Renal tubulointerstitial hypoxia: cause and consequence of kidney dysfunction.
1. Intrarenal oxygen availability is the balance between supply, mainly dependent on renal blood flow, and demand, determined by the basal metabolic demand and the energy-requiring tubular electrolyte transport. Renal blood flow is maintained within close limits in order to sustain stable glomerular filtration, so increased intrarenal oxygen consumption is likely to cause tissue hypoxia. 2. The increased oxygen consumption is closely linked to increased oxidative stress, which increases mitochondrial oxygen usage and reduces tubular electrolyte transport efficiency, with both contributing to increased total oxygen consumption. 3. Tubulointerstitial hypoxia stimulates the production of collagen I and α-smooth muscle actin, indicators of increased fibrogenesis. Furthermore, the hypoxic environment induces epithelial-mesenchymal transdifferentiation and aggravates fibrosis, which results in reduced peritubular blood perfusion and oxygen delivery due to capillary rarefaction. 4. Increased oxygen consumption, capillary rarefaction and increased diffusion distance due to the increased fibrosis per se further aggravate the interstitial hypoxia. 5. Recently, it has been demonstrated that hypoxia simulates the infiltration and maturation of immune cells, which provides an explanation for the general inflammation commonly associated with the progression of chronic kidney disease. 6. Therapies targeting interstitial hypoxia could potentially reduce the progression of chronic renal failure in millions of patients who are otherwise likely to eventually present with fully developed end-stage renal disease.


Comprehensive study of endocrine disrupting compounds using grab and passive sampling at selected wastewater treatment plants in South East Queensland, Australia.
Chemical (gas chromatography-mass spectrometry, GC-MS) and biological (E-Screen assay) analyses were used to determine the concentrations of 15 endocrine disrupting compounds (EDCs) and estrogen equivalent (EEq) in grab and passive samples from five municipal wastewater treatment plants (WWTPs) in South East Queensland, Australia. EEq concentrations derived by E-Screen assays for the grab samples were between 108-356 ng/L for the influents and < 1-14.8 ng/L for the effluents with the exception of one effluent sample which was at 67.8 ng/L EEq. The EDC concentrations and EEq values for the passive samples were several times lower than those of the grab samples: a decrease probably caused by, but not limited to biofouling, low flow rate, biodegradation and temperature which can progressively reduce the uptake of compounds into the sampler. At this stage, grab sampling is the most reliable method for field monitoring; nevertheless, passive sampler is a useful sampling tool but the method requires more research to ensure that the information obtained can be interpreted appropriately. Although alkylphenols and phthalates were detected at higher concentrations in the wastewater samples as compared to natural hormones, the environmental risk may be negligible as their estrogenic potencies are several orders of magnitude lower than that of the natural estrogens. In most wastewater samples, the natural estrogens contributed to 60% or more of the EEq value. Removal efficacy of most estrogenic and xenoestrogenic compounds from the conventional activated sludge or biological nutrient removal (BNR) WWTPs monitored in this study was in the range of 80-> 99%. The efficiency of the WWTPs in removing estrogenic activity was > 95%. The EEqs of the E-Screen and those calculated from the results of extensive chemical analyses using the estradiol equivalency factors were comparable for most of the WWTPs samples.


Summer ice and carbon dioxide.
The extent of Antarctic pack ice in the summer, as charted from satellite imagery, decreased by 2.5 million square kilometers between 1973 and 1980. The U.S. Navy and Russian atlases and whaling and research ship reports from the 1930's indicate that summer ice conditions earlier in this century were heavier than the current average. Surface air temperatures along the seasonally shifting belt of melting snow between 55 degrees and 80 degrees N during spring and summer were higher in 1974 to 1978 than in 1934 to 1938. The observed departures in the two hemispheres qualitatively agree with the predicted impact of an increase in atmospheric carbon dioxide. However, since it is not known to, what extent the changes in snow and ice cover and in temperature can be explained by the natural variability of the climate system or by other processes unrelated to carbon dioxide, a cause-and-effect relation cannot yet be established.


Cell proliferation and cytogenesis in the mouse hippocampus.
In the present work, processes of cell proliferation, cell death, neurogenesis, and gliogenesis in the mouse hippocampus were studied. The mapping of distribution of hippocampal mitoses and counting of their number allowed a more precise definition of the data concerning the disposition and age reduction of proliferative sites in Ammon's horn and the dentate gyrus in the mouse. As a result, the following generalized scheme of development and age reduction of the germinal zones in the mouse hippocampus has been suggested. 1. Ammon's horn a) The ventricular zone, from the beginning of formation of the hippocampus (E11) until E20 b) The suprafimbrial zone, from E16 until P7 2. Dentate gyrus a) The prime germinal zone ("the anlage of the dentate gyrus" of Stanfield and Cowan1979b), from E15 until P3 b) The proliferative zone of the hilus, from P3 until P14 c) The subgranular zone, from P3 until adult age The adduced scheme needs some comments: 1. In the hippocampus (as well as in other formations of the developing brain), primary precursors of all types of cells of neuroectodermal origin are represented by cells of the ventricular zone. They give rise to cells of secondary germinal zones in the dentate gyrus and Ammon's horn and are direct precursors of the majority (if not of all) neuronal cells in Ammon's horn, the earliest originating generations of neurons in the dentate gyrus, hippocampal radial glial cells, and, evidently, of a considerable part of astroblasts and oligodendroblasts in Ammon's horn. 2. In contrast to the subiculum, Ammon's horn in the mouse lacks a subventricular (subependymal) zone. These data differ from the results obtained in the rat, where the subventricular zone is described both in the subiculum and Ammon's horn (Bayer 1980b) and from the study in the monkey, where the subiculum and Ammon's horn lack the subventricular zone (Nowakowski and Rakic 1981). Thus, in the case of the mouse, the subventricular zone can serve as a cytoarchitectural characteristic allowing detection of a border between the developing subiculum and Ammon's horn. (Another developmental feature which also identifies the border between Ammon's horn and the subiculum is a considerable number of dying cells in the region of the joint of these structures on E20-P3). 3. In the developing dentate gyrus and Ammon's horn, several extraventricular (i.e., located at a distance from brain ventricles) secondary germinal zones exist. Thus, during embryonic and early postnatal periods of the mouse development, the prime germinal zone occupies a larger part of the presumptive dentate gyrus.(ABSTRACT TRUNCATED AT 400 WORDS)


Negative regulation of nitrate reductase gene expression by glutamine or asparagine accumulating in leaves of sulfur-deprived tobacco.
Tobacco (Nicotiana tabacum L.) plants were subjected to a prolonged period of sulfur-deprivation to characterize molecular and metabolic mechanisms that permit control of primary N-metabolism under these conditions. Prior to the appearance of chlorotic lesions, sulfur-deprived tobacco leaves showed a strong decrease in the sulfate content and changes in foliar enzyme activities, mRNA accumulation and amino-acid pools. The basic amino acids glutamine, asparagine and arginine accumulated in the leaves of sulfur-deprived plants, while the foliar concentrations of aspartate, glutamate, serine or alanine remained fairly unchanged. Maximal extractable nitrate reductase (NR; EC 184.108.40.206) activity decreased strongly in response to sulfur-deprivation. The decrease in maximal extractable NR activity was accompanied by a decline in NR transcripts while the mRNAs of the plastidic glutamine synthetase (EC 126.96.36.199) or the beta-subunit of the mitochondrial ATP synthase were much less affected. Nitrate first accumulated in leaves of tobacco during sulfur-deprivation but then declined. An appreciable amount of nitrate was, however, present in severely sulfur-depleted leaves. The repression of NR gene expression is, therefore, not related to the decrease in the leaf nitrate level. However, glutamine- and/or asparagine-mediated repression of NR gene transcription is a possible mechanism of control in situations when glutamine and asparagine accumulate in leaves and provides a feasible explanation for the reduction in NR activity during sulfur-deprivation. The removal of reduced nitrogen from primary metabolism by redirection and storage as arginine, asparagine or glutamine combined with the down-regulation of nitrate reduction via glutamine- and/or asparagine-mediated repression of NR gene transcription may contribute to maintaining a normal N/S balance during sulfur-deprivation and indicate that the co-ordination of N- and S-metabolism is retained under these conditions.


Retinoschisin (RS1) interacts with negatively charged lipid bilayers in the presence of Ca2+: an atomic force microscopy study.
Retinoschisin (RS1) is a retina-specific secreted protein encoding a conserved discoidin domain sequence. As an adhesion molecule, RS1 preserves the retinal cell architecture and promotes visual signal transduction. In young males, loss-of-function mutations in the X-linked retinoschisis gene (RS1) cause X-linked retinoschisis, a form of progressive blindness. Neither the structure of RS1 nor the nature of its anchoring and organization on the plasma membranes is fully understood. The discoidin C2 domains of coagulation factors V and VIII are known to interact with extracellular phosphatidylserine (PS). In this study we have used atomic force microscopy (AFM) to study the interactions of murine retinoschisin (Rs1) with supported anionic lipid bilayers in the presence of Ca(2+). The bilayers consisting of a single lipid, PS, and mixtures of lipids with or without PS were used. Consistent with previous X-ray diffraction studies, AFM imaging showed two distinct domains in pure PS bilayers when Ca(2+) was present. Upon Rs1 adsorption, these PS and PS-containing mixed bilayers underwent fast and extensive reorganization. Protein localization was ascertained by immunolabeling. AFM imaging showed the Rs1 antibody bound exclusively to the calcium-rich ordered phase of the bilayers pointing to the sequestration of Rs1 within those domains. This was further supported by the increased mechanical strength of these domains after Rs1 binding. Besides, changes in bilayer thickness suggested that Rs1 was partially embedded into the bilayer. These findings support a model whereby the Rs1 protein binds to PS in the retinal cell plasma membranes in a calcium-dependent manner.


Exercise holds immediate benefits for affect and cognition in younger and older adults.
Physical activity is associated with improved affective experience and enhanced cognitive processing. Potential age differences in the degree of benefit, however, are poorly understood because most studies examine either younger or older adults. The present study examined age differences in cognitive performance and affective experience immediately following a single bout of moderate exercise. Participants (144 community members aged 19 to 93) were randomly assigned to one of two experimental conditions: (a) exercise (15 min of moderate intensity stationary cycling) or (b) control (15 min completing ratings of neutral IAPS images). Before and after the manipulation, participants completed tests of working memory and momentary affect experience was measured. Results suggest that exercise is associated with increased levels of high-arousal positive affect (HAP) and decreased levels of low-arousal positive affect (LAP) relative to control condition. Age moderated the effects of exercise on LAP, such that younger age was associated with a drop in reported LAP postexercise, whereas the effects of exercise on HAP were consistent across age. Exercise also led to faster RTs on a working memory task than the control condition across age. Self-reported negative affect was unchanged. Overall, findings suggest that exercise may hold important benefits for both affective experience and cognitive performance regardless of age.


Management of impacted maxillary central incisor and abnormality of labial frenulum in children: case report.
Deposition of the maxillary permanent central incisor is a rare occurrence in dental practice. It is a difficult condition to treat due to its importance to facial esthetics. If complications are to be avoided, early detection of such teeth is important. The present case report of impacted maxillary central incisor encased within an abnormally thickened labial frenulum. A 9-year-old boy, came with his parents to the Pediatric Dentistry Specialist, Dental and Oral Hospital, Airlangga University (UNAIR) with a chief complaint that his left maxillary front teeth did not grow while his right front teeth had grown perfectly. From the anamnesis, the patient had experienced a falling trauma when he was young, which caused the deciduous tooth to fall out. Good general health, no history of allergies, and no medical history of case management. This is a fixed orthodontic treatment with surgical exposure of impacted teeth and frenectomy of labial frenulum. After the crown of the impacted incisor was surgically exposed, eruption ball chain was bonded to traction the incisor. The left maxillary incisor fully erupted and normally to percussion, mobility, and sensitivity testing with good attached gingiva in the next 9 months. Management abnormality of labial frenulum in this case with frenectomy by using electrocautery for minimalized trauma in children. Fixed orthodontic therapy was continued to achieve proper alignment leading to good esthetic and functional rehabilitation. The treatment of an unerupted tooth will depend on its state, position, and presence of enough space in the dental arch to accommodate.


Preterm Birth: A Narrative Review of the Current Evidence on Nutritional and Bioactive Solutions for Risk Reduction.
Preterm birth (PTB) (<37 weeks of gestation) is the leading cause of newborn death and a risk factor for short and long-term adverse health outcomes. Most cases are of unknown cause. Although the mechanisms triggering PTB remain unclear, an inappropriate increase in net inflammatory load seems to be key. To date, interventions that reduce the risk of PTB are effective only in specific groups of women, probably due to the heterogeneity of its etiopathogenesis. Use of progesterone is the most effective, but only in singleton pregnancies with history of PTB. Thus, primary prevention is greatly needed and nutritional and bioactive solutions are a promising alternative. Among these, docosahexaenoic acid (DHA) is the most promising to reduce the risk for early PTB. Other potential nutrient interventions include the administration of zinc (possibly limited to populations with low nutritional status or poor zinc status) and vitamin D; additional preliminary evidence exists for vitamin A, calcium, iron, folic acid, combined iron-folate, magnesium, multiple micronutrients, and probiotics. Considering the public health relevance of PTB, promising interventions should be studied in large and well-designed clinical trials. The objective of this review is to describe, summarize, and discuss the existing evidence on nutritional and bioactive solutions for reducing the risk of PTB.


Pulmonary Neuroendocrine Cells Secrete γ-Aminobutyric Acid to Induce Goblet Cell Hyperplasia in Primate Models.
Mucus overproduction is a major contributor to morbidity and mortality in asthma. Mucus overproduction is induced by orchestrated actions of multiple factors that include inflammatory cytokines and γ-aminobutyric acid (GABA). GABA is produced only by pulmonary neuroendocrine cells (PNECs) in the mouse lung. Recent studies in a neonatal mouse model of allergic inflammation have shown that PNECs play an essential role in mucus overproduction by GABA hypersecretion. Whether PNECs mediate dysregulated GABA signaling for mucus overproduction in asthma is unknown. In this study, we characterized the cellular source of GABA in the lungs of nonhuman primates and humans and assessed GABA secretion and signaling in primate disease models. We found that like in mice, PNECs were the major source of GABA in primate lungs. In addition, an infant nonhuman primate model of asthma exhibited an increase in GABA secretion. Furthermore, subjects with asthma had elevated levels of expression of a subset of GABA type α (GABAα) and type β (GABAβ) receptors in airway epithelium compared with those of healthy control subjects. Last, employing a normal human bronchial epithelial cell model of preinduced mucus overproduction, we showed pharmaceutical blockade of GABAα and GABAβ receptor signaling reversed the effect of IL-13 on MUC5AC gene expression and goblet cell proliferation. Together, our data demonstrate an evolutionarily conserved intraepithelial GABA signaling that, in concert with IL-13, plays an essential role in mucus overproduction. Our findings may offer new strategies to ameliorate mucus overproduction in patients with asthma by targeting PNEC secretion and GABA signaling.


Immunohistochemical detection of autophagy-related microtubule-associated protein 1 light chain 3 (LC3) in the cerebellums of dogs naturally infected with canine distemper virus.
We investigated the expression of microtubule-associated protein 1 light chain 3 (LC3) protein in the cerebellums of dogs infected with canine distemper virus (CDV) using immunohistochemistry to detect autophagy. The cerebellums of 20 dogs infected with CDV were used. Specimens showing demyelination of white matter were considered to have an acute infection, whereas specimens showing signs of severe perivascular cuffing and demyelination of white matter were classified as having chronic CDV. Cerebellar sections were immunostained with CDV and LC3 antibodies. The cytoplasm of Purkinje cells, granular layer cells, motor neurons in large cerebellar ganglia and some neurons in white matter were positive for the LC3 antibody in both the control and CDV-infected dogs. In the infected cerebellums, however, white matter was immunostained more intensely, particularly the neurons and gemistocytic astrocytes in the demyelinated areas, compared to controls. Autophagy also was demonstrated in CDV-positive cells using double immunofluorescence staining. Our findings indicate that increased autophagy in the cerebellum of dogs naturally infected with CDV may play a role in transferring the virus from cell to cell.


ARE WITHIN-SEX MATING STRATEGY PHENOTYPES AN EVOLUTIONARY STABLE STRATEGY?
Humans have been found to display considerable variety in their pursuit of mating strategies, varying in their preference for short-term mating encounters versus established long-term relationships. While we know that differences in mating strategy exist between the two sexes (as predicted by parental investment theory), it has recently been shown that each sex may further exhibit two mating phenotypes. Here we explore the possibility that the presence of two phenotypes may be frequency dependent, thus comprising an Evolutionary Stable Strategy (ESS). We suggest that the presence of these phenotypes reflects a compromise between male preference for promiscuity and a female preference in favour of long-term mating by males.


Golgi protein FAPP2 tubulates membranes.
The Golgi-associated four-phosphate adaptor protein 2 (FAPP2) has been shown to possess transfer activity for glucosylceramide both in vitro and in cells. We have previously shown that FAPP2 is involved in apical transport from the Golgi complex in epithelial MDCK cells. In this paper we assign an unknown activity for the protein as well as providing structural insight into protein assembly and a low-resolution envelope structure. By applying analytical ultracentrifugation and small-angle x-ray scattering, we show that FAPP2 is a dimeric protein in solution, having a curved shape 30 nm in length. The purified FAPP2 protein has the capability to form tubules from membrane sheets in vitro. This activity is dependent on the phosphoinositide-binding activity of the PH domain of FAPP2. These data suggest that FAPP2 functions directly in the formation of apical carriers in the trans-Golgi network.


Prevalence of anxiety disorders among children who stutter.
Stuttering during adulthood is associated with a heightened rate of anxiety disorders, especially social anxiety disorder. Given the early onset of both anxiety and stuttering, this comorbidity could be present among stuttering children. Participants were 75 stuttering children 7-12 years and 150 matched non-stuttering control children. Multinomial and binary logistic regression models were used to estimate odds ratios for anxiety disorders, and two-sample t-tests compared scores on measures of anxiety and psycho-social difficulties. Compared to non-stuttering controls, the stuttering group had six-fold increased odds for social anxiety disorder, seven-fold increased odds for subclinical generalized anxiety disorder, and four-fold increased odds for any anxiety disorder. These results show that, as is the case during adulthood, stuttering during childhood is associated with a significantly heightened rate of anxiety disorders. Future research is needed to determine the impact of those disorders on speech treatment outcomes.


[Immunohistochemical study of small cell sarcomas and factors(nm23, CD44) predicting metastatic spreading in children].
An immunohistochemical study was made of 43 tumors from children, which were classified as small blue round cell tumors (13 neuroblastomas, 13 rhabdomyosarcomas, 14 Ewing sarcomas/PNET, 2 undifferentiated sarcoma, 1 rhabdoid tumor). The use of a wide panel of antibodies confirmed (in 74% of the cases) and corrected (in 26%) the diagnosis of a diversity of small cell sarcomas established by routine methods. In neuroblastoma, the occurrence of distal metastases was associated with the loss of nm 23 protein. Increase in CD44 expression was more frequently observed in neuroblastomas with favorable prognostic signs and embryonic rhabdosarcomas that are not accompanied by metastases. This needs further study.


Ethylene enhances transcriptions of asparagine biosynthetic genes in soybean (Glycine max L. Merr) leaves.
Soybean, a vital protein-rich crop, offers bioactivity that can mitigate various chronic human diseases. Nonetheless, soybean breeding poses a challenge due to the negative correlation between enhanced protein levels and overall productivity. Our previous studies demonstrated that applying gaseous phytohormone, ethylene, to soybean leaves significantly boosts the accumulation of free amino acids, particularly asparagine (Asn). Current studies also revealed that ethylene application to soybeans significantly enhanced both essential and non-essential amino acid contents in leaves and stems. Asn plays a crucial role in ammonia detoxification and reducing fatigue. However, the molecular evidence supporting this phenomenon remains elusive. This study explores the molecular mechanisms behind enhanced Asn accumulation in ethylene-treated soybean leaves. Transcriptional analysis revealed that ethylene treatments to soybean leaves enhance the transcriptional levels of key genes involved in Asn biosynthesis, such as aspartate aminotransferase (AspAT) and Asn synthetase (ASN), which aligns with our previous observations of elevated Asn levels. These findings shed light on the role of ethylene in upregulating Asn biosynthetic genes, subsequently enhancing Asn concentrations. This molecular insight into amino acid metabolism regulation provides valuable knowledge for the metabolic farming of crops, especially in elevating nutraceutical ingredients with non-genetic modification (GM) approach for improved protein content.


Opioid growth factor inhibition of a human squamous cell carcinoma of the head and neck in nude mice: dependency on the route of administration.
Opioid growth factor (OGF), [Met5]-enkephalin, interacts with the OGF receptor (OGFr) to inhibit the growth of human squamous cell carcinoma of the head and neck (SCCHN) in vitro. Administration of OGF by daily intraperitoneal injection (i.p.) to animals with xenografts of CAL-27, a poorly differentiated SCCHN, is known to repress tumorigenic events. In this study, the ubiquity of OGF action on SCCHN was investigated by examination of OGF activity on SCC-1 tumors; this human cell line is well-differentiated and highly invasive. Mice receiving daily i.p. injections (10 mg/kg) of OGF had more than a 3-day delay in tumor appearance, and decreases in tumor volume ranging from 51 to 64% in comparison to controls throughout the experimental period. Receptor binding analysis for OGFr showed that binding capacity (Bmax) was 2.2-fold greater than control values, but binding affinity (Kd) was comparable. Plasma OGF levels did not vary between OGF and control groups. Mice receiving OGF by continuous infusion using minipumps, or by daily intratumoral injection, had characteristics of tumorigenicity similar to their corresponding control animals, although the OGF levels in mice receiving the OGF by minipump were elevated 18-fold greater than the control group. These data indicate that: i) the inhibitory action of OGF may be ubiquitous for SCCHN, ii) OGF treatment alters the characteristics of the OGF receptor but not of plasma OGF levels, and iii) the magnitude of effects of OGF on SCCHN is dependent on the route of administration.


Costs and Economic Impacts of Physician Continuous Professional Development: A Systematic Scoping Review.
Nearly all health care professionals engage in continuous professional development (CPD), yet little is known about the cost and cost-effectiveness of physician CPD. Clarification of key concepts, comprehensive identification of published work, and determination of research gaps would facilitate application of existing evidence and planning for future investigations. The authors sought to systematically map study themes, methods, and outcomes in peer-reviewed literature on the cost and value of physician CPD. The authors conducted a scoping review, systematically searching MEDLINE, Embase, PsycInfo, and Cochrane Library databases for comparative economic evaluations of CPD for practicing physicians through April 2020. Two reviewers, working independently, screened all articles for inclusion. Three reviewers iteratively reviewed all included articles to inductively identify key features including participants, educational interventions, study designs, cost ingredients, and cost analyses. Two reviewers then independently reexamined all included articles to code these features. Of 3,338 potentially eligible studies, 111 were included. Physician specialties included internal, family, or general medicine (80 studies [72%]), surgery (14 studies [13%]), and medicine subspecialties (7 studies [6%]). Topics most often addressed general medicine (45 studies [41%]) or appropriate drug use (37 studies [33%]). Eighty-seven studies (78%) compared CPD with no intervention. Sixty-three studies (57%) reported the cost of training, and 79 (71%) evaluated the economic impact (money saved/lost following CPD). Training cost ingredients (median 3 itemized per study) and economic impact ingredients (median 1 per study) were infrequently and incompletely identified, quantified, or priced. Twenty-seven studies (24%) reported cost-impact expressions such as cost-effectiveness ratio or net value. Nineteen studies (17%) reported sensitivity analyses. Studies evaluating the costs and economic impact of physician CPD are few. Gaps exist in identification, quantification, pricing, and analysis of cost outcomes. The authors propose a comprehensive framework for appraising ingredients and a preliminary reference case for economic evaluations.


Variability of Presentation in Hereditary Neuropathy with Liability to Pressure Palsy Results in Underrecognition.
The genetic defect responsible for hereditary neuropathy with liability to pressure palsy (HNPP) is located in the same segment that is duplicated in Charcot-Marie-Tooth type 1A (CMT1A). HNPP had been presumed to be rare until an epidemiological study found a much higher incidence than was expected; the researchers suggested that HNPP was underrecognized because many affected persons have mild symptoms. We believe that another reason for underdiagnosis of HNPP is the marked phenotypic variability of the disease. We recommend, therefore, that DNA analysis for the 17p11.2 deletion be considered in patients with unexplained demyelinating neuropathy regardless of family history.


Non-contiguous extensive multifocal spinal tuberculosis- treating uncommon scenarios.
Noncontiguous multiple tuberculous spondylitis is not uncommon, and most of the reported cases have lesions only on 2 or 3 levels. To the best of our knowledge, multifocal extensive spinal TB involving the whole spine is rarely reported in the literature, which may be presented as asymptomatic and have a higher incidence of neurological complications. It is noticeable that the possibility of TB is considered for any skip lesions involving the spine cautiously. Diagnosis and treatment at early stages would resolve the neurological deficits without operation.


Effect of Neuromuscular Fatigue on the Countermovement Jump Characteristics: Basketball-Related High-Intensity Exercises.
Yoshida, N, Hornsby, WG, Sole, CJ, Sato, K, and Stone, MH. Effect of neuromuscular fatigue on the countermovement jump characteristics: basketball related high-intensity exercises. J Strength Cond Res 38(1): 164-173, 2024-The purpose of this study was to investigate basketball specific neuromuscular (NM) fatigue effect on countermovement jump (CMJ) force-time (F-T) curve characteristics. Eleven male college-level basketball athletes performed 6 CMJ trials at 3 baseline (pre) and 6 postexercise time points. The fatiguing protocol consisted of high-intensity basketball related exercises commensurate with basketball game or practice. Typical CMJ (CMJ-TYP) and phase-specific CMJ variables were derived from the F-T curve. Meaningful differences in CMJ performance were examined using effect size (ES) compared with baseline and previous postexercise time point. Baseline with 3 separated measurements demonstrated suitable CMJ variables reproducibility (CV, coefficient of variation). Most CMJ-TYP output and performance variables displayed substantial alterations immediately postexercise (0 hour) and returned to baseline at 24 hours postexercise, whereas the time and rate-related CMJ-TYP and CMJ-phase variables tended to display delayed decline peaked at 2 hours and delayed recovery to baseline at 48 hours postexercise. In conjunction with the return of the time and rate-related variables, CMJ performance displayed supercompensation at 72 hours postexercise. The results indicate altered NM functions with desired CMJ performance, such as jump height, which imply an altered movement strategy at early stage of recovery process. Full recovery may take 48-72 hours. Practitioners are, therefore, advised to monitor variables reflecting NM functions for precise manipulation of the intensity and volume of exercise to avoid prolonging the recovery from NM fatigue.


Medial rectus fadenoperation for esotropia only at near fixation.
The patient with a large esodeviation at near and a minimal if any deviation at distance who is unable to discontinue bifocal wear or demonstrates a decrease in binocular function presents a difficult therapeutic dilemma to the strabismologist. The medial rectus fadenoperation has been proposed as a method of reducing the accommodative convergence/accommodation (AC/A) ratio and may be a reasonable approach to correct a deviation that is present only at near fixation. We examined our results with this type of operation in these difficult patients. A review of consecutive patients who underwent a bilateral medial rectus fadenoperation (no recession) for high AC/A ratio esotropia with a significant esodeviation only at near was performed. Preoperative and postoperative motility and sensory status were evaluated. Sixteen patients meeting the criteria were identified. The preoperative mean distance deviation was 8.8 PD of esodeviation and the mean near deviation was 33 PD of esodeviation. The mean AC/A ratio (by the gradient method) preoperatively was 7.4 PD/D. The average age at surgery was 8 years. Six (38%) had undergone prior strabismus surgery. Average follow-up was 36 months (range 12 to 98 months). Postoperatively the mean distance deviation was 3.2 PD of esodeviation and the mean near deviation was 10.3 PD of esodeviation. The mean decrease in the near deviation was 22.8 PD. The mean postoperative AC/A ratio was 2.9. Ninety-five percent of the patients normalized their AC/A ratios and 70% attained 400 arc-sec of stereopsis postoperatively (vs 44% preoperatively. The majority of patients (86%) were able to maintain satisfactory near ocular alignment without bifocals. The fadenoperation appears to be useful in this patient population by reducing the distance-near disparity and the AC/A ratio, by decreasing the necessity for bifocal wear, and by improving binocular function at near.


Characterization of an E-box-dependent cis element in the smooth muscle alpha-actin promoter.
Identification of the regulators of smooth muscle specific gene expression is critical for understanding smooth muscle cell (SMC) differentiation and the alterations in SMC phenotype seen in vascular diseases. Previous studies have identified that a 2-bp mutation in a conserved cis-acting element (TGTTTATC) in the promoter of the chicken smooth muscle (SM) alpha-actin gene abolished nuclear factor binding and decreased transcriptional activity of a 271-bp SM alpha-actin promoter fragment when transfected into rat aortic SMC. However, the promoter region containing this conserved sequence has negative cis regulatory activity when studied in homologous systems. The goal of the present studies was to further characterize the transcriptional activity of the rat SM alpha-actin promoter region between -224 and -236 that is conserved across mammals. DNAse I analysis and electrophoretic mobility shift assays demonstrated that SMC nuclear proteins bound an extended sequence (TGTTTATCCCCATAA). Transient transfection experiments of wild-type and mutant rat SM alpha-actin promoter-luciferase constructs into rat aortic SMC revealed that promoter activity was enhanced by mutations of specific nucleotides in the TGTTTATCCCCA region. Interestingly, the TGTTTATCCCCA element in the rat SM alpha-actin promoter is centered between 2 canonical E-boxes. Mutations of the flanking E-boxes abolished the enhancement in promoter activity seen with mutation of the TGTTTATCCCCA element alone. Thus studies provide evidence for a regulatory cassette in the rat SM alpha-actin promoter that regulates gene expression via combinatorial interactions between 2 E-boxes and a newly described TGTTTATCCCCA element.


Gas spectroscopy through multimode self-mixing in a double-metal terahertz quantum cascade laser.
A multimode self-mixing terahertz-frequency gas absorption spectroscopy is demonstrated based on a quantum cascade laser. A double-metal device configuration is used to expand the laser's frequency tuning range, and a precision-micromachined external waveguide module is used to enhance the optical feedback. Methanol spectra are measured using two laser modes at 3.362 and 3.428 THz, simultaneously, with more than eight absorption peaks resolved over a 17 GHz bandwidth, which provide the noise-equivalent absorption sensitivity of 1.20×10-3 cm-1 Hz-1/2 and 2.08×10-3 cm-1 Hz-1/2, respectively. In contrast to all previous self-mixing spectroscopy, our multimode technique expands the sensing bandwidth and duty cycle significantly.


Evolution of herpesvirus thymidine kinases from cellular deoxycytidine kinase.
The thymidine kinases encoded by herpesviruses of higher vertebrates form a distinct group and are unrelated to the thymidine kinases (TKs) of other organisms. Their evolutionary source has not been identified, but our analysis has revealed a clear relationship with a sequence of human deoxycytidine kinase (dCK) published recently. We report the sequence of the putative TK of channel catfish virus, a herpesvirus of a lower vertebrate, and show that it is also related to dCK. We propose, therefore, that the TKs of herpesviruses of higher and lower vertebrates have evolved, either independently or successively, from a cellular dCK.


Neuropeptide Y and corticotropin-releasing factor bi-directionally modulate inhibitory synaptic transmission in the bed nucleus of the stria terminalis.
Neuropeptide Y (NPY) and corticotropin-releasing factor (CRF) have opposing effects on stress and anxiety. Both can modify synaptic activity through their binding to NPY receptors (YRs) and CRF receptors (CRFRs) respectively. The bed nucleus of the stria terminalis (BNST) is a brain region with enriched expression of both NPY and YRs and CRF and CRFRs. A component of the "extended amygdala", the BNST is anatomically well-situated to integrate stress and reward-related processing in the CNS, regulating activation of the hypothalamic-pituitary-adrenal (HPA) axis and reward circuits. Using whole-cell recordings in a BNST slice preparation, we found that NPY and CRF inhibit and enhance GABAergic transmission, respectively. Pharmacological experiments suggest that NPY depresses GABAergic transmission through activation of the Y2 receptor (Y2R), while both pharmacological and genetic experiments suggest that CRF and urocortin enhance GABAergic transmission through activation of the CRF receptor 1 (CRFR1). Further, the data suggest that NPY acts to regulate GABA release, while CRF enhances postsynaptic responses to GABA. These results suggest potential anatomical and cellular substrates for the robust behavioral interactions between NPY and CRF.


Effect of decreased levels of intrinsic tetrahydrobiopterin on endothelial function in anesthetized rats.
Tetrahydrobiopterin (BH4) deficiency has been suggested to be an important factor in vascular endothelial dysfunction. In this study, we investigated the influence of decreased BH4 level produced by administration of 2,4-diamino-6-hydroxypyrimidine (DAHP), a specific inhibitor of the rate-limiting enzyme of BH4 synthesis, on vascular endothelial function in anesthetized rats. Wistar rats were given DAHP (0.1 - 1.0 g/kg, i.p.) or the vehicle 5 h before the experiment. Depressor responses to the endothelium-dependent vasodilator acetylcholine and the endothelium-independent vasodilator sodium nitroprusside were tested. After the experiment, blood and thoracic aorta were taken for estimating their BH4 levels and plasma concentrations of nitrite plus nitrate. DAHP produced marked decreases in BH4 levels in plasma and aorta in a dose-related manner. Baseline values for hemodynamics were not affected by DAHP. Depressor responses to acetylcholine were attenuated with the highest dose of DAHP (1.0 g/kg) but not with DAHP (0.3 g/kg), although similar decreases in BH4 levels were seen with these two doses of DAHP. Treatment with DAHP at each dose did not decrease plasma concentrations of nitrite plus nitrate. These findings suggest that a decrease in BH4 levels by acute inhibition of de novo BH4 synthesis does not necessarily cause endothelial dysfunction.


Which factors can aid clinicians to identify a risk of pain during the following month in patients with bone metastases? A longitudinal analyses.
Explore clinical factors associated with higher pain intensity and future pain in patients with bone metastases to identify patients who can benefit from closer follow-up or pain-modifying interventions. This is a secondary analysis of 606 patients with bone metastases included in a multicenter longitudinal study. The dependent variables were "average pain" and "worst pain" in the last 24 h (0-10 NRS). Twenty independent variables with potential association to pain intensity were selected based on previous literature. Cross-sectional analyses were performed with multiple linear regression to explore factors associated with pain intensity at baseline. Longitudinal data were analyzed with a generalized equation models to explore current factors associated with pain intensity at the next visit in 1 month. Current pain intensity (p < 0.001), sleep disturbances (p 0.01 and 0.006), drowsiness (p 0.003 and 0.033) and male gender (p 0.045 and 0.001) were associated with higher average and worst pain intensity in 1 month. In addition, breakthrough pain was related to higher worst pain intensity (p 0.003) in 1 month. The same variables were also associated with higher average pain intensity at baseline. Higher current pain intensity, sleep disturbances, drowsiness, male gender, and breakthrough pain are factors associated with higher pain intensity in patients with bone metastases at the next follow-up in 1 month. These factors should be assessed in clinical practice and may aid clinicians in identifying patients that can benefit from closer follow-up or interventions to prevent lack of future pain control. TRIAL REGISTRATION IN CLINICALTRIALS.GOV : NCT01362816.


Emotional orienting during interoceptive threat in orthostatic intolerance: Dysautonomic contributions to psychological symptomatology in the postural tachycardia syndrome and vasovagal syncope.
Cognitive and emotional processes are influenced by interoception (homeostatic somatic feedback), particularly when physiological arousal is unexpected and discrepancies between predicted and experienced interoceptive signals may engender anxiety. Due to the vulnerability for comorbid psychological symptoms in forms of orthostatic intolerance (OI), this study investigated psychophysiological contributions to emotional symptomatology in 20 healthy control participants (13 females, mean age 36 ± 8 years), 20 postural tachycardia syndrome (PoTS) patients (18 females, mean age 38 ± 13 years) and 20 vasovagal syncope (VVS) patients (15 females, mean age 39 ± 12 years). We investigated indices of emotional orienting responses (OR) to randomly presented neutral, pleasant and unpleasant images in the supine position and during the induced interoceptive threat of symptom provocation of head-up tilt (HUT). PoTS and VVS patients produced greater indices of emotional responsivity to unpleasant images and, to a lesser degree, pleasant images, during interoceptive threat. Our findings are consistent with biased deployment of response-focused emotion regulation (ER) while patients are symptomatic, providing a mechanistic underpinning of how pathological autonomic overexcitation predisposes to anxiogenic traits in PoTS and VVS patients. This hypothesis may improve our understanding of why orthostasis exacerbates cognitive symptoms despite apparently normal cerebral autoregulation, and offer novel therapeutic targets for behavioural interventions aimed at reducing comorbid cognitive-affective symptoms in PoTS and VVS.


Medical students' experience in practical skills is far from stakeholders' expectations.
This study compares medical graduates' experience in practical skills with a range of stakeholders' expectations. A questionnaire listing 58 practical skills was sent out to a group of graduating medical students. The medical students were asked to indicate their experience in each skill during medical school. A similar questionnaire was sent out to five groups of stakeholders asking for their expectations regarding graduates' experience. The stakeholders were: faculty members; consultants at clinical departments with interns in training; general practitioners; nurses; recently graduated junior doctors. A total of 472 questionnaires were sent out and 315 (67%) were returned. Medical graduates showed substantial variation in level of experience, and their experience was substantially lower than the expectations of the stakeholders. Nurses and junior doctors tended to have higher expectations compared with faculty members and consultants. Differences between our results and reports in the literature from elsewhere emphasize the importance of performing local needs assessments, and in this process stakeholders apart from faculty members should be involved.


Local transposition of a hobo element within the decapentaplegic locus of Drosophila.
We have efficiently mobilized a phenotypically silent hobo transgene inserted within the cis-regulatory heldout region of the decapentaplegic (dpp) locus in Drosophila melanogaster. The goal of our experiment was to identify germline transmission of a local transposition event within the dpp locus that meets two specific criteria. First, excision of the hobo construct does not generate an adult mutant phenotype, suggesting minimal alteration to the original site of insertion. Second, we required a new insertion of the hobo transgene into the Haploinsufficient region of the locus approximately 25 kb away. Genetic and molecular criteria are used to evaluate candidate germlines. In a pilot study, this local transposition event occurred independently in two individuals. Both of the transposition events appear to be new insertions into the dpp transcription unit. One insertion is between the two protein-coding exons, and the other is in the 3'-untranslated region of exon three. Strains carrying these insertions are valuable new reagents for the analysis of dpp function and molecular evolution. These results further support the use of the hobo system as an important tool in Drosophila genetics.


Asthma and risk of non-respiratory tract infection: a population-based case-control study.
Asthmatics have increased risks of airway-related infections. Little is known about whether this is true for non-airway-related serious infections such as Escherichia coli bloodstream infection (BSI). We assessed whether asthma is associated with a risk of developing community-acquired E coli BSI. The study was designed as a population-based retrospective case-control study. This population-based study was conducted in Olmsted County, Minnesota. The study included 259 all eligible community-acquired E coli BSI cases in Olmsted County, MN between 1998 and 2007 and 259 birthday-matched, gender-matched and residency-matched controls. Only community-acquired E coli BSI cases as the primary outcome was included. Asthma status as an exposure was ascertained by predetermined criteria. An adjusted OR and 95% CI for the association between asthma and risk of community-acquired E coli BSI was calculated using conditional logistic regression. Of 259 eligible cases, 179 (69%) were women and mean age was 61±22 years. Of the 259 cases 37 (14%) and 16 (6%) of 259 controls had a prior history of asthma (adjusted OR 2.74; 95% CI 1.11 to 6.76; p=0.029). The population attributable risk of asthma for community-acquired E coli BSI was 9%. Although not statistically significant, there was a borderline association between having a history of food allergy and increased risk of community-acquired E coli BSI (6% vs 2%; adjusted OR 3.51; 95% CI 0.94 to 13.11; p=0.062). Based on the findings of the current population-based, case-control investigation, a history of asthma may be associated with risk of community-acquired E coli BSI. The impact of asthma on risk of microbial infections may go beyond airways.


Computer-assisted implant placement. A case report: treatment of the mandible.
The authors present a case of immediate loading of mandibular implants using a 5-step procedure. The first step consists of building a scannographic template, the second step consists of taking a computerized tomographic (CT) scan, and the third step consists of implant planning using SurgiCase software. The final 2 steps consist of implant placement using a drill guide created by stereolithography and placement of the prosthesis. Using a CT scan-based planning system, the surgeon is able to select the optimal locations for implant placement. By incorporating the prosthetic planning using a scannographic template, the treatment is optimized from a prosthetic point of view. Furthermore, the use of a stereolithographic drill guide allows a physical transfer of the implant planning to the patient's mouth. The scannographic template is designed so that it can be transformed into a temporary fixed prosthesis for immediate loading, and the definitive restoration is placed 3 months later.


Contextual Factors to Improve Implementation of Malaria Chemoprevention in Children: A Systematic Review.
Malaria remains a leading cause of childhood morbidity and mortality in sub-Saharan Africa, particularly among children under 5 years of age. To help address this challenge, the WHO recommends chemoprevention for certain populations. For children and infants, the WHO recommends seasonal malaria chemoprevention (SMC), perennial malaria chemoprevention (PMC; formerly intermittent preventive treatment in infants [IPTi]), and, more recently, intermittent preventive treatment in school children (IPTsc). This review describes the contextual factors, including feasibility, acceptability, health equity, financial considerations, and values and preferences, that impact implementation of these strategies. A systematic search was conducted on July 5, 2022, and repeated April 13, 2023, to identify relevant literature. Two reviewers independently screened titles for eligibility, extracted data from eligible articles, and identified and summarized themes. Of 6,295 unique titles identified, 65 were included. The most frequently evaluated strategy was SMC (n = 40), followed by IPTi (n = 18) and then IPTsc (n = 6). Overall, these strategies were highly acceptable, although with IPTsc, there were community concerns with providing drugs to girls of reproductive age and the use of nonmedical staff for drug distribution. For SMC, door-to-door delivery resulted in higher coverage, improved caregiver acceptance, and reduced cost. Lower adherence was noted when caregivers were charged with giving doses 2 and 3 unsupervised. For SMC and IPTi, travel distances and inclement weather limited accessibility. Sensitization and caregiver education efforts, retention of high-quality drug distributors, and improved transportation were key to improving coverage. Additional research is needed to understand the role of community values and preferences in chemoprevention implementation.


Discovery and characterization of novel small molecule inhibitors of human Cdc25B dual specificity phosphatase.
Cdc25A and Cdc25B dual-specificity phosphatases are key regulators of cell cycle transition and proliferation. They have oncogenic properties and are overexpressed in many human tumors. Because selective Cdc25 phosphatase inhibitors would be valuable biological tools and possible therapeutic agents, we have assayed a small molecule library for in vitro inhibition of Cdc25. We now report the identification of two new structurally distinct classes of Cdc25 inhibitors with cellular activity. The cyclopentaquinoline 3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinoline-4,8-dicarboxylic acid (5661118) and the naphthofurandione 3-benzoyl-naphtho[1,2-b]furan-4,5-dione (5169131) had in vitro IC50 values of 2.5 to 11 microM against recombinant Cdc25 and were less potent inhibitors of other phosphatases. Unlike 5661118, 5169131 caused reversible inhibition of Cdc25B and displayed competitive inhibitor kinetics. No growth inhibitory activity was seen with 5661118, whereas 10 to 30 microM 5169131 caused G1/S and G2/M arrest. We also found that 5169131 inhibited human PC-3 prostate and MDA-MB-435 breast cancer cell proliferation. Concentration-dependent Tyr15 hyperphosphorylation was seen on cyclin-dependent kinase with a 1-h 5169131 treatment, consistent with Cdc25 inhibition. Cells resistant to DNA toposiomerase II inhibitors were as sensitive to 5169131 as parental cells, indicating that this quinone compound does not inhibit topoisomerase II in vivo. Molecular modeling was used to predict a potential interaction site between the inhibitor and Cdc25B and to provide insights as to the molecular origins of the experimental observations. Based on its kinetic profile and cellular activity, we suggest that 5169131 could be an excellent tool for further studies on the cellular roles of Cdc25.


Calibrated mitotic oscillator drives motile ciliogenesis.
Cell division and differentiation depend on massive and rapid organelle remodeling. The mitotic oscillator, centered on the cyclin-dependent kinase 1-anaphase-promoting complex/cyclosome (CDK1-APC/C) axis, spatiotemporally coordinates this reorganization in dividing cells. Here we discovered that nondividing cells could also implement this mitotic clocklike regulatory circuit to orchestrate subcellular reorganization associated with differentiation. We probed centriole amplification in differentiating mouse-brain multiciliated cells. These postmitotic progenitors fine-tuned mitotic oscillator activity to drive the orderly progression of centriole production, maturation, and motile ciliation while avoiding the mitosis commitment threshold. Insufficient CDK1 activity hindered differentiation, whereas excessive activity accelerated differentiation yet drove postmitotic progenitors into mitosis. Thus, postmitotic cells can redeploy and calibrate the mitotic oscillator to uncouple cytoplasmic from nuclear dynamics for organelle remodeling associated with differentiation.


Modernization, stress, and blood pressure: new directions in research.
The relationship between modernization and blood pressure has been formally examined in anthropology for some 3 decades. A prominent hypothesis to account for the increase in blood pressure in more modernized (or economically developed) communities is the stressful nature of cultural and social change. Research has progressed from hypothesizing that culture change is stressful to trying to operationalize theoretical models of what it is about culture change that is stressful and in turn relating those more precise variables to blood pressure variability within and between communities. Here, I selectively review the literature on modernization and blood pressure, especially the research literature that explicitly uses models of the stress process to guide that research. The most interesting results have been obtained when the use of the stress model has been informed by careful ethnographic work. This has enabled researchers to adapt models of the stress process to be culturally appropriate in local populations. In addition, incorporating an explicit model of culture, especially one that is sensitive to intracultural diversity, has led to new hypotheses regarding the modification of the effects of stressors by social and cultural context. I conclude with a discussion of recent innovations in ethnographic methods, specifically the cultural consensus model, and the use of those methods in operationalizing relevant variables in culturally appropriate and sensitive ways. The utility of combining these methods in the study of modernization and blood pressure is illustrated by recent research in Brazil.


Kaposi's sarcoma-associated herpesvirus oncoprotein K13 protects against B cell receptor-induced growth arrest and apoptosis through NF-κB activation.
Kaposi's sarcoma-associated herpesvirus (KSHV) has been linked to the development of Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman's disease (MCD). We have characterized the role of KSHV-encoded viral FLICE inhibitory protein (vFLIP) K13 in the modulation of anti-IgM-induced growth arrest and apoptosis in B cells. We demonstrate that K13 protects WEHI 231, an immature B-cell line, against anti-IgM-induced growth arrest and apoptosis. The protective effect of K13 was associated with the activation of the NF-κB pathway and was deficient in a mutant K13 with three alanine substitutions at positions 58 to 60 (K13-58AAA) and a structural homolog, vFLIP E8, both of which lack NF-κB activity. K13 upregulated the expression of NF-κB subunit RelB and blocked the anti-IgM-induced decline in c-Myc and rise in p27(Kip1) that have been associated with growth arrest and apoptosis. K13 also upregulated the expression of Mcl-1, an antiapoptotic member of the Bcl2 family. Finally, K13 protected the mature B-cell line Ramos against anti-IgM-induced apoptosis through NF-κB activation. Inhibition of anti-IgM-induced apoptosis by K13 may contribute to the development of KSHV-associated lymphoproliferative disorders.


Tobacco Product Use Among Lesbian, Gay, and Bisexual Adolescents.
Sexual minority youth face health disparities and a high risk of substance abuse. This study sought to fully characterize the disparity of tobacco use and risk factors in this high-risk subpopulation. Using data from the 2015 Youth Risk Behavior Survey, the current use of various tobacco products (cigarettes, smokeless tobacco, cigars, and e-cigarettes) was compared according to sex and distinct sexual identities (lesbian, gay, bisexual, and unsure). Heterosexual/straight adolescents served as the control group. Of 14 703 respondents, 88.8% were heterosexual/straight, 2.0% were lesbian or gay, 6.0% were bisexual, and 3.2% were unsure about their sexual identity. Sexual minorities had a higher prevalence of tobacco product use than their heterosexual/straight counterparts. Sex had a significant effect on the disparities of tobacco use. Lesbian and bisexual girls had higher odds of reporting current use of any tobacco product, cigarettes, cigars, and e-cigarettes than did straight girls, whereas sexual minority boys had similar smoking behaviors compared with straight boys. Substance use, including marijuana use, drinking, and binge drinking, was significantly associated with any tobacco use. Tobacco use differs among subgroups of sexual minority youth, with lesbians and bisexual girls having a higher prevalence of tobacco use than their straight peers. Heterogeneity of tobacco use across distinct sexual identity groups underscores the need to develop evidence-based tobacco control strategies for sexual minority youth.


L-Asparaginase production by Streptomyces griseus.
Streptomyces griseus ATCC 10137 synthesizes about 1 IU of L-asparaginase/100 ml of a 4% peptone medium. The enzyme has a pH optimum of 8.5 which is comparable to that of the L-asparaginase derived from Escherichia coli which has antitumor properties.


Use of natural coralline biomaterials as reinforcing and gas-forming agent for developing novel hybrid biomatrices: microarchitectural and mechanical studies.
This paper describes the first attempt in fabrication of three-dimensional macroporous composites of chitosan and natural coralline material with pore sizes of 300-400 microm, exceeding the upper pore size limit of 250 microm obtained with freeze-dried chitosan-based scaffolds. Natural coral particulates of less than 20 microm, which is mainly composed of calcium carbonate (CaCO3), was simultaneously used as reinforcing phase and gas-forming agent to obtain a structure with large pores and improved mechanical and biological properties. The reaction between the coralline material and the acidic chitosan polymer solvent, which produced carbon dioxide, was rapidly stopped by the subsequent thermally induced phase separation technique, leaving coralline particulates in the polymeric structure. Scaffolds containing five different proportions of coralline material (0, 25, 50, 75, and 100 wt%) were investigated. The coralline-chitosan weight ratio was studied for its effects on the physical properties of the scaffolds. The relation between scaffold microarchitecture and mechanical properties was assessed with scanning electron microscope (SEM), along with micro-CT imaging and compression testing. The scaffolds were used in bone marrow cell culturing experiments to assess the effect of composition on cell behavior through cell-material interaction and morphological observation by SEM. Higher coralline concentration increased the pore wall thickness and favored large pore formation. Varying the coralline particulate to chitosan polymer ratio from 0 to 75 wt% increased the average pore size from 80 microm to 400 microm while the porosity decreased from 91% to 78%. The compressive modulus was improved proportionally with the coralline content, and the 75 wt% composites had a significantly higher modulus than other chitosan-based scaffold groups. More cells were observed on scaffolds with higher coralline content. The cell culture experiments indicated that the scaffolds containing coralline material might have a high cell affinity, since it allowed fast cell attachment and spreading.


A Food System Approach for Sustainable Food-Based Dietary Guidelines: An Exploratory Scenario Study on Dutch Animal Food Products.
This study explores interconnections between food consumption and production of animal (by-)products in different food system scenarios within the scope of Dutch Food-Based Dietary Guidelines (FBDG). For this scenario study, a Microsoft Excel model was created that include seven scenarios with different quantities of eggs, milk, cheese, beef cattle, broilers, and pigs as input. Number of animals, intake of energy, animal protein, saturated fatty acids (SFAs), trans-fatty acids (TFAs), salt, greenhouse gas emissions (GHGEs), and land use (LU) were calculated and compared with current consumption and reference values. Based on the concept of eating the whole animal, every recommended lean, unprocessed portion of beef comes along with a non-recommended portion of beef (two portions for pork, 0.5 portion for broilers). The reference values for SFAs, TFAs, and salt were not exceeded if the intake of meat is limited to 410 g/week. The scenarios with recommended 450 mL semi-skimmed milk and 40 g/day low-fat cheese results in 36 g/day of butter as by-product, exceeding its acceptable intake three times. The near-vegetarian scenario with recommended amounts of eggs, milk, and cheese, includes only a portion of beef/calf per 6 days and a portion of chicken per 9 weeks as by-products. This scenario more than halves the GHGE and LU. Finally, the scenario that included the maximum recommended amounts of animal products is reachable with half the current size of Dutch livestock. This conceptual framework may be useful in the discussion on how future sustainable FBDG can incorporate a more food system-based approach.


The roles of nucleoid-associated proteins and topoisomerases in chromosome structure, strand segregation, and the generation of phenotypic heterogeneity in bacteria.
How to adapt to a changing environment is a fundamental, recurrent problem confronting cells. One solution is for cells to organize their constituents into a limited number of spatially extended, functionally relevant, macromolecular assemblies or hyperstructures, and then to segregate these hyperstructures asymmetrically into daughter cells. This asymmetric segregation becomes a particularly powerful way of generating a coherent phenotypic diversity when the segregation of certain hyperstructures is with only one of the parental DNA strands and when this pattern of segregation continues over successive generations. Candidate hyperstructures for such asymmetric segregation in prokaryotes include those containing the nucleoid-associated proteins (NAPs) and the topoisomerases. Another solution to the problem of creating a coherent phenotypic diversity is by creating a growth-environment-dependent gradient of supercoiling generated along the replication origin-to-terminus axis of the bacterial chromosome. This gradient is modulated by transcription, NAPs, and topoisomerases. Here, we focus primarily on two topoisomerases, TopoIV and DNA gyrase in Escherichia coli, on three of its NAPs (H-NS, HU, and IHF), and on the single-stranded binding protein, SSB. We propose that the combination of supercoiling-gradient-dependent and strand-segregation-dependent topoisomerase activities result in significant differences in the supercoiling of daughter chromosomes, and hence in the phenotypes of daughter cells.


Does nedocromil sodium have a steroid sparing effect in adult asthmatic patients requiring maintenance oral corticosteroids?
A randomised, double blind, placebo controlled trial of nedocromil sodium was undertaken to assess its corticosteroid sparing effect in 50 adults with asthma who had required an oral corticosteroid dose of (or equivalent to) at least 5 mg prednisolone a day continuously during the preceding year, in addition to inhaled beclomethasone dipropionate and bronchodilators. Patients having corticosteroids other than prednisolone were changed to prednisolone. A four week baseline period was followed by 20 weeks of inhaled nedocromil sodium (16 mg daily) or placebo. After four weeks of the treatment phase an attempt was made to reduce the oral prednisolone maintenance dose by 2.5 mg a fortnight until a dose of 5 mg daily was reached and thereafter by 1 mg a fortnight, provided that there was no significant clinical deterioration as judged by clinic assessments and daily diary cards. Of 50 patients recruited, 47 entered the treatment phase (age range 16-64 years), 24 receiving nedocromil sodium and 23 placebo. The total steroid reduction achieved was 2.5 mg in the nedocromil group and 3 mg in the placebo group, which did not differ significantly. There was no significant change in symptoms, lung function or inhaler use in either group during the study. The number of patient requiring short term upward adjustment of booster doses of oral prednisolone for exacerbations of asthma was similar in the two groups (26 with placebo, 28 with nedocromil). Thus nedocromil sodium does not appear to provide an oral corticosteroid sparing effect in chronic steroid dependent asthma.


Perioperative posterior reversible encephalopathy syndrome in 2 pediatric neurosurgery patients with brainstem ependymoma.
Posterior reversible encephalopathy syndrome (PRES) has been described in pediatric neurooncology patients, although it has not been documented perioperatively in pediatric neurosurgery patients not actively receiving chemotherapy. Recently at the authors' facility, 2 cases of PRES were diagnosed perioperatively in children with brainstem ependymoma. Both patients had presented with hypertension, altered mental status, and seizures and demonstrated MR imaging features consistent with PRES. The patients were treated with antiseizure and antihypertension medications, leading to improvement in both clinical symptoms and neuroimaging findings. These cases are the first to document PRES in perioperative pediatric neurosurgery patients not actively receiving chemotherapy. Both patients had ependymoma involving the brainstem, which may have led to intra- and perioperative hemodynamic instability (including hypertension) and predisposed them to this syndrome. An awareness of PRES in similar scenarios will aid in the prevention, diagnosis, and treatment of pediatric neurosurgery patients with this syndrome.


Commercial fishing deaths - United States, 2000-2009.
Commercial fishing is one of the most dangerous occupations in the United States. During 1992-2008, an annual average of 58 reported deaths occurred (128 deaths per 100,000 workers), compared with an average of 5,894 deaths (four per 100,000 workers) among all U.S. workers. During the 1990s, safety interventions addressing specific hazards identified in Alaska resulted in a significant decline in the state's commercial fishing fatality rate. During 2007-2010, CDC expanded surveillance of commercial fishing fatalities to the rest of the country's fishing areas. To review the hazards and risk factors for occupational mortality in the U.S. commercial fishing industry, and to explore how hazards and risk factors differ among fisheries and locations, CDC collected and analyzed data on each fatality reported during 2000-2009. This report summarizes the results, which showed that, among the 504 U.S. commercial fishing deaths, the majority occurred after a vessel disaster (261 deaths, 52%) or a fall overboard (155 deaths, 31%). By region, 133 (26%) deaths occurred off the coast of Alaska, 124 (25%) in the Northeast, 116 (23%) in the Gulf of Mexico, 83 (16%) off the West Coast, and 41 (8%) in the Mid- and South Atlantic. Type of fishing was known in 478 deaths; shellfish (226, 47%) was the most common, followed by groundfish (144, 30%) and pelagic fish (97, 20%). To reduce fatalities in this industry, additional prevention measures tailored to specific high-risk fisheries and focusing on prevention of vessel disasters and falls overboard are needed.


Biophysical characterization of lynx-nicotinic receptor interactions using atomic force microscopy.
Nicotinic acetylcholine receptors (nAChRs) are broadly expressed in the central and peripheral nervous systems, playing essential roles in cholinergic neurotransmission. The lynx family proteins, a subset of the Ly6/uPAR superfamily expressed in multiple brain regions, have been shown to bind to nAChRs and modulate their function via allosteric regulation. The binding interactions between lynx and nAChRs, however, have not been systematically quantified and compared. In this work, we characterized the interactions between lynx1 or lynx2 and α3β4- or α7-nAChRs using single-molecule atomic force microscopy (AFM). The AFM technique allows the quantification of the off-rate of lynx-nAChR binding and of the energetic barrier width between the bound state and transition state, providing a biophysical means to compare the selectivity of lynx proteins for nAChR subtypes. Results indicate that lynx1 has a marginal preference for α7- over α3β4-nAChRs. Strikingly, lynx2 exhibits a two order of magnitude stronger affinity for α3β4- compared to α7-nAChRs. Together, the AFM assay serves as a valuable tool for the biophysical characterization of lynx-nAChR binding affinities. Revealing the differential affinities of lynx proteins for nAChR subtypes will help elucidate how lynx regulates nAChR-dependent functions in the brain, including nicotine addiction and other critical pathways.


A Multi-Site Analysis of the Prevalence of Food Insecurity in the United States, before and during the COVID-19 Pandemic.
The coronavirus disease 2019 (COVID-19) pandemic profoundly affected food systems including food security. Understanding how the COVID-19 pandemic impacted food security is important to provide support and identify long-term impacts and needs. The National Food Access and COVID research Team (NFACT) was formed to assess food security over different US study sites throughout the pandemic, using common instruments and measurements. This study presents results from 18 study sites across 15 states and nationally over the first year of the COVID-19 pandemic. A validated survey instrument was developed and implemented in whole or part through an online survey of adults across the sites throughout the first year of the pandemic, representing 22 separate surveys. Sampling methods for each study site were convenience, representative, or high-risk targeted. Food security was measured using the USDA 6-item module. Food security prevalence was analyzed using ANOVA by sampling method to assess statistically significant differences. Respondents (n = 27,168) indicate higher prevalence of food insecurity (low or very low food security) since the COVID-19 pandemic, compared with before the pandemic. In nearly all study sites, there is a higher prevalence of food insecurity among Black, Indigenous, and People of Color (BIPOC), households with children, and those with job disruptions. The findings demonstrate lingering food insecurity, with high prevalence over time in sites with repeat cross-sectional surveys. There are no statistically significant differences between convenience and representative surveys, but a statistically higher prevalence of food insecurity among high-risk compared with convenience surveys. This comprehensive study demonstrates a higher prevalence of food insecurity in the first year of the COVID-19 pandemic. These impacts were prevalent for certain demographic groups, and most pronounced for surveys targeting high-risk populations. Results especially document the continued high levels of food insecurity, as well as the variability in estimates due to the survey implementation method.


Effectiveness of Ergonomic Training to Decrease Awkward Postures during Dental Scaling Procedures: A Randomized Clinical Trial.
Studies demonstrate that there is a lack of effective ergonomic principles for adopting a neutral posture during the execution of dental procedures. ISO 11.226:2000 Standard, Corr. 1:2006 has been thoroughly evaluated and adapted to the way that dentists work by the European Society of Dental Ergonomics (ESDE). However, after 15 years, no studies that showed strong evidence of effectiveness in reducing the prevalence of awkward posture in applying its parameters within the scope of dental practice were found. The aim of this study was to verify the effectiveness of applying the ergonomic parameters proposed by the European Society of Dental Ergonomics (ESDE) and ISO 11226 in reducing the prevalence of the main awkward postures adopted by female dental surgeons during the execution of dental scaling on a dental mannequin. A randomized clinical trial was carried out with sixty dental surgeons randomly assigned to two groups: the intervention group, who received instructions and theoretical and practical ergonomic training; and the control group, who received the same training only at the end of the study. For data analysis, Software IBM SPSS 27 and RStudio was used. Descriptive statistics were performed to verify the effectiveness of the intervention, and generalized linear models (specifically, generalized estimated equation models) were used. Poisson distribution was carried out with log link function and network analyses. Sixty female dental surgeons participated in the study. Twenty-two were distributed in the intervention group and thirty-eight in the control group. It was found that ergonomic training enabled a 63% reduction in the prevalence of awkward postures and that there was a statistically significant difference (p < 0.001) only in the intervention group. The analyses showed that the estimated marginal means of postures not recommended in the groups' initial control, final control, initial intervention, and final intervention were 8.6, 8.2, 9.0, and 3.4, respectively. The relationship of networks analyses of the variables is shown with different profiles in the control and intervention groups, but the same pattern between the groups only vary in the strength and direction of the correlations. It was concluded that the ergonomic training based on the parameters of ISO 11226 and DIN EN 1005-4, and its adaptations to the dental practice provided by the European Society of Dental Ergonomics, as well as recent studies, contributed significantly to reducing the prevalence of awkward postures adopted by female dentists during the simulation of the basic periodontal procedures; however, it was not effective enough to improve the posture of the head and neck.


Isolation and sequencing of cDNA clones coding for the catalytic unit of glucose-6-phosphatase from two haplochromine cichlid fishes.
Complementary DNA clones coding for the catalytic unit of the enzyme glucose-6-phosphatase (G6Pase) were obtained from Haplochromis nubilus and Haplochromis xenognathus, two cichlid fish species from Lake Victoria. The translated sequence of these two cDNAs identifies a polypeptide consisting of 352 amino acid residues and showing a 54.4% similarity to the human form of G6Pase. The amino acid sequences of the two fish species are identical. The comparison of the fish amino acid sequence with the corresponding sequences of rat, mouse, and human G6Pase revealed that the amino acid residues, which are involved in G6Pase catalysis in humans, are also conserved in fish G6Pase. Northern blot analysis showed that G6Pase is expressed at the same level in 6- and 10-day-old fish. A three base pair insertion/deletion polymorphism was found in the 3'-untranslated region of the fish G6Pase gene. The polymorphism will be a useful marker in a phylogenetic study of Lake Victoria cichlids.


Impacts of cognitive impairment for different levels and causes of traumatic brain injury, and education status in TBI patients.
Traumatic brain injury (TBI) is one of main causes of death and disability among many young and old populations in different countries. The aim of this study were to consider and predict the cognitive impairments according to different levels and causes of TBI, and education status. The study was performed using the Mini-Mental State Examination (MMSE) to estimate cognitive impairment in patients at a trauma center in Zahedan city. Individuals were considered eligible if 18 years of age or older. This investigation assessed a subset of patients from a 6-month pilot study. The study participants comprised 66% males and 34% females. Patient mean age was 32.5 years and SD was 12.924 years. One-way analysis of variance between groups indicated cognitive impairment related to different levels and causes of TBI, and education status in patients. There was a significant difference in the dimensions of cognitive impairments for different levels and causes of TBI, and education status. A regression test showed that levels of traumatic brain injury (b=.615, p=.001) and education status (b=.426, p=.001) predicted cognitive impairment. Different levels of TBI and education status were useful for predicting cognitive impairment in patients. Severe TBI and no education were associated with worse cognitive performance and higher disability. These data are essential in terms of helping patients understand their needs. Therefore, the factors identified can help plan effective rehabilitation programs.


Impact of culture and cryopreservation on MHC class II antigen expression in canine and porcine islets.
Canine and porcine islets were either cultured for 10 days at 37 degrees C or cryopreserved. The effect of these treatments on MHC class II antigen expression was examined by indirect immunofluorescence test using the class II monoclonal antibody 2MC3. Within untreated canine islets exclusively round shaped cells as leukocytes and monocyte/macrophage like cells with dendritic branches were positive with 2MC3. Porcine islets additionally exhibited strong immunofluorescence of the vascular endothelium. Tissue culture significantly reduced the class II antigen expression in both species. The majority of the untreated canine islets had between 5 and 19 2MC3 positive cells. Cell culture reduced the number of class II positive cells to a maximum of 3 and even 84.9% of the islets were completely negative for class II antigens. Porcine islets showed a total class II antigen loss of the vascular endothelium, whereas leukocytes and monocytes/macrophages remained class II antigen positive. Cryopreservation did not have clear-cut effects on the MHC expression in both species.


Hormonal contraception and HIV disease progression.
The majority of the 15.4 million human immunodeficiency virus (HIV)-infected women worldwide are of child-bearing age and need access to contraception. Hormonal methods of contraception are safe, acceptable, and effective in preventing unwanted pregnancies. Many published studies have examined the impact of hormonal contraception on HIV disease acquisition and transmissibility. Far fewer have investigated the relationship between hormonal contraception and HIV disease progression. This review examines available data on this relationship from clinical, animal, and immunological studies. Several clinical studies suggest an overall effect but are not definitive, and the mechanisms behind HIV disease progression are unclear. Animal and immunological data suggest that immunomodulation by hormonal contraceptive methods may affect the immune response to HIV infection. Additional work is needed in this area to elucidate the possible relationship between hormonal methods for birth control and progression to acquired immunodeficiency syndrome in HIV-infected women.


In vivo evidence for the resistance of Theileria annulata to buparvaquone.
The present study describes an outbreak of tropical theileriosis cases refractory to buparvaquone treatment, which occurred in a small-size dairy farm in Tunisia. Out of seven treated cows, four died in spite of repeated buparvaquone injections (2.5 and 5 mg kg(-1)) and the monitoring of the affected cows showed no improvement of the course of the disease with a consistent decrease in the haematocrit, the persistence of fever and an increased parasitaemia after treatment. Ticks were fed on a calf experimentally infected with one isolate established in culture from one of the cases and the resultant infected ticks ground up to generate a supernatant infected with the potentially resistant stock. This was used to experimentally infect three calves, and the clinical observations, post-buparvaquone treatment, showed an absence of the usual effect of buparvaquone treatment on the parasite Theileria annulata, such as the rapid decline of schizont index and parasitaemia and a rapid recovery from the disease. These results confirmed for the first time the occurrence of resistance to buparvaquone in the protozoan T. annulata.


[Anterior approaches to the acetabulum].
Since the 1960s the ilioinguinal approach by Letournel with the three anatomic windows has been successfully established for the treatment of acetabular fractures involving predominantly the anterior column. The previous standard approach, the iliofemoral approach by Smith-Petersen, is still used for the therapy of anterior wall or isolated femoral head fractures. The increase in acetabular fractures in the elderly with lateral compression fractures after lateral falls, characterized by medial displacement of the quadrilateral plate and superomedial dome impaction, led to the use of the intrapelvic modified Stoppa approach with or without the first window of the ilioinguinal approach in the 1990s. To combine the advantages of the second and third windows of the ilioinguinal approach and the medial view of the modified Stoppa approach the Berne research group recently introduced the pararectus approach in acetabular surgery, which can be used as a less invasive acetabular surgical (LIAS) technique especially in the elderly.


A comparison of trabecular meshwork sphingolipids and ceramides of ocular normotensive and hypertensive states of DBA/2J mice.
To determine the differential profiles of sphingomyelin, sphingoid base, sphingoid base-1-phosphate, and ceramide and their quantitative differences between trabecular meshwork (TM) derived from normotensive and hypertensive intraocular pressure states of DBA/2J mice. Normotensive and hypertensive state TM were collected from mice and analyzed. Lipid extraction was performed using the Bligh and Dyer method, and the protein concentrations were determined using the Bradford method. The lipids were identified and quantified using appropriate standards with a TSQ Quantum Access Max triple quadrupole mass spectrometer applying class-specific lipid identification settings. The comparative profiles of sphingomyelin, sphingoid base, sphingoid base-1-phosphate, and ceramide between normotensive and hypertensive TM showed several species unique to a phase and as well common between states. The presence or absence of several sphingolipids and ceramides in the normotensive or hypertensive states may contribute to better understanding of the glaucomas.


Nuclear localization signal peptides for the import of plasmid DNA in gene therapy (review).
Failure of cells to import specific proteins into nuclei can lead to carcinogenesis. Trafficking of nuclear proteins from the site of their synthesis in the cytoplasm to the sites of function in the nucleus through pore complexes is mediated by the presence of nuclear localization signals (NLSs) on proteins to be imported into nuclei. mRNA is exported through the same route as a complex with nuclear proteins. During gene therapy the foreign DNA needs to enter nuclei for its transcription in order to express the therapeutic protein; a pathway is proposed involving the complexation of plasmids and oligonucleotides with nascent nuclear proteins possessing NLSs as a prerequisite for their nuclear import. Covalent linkage of NLS peptides to oligonucleotides and plasmids or formation of complexes of plasmids with proteins possessing multiple NLS peptides is proposed to increase their import rates and the efficiency of gene expression. We suggest that cancer cells import more efficiently foreign DNA into nuclei compared with terminally differentiated cells because of their increased rates of proliferation and protein import and are easier targets for expression of foreign genes.


Metabolic engineering with ATP-citrate lyase and nitrogen source supplementation improves itaconic acid production in Aspergillus niger.
Bio-based production of organic acids promises to be an attractive alternative for the chemicals industry to substitute petrochemicals as building-block chemicals. In recent years, itaconic acid (IA, methylenesuccinic acid) has been established as a sustainable building-block chemical for the manufacture of various products such as synthetic resins, coatings, and biofuels. The natural IA producer Aspergillus terreus is currently used for industrial IA production; however, the filamentous fungus Aspergillus niger has been suggested to be a more suitable host for this purpose. In our previous report, we communicated the overexpression of a putative cytosolic citrate synthase citB in an A. niger strain carrying the full IA biosynthesis gene cluster from A. terreus, which resulted in the highest final titer reported for A. niger (26.2 g/L IA). In this research, we have attempted to improve this pathway by increasing the cytosolic acetyl-CoA pool. Additionally, we have also performed fermentation optimization by varying the nitrogen source and concentration. To increase the cytosolic acetyl-CoA pool, we have overexpressed genes acl1 and acl2 that together encode for ATP-citrate lyase (ACL). Metabolic engineering of ACL resulted in improved IA production through an apparent increase in glycolytic flux. Strains that overexpress acl12 show an increased yield, titer and productivity in comparison with parental strain CitB#99. Furthermore, IA fermentation conditions were improved by nitrogen supplementation, which resulted in alkalization of the medium and thereby reducing IA-induced weak-acid stress. In turn, the alkalizing effect of nitrogen supplementation enabled an elongated idiophase and allowed final titers up to 42.7 g/L to be reached at a productivity of 0.18 g/L/h and yield of 0.26 g/g in 10-L bioreactors. Ultimately, this study shows that metabolic engineering of ACL in our rewired IA biosynthesis pathway leads to improved IA production in A. niger due to an increase in glycolytic flux. Furthermore, IA fermentation conditions were improved by nitrogen supplementation that alleviates IA induced weak-acid stress and extends the idiophase.


Isolation of two distinct epithelial cell lines from a single feline mammary carcinoma with different tumorigenic potential in nude mice and expressing different levels of epidermal growth factor receptors.
From a single spontaneous feline mammary carcinoma, two subpopulations of epithelial tumor cells have been isolated. The variant cells were established as cell lines designated K248C and K248P. DNA ploidy analysis showed that the two cell lines represented cell populations already present in the original tumor. Chromosome analysis confirmed the feline origin of K248C and K248P and demonstrated that in addition to unique marker chromosomes characteristic for each cell line, both cell lines had several marker chromosomes in common. These data suggest that the two cell populations arose from a hypothetical single ancestor which diverged during tumor progression. The K248C and K248P cell lines differed from one another with respect to their tumorigenicity in athymic mice and epidermal growth factor (EGF) receptor content. The K248C cells were highly tumorigenic as indicated by a short latency period and high take rate. The K248P cells were poorly tumorigenic. Southern blot analysis revealed that the K248C cells contained an amplified EGF receptor gene that was accompanied by elevated levels of EGF receptor RNA and protein. The K248C cells were growth inhibited in vitro at EGF concentrations that stimulated growth of K248P cells. The amplification of the EGF receptor gene could be detected only in DNA derived from K248C cells at high passage numbers and not in DNA derived from the original tumor and K248C cells at low passage numbers. These data suggest that amplification of the EGF receptor gene occurred during establishment of the K248C cell line.


EphA5 and ephrin-A2 expression during optic nerve regeneration: a 'two-edged sword'.
During development, gradients of EphA receptors (nasal(low)-temporal(high)) and their ligands ephrin-As (rostral(low)-caudal(high)) are involved in establishing topography between retinal ganglion cells (RGCs) and the superior colliculus (SC). EphA5-expressing RGC axons are repulsed by ephrin-A2-expressing SC neurones. In adult rats RGCs maintain graded EphA5 expression but ephrin-A2 expression is down-regulated in the SC to a weak gradient. At 1 month after optic nerve transection, EphA5 expression is reduced in the few remaining RGCs and is no longer graded; by contrast, SC ephrin-A2 is up-regulated to a rostral(low)-caudal(high) gradient. Here we examined expression in adult rat 1 month after bridging the retina and SC with a peripheral nerve graft, a procedure that enhances RGC survival and permits RGC axon regeneration. Double labelling with cell markers revealed preservation of a nasal(low)-temporal(high) EphA5 gradient in RGCs and establishment of a rostral(low)-caudal(high) ephrin-A2 gradient within neurones of the SC. The results suggest a potential for guidance cues to restore the topography of RGC axons in the SC. However, high ephrin-A2 levels were also found in astrocytes surrounding the peripheral nerve graft insertion site. The repulsive ephrin-A2 environment offers at least a partial explanation for the observation that only a limited number of RGC axons can exit the graft to enter target central nervous system tissue.


A two-dimensional immunomagnetic nano-net for the efficient isolation of circulating tumor cells in whole blood.
An immunomagnetic "nano-net" was designed and synthesized for specifically capturing rare cells of interest from mixtures. The nano-net, Ab@Lipo-MNP-GO, consists of conjugated antibody molecules on a lipid coated magnetic nanoparticle-graphene oxide sheet complex. The magnetism, chemical composition, and the morphology of the construct and its precursors were characterized by SQUID, FTIR, TGA, DLS and SEM, to confirm the feasibility of the synthetic steps and the resulting properties suitable for solution phase immuno-recognition for cell capture. When applied to capturing circulating tumor cells (CTCs) in oral, colon and lung cancer clinical patients' blood samples, the nano-net construct exhibited far superior ability whereas conventional immunomagnetic beads in some cases were unable to capture any CTCs, even by increasing the bead concentration. Confocal images showed that the nano-net wrapped around the CTCs while the immunomagnetic beads attached them with point contacts. A stable, patch-like multivalent matrix nano-net was demonstrated to tackle the shortcomings of single point contact of immunomagnetic beads to the target cell. This strategy is universal for any cell separation in complex fluids.


Quantitative proteomic analysis in symptomatic and asymptomatic apical periodontitis.
To quantitatively and qualitatively compare the host proteomic profile in samples of symptomatic and asymptomatic apical periodontitis (AP) using nano-liquid chromatography-electron spray tandem mass spectrometry. Samples were obtained from 18 patients with radiographically evident AP, divided into symptomatic and asymptomatic groups (nine per group) according to clinical characteristics. After sample collection, protein extraction, purification and quantification of the samples were performed, which were analysed by reverse-phase liquid chromatography coupled to mass spectrometry. Label-free quantitative proteomic analysis was performed by Protein Lynx Global Service software. Differences in expression of proteins between the groups were calculated using the Monte Carlo algorithm, considering P < 0.05 for down-regulated proteins and 1 - P > 0.95 for up-regulated proteins. Proteins were identified with the embedded ion accounting algorithm in the software and a search of the Homo sapiens UniProt database. A total of 853 individual human proteins were identified. In the quantitative analysis, common proteins to both groups accounted for 143 proteins. Differences in expression between groups resulted in 51 up-regulated proteins (1 - P > 0.95) in the symptomatic group, including alpha-1-antitrypsin, protein S100-A8, myeloperoxidase, peroxiredoxin and lactotransferrin. This group also had 43 down-regulated proteins (P < 0.05), comprising immunoglobulin, neutrophil defensin, pyruvate kinase and alpha-enolase. The qualitative analysis considered only the exclusive proteins of each group. For the symptomatic group, 318 complete proteins and 29 fragments were identified, such as dedicator of cytokinesis protein, intersectin, prostaglandin, phospholipase DDHD2 and superoxide dismutase. For the asymptomatic group, 326 complete proteins and 37 fragments were identified, including azurocidin, C-reactive protein, collagen alpha, cathepsin, heat shock and laminin. Quantitative differences in the expression of common proteins in cases of symptomatic and asymptomatic AP were found, which were mostly related to host immune response in both groups. Exclusive proteins in the symptomatic group were mainly related to the host response to the presence of viruses in endodontic infections, oxidative stress and proteolytic enzymes. The results provide a basis for a better understanding of cellular and molecular pathways involved in AP, establishing specific proteomic profiles for symptomatic and asymptomatic conditions.


[Comparison of postoperative complications following bone harvesting from two different donor sites for autologous bone replacement (hip bone and proximal epiphysis of the tibia)].
Whereas autologous bone replacement was earlier applied in maxillofacial surgery virtually only for the restoration of mandibular defects and for the osteoplasty of patients with cleft alveolar process, the free transplantation of autologous bone (spongiosa or cortical bone or both) is nowadays primarily used for implantation purposes. Autologous bone is still the gold standard for bone replacement. This is the case even though a wide selection of bone substitutes is currently available, with which new bone equivalent to autologous bone can be produced in certain cases. Autologous bone is often obtained from intraoral sources, but if a larger quantity of spongiosa is required, these sites (the chin, the retromolar area of the mandible, the muscular process, etc.) are not suitable. Of the extraoral donor sites, the most frequently used site is the iliac crest, but the proximal epiphysis of the tibia is also appropriate for this purpose since we have recently performed bone transplantations on appreciable numbers of patients, we decided to compare the morbidity data relating to the two donor sites. In the 9 months between March and November 2007, sinus elevations were carried out on 14 patients with bone taken from the tibia, while in 38 patients bone was taken from the iliac crest for osteoplasty on clefted alveolar process. The comparison was based on postoperative clinical examinations, the complaints of the patients and objective study of the morbidity relating to the two donor sites. Clinically the patients tolerated both interventions well. Mobilization took place on the day of intervention. There were no major complications; one minor haematoma was observed after each type of surgery. The postoperative complaints of the patients did not reveal any essential difference. Following bone harvesting from the iliac crest, the gait of the patients slightly hampered for up to 10 to 14 days. In the tibia cases, the patients experienced no pain on walking by the second day. As regards donor site morbidity, protracted (1-2 weeks) oedema was observed after hip surgery, with paraesthesia of the area of innervation of the n. cutaneous femoris lateralis in 1 case, while there was a minor seroma following tibia surgery in 1 case. Our clinical experience suggests that, if 10-15 cm(3) spongiosa is required for augmentation purposes and there is no need for cortical bone, the patient is exposed to less stress when bone is taken from the proximal epiphysis of the tibia.


Cost optimal allocation of amplifiers and DCMs in WDM ring networks.
Designing metropolitan wavelength division multiplexing (WDM) ring networks with minimum deployment cost is a demanding issue in Telecommunication Network planning . We have already presented amplifier placement methods to minimize the number of amplifiers in WDM rings for the case all amplifiers follow a unique gain model. In this paper, we take into account different types of amplifiers with predefined fixed characteristics and costs. We also formulate fiber dispersion limitations on the ring design, and present two efficient methods for placing amplifiers and Dispersion Compensation Modules (DCMs) in WDM rings to minimize the total deployment cost of the system. The first method deals with both linear and nonlinear equations and uses a mixed integer nonlinear programming (MINLP) solver where the second method applies the linear approximation of nonlinear constraints, and uses a mixed integer linear programming (MILP) solver to minimize the total cost of the system. We carry out Simulation experiments to confirm the applicability of the methods and compare the results for various network configurations.