Effect of Otoconial Proteins Fetuin A, Osteopontin, and Otoconin 90 on the Nucleation and Growth of Calcite.
We investigated the roles of three proteins associated with the formation of otoconia including fetuin A, osteopontin (OPN), and otoconin 90 (OC90). In situ atomic force microscopy (AFM) studies of the effects of these proteins on the growth of atomic steps on calcite surfaces were performed to obtain insight into their effects on the growth kinetics. We also used scanning electron microscopy to examine the effects of these proteins on crystal morphology. All three proteins were found to be potent inhibitors of calcite growth, although fetuin A promoted growth at concentrations below about 40 nM and only became an inhibitor at higher concentrations. We then used in situ optical microscopy to observe calcite nucleation on films of these proteins adsorbed onto mica surfaces. By measuring the calcite nucleation rate as a function of supersaturation, the value of the interfacial energy that controls the free energy barrier to heterogeneous nucleation was determined for each protein. OPN and OC90 films led to significantly reduced interfacial energies as compared to the value for homogeneous calcite nucleation in bulk solution. The value for fetuin A was equal to that for bulk solution within experimental error. Zeta potential measurements showed all of the proteins possessed negative surface charge and varied in magnitude according to sequence fetuin A > OC90 > OPN. In addition, the interfacial energies exhibited an inverse scaling with the zeta potential. In analogy to previous measurements on polysaccharide films, this scaling indicates the differences between the proteins arise from the effect of protein surface charge on the solution-substrate interfacial energy.


Plastic surgical delivery systems for targeted gene therapy.
The expansion of gene therapy applications from inherited disorders to acquired conditions has been mirrored by an exponential rise in both experimental work and clinical trials. This review highlights current plastic surgical delivery systems and clinical applications for targeted gene therapy. We revisit some of the vectors used both experimentally and in clinical gene therapy trials, with an emphasis on developments in plastic surgical delivery systems resulting in improved targeting of therapeutic genes. In addition, we discuss a novel technique for the delivery of gene therapy using the ex vivo transduction of free flaps, developed in our laboratory. This delivery system achieves targeted high-level transgene expression with minimal demonstrable systemic toxicity. Advances in delivery systems are essential for translating basic research into clinical therapeutics.


COVID-19 risk perceptions and depressive symptoms in South Africa: Causal evidence in a longitudinal and nationally representative sample.
Studies worldwide have highlighted the acute and long-term depressive impacts of psychosocial stressors due to the 2019 coronavirus disease (COVID-19) pandemic, particularly in low- and middle-income countries. Among the wide range of risk factors for depression that transpired during pandemic, greater perceptions of individual vulnerability to the COVID-19 have emerged as a major predictor of increased depressive risk and severity in adults. We estimated the extent to which COVID-19 risk perceptions affected adult depressive symptoms in a longitudinal, nationally representative sample in South Africa. We used covariate balanced propensity scores to minimize the bias from treatment assignment to estimate average causal effects of COVID-19 risk perceptions. The point prevalence of perceived COVID-19 infection risk increased between the third and fifth months of the pandemic, which corresponded with elevations in national COVID-19 infection rates. Approximately 33% of adults met or surpassed the PHQ-2 cut-off score of 2. An increase in perceived risk of COVID-19 infection predicted worse depressive symptoms in adults four months later. Our findings highlight the widespread mental health burdens of the COVID-19 pandemic and emphasize the importance of greater psychological resources and structural changes to promote equitable access to COVID-19 risk mitigation policies.


The effects of supervised learning on event-related potential correlates of music-syntactic processing.
Humans process music even without conscious effort according to implicit knowledge about syntactic regularities. Whether such automatic and implicit processing is modulated by veridical knowledge has remained unknown in previous neurophysiological studies. This study investigates this issue by testing whether the acquisition of veridical knowledge of a music-syntactic irregularity (acquired through supervised learning) modulates early, partly automatic, music-syntactic processes (as reflected in the early right anterior negativity, ERAN), and/or late controlled processes (as reflected in the late positive component, LPC). Excerpts of piano sonatas with syntactically regular and less regular chords were presented repeatedly (10 times) to non-musicians and amateur musicians. Participants were informed by a cue as to whether the following excerpt contained a regular or less regular chord. Results showed that the repeated exposure to several presentations of regular and less regular excerpts did not influence the ERAN elicited by less regular chords. By contrast, amplitudes of the LPC (as well as of the P3a evoked by less regular chords) decreased systematically across learning trials. These results reveal that late controlled, but not early (partly automatic), neural mechanisms of music-syntactic processing are modulated by repeated exposure to a musical piece. This article is part of a Special Issue entitled SI: Prediction and Attention.


Hemoglobin Induces Early and Robust Biofilm Development in Streptococcus pneumoniae by a Pathway That Involves comC but Not the Cognate comDE Two-Component System.
Streptococcus pneumoniae grows in biofilms during both asymptomatic colonization and infection. Pneumococcal biofilms on abiotic surfaces exhibit delayed growth and lower biomass and lack the structures seen on epithelial cells or during nasopharyngeal carriage. We show here that adding hemoglobin to the medium activated unusually early and vigorous biofilm growth in multiple S. pneumoniae serotypes grown in batch cultures on abiotic surfaces. Human blood (but not serum, heme, or iron) also stimulated biofilms, and the pore-forming pneumolysin, ply, was required for this induction. S. pneumoniae transitioning from planktonic into sessile growth in the presence of hemoglobin displayed an extensive transcriptome remodeling within 1 and 2 h. Differentially expressed genes included those involved in the metabolism of carbohydrates, nucleotides, amino acid, and lipids. The switch into adherent states also influenced the expression of several regulatory systems, including the comCDE genes. Inactivation of comC resulted in 67% reduction in biofilm formation, while the deletion of comD or comE had limited or no effect, respectively. These observations suggest a novel route for CSP-1 signaling independent of the cognate ComDE two-component system. Biofilm induction and the associated transcriptome remodeling suggest hemoglobin serves as a signal for host colonization in pneumococcus.


Effects of ankle-foot orthoses on mediolateral foot-placement ability during post-stroke gait.
Accurate and precise mediolateral foot placement is important for balance during gait, but is impaired post stroke. Mediolateral foot placement may be improved with ankle-foot orthosis use. The purpose of this study was to determine whether an ankle-foot orthosis improves mediolateral foot-placement ability during post-stroke ambulation. Crossover trial with randomized order of conditions tested. The accuracy and precision of mediolateral foot placement was quantified while subjects targeted four different randomized step widths. Subjects were tested with and without their regular non-rigid ankle-foot orthosis in two separate visits (order randomized). While ankle-foot orthosis use corrected foot and ankle alignment (i.e. significantly decreased mid-swing plantar flexion, p = 0.000), effects of ankle-foot orthosis use on hip hiking (p = 0.545), circumduction (p = 0.179), coronal plane hip range of motion (p = 0.06), and mediolateral foot-placement ability (p = 0.537) were not significant. While ankle-foot orthosis-mediated equinovarus correction of the affected foot and ankle was not associated with improved biomechanics of walking (i.e. proximal ipsilateral hip kinematics or mediolateral foot-placement ability), it may affect other aspects of balance that were not tested in this study (e.g. proprioception, cerebellar, vestibular, and cognitive mechanisms). Studies that investigate the effect of ankle-foot orthosis on gait can help advance stroke rehabilitation by documenting the specific gait benefits of ankle-foot orthosis use. In this study, we investigated the effect of ankle-foot orthosis use on mediolateral foot-placement ability, an aspect of gait important for maintaining balance.


Control of transplant tolerance and intragraft regulatory T cell localization by myeloid-derived suppressor cells and CCL5.
Myeloid-derived suppressor cells (MDSC) are a heterogeneous population of immature cells that are believed to inhibit immune responses in the contexts of cancer and organ transplantation, in association with regulatory T cells (Treg). However, the way in which MDSC cooperate with Treg remains elusive. In this study, we used DNA microarrays to analyze gene expression in blood-derived MDSC from rat recipients of kidney allografts. We found CCL5 (Rantes), a chemotactic C-C motif 5 chemokine, to be strongly downregulated after treatment with a tolerizing regimen. The amount of CCL5 protein was also lower in the plasma of tolerant recipients, whereas intragraft CCL5 was unchanged. Because CCL5 is chemotactic for Treg, we hypothesized that a gradient of CCL5 between the graft and peripheral blood might contribute to the intragraft localization of Treg in tolerant animals. To test this hypothesis, we treated tolerant rat recipients of kidney allografts with recombinant rat CCL5 to restore normal plasma concentrations. This led to a strong reduction in intragraft Treg monitored by immunohistofluorescence and by quantitative real-time PCR measurement of Foxp3 mRNA. Ultimately, this treatment led to an increase in serum creatinine concentrations and to kidney graft rejection after about a month. The kidney function of syngeneic grafts was not affected by a similar administration of CCL5. These data highlight the contribution of MDSC to the establishment of a graft-to-periphery CCL5 gradient in tolerant kidney allograft recipients, which controls recruitment of Treg to the graft where they likely contribute to maintaining tolerance.


Pharmacology through Play: using Lego® to revise core concepts for undergraduates.
This article was migrated. The article was marked as recommended. Background: Pharmacology, while critical knowledge for healthcare professionals, is often viewed by students as dry and difficult to understand. We sought to examine the student acceptability of a Lego®-based learning session, in an attempt to improve pharmacology learning. Methods: In line with constructivist theories, students were facilitated to build, in small groups, their own Lego® shape to represent an area of core pharmacology and to use this to explain the concept to other students (e.g. agonist-receptor interactions). The validated Course Experience Questionnaire (CEQ) was used to gauge students' ideas on the session. Multiple choice questions were used before and after the session to evaluate knowledge. Results: Most students were positive regarding the session, finding it enjoyable, relevant for their learning and even recommending it be used to explore more complex areas of pharmacology. In addition, there was a significant increase in the MCQ scores following the session. Conclusions: This study used constructivist theory to develop a novel teaching intervention to create a more student-centred, active learning environment. This effective low-cost method could be applied to other teaching programmes to enhance student learning.


Predictors of type of vascular access in hemodialysis patients.
Complications from vascular access account for 15% of hospital admissions among US hemodialysis patients. Complications are less frequent with arteriovenous fistulas than with synthetic grafts. We assessed clinical and nonclinical predictors of whether patients with end-stage renal disease (ESRD) starting hemodialysis receive a fistula or graft. We also investigated changes in practice between 1986-1987 and 1990. Cross-sectional study. United States hemodialysis population. Random, national samples of ESRD patients who started hemodialysis in 1986-1987 (n=2741) or 1990 (n=1409) from United States Renal Data System Special Studies. Type of permanent vascular access (arteriovenous fistula vs synthetic graft), analyzed using multivariate logistic regression. Clinical and demographic factors as well as socioeconomic status, region of residence, and year starting hemodialysis predicted the type of vascular access. Overall, 56% of patients had grafts 30 days after starting dialysis, but graft use increased from 51% in 1986-1987 to 65% in 1990 (adjusted odds ratio [AOR], 1.67 for 1990 vs 1986-1987; 95% confidence interval [CI], 1.43-1.95; P<.001). Graft use (relative to fistula) varied by region of residence (ranging from AOR, 0.20; 95% CI, 0.14-0.28; P<.001 [New England], to AOR, 2.69; 95% CI, 2.03-3.58; P<.001 [East South Central]; both relative to the national average). This national study documents large variations in the relative use of fistulas and grafts and a trend away from fistulas. The prevalence of comorbid conditions fails to explain these findings. Presentation and referral of patients early in the process of their ESRD, teaching surgeons to place fistulas, and training dialysis nurses to access fistulas may increase their use.


Preparation, X-ray structure, and reactivity of 2-iodylpyridines: recyclable hypervalent iodine(V) reagents.
2-Iodylpyridine and four examples of 3-alkoxy-2-iodylpyridines were prepared by oxidation of the respective 2-iodopyridines with 3,3-dimethyldioxirane. Structures of 2-iodylpyridine, 2-iodyl-3-isopropoxypyridine, and 2-iodyl-3-propoxypyridine were established by single-crystal X-ray diffraction analysis. 2-Iodyl-3-propoxypyridine has moderate solubility in organic solvents (e.g., 1.1 mg/mL in acetonitrile) and can be used as a recyclable reagent for oxidation of sulfides and alcohols. The reduced form of this reagent, 2-iodo-3-propoxypyridine, can be effectively separated from the reaction mixture by treatment with diluted sulfuric acid and recovered from the acidic aqueous solution by adding aqueous sodium hydroxide.


Gene transfer into hematopoietic stem cells of nonhuman primates.
Nonhuman primates provide an appropriate preclinical large-animal model to test the efficacy of bone marrow gene therapy procedures. Successful retroviral vector-mediated gene transfer into monkey pluripotent hematopoietic stem cells (PHSC) has closed the gap between gene transfer experiments in mouse models and clinical application of bone marrow gene therapy. After initial bone marrow transplant failures, ex vivo bone marrow culture conditions were found that sufficiently supported maintenance of the long-term repopulating ability of genetically modified autologous monkey grafts. The efficiency of gene transfer into primate PHSC has, however, remained at least one order of magnitude lower than has been achieved in mice. Similar gene transfer efficiencies have been obtained with total bone marrow grafts, CD34+ bone marrow grafts, and mobilized peripheral blood progenitor cell grafts; however, various attempts to increase the transduction efficiency have been without significant success. Primate PHSC seem to require quite different culture conditions for their maintenance and transduction than mouse PHSC, in particular regarding hematopoietic growth factor addition. In contrast to observations in other species, some form of conditioning appeared essential for engraftment of transduced PHSC in monkeys. Although it has been shown that mouse retroviruses can replicate in monkeys and are capable of inducing neoplasms, experiments in monkeys have sufficiently confirmed the safety of current gene transfer procedures to allow their clinical application.


Disruption of zinc and copper interactions with Aβ(1-40) by a non-toxic, isoniazid-derived, hydrazone: a novel biometal homeostasis restoring agent in Alzheimer's disease therapy?
Disruptions of biometal-Aβ(1-40) interactions by an isoniazid-derived hydrazone, INHHQ, were demonstrated via in vitro NMR titrations. The compound has adequate theoretical BBB absorption properties, assessed by in silico studies. In vivo acute toxicity assays indicate that INHHQ is innocuous up to 300 mg kg(-1), showing potential as an anti-Alzheimer's drug.


Cofilin-mediated F-actin severing is regulated by the Rap GTPase and controls the cytoskeletal dynamics that drive lymphocyte spreading and BCR microcluster formation.
When lymphocytes encounter APCs bearing cognate Ag, they spread across the surface of the APC to scan for additional Ags. This is followed by membrane contraction and the formation of Ag receptor microclusters that initiate the signaling reactions that lead to lymphocyte activation. Breakdown of the submembrane cytoskeleton is likely to be required for the cytoskeleton reorganization that drives cell spreading and for removing physical barriers that limit Ag receptor mobility. In this report, we show that Ag receptor signaling via the Rap GTPases promotes the dephosphorylation and activation of the actin-severing protein cofilin and that this results in increased severing of cellular actin filaments. Moreover, we show that this cofilin-mediated actin severing is critical for the changes in actin dynamics that drive B and T cell spreading, for the formation of BCR microclusters, and for the increased mobility of BCR microclusters within the plasma membrane after BCR engagement. Finally, using a model APC, we show that activation of this Rap-cofilin signaling module controls the amount of Ag that is gathered into BCR microclusters and that this is directly related to the magnitude of the resulting BCR signaling that is initiated during B cell-APC interactions. Thus, Rap-dependent activation of cofilin is critical for the early cytoskeletal changes and BCR reorganization that are involved in APC-dependent lymphocyte activation.


PPAR gamma is required for placental, cardiac, and adipose tissue development.
The nuclear hormone receptor PPAR gamma promotes adipogenesis and macrophage differentiation and is a primary pharmacological target in the treatment of type II diabetes. Here, we show that PPAR gamma gene knockout results in two independent lethal phases. Initially, PPAR gamma deficiency interferes with terminal differentiation of the trophoblast and placental vascularization, leading to severe myocardial thinning and death by E10.0. Supplementing PPAR gamma null embryos with wild-type placentas via aggregation with tetraploid embryos corrects the cardiac defect, implicating a previously unrecognized dependence of the developing heart on a functional placenta. A tetraploid-rescued mutant surviving to term exhibited another lethal combination of pathologies, including lipodystrophy and multiple hemorrhages. These findings both confirm and expand the current known spectrum of physiological functions regulated by PPAR gamma.


Gold Nanoparticles Using Ecklonia stolonifera Protect Human Dermal Fibroblasts from UVA-Induced Senescence through Inhibiting MMP-1 and MMP-3.
The effect of gold nanoparticles (GNPs) synthesized in marine algae has been described in the context of skin, where they have shown potential benefit. Ecklonia stolonifera (ES) is a brown algae that belongs to the Laminariaceae family, and is widely used as a component of food and medicine due to its biological activities. However, the role of GNPs underlying cellular senescence in the protection of Ecklonia stolonifera gold nanoparticles (ES-GNPs) against UVA irradiation is less well known. Here, we investigate the antisenescence effect of ES-GNPs and the underlying mechanism in UVA-irradiated human dermal fibroblasts (HDFs). The DPPH and ABTS radical scavenging activity of ES extracts was analyzed. These analyses showed that ES extract has potent antioxidant properties. The facile and optimum synthesis of ES-GNPs was established using UV-vis spectra. The surface morphology and crystallinity of ES-GNPs were demonstrated using high resolution transmission electron microscopy (HR-TEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD), and Fourier-transform infrared spectroscopy (FT-IR). ES-GNPs presented excellent photocatalytic activity, as shown by the photo-degradation of methylene blue and rhodamine B. A cellular senescence model was established by irradiating HDFs with UVA. UVA-irradiated HDFs exhibited increased expression of senescence-associated β-galactosidase (SA-β-galactosidase). However, pretreatment with ES-GNPs resulted in reduced SA-β-galactosidase activity in UVA-irradiated HDFs. Intracellular ROS levels and G1 arrest in UVA-irradiated HDFs were checked against the background of ES-GNP treatment to investigate the antisenescence effects of ES-GNPs. The results showed that ES-GNPs significantly inhibit UVA-induced ROS levels and G1 arrest. Importantly, ES-GNPs significantly downregulated the transcription and translation of MMP (matrix metalloproteinases)-1/-3, which regulate cellular senescence in UVA-irradiated HDFs. These findings indicate that our optimal ES-GNPs exerted an antisenescence effect on UVA-irradiated HDFs by inhibiting MMP-1/-3 expression. Collectively, we posit that ES-GNPs may potentially be used to treat photoaging of the skin.


Contradictions between perceptions and practices of caring in long-term care of elderly people.
Care for elderly people in nursing homes is a subject of increasing interest as populations around the world age. Numerous studies have been reported in relation to caring, how it is defined and what it means to nurses and clients in different contexts. Nursing is equated with caring and yet reports of nursing home care frequently cite such uncaring practices as neglect, fostered dependency, infantilization and depersonalization. The contradictions are explored with the aim of improving care of elderly people through greater understanding of how caring is perceived and practised.


Cancer-Cachexia-Induced Human Skeletal Muscle Myotube Degeneration Is Prevented via Cannabinoid Receptor 2 Agonism In Vitro.
Cachexia syndrome, leading to reduced skeletal muscle and fat mass, is highly prevalent in cancer patients, resulting in further negative implications for these patients. To date, there is no approved therapy for cachexia syndrome. The objective of this study was to establish an in vitro model of cancer cachexia in mature human skeletal muscle myotubes, with the intention of exploiting the cell model to assess potential cachexia therapeutics, specifically cannabinoid related drugs. Having cultured and differentiated primary human muscle myoblasts to mature myotubes, we successfully established two cancer cachexia models using conditioned media (CM) from human colon adenocarcinoma (SW480) and from non-small-cell lung carcinoma (H1299) cultured cells. The cancer-CM-induced extensive myotube degeneration, demonstrated by a significant reduction in mature myotube diameter, which progressed over the period studied. Myotube degeneration is a characteristic feature of cancer cachexia and was used in this study as an index of cachexia. Expression of cannabinoid 1 and 2 receptors (CB1R and CB2R) was confirmed in the mature human skeletal muscle myotubes. Subsequently, the effect of cannabinoid compounds on this myotube degeneration were assessed. Tetrahydrocannabinol (THC), a partial CB1R/CB2R agonist, and JWH133, a selective CB2R agonist, proved efficacious in protecting mature human myotubes from the deleterious effects of both (SW480 and H1299) cancer cachexia conditions. ART27.13, a full, peripherally selective CB1R/CB2R agonist, currently being trialled in cancer cachexia (IRAS ID 278450, REC 20/NE/0198), was also significantly protective against myotube degeneration in both (SW480 and H1299) cancer cachexia conditions. Furthermore, the addition of the CB2R antagonist AM630, but not the CB1R antagonist Rimonabant, abolished the protective effect of ART27.13. In short, we have established a convenient and robust in vitro model of cancer-induced human skeletal muscle cachexia. The data obtained using the model demonstrate the therapeutic potential of ART27.13 in cancer-induced cachexia prevention and provides evidence indicating that this effect is via CB2R, and not CB1R.


Performance of "standardized examinees" in a standardized-patient examination of clinical skills.
As a first step in testing the utility of using trained "standardized examinees" (SEs) as a quality-assurance measure for the scoring process in a standardized-patient (SP) examination, to test whether medical residents could simulate students in an SP examination and perform consistently to specified levels under test conditions. Fourth-year students from the Baltimore-Washington Consortium for SPs participated in a National Board of Medical Examiners Prototype Examination of clinical skills consisting of twelve 15-minute student-patient encounters in 1994-95. For this examination, internal medicine residents were trained to act as ordinary candidates and to achieve target scores by performing to a set level on specific checklist items used by SPs for recording interviewing, physical-examination, and communication skills. The "strong SEs" were trained to score 80% correct on six of the examination's 12 cases (study cases), and the "weak SEs" were trained to score 40% correct on the same six cases. The strong and weak SEs' checklist scores on the study cases were compared through independent, two-tailed t-tests. When there was less than 85% agreement on specific checklist items in each case between the SE training and the SP recording, videotapes of the cases were reviewed; in such cases an SE's performance was the final score agreed upon after review. Seven SEs took the SP examination and were not detected by the SPs. There was a total of 84 discrepancies between predicted and recorded checklist scores across 659 checklist items in 40 encounters scored by the SPs. After correcting the discrepancies based on videotape review, the estimated actual mean score was 77.3% for the strong SEs and 44.0% for the weak SEs, and was higher for the strong SEs in each study case. The overall fidelity of the SEs to their training was estimated to be 97%, and the overall SP accuracy was estimated to be 91%. The videotape review revealed 47 training-scoring discrepancies, most in the area of communication skills. This study suggests that SEs can be trained to specific performance levels and may be an effective internal control for a high-stakes SP examination. They may also provide a mechanism for refining scoring checklists and for exploring the validity of SP examinations.


A preliminary fertility appraisal of exotic cattle herds.
The fertility of 754 cows and 13 heifers in 4 government and several private herds of exotic milk and beef breeds in Uganda were studied using information originated from the records of genital examinations conducted during a period of 15 months. The pregnancy or in-calf rate in these animals ranged between 59.2% (private farms) and 85.3% (government farms). The proportion of cows not inseminated by three months after parturition varied between 50% (government farms) and 92.9% (private farms). The average number of days open was 119.07 to 249.51 in government farms and 187.43 days in private farms, the index of artificial insemination ranged between 1.5 and 3.18. Clinical examination revealed that inactive ovaries (52.9%) and normal cyclic changes (24.5%) were the main disturbances in cows with fertility problems. A smaller percentage was attributed to cystic follicles, endometritis, and others.


Empetrum nigrum var. japonicum Extract Suppresses Ultraviolet B-Induced Cell Damage via Absorption of Radiation and Inhibition of Oxidative Stress.
This study focused on the protective actions of Empetrum nigrum against ultraviolet B (UVB) radiation in human HaCaT keratinocytes. An ethyl acetate extract of E. nigrum (ENE) increased cell viability decreased by exposure to UVB rays. ENE also absorbed UVB radiation and scavenged UVB-induced intracellular reactive oxygen species (ROS) in HaCaT keratinocytes. In addition, ENE shielded HaCaT keratinocytes from damage to cellular components (e.g., peroxidation of lipids, modification of proteins, and breakage of DNA strands) following UVB irradiation. Furthermore, ENE protected against UVB-induced apoptotic cell death, as determined by a reduction in the numbers of apoptotic bodies and sub-G1 hypodiploid cells, as well as by the recovery of mitochondrial membrane potential. The results of the current study therefore suggest that ENE safeguards human keratinocytes against UVB-induced cellular damage via the absorption of UVB ray and scavenging of UVB-generated ROS.


Effects of gestational maternal undernutrition on growth, carcass composition and meat quality of rabbit offspring.
An experiment was conducted in order to evaluate the effects of gestational undernutrition of rabbit does on growth, carcass composition and meat quality of the offsprings. Thirty primiparous non lactating rabbit does were artificially inseminated and randomly divided in three treatment groups: Control (C; fed to 100% of maintenance requirements throughout gestation, n = 10), early undernourished (EU; fed to 50% of maintenance requirements during days 7-19 of gestation, n = 10) and late undernourished (LU; fed to 50% of maintenance requirements during days 20-27 of gestation, n = 10). During the 4th week of the gestation period, LU does significantly lost weight compared to C and EU groups (P<0.05). At kindling, C does produced litters with higher proportions of stillborn kits (P<0.05) while the total litter size (alive and stillborn kits) was not different among groups (10.7, 12.8 and 12.7 kits in C, EU and LU groups, respectively). Kit birth weight tended to be lower in the LU group. During fattening, body weight and feed intake were not different among offsprings of the three experimental groups. Moreover, the maternal undernutrition did not have any impact on carcass composition of the offsprings in terms of carcass parts and internal organs weights as well as meat quality of L. lumborum muscle (pH24, colour, water holding capacity and shear values) at slaughter (70 days of age). Therefore, it can be concluded that the gestational undernutrition of the mother does not have detrimental effects on the productive and quality traits of the offsprings.


Cytoplasmic Ca2+ at low submicromolar concentration stimulates mitochondrial metabolism in rat luteal cells.
The cytoplasmic Ca2+ signal is transferred to the mitochondrial matrix and activates mitochondrial dehydrogenases. The requirement for supramicromolar cytoplasmic [Ca2+] ([Ca2+]i) in perimitochondrial microdomains in this response has been suggested. We studied the correlation between [Ca2+]i, mitochondrial [Ca2+] ([Ca2+]m) and mitochondrial formation of reduced nicotinamide adenine dinucleotide (phosphate) [NAD(P)H] in the presence of submicromolar [Ca2+]i in cultured rat "large" luteal cells. [Ca2+]i was monitored fluorimetrically with fura-PE3, [Ca2+]m with rhod-2 and NAD(P)H with autofluorescence. In intact cells, prostaglandin F2alpha, which induces both intracellular Ca2+ release and Ca2+ entry, stimulated mitochondrial NAD(P)H formation. Thapsigargin-induced Ca2+ release and subsequent capacitative Ca2+ entry, both resulting in Ca2+ responses not exceeding 150-200 nM, also enhanced the reduction of pyridine nucleotides. As shown in inhibitor studies, the increased steady-state NAD(P)H level was due to activation of Ca2+-dependent dehydrogenases. [Ca2+]m, measured in permeabilized cells, increased moderately, but significantly, following elevation of [Ca2+]i from 50 to 180 nM, showed a further gradual increase at higher submicromolar [Ca2+]i values and rose steeply at supramicromolar [Ca2+]i. In summary, our results demonstrate that, in a steroid-producing cell type, net mitochondrial Ca2+ uptake and mitochondrial dehydrogenation can be activated even by low submicromolar increases of [Ca2+]i.


Genomic profiles of colorectal cancers differ based on patient smoking status.
Human sporadic colorectal cancer is the result of a lengthy somatic evolutionary process facilitated by various forms of genomic instability. Such instability arises endogenously from mutations in genes whose role is to preserve genomic integrity, and exogenously from environmental agents that generate genomic damage. We have found that cigarette smoking shifts the genomic profiles and genomic instability patterns of colorectal carcinomas. The genomic profiles of 57 consecutive cancers were examined; 31 cases were current or former smokers and 26 were nonsmokers. Genome-wide allelotypes of 348 markers were examined, along with comparative genomic hybridization (CGH) on ordered BAC microarrays, microsatellite instability, and inter-(simple sequence repeat) polymerase chain reaction instability. Tumors from nonsmokers exhibited losses of heterozygosity, particularly on chromosomes 14 and 18, whereas tumors from smokers exhibited a more diffuse pattern of allelic losses. Tumors from smokers exhibited higher overall rates of loss of heterozygosity, but showed lower rates of background microsatellite instability (MSI-L). On BAC array CGH, higher levels of generalized amplifications and deletions were observed in tumors from smokers, differentially affecting male smokers. In the transforming growth factor-beta signaling pathway, MADH4 mutations were more common in tumors from smokers, whereas transforming growth factor-beta RII mutations were more common among nonsmokers.


Randomized controlled trial of the CGRP receptor antagonist MK-3207 in the acute treatment of migraine.
This study evaluated the CGRP receptor antagonist MK-3207 for acute treatment of migraine. Multicenter, double-blind, randomized, placebo-controlled, parallel-group, two-stage adaptive study with two interim efficacy analyses to facilitate optimal dose selection. Migraine patients were initially randomized to MK-3207 2.5, 5, 10, 20, 50 and 100 mg or placebo to treat a moderate/severe migraine. One or more doses were to be discontinued based on the first interim analysis and a lower or higher dose could be added based on the second interim analysis. The primary endpoint was two-hour pain freedom. A total of 547 patients took study medication. After the first interim analysis, the two lowest MK-3207 doses (2.5, 5 mg) were identified as showing insufficient efficacy. Per the pre-specified adaptive design decision rule, only the 2.5-mg group was discontinued and the five highest doses (5, 10, 20, 50, 100 mg) were continued into the second stage. After the second interim efficacy analysis, a 200 mg dose was added due to insufficient efficacy at the top three (20, 50, 100 mg) doses. A positive dose-response trend was demonstrated when data were combined across all MK-3207 doses for two-hour pain freedom (p < .001). The pairwise difference versus placebo for two-hour pain freedom was significant for 200 mg (p < .001) and nominally significant for 100 mg and 10 mg (p < .05). The incidence of adverse events appeared comparable between active treatment groups and placebo, and did not appear to increase with increasing dose. MK-3207 was effective and generally well tolerated in the acute treatment of migraine.


Fabrication of three-dimensional ordered macroporous spinel CoFe2O4 as efficient bifunctional catalysts for the positive electrode of lithium-oxygen batteries.
Three-dimensionally ordered macroporous (3DOM) CoFe2O4 (CFO) catalysts were prepared by using the colloidal crystal templating method to be used as bifunctional catalysts of Li-O2 battery positive electrodes. In order to study the relationship between the macropore diameter and charge/discharge behavior, 3DOM CFO catalysts with two different pore diameters of 140 and 60 nm were prepared. The physicochemical properties of the 3DOM CFO catalysts were investigated by scanning electron microscopy, X-ray diffraction, transmission electron microscopy, X-ray photoelectron spectroscopy, and X-ray absorption spectroscopy. When the 3DOM CFO catalyst with a pore diameter of 140 nm (CFO@140) was used in the O2-electrode of Li-O2 batteries, it exhibited a substantially enhanced discharge capacity (ca. 11 658.5 mA h g-1) in the first cycle. Moreover, the Li-O2 cells with the CFO@140 catalyst showed cycling stability over 47 cycles at a limited capacity of 500 mA h g-1 with a reduced potential polarization of 1.13 V, as compared with that with Ketjen Black carbon and the 3DOM CFO of 60 nm pore diameter (CFO@60). Their high cycling stability, low overpotential, high round-trip efficiency, and high rate performance suggest that these 3DOM CFO catalysts could be promising O2-electrode catalysts for next-generation lithium-oxygen batteries.


Case Report: Acute Macular Neuroretinopathy Which Led to COVID-19 Infection Diagnosis.
We present a case of Acute Macular Neuroretinopathy (AMN), which led to the coronavirus disease of 2019 (COVID-19) infection diagnosis. A 27-year-old female patient with flu-like symptoms later proven to be COVID-19 infection presented with acute-onset bilateral gray-like paracentral scotomas. Fundus examination showed a hypopigmented, wedge-like lesion on the superior temporal juxtafoveal area in the right eye, while no significant finding was found in the left eye. Infrared reflectance images demonstrated bilateral hyporeflective lesions in the parafoveal regions of the macula. Spectral-domain optical coherence tomography scans over the corresponding areas detected focal hyperreflectivity in the outer nuclear layer with disruption in the ellipsoid zone and retinal pigment epithelium layers. Based on these findings, the AMN diagnosis was considered. The COVID-19 infection diagnosis was confirmed by a polymerase chain reaction test. COVID-19 disease may cause retinal vascular complications such as AMN. AMN, which shares common viral prodromal symptoms with COVID-19 infection, may be a presenting sign of COVID-19 infection.


Role of endogenous brain-derived neurotrophic factor and sortilin in B cell survival.
Brain-derived neurotrophic factor (BDNF), a major neuronal growth factor, is also known to exert an antiapoptotic effect in myeloma cells. Whereas BDNF secretion was described in B lymphocytes, the ability of B cells to produce sortilin, its transport protein, was not previously reported. We studied BDNF production and the expression of its receptors, tyrosine protein kinase receptor B and p75 neurotrophin receptor in the human pre-B, mature, and plasmacytic malignant B cell lines under normal and stress culture conditions (serum deprivation, Fas activation, or their combination). BDNF secretion was enhanced by serum deprivation and exerted an antiapoptotic effect, as demonstrated by neutralization experiments with antagonistic Ab. The precursor form, pro-BDNF, also secreted by B cells, decreases under stress conditions in contrast to BDNF production. Stress conditions induced the membranous expression of p75 neurotrophin receptor and tyrosine protein kinase receptor B, maximal in mature B cells, contrasting with the sequestration of both receptors in normal culture. By blocking Ab and small interfering RNA, we evidenced that BDNF production and its survival function are depending on sortilin, a protein regulating neurotrophin transport in neurons, which was not previously described in B cells. Therefore, in mature B cell lines, an autocrine BDNF production is up-regulated by stress culture conditions and exerts a modulation of apoptosis through the sortilin pathway. This could be of importance to elucidate certain drug resistances of malignant B cells. In addition, primary B lymphocytes contained sortilin and produced BDNF after mitogenic activation, which suggests that sortilin and BDNF might be implicated in the survival and activation of normal B cells also.


Role of presynaptic kainate receptors at parallel fiber-purkinje cell synapses in induction of cerebellar LTD: interplay with climbing fiber input.
Until recently, except for A1 adenosine, N-methyl-d-aspartate, and cannabinoid receptors, little effort has been made to unravel possible roles of parallel fiber (PF) presynaptic receptors in long-term depression (LTD) of synaptic transmission at PF-Purkinje cell (PC) synapses. Presynaptic kainate (KA) receptors are also present on PFs and might also influence LTD induction by modulating glutamate (Glu) release at PF-PC synapses. This hypothesis was tested by comparing the efficacy of two pairing protocols in inducing LTD in adult wild-type and knock-out mice for the Glu receptor 6 (GluR6) subunit of KA receptors. Activation of presynaptic KA receptors was unnecessary for LTD induction when PF inputs were paired with climbing fiber (CF) stimulation but became crucial when CF input was replaced by direct depolarization of PCs. By comparing basal paired-pulse facilitation of PF-excitatory postsynaptic currents (EPSCs) and depolarization-induced suppression of excitation in adult wild-type and GluR6 knock-out mice, it was shown that the participation of PF presynaptic KA receptors in LTD induction was likely to mainly result from their tonic activation by basal extracellular Glu, rather than from their activation by retrograde release of Glu by PCs during pairing protocols. Finally, this study suggests that, in adult mice, CFs not only participate in LTD induction by depolarizing postsynaptic cells but also by activating postsynaptic mGluR1alpha metabotropic glutamate receptors at CF-PC synapses.


Expression of fatty acid synthesis genes and fatty acid accumulation in haematococcus pluvialis under different stressors.
Biofuel has been the focus of intensive global research over the past few years. The development of 4th generation biofuel production (algae-to-biofuels) based on metabolic engineering of algae is still in its infancy, one of the main barriers is our lacking of understanding of microalgal growth, metabolism and biofuel production. Although fatty acid (FA) biosynthesis pathway genes have been all cloned and biosynthesis pathway was built up in some higher plants, the molecular mechanism for its regulation in microalgae is far away from elucidation. We cloned main key genes for FA biosynthesis in Haematococcus pluvialis, a green microalga as a potential biodiesel feedstock, and investigated the correlations between their expression alternation and FA composition and content detected by GC-MS under different stress treatments, such as nitrogen depletion, salinity, high or low temperature. Our results showed that high temperature, high salinity, and nitrogen depletion treatments played significant roles in promoting microalgal FA synthesis, while FA qualities were not changed much. Correlation analysis showed that acyl carrier protein (ACP), 3-ketoacyl-ACP-synthase (KAS), and acyl-ACP thioesterase (FATA) gene expression had significant correlations with monounsaturated FA (MUFA) synthesis and polyunsaturated FA (PUFA) synthesis. We proposed that ACP, KAS, and FATA in H. pluvialis may play an important role in FA synthesis and may be rate limiting genes, which probably could be modified for the further study of metabolic engineering to improve microalgal biofuel quality and production.


Predicting Anti-inflammatory Peptides by Ensemble Machine Learning and Deep Learning.
Inflammation is a biological response to harmful stimuli, aiding in the maintenance of tissue homeostasis. However, excessive or persistent inflammation can precipitate a myriad of pathological conditions. Although current treatments such as NSAIDs, corticosteroids, and immunosuppressants are effective, they can have side effects and resistance issues. In this backdrop, anti-inflammatory peptides (AIPs) have emerged as a promising therapeutic approach against inflammation. Leveraging machine learning methods, we have the opportunity to accelerate the discovery and investigation of these AIPs more effectively. In this study, we proposed an advanced framework by ensemble machine learning and deep learning for AIP prediction. Initially, we constructed three individual models with extremely randomized trees (ET), gated recurrent unit (GRU), and convolutional neural networks (CNNs) with attention mechanism and then used stacking architecture to build the final predictor. By utilizing various sequence encodings and combining the strengths of different algorithms, our predictor demonstrated exemplary performance. On our independent test set, our model achieved an accuracy, MCC, and F1-score of 0.757, 0.500, and 0.707, respectively, clearly outperforming other contemporary AIP prediction methods. Additionally, our model offers profound insights into the feature interpretation of AIPs, establishing a valuable knowledge foundation for the design and development of future anti-inflammatory strategies.


Objective determination of Fitzpatrick skin type.
The overall aim of this Ph.D. project was to clarify what the subjective Fitzpatrick skin type represents with regard to the skin's reaction to UVR. Fitzpatrick skin type is used as an expression of the constitutive UV-sensitivity. It has been used for guiding dose-levels in phototherapy and is an important risk factor for skin cancer. The subjective Fitzpatrick skin type and the measured skin type PPF (pigment protection factor, calculated based on a skin reflectance measurement, predicts the UV-dose (SED) to give 1 MED) were investigated parallelly in relation to the clinically determined dose to erythema (MED) and/or pigmentation (MMD) to determine which one related best. PPF is an established method for assessing UV-sensitivity by predicting SED to MED. UV-dose to MED and/or MMD was determined after single UV-exposure to Solar Simulator on nates (n= 84) and after single and multiple (5, 6 or 12) UV-exposures (n = 24-62) on the back to four UV-sources (nUVB, Solar, bUVA and UVA1). SED to MMD was also related to wavelength. MED was only determined after a single and four UV-exposures to narrowband UVB (nUVB) and Solar Simulator (Solar). Volunteers with a broad range of constitutive pigmentation (skin types I-V) were included. Equal MMD doses (predetermined after a single UV-exposure) were used at the multiple exposures. The absolute increase in pigmentation after 6 and 12 UV-exposures, where steady-state pigmentation was reached, was independent of skin type and therefore could not enter into the calculations. But it proved that the MMD determinations after single exposure were correct and could be used at multiple UV-exposures. In contrary to what we expected, our results indicate that people may refer to the constitutive pigmentation, when they reply to the question of Fitzpatrick skin type. This applied to both erythema and pigmentation response as both dose to MED and MMD showed a better correlation to nates than to the back. As expected, our results from the back indicate that people seem to refer to sun sensitivity after multiple exposures to the sun rather than a single sun exposure, when they reply to the question of Fitzpatrick skin type. Hence, both SED to MED and SED to MMD are better correlated to skin type after respectively 4 and 5 exposures to Solar Simulator and nUVB compared to 1 exposure. Only when tanning is preceded by erythema there is a relation between SED to MMD and skin type/PPF. Thus only after nUVB and Solar. For nUVB and Solar there was a linear relation between erythema and tanning ability with the intercept different from zero. In spite of what we expected based on the literature, the correlation was better between SED to MED and skin type than between SED to MMD and skin type. This applied to single and multiple exposures and to single calculations and multiple regression analyses. The long-waved UVA1 and broadband UVA should definitely not be used for skin type determination, as there was no relation between MMD and skin type/PPF. Both nUVB and Solar can be considered. Finally, based on the objective parameters: pre-exposure pigmentation measured by skin reflectance, MED and MMD we tried to predict the Fitzpatrick skin type by multinominal logistic regression analyses to evaluate the significance of the different parameters for the subjective skin type classification and thereby hopefully enlighten what Fitzpatrick skin type represents. For single UV-exposure only the pre-exposure pigmentation worked as a predictor of Fitzpatrick skin type, and that is what PPF is based on. When this parameter was removed, only SED to MED was significant. Our model succeeds better to classify people correct after multiple UV-exposure compared to a single UV-exposure. PPF was predicted likewise and was highly correlated to SED to MED, as expected, and even higher correlated to Fitzpatrick skin type. SED to MMD was not significant. This study confirms that Fitzpatrick skin type is an unreliable predictor of UV-sensitivity with regard to MED- and MMD test. Fitzpatrick skin type in epidemiological context (risk for skin cancer) stands for burns and ability to tan may represent "cumulative" dose. SED to MED is equivalent to burns. PPF may also indirectly represent cumulative dose--the less pigmented the skin the more UVR penetrates the epidermis and will be able to accumulate and induce skin cancer. Our results indicate that Fitzpatrick skin type predominantly is determined by the skin pigmentation and that the second most important objective parameter is SED to MED (and not SED to MMD). This explains why Fitzpatrick skin type, eventhough being an unreliable predictor of UV-sensitivity, still plays an important role in epidemiology with regard to estimation of risk of skin cancer. This study showed that PPF can predict the UV-sensitivity also with regard to the tanning ability (MMD), can be applied to multiple UV-exposures and to a broader pigmentation spectrum. PPF is preferred to predict the individual UV-sensitivity rather than the subjective Fitzpatrick skin type, confirmed for both nates and back, single as well as repetitive UV-exposures. It should therefore be considered to concentrate on skin reflectance measurements.


Alpha and beta replication origins of plasmid R6K show similar distortions of the DNA helix in vivo.
Plasmid R6K contains inverted repeats of an approximately 100-base-pair sequence separated by 5.5 kilobases. These long inverted repeats (LIRs) occur within the alpha and beta origins of replication and are essential for origin function. In this study, primer-extension analysis of DNA modified in vivo by dimethyl sulfate or KMnO4 revealed that both alpha and beta LIRs acquire similar structural distortions of the DNA helix in a functional R6K replicon. These distortions were not seen in plasmids containing isolated LIR sequences. In the functional replicon, the dimethyl sulfate and KMnO4 hyperreactive sites appear on complementary strands and are located to one side of an internal palindromic sequence within the LIRs. This asymmetry coincides with the primary direction of DNA replication from alpha and beta origins in vivo. We also observed two intermediate structures when certain R6K cis- or trans-acting elements are missing. Sequences near the alpha origin are required for generation of the dimethyl sulfate hyperreactive sites, whereas sequences near the beta origin are responsible for the appearance of KMnO4 hyperreactive sites. We suggest that these structures represent a hierarchy that leads to a "locked" preinitiation complex, which functions to synchronize and determine the direction of replication from the alpha and beta replication origins in vivo.


[Gerbil experiment in the flight of spacecraft "Foton-M3"].
The 12-d mission of Russian spacecraft Foton-M3 in September of 2007 was used as an opportunity to fly an experiment with 12 male gerbils (Meriones unguiculatus) at the age of 4-4.5 mos. and mean body weight of 51.6 grams. Considering the behavior pattern of these animals, selection and preparation of the experimental groups continued in the course of 2.5-3 months. The flight animals were contained in module Kontur-L outfitted with a self-sustained system of life support. In orbit, the animals received a palletized hydrogenous feed. The physiological and hygienic parameters of the gerbil environment during the flight complied with the official standards. Analysis of the video recorded behavior of animals in microgravity showed that virtually throughout the flight they moved chaotically along the cage never attempting to stabilize position catching at the wire netting of the cage. The animals were decapitated in 21-24 hours after landing. The investigations showed that structural and functional changes in gerbil organs and tissues were generally of the same type as in rats following fights of comparable duration. However, some differences between the animals were attributed to the specifics of water turnover in gerbils.


Role of staphylococcal wall teichoic acid in targeting the major autolysin Atl.
Staphylococcal cell separation depends largely on the bifunctional autolysin Atl that is processed to amidase-R(1,2) and R(3)-glucosaminidase. These murein hydrolases are targeted via repeat domains (R) to the septal region of the cell surface, thereby allowing localized peptidoglycan hydrolysis and separation of the dividing cells. Here we show that targeting of the amidase repeats is based on an exclusion strategy mediated by wall teichoic acid (WTA). In Staphylococcus aureus wild-type, externally applied repeats (R(1,2)) or endogenously expressed amidase were localized exclusively at the cross-wall region, while in Delta tagO mutant that lacks WTA binding was evenly distributed on the cell surface, which explains the increased fragility and autolysis susceptibility of the mutant. WTA prevented binding of Atl to the old cell wall but not to the cross-wall region suggesting a lower WTA content. In binding studies with ConcanavalinA-fluorescein (ConA-FITC) conjugate that binds preferentially to teichoic acids, ConA-FITC was bound throughout the cell surface with the exception of the cross wall. ConA binding suggest that either content or polymerization of WTA gradually increases with distance from the cross-wall. By preventing binding of Atl, WTA directs Atl to the cross-wall to perform the last step of cell division, namely separation of the daughter cells.


[The local immunity of the urinary system in patients with recurrent urolithiasis].
51 patients with urolithiasis complicated by pyelonephritis in the active phase of inflammation were studied for the condition of local immunity by determining the urine content of secretory immunoglobulin A (SIgA) under conditions of combined treatment with making use of phlogenzyme, a drug of II-generation systemic enzymotherapy (SE). Recordable in this patients population was a marked increase in the urine level of SIgA. Incorporation in a combined treatment of phlogenzyme results in normalizing the status of the urinary system local immunity. Evidence has been obtained on the lack of parallelism in the dynamics between the serum IgA content and urine concentration of SIgA, which fact suggests independence of local immunity. Our theory is that an appreciable increase in the urine level of SIgA in patients with urolithiasis concurrent with pyelonephritis may have an important part to play in the genesis of nephrolithiasis.


Acupoints massage in improving the quality of sleep and quality of life in patients with end-stage renal disease.
Traditional Chinese acupressure is a noninvasive technique that employs pressure and massage to acupoints in order to stimulate the balance of life energy that promotes health and comfort. Sleep disturbance is common in patients with end-stage renal disease but no intervention studies have addressed this problem. Aim. The purpose of the present study was to test the effectiveness of acupoints massage for patients with end-stage renal disease and experiencing sleep disturbances and diminished quality of life. The study was a randomized control trial. A total of 98 end-stage renal disease patients with sleep disturbances were randomly assigned into an acupressure group, a sham acupressure group, and a control group. Acupressure and sham acupressure group patients received acupoints or no acupoints massage three times a week during haemodialysis treatment for a total of 4 weeks. The measures included the Pittsburgh Sleep Quality Index, Sleep Log, and the Medical Outcome Study - Short Form 36. The results indicated significant differences between the acupressure group and the control group in Pittsburgh Sleep Quality Index subscale scores of subjective sleep quality, sleep duration, habitual sleep efficiency, sleep sufficiency, and global Pittsburgh Sleep Quality Index scores. Sleep log data revealed that the acupressure group significantly decreased wake time and experienced an improved quality of sleep at night over the control group. Medical Outcome Study - Short Form 36 data also documented that acupressure group patients experienced significantly improved quality of life. This study supports the effectiveness of acupoints massage in improving the quality of sleep and life quality of end-stage renal disease patients, and offers a noninvasive therapy for sleep-disturbed patients.


Sub-cellular location of mercury in yeast grown in the presence of mercuric chloride.
The distribution of 203Hg in Saccharomyces cerevisiae grown in the presence of mercuric chloride has been examined by physical and chemical fractionation procedures and autoradiography. The major fraction of the bound mercury is tightly bound to the wall. A significant quantity of mercury penetrates to the cytoplasm but only a minor fraction is present as low molecular weight components. The wall associated mercury is not readily released by extraction with sodium hydroxide or ethylenediamine but a major fraction is solubilized by pronase and Helicase treatment. Isolated walls are capable of binding their own weight of mercury to high-affinity adsorption sites. The major role of the cell envelope in the in vivo binding of mercury and the penetration to the cytoplasm of mercury was confirmed by autoradiography.


[Effect of diet with high fat content on lipogenesis in the liver, lipid composition and lipoprotein spectrum of blood].
Content of lipids and lipoprotein composition in blood did not reflect the intensity of lipogenesis in liver tissue of rats, which were kept on diets with high content of vegetable or animal fats. Both diets with excess of fat increased the rate of acetylation reaction in synthesis of glycerides and esterification of free cholesterol in liver tissue. In this case secretion of synthesized lipids into blood was decreased. Excess of vegetable oil, as distinct from animal fat, decreased the content of glycerides, pre-beta and beta-lipoproteins in blood and activated the synthesis of free cholesterol in liver tissue.


Sleep Quality and Perinatal Depression in Pregnant Women Treated in a Primary Care Centre in Lima, Peru.
The aim is to evaluate the association between sleep quality and perinatal depression in pregnant women between the 12th week of gestation and the 36th week of gestation in a maternity and child centre in Lima. Analytical cross-sectional study in pregnant women cared for in a primary care centre between August and December 2019. The data were collected through a self-administered survey. The Pittsburgh Sleep Quality Index (PSQI) was used to assess sleep quality. The evaluation of perinatal depression was performed with the Edinburgh Postnatal Depression Scale (EPDS). For the multivariate analysis, Poisson regression with robust variances was used to calculate the crude and adjusted prevalence ratios and their respective confidence intervals. The sample was composed of 200 participants. The median [IQR] age was 26 [22-32] years, and 111 (55.5%) pregnancies were unplanned. 52% presented a poor quality of sleep and the risk of perinatal depression was 31.5%. Poor quality of sleep was associated with a significantly higher frequency of perinatal depression (aPR=4.8 for those with poor quality of sleep warranting medical attention, and aPR=6.6 for those with poor quality of sleep warranting medical attention and treatment). There is a possible association between poor sleep quality and perinatal depression in pregnant women between weeks 12 and 36 of gestation. Operational research should be promoted to assess whether interventions to improve sleep quality could have a positive impact on reducing perinatal depression.


Sensilla on the antenna of blow fly, Triceratopyga calliphoroides Rohdendorf (Diptera: Calliphoridae).
Blow fly, Triceratopyga calliphoroides Rohdendorf, is a common and synanthropic species of medical and forensic significance in eastern Asia. Field monitoring studies have indicated that olfaction system plays an important role in guiding the behavior of insect species. To further our understanding of fly olfaction, scanning electron microscopy (SEM) and laser scanning confocal microscopy (LSCM) are applied to examine the sensillar morphology of adults, with an emphasis on sensory pit and sacculus. Both microtrichiae and several mechanoreceptors are detected on antennal scape and pedicel. Except for these two structures, pedicellar buttons are also found in antennal pedicellar depression after the separation of antennal pedicel and funiculus. Eight types of antennal sensilla are observed on external surface of antennal funiculus (one type of trichoid sensilla, three types of basiconic sensilla, three types of coeloconic sensilla, and one type of clavate-like sensilla), while two types (one type of basiconic sensilla and one type of coeloconic-like sensilla) are detected at the inner surface of sensory pits or sacculus. As the first to investigate cuticular invaginations of blow flies via paraffin section, the internal structure of abundant sensory pits and an excessively complex sacculus on antennal funiculus are revealed. After comparison to other species previously studied, this phenomenon is proved to be the most unique feature of T. calliphoroides, armed with a discussion on its morphology, function, and possible evolutionary implications.


Chl1 DNA helicase regulates Scc2 deposition specifically during DNA-replication in Saccharomyces cerevisiae.
The conserved family of cohesin proteins that mediate sister chromatid cohesion requires Scc2, Scc4 for chromatin-association and Eco1/Ctf7 for conversion to a tethering competent state. A popular model, based on the notion that cohesins form huge ring-like structures, is that Scc2, Scc4 function is essential only during G1 such that sister chromatid cohesion results simply from DNA replisome passage through pre-loaded cohesin rings. In such a scenario, cohesin deposition during G1 is temporally uncoupled from Eco1-dependent establishment reactions that occur during S-phase. Chl1 DNA helicase (homolog of human ChlR1/DDX11 and BACH1/BRIP1/FANCJ helicases implicated in Fanconi anemia, breast and ovarian cancer and Warsaw Breakage Syndrome) plays a critical role in sister chromatid cohesion, however, the mechanism through which Chl1 promotes cohesion remains poorly understood. Here, we report that Chl1 promotes Scc2 loading unto DNA such that both Scc2 and cohesin enrichment to chromatin are defective in chl1 mutant cells. The results further show that both Chl1 expression and chromatin-recruitment are tightly regulated through the cell cycle, peaking during S-phase. Importantly, kinetic ChIP studies reveals that Chl1 is required for Scc2 chromatin-association specifically during S-phase, but not during G1. Despite normal chromatin enrichment of both Scc2 and cohesin during G1, chl1 mutant cells exhibit severe chromosome segregation and cohesion defects--revealing that G1-loaded cohesins is insufficient to promote cohesion. Based on these findings, we propose a new model wherein S-phase cohesin loading occurs during DNA replication and in concert with both cohesion establishment and chromatin assembly reactions--challenging the notion that DNA replication fork navigates through or around pre-loaded cohesin rings.


Development of the retinotectal system in the direct-developing frog Eleutherodactylus coqui in comparison with other anurans.
Frogs primitively have a biphasic life history with an aquatic larva (tadpole) and a usually terrestrial adult. However, direct developing frogs of the genus Eleutherodactylus have lost a free living larval stage. Many larval structures never form during development of Eleutherodactylus, while limbs, spinal cord, and an adult-like cranial musculoskeletal system develop precociously. Here, I compare growth and differentiation of the retina and tectum and development of early axon tracts in the brain between Eleutherodactylus coqui and the biphasically developing frogs Discoglossus pictus, Physalaemus pustulosus, and Xenopus laevis using morphometry, immunohistochemical detection of proliferating cell nuclear antigen (PCNA) and acetylated tubulin, biocytin tracing, and in situ hybridization for NeuroD. Findings of the present study indicate that retinotectal development was greatly altered during evolution of Eleutherodactlyus mostly due to acceleration of cell proliferation and growth in retina and tectum. However, differentiation of retina, tectum, and fiber tracts in the embryonic brain proceed along a conserved slower schedule and remain temporally coordinated with each other in E. coqui. These findings reveal a mosaic pattern of changes in the development of the central nervous system (CNS) during evolution of the direct developing genus Eleutherodactylus. Whereas differentiation events in directly interconnected parts of the CNS such as retina, tectum, and brain tracts remained coordinated presumably due to their interdependent development, they were dissociated from proliferation control and from differentiation events in other parts of the CNS such as the spinal cord. This suggests that mosaic evolutionary changes reflect the modular character of CNS development.


Bilateral parotid swelling caused by sarcoidosis.
Sarcoidosis is a multisystem granulomatous disease characteristically causing bilateral hilar lymphadenopathy. Salivary gland swellings also are seen, which mandate recognition by the dental profession. The authors describe a patient whose diagnosis of sarcoidosis was tenuous. Ocular involvement and an elevated serum angiotensin-converting enzyme level suggested a diagnosis of sarcoidosis. Confirmation with a tissue biopsy was achieved only when the clinicians recognized that salivary gland swelling probably represented a manifestation of the disease. Steroid therapy was instituted. It is important for the dental practitioner to be familiar with the salivary gland manifestation of sarcoidosis. Early recognition and intervention aborts or slows disease progression.


Staphylococcus aureus NrdH redoxin is a reductant of the class Ib ribonucleotide reductase.
Staphylococci contain a class Ib NrdEF ribonucleotide reductase (RNR) that is responsible, under aerobic conditions, for the synthesis of deoxyribonucleotide precursors for DNA synthesis and repair. The genes encoding that RNR are contained in an operon consisting of three genes, nrdIEF, whereas many other class Ib RNR operons contain a fourth gene, nrdH, that determines a thiol redoxin protein, NrdH. We identified a 77-amino-acid open reading frame in Staphylococcus aureus that resembles NrdH proteins. However, S. aureus NrdH differs significantly from the canonical NrdH both in its redox-active site, C-P-P-C instead of C-M/V-Q-C, and in the absence of the C-terminal [WF]SGFRP[DE] structural motif. We show that S. aureus NrdH is a thiol redox protein. It is not essential for aerobic or anaerobic growth and appears to have a marginal role in protection against oxidative stress. In vitro, S. aureus NrdH was found to be an efficient reductant of disulfide bonds in low-molecular-weight substrates and proteins using dithiothreitol as the source of reducing power and an effective reductant for the homologous class Ib RNR employing thioredoxin reductase and NADPH as the source of the reducing power. Its ability to reduce NrdEF is comparable to that of thioredoxin-thioredoxin reductase. Hence, S. aureus contains two alternative thiol redox proteins, NrdH and thioredoxin, with both proteins being able to function in vitro with thioredoxin reductase as the immediate hydrogen donors for the class Ib RNR. It remains to be clarified under which in vivo physiological conditions the two systems are used.


Heparin and Derivatives for Advanced Cell Therapies.
Heparin and its derivatives are saving thousands of human lives annually, by successfully preventing and treating thromboembolic events. Although the mode of action during anticoagulation is well studied, their influence on cell behavior is not fully understood as is the risk of bleeding and other side effects. New applications in regenerative medicine have evolved supporting production of cell-based therapeutics or as a substrate for creating functionalized matrices in biotechnology. The currently resurgent interest in heparins is related to the expected combined anti-inflammatory, anti-thrombotic and anti-viral action against COVID-19. Based on a concise summary of key biochemical and clinical data, this review summarizes the impact for manufacturing and application of cell therapeutics and highlights the need for discriminating the different heparins.


Extranuclear detection of histones and nucleosomes in activated human lymphoblasts as an early event in apoptosis.
To evaluate the presence of histones and nucleosomes in cell lysates of freshly isolated peripheral blood mononuclear cells (PBMC), fully activated lymphoblasts, or lymphoblasts after induction of apoptosis. Each histone class (H1, H2A, H2B, H3, and H4) was detected by western blot analysis with specific antibodies. Annexin V/propidium iodide (PI) staining was performed for each treatment to distinguish viable, early, and late apoptotic, and necrotic cells. DNA degradation was evaluated by PI staining in a hypotonic buffer. All five histones were detected in cell lysates of activated lymphoblasts in higher concentrations than in lysates of PBMC. An additional significant increase of H2A, H2B, H3, H4, and complete nucleosomes in cell lysates of lymphoblasts was found during interleukin (IL)2 deprivation for 8 or 24 hours. The content of these histones or nucleosomes in cell lysates decreased after delayed IL2 readdition. H1 content in cell lysates was not affected by IL2 deprivation or addition. Quantities of H2A, H2B, H3, and H4 in cell lysates correlated significantly with signs of early apoptosis. UV-B light exposure or etoposide treatment of lymphoblasts resulted in a distinct increase for each histone class in cell lysates compared with standard curves. No bands for post-translationally modified histones were detected in cell lysates in contrast with signals in nuclear preparations. Extranuclear accumulation of histones and nucleosomes is an early event of apoptosis in human lymphoblasts. Dysregulation of early apoptosis might support the induction of autoimmunity against nuclear components.


Beyond Genetics in Glioma Pathways: The Ever-Increasing Crosstalk between Epigenomic and Genomic Events.
Diffuse gliomas are the most frequent brain tumor in adults. This group of brain neoplasms, ranging from histologically benign to aggressive malignant forms, represents a challenge in modern neurooncology because of the diffuse infiltrative growth pattern and the inherent tendency to relapse as a more malignant tumor. Once the disease achieves the stage of glioblastoma multiforme (GBM), the prognosis of patients is dismal and the median survival time is 15 months. Exhaustive genetic analyses have revealed a variety of deregulated genetic pathways involved in DNA repair, apoptosis, cell migration/adhesion, and cell cycle. Recently, investigation of epigenetic alterations in gliomas has contributed to depict the complexity of the molecular lesions leading to these malignancies. Even though, the efficacy of the state-of-the-art form of chemotherapy in malignant gliomas with temozolomide is based on the methylation-associated silencing of the DNA repair gene MGMT. Nevertheless, the whole scenario including global DNA hypomethylation, aberrant promoter hypermethylation, histone modification, chromatin states, and the role of noncoding RNAs in gliomas has only been partially revealed. We discuss the repercussion of epigenetic alterations underlying deregulated molecular pathways in the pathogenesis and evolution of gliomas and their impact on management of patients.


Curcumin improves vascular function and alleviates oxidative stress in non-lethal lipopolysaccharide-induced endotoxaemia in mice.
Oxidative stress is implicated in various pathological conditions, including septic shock, and other diseases associated with local or systemic inflammation. Curcumin, a major component from turmeric (Curcuma longa), possesses diverse anti-inflammatory, anti-tumour and antioxidant properties. The aim of this study was to investigate the effect of curcumin on modulation of vascular dysfunction and oxidative stress induced by lipopolysaccharide (LPS) in mice. Male ICR mice were treated with curcumin (50 or 100 mg/kg), administered intragastrically, either before or after intraperitoneal injection of LPS (10 mg/kg). Fifteen hours after LPS administration, arterial blood pressure was measured and vascular response to vasoactive agents were assessed. Aortic tissues and blood samples were taken for assays of antioxidant and oxidative stress markers. LPS caused marked hypotension, tachycardia and vascular hyporeactivity. The mean arterial pressures in responses to phenylephrine, acetylcholine, and sodium nitroprusside of LPS-treated mice were significantly decreased when compared with the untreated controls. Curcumin modulated heart rate and restored arterial blood pressure in a dose-dependent manner in both protectively- and therapeutically-treated regimens. Furthermore, the vascular responsiveness of LPS-treated mice was improved by curcumin. Interestingly, the improvements of haemodynamics and vascular response during endotoxaemia were related to alleviation of oxidative stress by reducing aortic-derived superoxide production, suppression of lipid peroxidation and protein oxidation, and decrease in urinary nitric oxide metabolites with preservation of the ratio of glutathione/glutathione disulfide. This study provides the first evidence for the potential role of curcumin in prevention and treatment of vascular dysfunction in mice with endotoxaemia elicited by LPS.


[Diagnostic value of low versus high titers of MOG-IgG and their clinical implications in myelin oligodendrocyte glycoprotein antibody-associated disease: a retrospective singlecenter study].
To summarize the clinical characteristics of myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD) and explore the value of MOG-IgG titers for diagnosis and evaluating disease severity of MOGAD. This retrospective study was conducted among the patients positive for serum MOG-IgG admitted in our hospital from December, 2020 to December, 2022. With 1∶100 as the cutoff value of MOG-IgG titer, the patients were divided into low and high MOG-IgG titer groups for comparison of general demographic data, laboratory test results, imaging features, treatment, and prognosis. Of the 36 patients included, 28 were diagnosed with MOGAD (77.78%), including 15 with low and 13 with high MOG-IgG titers. Compared with the cases reported previously, the MOGAD patients included herein showed a higher prevalence of sensory abnormalities (52.78%) and a lower prevalence of vision loss (25%). Magnetic resonance imaging showed that the periventricular white matter was the most common lesion site (45.71%). The patients with high MOGIgG titers had significantly lower peripheral blood lymphocyte counts (P=0.025) with an increased neutrophil-to-lymphocyte ratio (P=0.045), higher serum free thyroxine levels (P=0.033) and higher cerebrospinal fluid glucose levels (P=0.007) as compared with those with low MOG-IgG titers. Low titers and high titers of MOG-IgG have similar diagnostic value for MOGAD. The serum titers of MOG-IgG are probably associated with the disease severity and stress levels of the patients with MOGAD.


Ecological niche and phylogeography elucidate complex biogeographic patterns in Loxosceles rufescens (Araneae, Sicariidae) in the Mediterranean Basin.
Understanding the evolutionary history of morphologically cryptic species complexes is difficult, and made even more challenging when geographic distributions have been modified by human-mediated dispersal. This situation is common in the Mediterranean Basin where, aside from the environmental heterogeneity of the region, protracted human presence has obscured the biogeographic processes that shaped current diversity. Loxosceles rufescens (Araneae, Sicariidae) is an ideal example: native to the Mediterranean, the species has dispersed worldwide via cohabitation with humans. A previous study revealed considerable molecular diversity, suggesting cryptic species, but relationships among lineages did not correspond to geographic location. Delimitation analyses on cytochrome c oxidase subunit I identified 11 different evolutionary lineages, presenting two contrasting phylogeographic patterns: (1) lineages with well-structured populations in Morocco and Iberia, and (2) lineages lacking geographic structure across the Mediterranean Basin. Dating analyses placed main diversification events in the Pleistocene, and multiple Pleistocene refugia, identified using ecological niche modeling (ENM), are compatible with allopatric differentiation of lineages. Human-mediated transportation appears to have complicated the current biogeography of this medically important and synanthropic spider. We integrated ecological niche models with phylogeographic analyses to elucidate the evolutionary history of L. rufescens in the Mediterranean Basin, with emphasis on the origins of mtDNA diversity. We found support for the hypothesis that northern Africa was the center of origin for L. rufescens, and that current genetic diversity originated in allopatry, likely promoted by successive glaciations during the Pleistocene. We corroborated the scenario of multiple refugia within the Mediterranean, principally in northern Africa, combining results from eight atmosphere-ocean general circulation models (AOGCMs) with two different refugium-delimitation methodologies. ENM results were useful for providing general views of putative refugia, with fine-scale details depending on the level of stringency applied for agreement among models.


Evaluation of tear evaporation from ocular surface by functional infrared thermography.
A novel technique was developed to measure tear evaporation and monitor its variation with respect to time, for the studying of ocular physiology based on dynamic functional infrared thermography and the first law of thermodynamics using the measured ocular surface temperatures (OSTs). This is a noninvasive, noncontact temperature measuring method that is widely applied in the field of biomedicine. A simple method based on the ocular thermal data was proposed to measure the rate of tear evaporation. The OST of 60 normal subjects were recorded in the form of sequential thermal images. For each thermal sequence, the ocular region was selected and warped to a standard form. Thermal data within the regions were processed, on the basis of the first law of thermodynamics to derive the evaporation rate. For elder subjects (aged above 35), the rate was determined to be 55.82 Wm(-2) and for younger subjects, the rate was 58.9 Wm(-2). The corneal rate of evaporation in elder subjects was found statistically (p < 0.11) larger than their younger counterparts. The rate of blinking was observed to be related to the variation of evaporation rate. The authors have measured the evaporation rate on a sequence of thermographic images. A region of interest was selected at first and the same region on all the images were warped into a standard form. Calculations were performed based on the thermal data in those regions to obtain the values of interest. The authors found that the tear evaporation rate for subjects of all age groups was 57.36 +/- 12.73 Wm(-2) and the corneal tear evaporation was higher in elder subjects. The corneal rate of evaporation fluctuated in a larger magnitude in subjects who blinked more than average.


Melatonin alleviates brain injury in mice subjected to cecal ligation and puncture via attenuating inflammation, apoptosis, and oxidative stress: the role of SIRT1 signaling.
Sepsis is a systemic inflammatory response to infection that causes severe neurological complications. Previous studies have suggested that melatonin is protective during sepsis. Additionally, silent information regulator 1 (SIRT1) was reported to be beneficial in sepsis. However, the role of SIRT1 signaling in the protective effect of melatonin against septic encephalopathy remains unclear. This study aimed to investigate the role of SIRT1 in the protective effect of melatonin. EX527, a SIRT1 inhibitor, was used to reveal the role of SIRT1 in melatonin's action. Cecal ligation and puncture or sham operation was performed in male C57BL/6J mice. Melatonin was administrated intraperitoneally (30 mg/kg). The survival rate of mice was recorded for the 7-day period following the sham or CLP operation. The blood-brain barrier (BBB) integrity, brain water content, levels of inflammatory cytokines (TNF-α, IL-1β, and HMGB1), and the level of oxidative stress (superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA)) and apoptosis were assessed. The expression of SIRT1, Ac-FoxO1, Ac-p53, Ac-NF-κB, Bcl-2, and Bax was detected by Western blot. The results suggested that melatonin improved survival rate, attenuated brain edema and neuronal apoptosis, and preserved BBB integrity. Melatonin decreased the production of TNF-α, IL-1β, and HMGB1. Melatonin increased the activity of SOD and CAT and decreased the MDA production. Additionally, melatonin upregulated the expression of SIRT1 and Bcl-2 and downregulated the expression of Ac-FoxO1, Ac-p53, Ac-NF-κB, and Bax. However, the protective effects of melatonin were abolished by EX527. In conclusion, our results demonstrate that melatonin attenuates sepsis-induced brain injury via SIRT1 signaling activation.


Alpha-amanitin resistance in three wild strains of Drosophila melanogaster.
Three wild strains of D. melanogaster have been identified which are resistant to normally lethal levels of the fungal toxin, alpha-amanitin. Dietary LD50S for Oregon-R, the reference strain, and Ama-KTT, Ama-MI and Ama-KLM, the resistant strains, are 1.2, 35, 30 and 10 micrograms alpha-amanitin/vial, respectively. Resistance in all three strains is a digenic trait, being determined by two independently acting dominant genes, Ama-1, Ama-2, either of which is sufficient to confer resistance and which are located at approximately 18.8 and 100.7, respectively, on chromosome 3. Resistance to alpha-amanitin in all three strains is apparently not mediated by modification of RNA polymerase form II nor by failure of toxin transport or by toxin inactivation.


Ethanol: modifications of acute intoxication by divalent cations.
Calcium, other divalent cations, and calcium antagonists were tested for their ability to alter ethanol-induced sleeping time, hypothermia, and behavioral intoxication in mice and rats. Calcium given intraventricularly significantly enhanced sleeping time and behavioral intoxication in a dose-related manner. The ionophores X537A and A23187 accentuated the effect of a low dose of calcium, whereas the calcium chelators EDTA and EGTA decreased sleeping time. Calcium also enhanced tertiary butanol- and chloral hydrate-induced sleeping time. The effects of cations on ethanol-induced hypothermia were less significant. The results suggest the existence of a central calcium pool that is involved in ethanol intoxication in rodents.


Decision making with clinical diagnostic tests in terms of statistical decision theory and information theory.
The value of a diagnostic test for an individual patient is expressed in terms of the test's ability to determine whether the patient does or does not have a disease. For diagnostic tests with a continuous diagnostic variable, various decision thresholds can be selected to adjust the sensitivity and specificity of the diagnostic test to conform with the clinician's concerns (e.g., health costs, financial costs, and diagnostic information of the test). Three methods of statistical decision theory and information theory are presented for critically evaluating the value of a clinical diagnostic test. To provide an example of the application of the three methods we critically evaluate the value of using intraocular pressure (IOP) data from three populations: open angle glaucoma, open angle glaucoma suspect, and no open angle glaucoma. Based on these methods, IOP data are shown to provide a means of indicating the higher probability of the presence of open angle glaucoma (whereupon the clinician would need to conduct further diagnostic tests), but on average IOP data are shown to have very little value as a diagnostic variable.


The cell surface of Trypanosoma cruzi: cytochemistry and freeze-fracture.
The ultrastructure of epimastigotes of Trypanosoma cruzi, obtained from acellular cultures, and bloodstream trypomastigotes, isolated from infected mice, were studied by thin-sectioning and freeze-fracturing techniques. Epimastigotes showed a thin (5 nm) surface coat when stained with ruthenium red, while the surface coat of trypomastigotes was more prominent (15 nm thick). Both P and E faces of the plasma membrane of T. cruzi had roughly the same number of intramembranous particles (IMP) as seen by freeze-fracture. The plasma membrane of bloodstream trypomastigotes had less IMP than epimastigotes. Several differentiations of the plasma membrane was observed. In epimastigotes a cytostome appears as a particle-poor region delimited by a pallisade-like row of adjacent IMP. Bloodstream trypomastigotes did not have a cytostome. Instead, abundant pinocytic vesicles were observed. At the base of the flagellum of epimastigotes a ciliary necklace was found. At this region, the surface coat was differentiated as long, hair-like projections after staining with ruthenium red. The flagellar membrane had less IMP than the body membrane. Clusters of IMP were present on both faces of the flagellar membrane at the flagellar-body adhesion zone of epimastigotes. Linear arrays of IMP were also seen. In bloostream trypomastigotes clusters of particles were observed both on the flagellar and cell body membranes. Our observations demonstrate the presence of considerable structural variations of the T. cruzi plasma membrane at the two stages of the life cycle studied.


Problem reduction representation for the linguistic analysis of waveforms.
This paper shows how the nondirectional structural analysis of pattern data can be performed by matching a problem reduction representation (PRR) of pattern structure with sample data, using a best-first state space search algorithm called SSS*. The end result of the matching algorithm is a tree whose nodes represent recognized structures in the data. Tip nodes of the tree structure correspond to primitives which are recognized in the raw data by curve fitting routines. The operators of the algorithm allow the tree to be constructed with a combination of top-down or bottom-up steps. The matching of the structure tree to waveform segments need not be done in a left-right sequence. Moreover ambiguous matches are pursued in a best first order by using state space search with partial parse trees as states. A software system called WAPSYS (for waveform parsing system) is described, which implements this structural analysis paradigm. Experience using WAPSYS to analyze carotid pulse waves is also discussed.


Substitutions of the highly conserved M2 leucine create spontaneously opening rho1 gamma-aminobutyric acid receptors.
All members of the receptor-operated ion channel family that includes gamma-aminobutyric acid (GABA), glycine, nicotinic acetylcholine, and serotonin type 3 receptors have a conserved leucine near the center of the presumed second membrane-spanning domain. This leucine has been postulated to play a role in the gating of the pore. In this study, we examined the effects of mutating this leucine (L301) on the function of human homomeric rho1 GABA receptors. Oocytes expressing rho1 GABA receptors in which this leucine was substituted with alanine (A), glycine (G), serine (S), threonine (T), valine, or tyrosine, but not isoleucine or phenylalanine, demonstrated larger-than-normal resting conductances in the absence of GABA. This resting conductance had a reversal potential (and shifted reversal potential with chloride substitution) indistinguishable from that of the wild-type rho1 GABA-activated current. This resting conductance was antagonized by picrotoxin and, in the case of the A, G, S, and T substitutions, by GABA itself. Although the rho1 competitive antagonist 3-aminopropyl(methyl)-phosphinic acid did not block the resting conductance, this compound did competitively inhibit the GABA-mediated antagonism of the resting conductance. At higher concentrations, both 3-aminopropyl(methyl)-phosphinic acid and GABA directly activated the A, G, S, and T mutant receptors. Taken together, these data suggest that substitution of this highly conserved leucine with either small or polar residues produced rho1 GABA receptors that can open in the absence of GABA and support the hypothesis that this leucine may play a key role in the gating of the pore.


Hepatorenal syndrome associated with obstructive jaundice.
In almost all cases of acute renal failure associated with cholestatic jaundice, the occurrence of renal failure is preceded by episodes of shock, hypotension, sepsis, or surgical intervention. The pathologic finding is usually that of acute tubular necrosis. A patient with obstructive jaundice developed renal failure; the clinical and pathologic features were consistent with those found in the hepatorenal syndrome. No episodes of shock or sepsis preceded the onset of that renal failure. At autopsy, the findings were normal.


Integral Characterization of Defective BDNF/TrkB Signalling in Neurological and Psychiatric Disorders Leads the Way to New Therapies.
Enhancement of brain-derived neurotrophic factor (BDNF) signalling has great potential in therapy for neurological and psychiatric disorders. This neurotrophin not only attenuates cell death but also promotes neuronal plasticity and function. However, an important challenge to this approach is the persistence of aberrant neurotrophic signalling due to a defective function of the BDNF high-affinity receptor, tropomyosin-related kinase B (TrkB), or downstream effectors. Such changes have been already described in several disorders, but their importance as pathological mechanisms has been frequently underestimated. This review highlights the relevance of an integrative characterization of aberrant BDNF/TrkB pathways for the rational design of therapies that by combining BDNF and TrkB targets could efficiently promote neurotrophic signalling.


ATP hydrolysis cycles and mechanism in P-glycoprotein and CFTR.
P-glycoprotein and CFTR are prominent members of the ABC Transporter family. Both use ATP, the former to drive extrusion of drugs from cells and confer multidrug-resistance, the latter to drive opening and closing of anion channels. We compare current working models of catalytic cycle and mechanism of the two proteins. In Pgp the NBDs appear functionally equivalent, in CFTR they appear functionally distinct. In both proteins, ATP hydrolysis occurs in both NBDs, and it is proposed that the two NBDs alternate in catalysis. Other differences and similarities are noted, fostering ideas for future developments.


Analysis of a new family of widely distributed metal-independent alpha-mannosidases provides unique insight into the processing of N-linked glycans.
The modification of N-glycans by α-mannosidases is a process that is relevant to a large number of biologically important processes, including infection by microbial pathogens and colonization by microbial symbionts. At present, the described mannosidases specific for α1,6-mannose linkages are very limited in number. Through structural and functional analysis of two sequence-related enzymes, one from Streptococcus pneumoniae (SpGH125) and one from Clostridium perfringens (CpGH125), a new glycoside hydrolase family, GH125, is identified and characterized. Analysis of SpGH125 and CpGH125 reveal them to have exo-α1,6-mannosidase activity consistent with specificity for N-linked glycans having their α1,3-mannose branches removed. The x-ray crystal structures of SpGH125 and CpGH125 obtained in apo-, inhibitor-bound, and substrate-bound forms provide both mechanistic and molecular insight into how these proteins, which adopt an (α/α)(6)-fold, recognize and hydrolyze the α1,6-mannosidic bond by an inverting, metal-independent catalytic mechanism. A phylogenetic analysis of GH125 proteins reveals this to be a relatively large and widespread family found frequently in bacterial pathogens, bacterial human gut symbionts, and a variety of fungi. Based on these studies we predict this family of enzymes will primarily comprise such exo-α1,6-mannosidases.


[Fever of unknown origin in patients infected with the human immunodeficiency virus].
To describe the clinical features and the final diagnosis of patients infected with human immunodeficiency virus who presented fever of unknown origin. Retrospective study, from November 1989 to January 1994, of all patients infected with HIV who had fever of unknown origin and who were admitted to a community hospital in a Mediterranean area in Alicante (Spain). Fever of unknown origin was defined as fever exceeding 38.3 degrees C lasting for at least three weeks with no diagnosis in the first three days of hospitalization after fulfilling clinical exam, three blood cultures, acid-fast bacilli stain in sputum and chest-X-ray. Of a cohort of 231 patients, 27 (12%) were evaluated because of fever of unknown origin during their follow-up. Patients' mean age was 31 years (interval, 22-75) and intravenous drug use (81%) was the most common risk factor for HIV infection. A total of 31 episodes of fever of unknown origin were reviewed. Twenty-three (74%) episodes occurred in patients with less than 200 CD4 lymphocytes/mm3. A final diagnosis of fever of unknown origin was achieved in 24 (77%) episodes: visceral leishmaniasis (n = 11), tuberculosis (n = 9), non-Hodgkin's lymphoma (n = 1), CNS toxoplasmosis (n = 1), cryptococcal meningitis (n = 1) and drug adverse reaction (n = 1). HIV-infected patients with fever of unknown origin very often show severe immunodeficiency. Cryptococcal antigen testing should be carried out in the initial evaluation of fever of unknown origin in HIV-infected patients. In our area, 64% of episodes of fever of unknown origin were caused by visceral leishmaniasis or tuberculosis.


Diagnostic performance and cost effectiveness of measurements of plasma N-terminal pro brain natriuretic peptide in patients presenting with acute dyspnea or peripheral edema.
The purpose of this study was to determine the diagnostic power of a newly available assay for amino-terminal pro-brain natriuretic peptide (NT-proBNP) to identify patients with acute heart failure. In addition, the influence of initial NT-proBNP measurements on economic consequences, diagnostic procedures and staff involvement was evaluated. 401 patients presenting with acute dyspnea or peripheral edema in the emergency department were enrolled. NT-proBNP was measured after initial clinical evaluation. Clinical routine care and diagnostic assessment were blinded to NT-proBNP results. Two cardiologists independently validated the period of hospitalization, clinical examinations and medical therapies of each patient considering NT-proBNP results. The median NT-proBNP level among patients with acute congestive heart failure (CHF) (n=122) was 3497 pg/ml as compared to 320 pg/ml in patients without (n=279) (p<0.0001). An NT-proBNP cutoff level <300 pg/ml was optimal to rule out acute CHF (negative predictive value 96%; sensitivity 96%). NT-proBNP >or=300 pg/ml could strongly predict acute CHF when compared to patients' history or physical examination (odds ratio 9.5; p<0.0001) and diagnostic technical findings (odds ratio 14.7; p<0.05). In patients with NT-proBNP<300 pg/ml, 14% of the period of hospitalization could be saved, corresponding to savings of US $481 per patient. In addition, 9% of the number and time of staff involvement of clinical examinations and therapies could be saved, 10% of the costs of clinical examinations. Chest X-rays were saved in 34%, echocardiography in 9%. Measurement of NT-proBNP leads to multiple saving amounts and optimizes diagnostic pathways and resource allocation.