy of boiling histotripsy for fractionating solid tumors, the exact mechanisms underpinning this phenomenon are not yet well understood, particularly the interaction of a boiling vapor bubble with incoming incident shockwaves. To investigate the mechanisms involved in boiling histotripsy, a high-speed camera with a passive cavitation detection system was used to observe the dynamics of bubbles produced in optically transparent tissue-mimicking gel phantoms exposed to the field of a 2.0-MHz high-intensity focused ultrasound (HIFU) transducer. We observed that boiling bubbles were generated in a localized heated region and cavitation clouds were subsequently induced ahead of the expanding bubble. This process was repeated with HIFU pulses and eventually resulted in a tadpole-shaped lesion. A simplified numerical model describing the scattering of the incident ultrasound wave by a vapor bubble was developed to help interpret the experimental observations. Together with the numerical results, these observations suggest that the overall size of a lesion induced by boiling histotripsy is dependent on the sizes of (i) the heated region at the HIFU focus and (ii) the backscattered acoustic field by the original vapor bubble.


Paediatric diabetes subtypes in a consanguineous population: a single-centre cohort study from Kurdistan, Iraq.
Monogenic diabetes is estimated to account for 1-6% of paediatric diabetes cases in primarily non-consanguineous populations, while the incidence and genetic spectrum in consanguineous regions are insufficiently defined. In this single-centre study we aimed to evaluate diabetes subtypes, obtain the consanguinity rate and study the genetic background of individuals with syndromic and neonatal diabetes in a population with a high rate of consanguinity. Data collection was carried out cross-sectionally in November 2021 at the paediatric diabetic clinic, Dr Jamal Ahmad Rashed Hospital, in Sulaimani, Kurdistan, Iraq. At the time of data collection, 754 individuals with diabetes (381 boys) aged up to 16 years were registered. Relevant participant data was obtained from patient files. Consanguinity status was known in 735 (97.5%) participants. Furthermore, 12 families of children with neonatal diabetes and seven families of children with syndromic diabetes consented to genetic testing by next-generation sequencing. Prioritised variants were evaluated using the American College of Medical Genetics and Genomics guidelines and confirmed by Sanger sequencing. A total of 269 of 735 participants (36.5%) with known consanguinity status were offspring of consanguineous families. An overwhelming majority of participants (714/754, 94.7%) had clinically defined type 1 diabetes (35% of them were born to consanguineous parents), whereas only eight (1.1%) had type 2 diabetes (38% consanguineous). Fourteen (1.9%) had neonatal diabetes (50% consanguineous), seven (0.9%) had syndromic diabetes (100% consanguineous) and 11 (1.5%) had clinically defined MODY (18% consanguineous). We found that consanguinity was significantly associated with syndromic diabetes (p=0.0023) but not with any other diabetes subtype. The genetic cause was elucidated in ten of 12 participants with neonatal diabetes who consented to genetic testing (homozygous variants in GLIS3 [sibling pair], PTF1A and ZNF808 and heterozygous variants in ABCC8 and INS) and four of seven participants with syndromic diabetes (homozygous variants in INSR, SLC29A3 and WFS1 [sibling pair]). In addition, a participant referred as syndromic diabetes was diagnosed with mucolipidosis gamma and probably has type 2 diabetes. This unique single-centre study confirms that, even in a highly consanguineous population, clinically defined type 1 diabetes is the prevailing paediatric diabetes subtype. Furthermore, a pathogenic cause of monogenic diabetes was identified in 83% of tested participants with neonatal diabetes and 57% of participants with syndromic diabetes, with most variants being homozygous. Causative genes in our consanguineous participants were markedly different from genes reported from non-consanguineous populations and also from those reported in other consanguineous populations. To correctly diagnose syndromic diabetes in consanguineous populations, it may be necessary to re-evaluate diagnostic criteria and include additional phenotypic features such as short stature and hepatosplenomegaly.


A three-barrier model for the hemocyanin channel.
Keyhole limpet hemocyanin forms ion-conducting channels in planar lipid bilayer membranes. Ionic current through the open hemocyanin channel presents the following characteristics: (a) it is carried mainly by cations; (b) it is a nonlinear function of membrane potential; (c) channel conductance is a saturating function of ion activity; (d) it shows ionic competition. A model for the open hemocyanin channel is developed from absolute reaction rate theory. The model calls for three energy barriers in the channel. Two energy barriers represent the entrance and exit of the ion into and out of the channel. The third barrier separates two energy minima that represent two binding sites. Furthermore, only one ion is allowed inside the channel at a given time. This model is able to recreate all the hemocyanin characteristics found experimentally in negatively charged and neutral membranes.


Processing of the human heparanase precursor and evidence that the active enzyme is a heterodimer.
Human platelet heparanase has been purified to homogeneity and shown to consist of two, non-covalently associated polypeptide chains of molecular masses 50 and 8 kDa. Protein sequencing provided the basis for determination of the full-length cDNA for this novel protein. Based upon this information and results from protein analysis and mass spectrometry, we propose a scheme to define the structural organization of heparanase in relation to its precursor forms, proheparanase and pre-proheparanase. The 8- and 50-kDa chains which make up the active enzyme reside, respectively, at the NH(2)- and COOH-terminal regions of the inactive precursor, proheparanase. The heparanase heterodimer is produced by excision and loss of an internal linking segment. This paper is the first to suggest that human heparanase is a two-chain enzyme.


Mycobacterium avium with the bird type IS1245 RFLP profile is commonly found in wild and domestic animals, but rarely in humans.
Cervical lymphadenitis is the main manifestation of Mycobacterium avium infection in immunocompetent children. Exposure to birds has been discussed as a source of infection. To clarify from where children acquire the infection, M. avium isolates from different origins were analysed with restriction fragment length polymorphism (RFLP) on insertion sequence IS1245, and compared by computer cluster correlation analysis. This molecular epidemiological tool has previously revealed a distinction between multiband profiles found mainly in strains from humans, and a 3-band/bird type profile in strains isolated mainly from birds. 32 isolates from children were compared with 28 isolates from adults and 45 isolates from animals. We found that 67% of the animal isolates had the bird type profile, also found in 1 sputum isolate from an adult. Strains from children showed only multiband profiles that did not differ significantly from profiles of isolates from adults. All but 2 bird isolates showed the bird type profile. Neither of the remaining 2, which had multiband profiles, clustered with the isolates from children. Our results indicate that the true reservoir of M. avium is unknown. Thus the question of whether or not M. avium can be incriminated as a zoonotic disease remains unanswered.


Intravenous Caffeine Rescue for Postoperative Hypoventilation in a 16-Year-Old With Trisomy 10: A Case Report.
Trisomy 10 is a rare disorder, with only 35 cases reported in the literature. Anesthetic management may be challenging in this patient population because of craniofacial, cardiac, and renal abnormalities commonly seen in the disorder. We describe a 16-year-old male with an anesthetic history notable for prolonged emergence, postoperative hypoxia, postoperative reintubation, and unexpected hospital admission presenting for dental extraction of impacted teeth. We utilized intravenous caffeine, a respiratory stimulant used in preterm infants, to facilitate recovery from anesthesia.


'Factors affecting services offered to older adults with psychological morbidity: an exploration of health professional attitudes'.
Poor collaboration between the multiple services involved in hospital discharge planning may contribute to suboptimal patient outcomes post discharge. This study aimed to explore clinician (medical, allied health and nursing) attitudes towards the management of the older patient with psychological morbidity during and following hospitalization. Focus groups were held with 54 health professionals comprising of 7 from acute, 20 from subacute (geriatric assessment and rehabilitation), and 27 from community care settings. A qualitative study using focus groups of clinicians from a range of disciplines working within a large Australian health care service. Data were analysed using an inductive thematic approach with constant comparison. Key themes included: (1) Clinician decision making towards psychological morbidity; (2) Supply of people with specialised skills dealing with psychological morbidity; (3) Confidence and capability; (4) Facilitating continuity of care; and (5) Perception of depression and aging. Clinicians across healthcare settings are uniquely placed to identity psychological morbidity in older patients and make appropriate referrals for support. Management and referral making for older patients with psychological morbidity can be enhanced by routine education for clinicians and the introduction of clinical pathways. This has potential to improve management of psychological morbidity; however, evaluation of impact on patient outcome is required. Specifically, there is a need for greater access for counselling services.


[Remineralization of demineralized dentin induced by bioactive glass NovaMin].
To explore the remineralization effect of bioactive glass NovaMin on demineralized dentin specimens, and to study the physical and chemical properties of formed structure at dentin surface. Methods: One mm-thickness coronal dentin slices were soaked in ethylene diamine tetraacetic acid (EDTA) for 48 h to prepare the completely demineralized dentin specimens and they were divided into 2 groups: an artificial saliva group (control group) and a NovaMin powder group. The specimens were treated with artificial saliva or NovaMin powder for 2 min (2 times every day), and the interval was 8 hours. Then, the specimens were soaked in the remineralization solution. After 7 days, the scanning electron microscope (SEM), energy dispersive X-ray (EDX), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and X-ray diffraction (XRD) were used to detect dentin morphology, the physical and chemical properties of the formed structure at dentin surface. Results: The results of SEM showed that a defined surface layer in the NovaMin powder group could be observed in the SEM imaging at the 7th day, which completely occluded dentinal tubules; the EDX, ATR-FTIR and XRD analysis found that the mineralized layer formed at dentin surface was mainly composed of calcium and phosphate elements, which was similar to the hydroxyapatite-like crystal. However, there were no materials formed at the dentin surface in the control group, and the dentinal tubules were still open. Conclusion: NovaMin can remineralize the demineralized dentin specimens and occlude the dentinal tubules in hydroxyapatite-like crystal structure.


The Uto-Aztecan premolar among North and South Amerindians: Geographic variation and genetics.
The Uto-Aztecan premolar (UAP) is a dental polymorphism characterized by an exaggerated distobuccal rotation of the paracone in combination with the presence of a fossa at the intersection of the distal occlusal ridge and distal marginal ridge of upper first premolars. This trait is important because, unlike other dental variants, it has been found exclusively in Native American populations. However, the trait's temporal and geographic variation has never been fully documented. The discovery of a Uto-Aztecan premolar in a prehistoric skeletal series from northern South America calls into question the presumed linguistic and geographic limits of this trait. We examined published and unpublished data for this rare but highly distinctive trait in samples representing over 5,000 Native Americans from North and South America. Our findings in living Southwest Amerindian populations corroborate the notion that the variable goes beyond the bounds of the Uto-Aztecan language family. It is found in prehistoric Native Americans from South America, eastern North America, Northern and Central Mexico, and in living and prehistoric populations in the American Southwest that are not members of the Uto-Aztecan language stock. The chronology of samples, its geographic distribution, and trait frequencies suggests a North American origin (Southwest) for UAP perhaps between 15,000 BP and 4,000 BP and a rapid and widespread dispersal into South America during the late Holocene. Family data indicate that it may represent an autosomal recessive mutation that occurred after the peopling of the Americas as its geographic range appears to be limited to North and South Amerindian populations.


Empagliflozin targets Mfn1 and Opa1 to attenuate microglia-mediated neuroinflammation in retinal ischemia and reperfusion injury.
Neuroinflammation and mitochondrial dysfunction play crucial roles in retinal ischemia and reperfusion (IR) injury. Recent studies have identified mitochondrial function as a promising target for immunomodulation. Empagliflozin (EMPA), an anti-diabetic drug, has exhibited great potential as both an anti-inflammatory agent and a protector of mitochondrial health. This study aimed to assess the therapeutic efficacy of EMPA in retinal IR injury. To evaluate the protective effects of EMPA, the drug was injected into the vitreous body of mice post-retinal IR. Single-cell RNA sequencing (scRNA-seq) analysis was conducted to uncover the underlying mechanisms, and the results were further validated through in vivo and in vitro experiments. EMPA effectively protected retinal ganglion cells (RGCs) from IR injury by attenuating local retinal inflammation. The scRNA-seq analysis revealed that EMPA downregulated the nucleotide-binding domain and leucine-rich repeat containing protein 3 (NLRP3) signaling pathway and restored mitochondrial dynamics by upregulating the expression of mitochondrial fusion-related genes, Mitofusin 1 (Mfn1) and optic atrophy 1 (Opa1). These findings were further corroborated by Western blotting. In vitro experiments provided additional insights, demonstrating that EMPA suppressed lipopolysaccharide (LPS)-induced cell inflammation and NLRP3 inflammasome activation. Moreover, EMPA enhanced mitochondrial fusion, neutralized mitochondrial reactive oxygen species (mtROS), and restored mitochondrial membrane potential (MMP) in BV2 microglia. Notably, genetic ablation of Mfn1 or Opa1 abolished the anti-inflammatory effects of EMPA. Our findings highlight the positive contribution of Mfn1 and Opa1 to the anti-inflammatory therapeutic effect of EMPA. By restoring mitochondrial dynamics, EMPA effectively mitigates microglia-mediated neuroinflammation and prevents RGC loss in retinal IR injury.


Hydrolysis and acidogenesis of particulate organic material in mesophilic and thermophilic anaerobic digestion.
The purpose of this study was to evaluate the effect of pH and inorganic nutrient supplementations for anaerobic hydrolysis and acidogenesis of particulate organic materials at both mesophilic (35 degrees C) and thermophilic (55 degrees C) temperatures. Hydrolysis and acidogenesis of a synthetic sludge was observed in batch operation for the evaluation of the pH effect. pH was uncontrolled in one reactor and controlled at 4.5, 5.5, and 6.5 in the other three reactors at both temperatures. The greatest degree of hydrolysis and acidogenesis occurred when the pH was controlled at 6.5. The pH of the uncontrolled reactor dropped to 3.4 at both temperatures severely retarding hydrolysis and acidogenesis. Concentrations of acetic and n-butyric acids predominated with lower concentrations of propionic acid at both temperatures in all reactors. Lactic acid was produced as the earliest intermediate but as the reaction proceeded, short chain VFAs were produced as final end products with a decrease in lactic acid. The higher the pH, the earlier this trend was observed. For the controlled reactors at pH 6.5, the soluble COD production and the VSS reduction peaked in 4 days at 55 degrees C whereas it took about 11 days at 35 degrees C to obtain the same result. During the linear SCOD production period at a pH of 6.5 the hydrolysis rate of the thermophilic reactor was greater than that for mesophilic. Thermophilic conditions appeared to be more sensitive to pH than mesophilic ones for both hydrolysis and acidogenesis. Additional experiments were conducted to establish the effect of inorganic nutrient (Ca, Fe, Co, and Ni) supplementation on hydrolysis and acidogenesis at both temperatures. It has, prior to this, been assumed that only methanogenesis benefited from trace metal supplementation. However, the results demonstrated the importance of inorganic nutrient supplementation to optimize hydrolysis and acidogenesis at both temperatures.


Do International Commission of Agricultural and Biosystems Engineering (CIGR) dimension recommendations for loose housing of cows improve animal welfare?
Design of cubicles and self-locking barriers can affect cow skin alterations, lameness, and dirtiness. We investigated whether the International Commission of Agricultural and Biosystems Engineering (CIGR, Gainesville, FL)-recommended cattle housing design and dimensions actually improve cow welfare. We recorded individual cow body dimensions and assessed skin alterations, dirtiness, and lameness on 3,841 cows from 131 loose housing dairy farms (76 farms with cubicles and 55 straw-yard systems). We recorded the dimensions of cubicles (e.g., width, length, and so on) and of the self-locking barrier (e.g., top rail height and so on) for each farm. We then compared whether these dimensions would match with the individual cow body dimensions and whether compliance was associated with the occurrence of skin alterations, lameness, and dirtiness. Most cows (69.2%) had at least one skin alteration, on the tarsus (41.2%); neck, shoulder, or back (28.2%); hindquarters (22.0%); carpus (21.2%); and flank, side, or udder (11.6%). Lameness prevalence reached 17.9%. Most cows (83.1%) were scored dirty in at least one body region, mainly on the lower hind legs including hocks (81.4%), hindquarters (41.8%), and udder (21.0%). Cubicle recommendations were mostly met for cubicle resting length (75.9% of cubicles) and neck rail distance (NRD, 60.7%) but less so for overall cubicle length (CL, 38.8%), cubicle width (CW, 30.9%), neck rail height (NRH, 22.5%), head space (HS, 15.8%), partition zone for controlling lying position (ii zone, 15.7%), head and lunging space (HLS, 10.6%), partition head zone (i zone, 9.4%), and partition zone for pelvis freedom (iii zone, 6.0%). Compliance with design recommendations was associated with fewer skin alterations on neck (recommendation met for CW, CL, HS, and NRD and i zone), hindquarters (CW, CL, NRD), tarsus (CW, i and ii zones), and carpus (CW, HS, i and ii zones); less lameness (CW, NRH, i zone); and less dirtiness on the hindquarters (CW, HLS, NRD), lower hind legs (iii zone), and udder (CW, CL, HLS, NRD). Compliance with recommended i zone and ii zone design was associated with less injury and lameness but more dirtiness, whereas the opposite was true for the iii zone. Self-locking barrier recommendations were mostly met for bottom rail height (68.2%), separation wall width (SWW, 68.3%), and top rail height (TRH, 56.9%) and less often met for separation wall height (36.3%) and height difference between feeding floor and walking alley (26.5%). Risk for skin alterations on carpus and neck only decreased when SWW and TRH met recommendations. In conclusion, compliance with CIGR recommendations for some cubicle dimensions and neck rail position was associated with lower risk for cow welfare. However, the CIGR recommendations on cubicle partitions and self-locking barriers still leave welfare at risk and so need to be refined through further research with close observation of cow lying and feeding behavior.


Effects of aging on mnemonic discrimination of emotional information.
Episodic memory loss is one of the hallmarks of age-related cognitive decline and a major symptom of Alzheimer's disease. The persistence and strength of memories is determined by modulatory factors such as emotional arousal. Whether emotional memories are preserved with age or if these memories are just as susceptible to loss and forgetting is not well understood. We have recently shown that emotion alters how similar memories are stored using nonoverlapping representations (i.e., pattern separation) in an emotional mnemonic discrimination task. Here, we extend this work to testing young and older adults at 2 time points (immediately after encoding and 24 hr later). Overall, older adults performed worse than young adults, a memory deficit that was not secondary to perceptual or attentional deficits. When tested immediately, older adults were impaired on neutral target recognition but intact on emotional target recognition. We also found that a pattern we previously reported in young adults (reduced emotional compared to neutral discrimination of similar items) was reversed in older adults. When tested after 24 hr, young adults exhibited less forgetting of emotional targets compared to neutral, while older adults exhibited more forgetting of emotional targets. Finally, discrimination of highly similar positive items was preserved in older adults. These results suggest that emotional modulation of memory interacts with age in a complex manner such that the emotion-induced memory trade-off reported in young adults is reversed in older adults. These findings shed light on how emotion and memory interact in the aging brain.


[Study of the possibilities of inactivating adeno-associated virus type 4].
Adeno-associated type 4 virus (AAV-4) is inactivated by formalin dilutions 1 : 400, 1 : 1000 and 1 : 2000 for 24 hours at 37 degrees C. Under these conditions hemogglutinins (HA) and complement-fixing antigen (CFA) are retained. The residual formalin must be neutralized with sodium bisulphite. AAV-4 can also be inactivated by 3% hydrogen peroxide and HA and CFA are retained too. Hydrogen peroxide may be neutralized with catalase. No inactivation of AAV-4 could be achieved with lowere concentrations of hydrogen peroxide, even in the presence of copper sulphate.


Myofiber length and three-dimensional localization of NMJs in normal and botulinum toxin treated adult extraocular muscles.
The density and three-dimensional localization of neuromuscular junctions (NMJs) of normal and botulinum toxin-treated normal adult rabbit and monkey extraocular muscles (EOMs) were analyzed. To demonstrate average myofiber length, randomly selected individual myofibers were reconstructed and compared with total muscle length. Normal adult rabbit and monkey EOM and normal adult rabbit tibialis anterior were dissected in their entirety, frozen, sectioned longitudinally, and immunostained for NMJ localization. In addition, adult rabbit EOMs were injected with 5 U botulinum toxin, and NMJ density was determined after 2 weeks. NMJ locations for the three groups of EOM were reconstructed, and density of NMJ was determined. Individual myofibers were reconstructed from the orbital and global layers to determine mean fiber length. NMJs were dispersed throughout the entire length of all EOMs examined from adult rabbits and monkeys and were visualized by alpha-bungarotoxin staining and three-dimensional reconstruction of serial sections. In leg muscle, two relatively tight bands of NMJs were seen. Botulinum toxin significantly increased total NMJ density. Mean fiber lengths were 1.9 and 4.83 mm in the orbital and global layers, respectively, approximately 10% and 24% of the total origin-to-insertion muscle lengths. In addition, individual myofibers continuously changed their intrafascicular relationships over their lengths. The density and distribution of NMJs in normal EOMs are more extensive than previously described. Individual myofibers are significantly shorter than the tendon-to-tendon muscle length in both muscle layers. Botulinum toxin results in a doubling of NMJ density. NMJ localization in normal EOMs has ramifications for understanding eye movement control, but it is also important when surgical or pharmacologic intervention is used for the treatment of strabismus, nystagmus, or other eye muscle disorders.


Hereditary disorders of the glomerular basement membrane.
Increased knowledge of the biochemical composition of the glomerular basement membrane (GBM) and the introduction of molecular genetics has shed new light on the hereditary disorders of the GBM. In this review three disorders are highlighted. About 85% of the cases reported as Alport syndrome are transmitted as the X-linked form and are due to mutations of the COl4A5 chain localized at Xq22. The autosomal recessive form can be explained by mutations in the COl4A3 and COl4A4 gene. Anti-GBM nephritis leading to loss of the renal allograft in about 1%-5% of transplanted Alport patients can be the tragic consequence of this disorder. Some patients with familial benign hematuria have an abnormality of COl4A4. The nail-patella syndrome is a rare autosomal dominant disorder defined by the association of nail dysplasia, bone abnormalities, and frequently renal disease. The gene is localized in region 9q34.1, COl5A1 is not involved. The Finnish type is the best known of the different forms of congenital nephrotic syndrome. The gene has been mapped to the long arm of chromosome 19. Diffuse mesangial sclerosis occurs in the isolated form and as part of the Denys Drash syndrome. Disturbances of the WT1 function in the epithelial cells can have a role in the renal abnormalities of the Denys Drash syndrome.


Effects of magnesium on growth and proliferation of human embryonic stem cells.
The effects of magnesium on the growth and proliferation of human embryonic stem cells (hESCs) was explored to advance magnesium as an implant biomaterial. When magnesium ions from magnesium salt were added to the culture media at 10, 100, 250, 500, 750, and 1000 ppm (0.4, 4, 10, 20, 30, 40 mM) the rate of increase in viable cell coverage over time was higher for the larger doses of magnesium salt. Thus, the addition of magnesium ions exerted a positive effect on viable cell coverage. When hESCs were cultured with pure magnesium metal strips through transwell inserts, the cells underwent an initial increase in viable cell coverage, followed by rapid cell death within the first 24 hours. This initial increase in viable cell coverage corresponded to the colonies dispersing and losing their tightly packed morphologies. The cell death may be attributed to an increased alkalinity in the culture media incubated with the magnesium metal strips. In conclusion, since the degradation of magnesium results in both magnesium ions and OH- ions (an increase of pH), controlling the degradation of magnesium to obtain the perfect balance of ions is critical for advancing magnesium as an implant biomaterial.


Dural sac shrinkage signs on magnetic resonance imaging at the thoracic level in spontaneous intracranial hypotension-its clinical significance.
Spontaneous intracranial hypotension (SIH) is secondary to a cerebrospinal fluid leak at the spinal level without obvious causative events. Several signs on brain and cervical spine magnetic resonance (MR) imaging (MRI) have been associated with SIH but can be equivocal or negative. This retrospective study sought to identify characteristic SIH signs on thoracic spinal MRI. Cranial and spinal MR images of 27 consecutive patients with classic SIH symptoms, who eventually received epidural autologous blood patches (EBPs), were analyzed. The most prevalent findings on T2-weighted MRI at the thoracic level were anterior shift of the spinal cord (96.3%) and dorsal dura mater (81.5%), probably caused by dural sac shrinkage. These dural sac shrinkage signs (DSSS) were frequently accompanied by cerebrospinal fluid collection in the posterior epidural space (77.8%) and a prominent epidural venous plexus (77.8%). These findings disappeared in all six patients who underwent post-EBP spinal MRI. Dural enhancement and brain sagging were minimum or absent on the cranial MR images of seven patients, although DSSS were obvious in these seven patients. For 23 patients with SIH and 28 healthy volunteers, a diagnostic test using thoracic MRI was performed by 13 experts to validate the usefulness of DSSS. The median sensitivity, specificity, positive-predictive value, negative-predictive value, and accuracy of the DSSS were high (range, 0.913-0.931). Detection of DSSS on thoracic MRI facilitates an SIH diagnosis without the use of invasive imaging modalities. The DSSS were positive even in patients in whom classic cranial MRI signs for SIH were equivocal or minimal.


Sepsis as a cause and consequence of acute kidney injury: Program to Improve Care in Acute Renal Disease.
Sepsis commonly contributes to acute kidney injury (AKI); however, the frequency with which sepsis develops as a complication of AKI and the clinical consequences of this sepsis are unknown. This study examined the incidence of, and outcomes associated with, sepsis developing after AKI. We analyzed data from 618 critically ill patients enrolled in a multicenter observational study of AKI (PICARD). Patients were stratified according to their sepsis status and timing of incident sepsis relative to AKI diagnosis. We determined the associations among sepsis, clinical characteristics, provision of dialysis, in-hospital mortality, and length of stay (LOS), comparing outcomes among patients according to their sepsis status. Among the 611 patients with data on sepsis status, 174 (28%) had sepsis before AKI, 194 (32%) remained sepsis-free, and 243 (40%) developed sepsis a median of 5 days after AKI. Mortality rates for patients with sepsis developing after AKI were higher than in sepsis-free patients (44 vs. 21%; p < 0.0001) and similar to patients with sepsis preceding AKI (48 vs. 44%; p = 0.41). Compared with sepsis-free patients, those with sepsis developing after AKI were also more likely to be dialyzed (70 vs. 50%; p < 0.001) and had longer LOS (37 vs. 27 days; p < 0.001). Oliguria, higher fluid accumulation and severity of illness scores, non-surgical procedures after AKI, and provision of dialysis were predictors of sepsis after AKI. Sepsis frequently develops after AKI and portends a poor prognosis, with high mortality rates and relatively long LOS. Future studies should evaluate techniques to monitor for and manage this complication to improve overall prognosis.


Perceived duration of brief visual events is mediated by timing mechanisms at the global stages of visual processing.
There is a growing body of evidence pointing to the existence of modality-specific timing mechanisms for encoding sub-second durations. For example, the duration compression effect describes how prior adaptation to a dynamic visual stimulus results in participants underestimating the duration of a sub-second test stimulus when it is presented at the adapted location. There is substantial evidence for the existence of both cortical and pre-cortical visual timing mechanisms; however, little is known about where in the processing hierarchy the cortical mechanisms are likely to be located. We carried out a series of experiments to determine whether or not timing mechanisms are to be found at the global processing level. We had participants adapt to random dot patterns that varied in their motion coherence, thus allowing us to probe the visual system at the level of motion integration. Our first experiment revealed a positive linear relationship between the motion coherence level of the adaptor stimulus and duration compression magnitude. However, increasing the motion coherence level in a stimulus also results in an increase in global speed. To test whether duration compression effects were driven by global speed or global motion, we repeated the experiment, but kept global speed fixed while varying motion coherence levels. The duration compression persisted, but the linear relationship with motion coherence was absent, suggesting that the effect was driven by adapting global speed mechanisms. Our results support previous claims that visual timing mechanisms persist at the level of global processing.


Membrane curvature induction and tubulation are common features of synucleins and apolipoproteins.
Synucleins and apolipoproteins have been implicated in a number of membrane and lipid trafficking events. Lipid interaction for both types of proteins is mediated by 11 amino acid repeats that form amphipathic helices. This similarity suggests that synucleins and apolipoproteins might have comparable effects on lipid membranes, but this has not been shown directly. Here, we find that α-synuclein, β-synuclein, and apolipoprotein A-1 have the conserved functional ability to induce membrane curvature and to convert large vesicles into highly curved membrane tubules and vesicles. The resulting structures are morphologically similar to those generated by amphiphysin, a curvature-inducing protein involved in endocytosis. Unlike amphiphysin, however, synucleins and apolipoproteins do not require any scaffolding domains and curvature induction is mediated by the membrane insertion and wedging of amphipathic helices alone. Moreover, we frequently observed that α-synuclein caused membrane structures that had the appearance of nascent budding vesicles. The ability to function as a minimal machinery for vesicle budding agrees well with recent findings that α-synuclein plays a role in vesicle trafficking and enhances endocytosis. Induction of membrane curvature must be under strict regulation in vivo; however, as we find it can also cause disruption of membrane integrity. Because the degree of membrane curvature induction depends on the concerted action of multiple proteins, controlling the local protein density of tubulating proteins may be important. How cellular safeguarding mechanisms prevent such potentially toxic events and whether they go awry in disease remains to be determined.


Characterizing the PrEP Continuum for Black and Latinx Sexual and Gender Minority Youth.
Pre-exposure prophylaxis (PrEP) rollout efforts thus far have inadequately reached young people from underrepresented backgrounds. This study explores PrEP engagement among young Black and Latinx men who have sex with men, transgender women, and gender diverse individuals in three U.S. cities using an adapted PrEP continuum measure. We analyze data from the recruitment phase of an ongoing PrEP engagement intervention (n = 319) using partial proportional odds logistic regression. Participants reported high willingness and intention to use PrEP, yet most (82%) were not currently taking PrEP. Being insured (aOR = 2.95, 95%-CI = 1.60-5.49), having one or more PrEP users in one's sexual network (aOR = 4.19, 95%-CI = 2.61-6.79), and higher individual HIV risk scores (aOR = 1.62, 95%-CI = 1.34-1.97) were each associated with being further along the PrEP continuum. Strategies are needed to address barriers to healthcare access and leverage connections within social and sexual networks in order to bolster PrEP engagement among marginalized young people from diverse backgrounds.


Issues of consent and feedback in a genetic epidemiological study of women with breast cancer.
Women (N=21) who had had breast cancer and had been enrolled in a large genetic breast cancer epidemiological study were interviewed about their experience of participation in the study, their attitudes to the confidentiality of data, and the feedback of personal and general research results. Collection of family history information seemed more salient in indicating the genetic nature of the study than the enrolment information sheet. There were no concerns about confidentiality. While participants would have welcomed general feedback about the results of the study and were critical that this had not been provided, the feedback of personal information proved complicated and, sometimes, difficult. It is suggested that individual feedback of genetic test information in epidemiological studies should be undertaken only when there are specific reasons.