Altered immune phenotype and DNA methylation in panic disorder.
Multiple studies have related psychiatric disorders and immune alterations. Panic disorder (PD) has been linked with changes in leukocytes distributions in several small studies using different methods for immune characterization. Additionally, alterations in the methylation of repetitive DNA elements, such as LINE-1, have been associated with mental disorders. Here, we use peripheral blood DNA methylation data from two studies and an updated DNA methylation deconvolution library to investigate the relation of leukocyte proportions and methylation status of repetitive elements in 133 patients with panic disorder compared with 118 controls. We used DNA methylation data to deconvolute leukocyte cell-type proportions and to infer LINE-1 element methylation comparing PD cases and controls. We also identified differentially methylated CpGs associated with PD using an epigenome-wide association study approach (EWAS), with models adjusting for sex, age, and cell-type proportions. Individuals with PD had a lower proportion of CD8T cells (OR: 0.86, 95% CI: 0.78-0.96, P-adj = 0.030) when adjusting for age, sex, and study compared with controls. Also, PD cases had significantly lower LINE-1 repetitive element methylation than controls (P < 0.001). The EWAS identified 61 differentially methylated CpGs (58 hypo- and 3 hypermethylated) in PD (Bonferroni adjusted P < 1.33 × 10-7). These results suggest that those with panic disorder have changes to their immune system and dysregulation of repeat elements relative to controls.


Negative regulation of Akt activity by p38alpha MAP kinase in cardiomyocytes involves membrane localization of PP2A through interaction with caveolin-1.
Cardiomyocyte-derived cell lines deficient in p38alpha are more resistant to apoptosis owing to lower expression of the pro-apoptotic proteins Bax and Fas and upregulation of the ERK survival pathway. Here, we show that increased Akt activity also contributes to the enhanced survival of p38alpha-deficient cardiomyocytes. We found that the serine/threonine phosphatase PP2A can be targeted to caveolae through interaction with caveolin-1 in a p38alpha-dependent manner. In agreement with this, PP2A activity associated with caveolin-1 was higher in wild type than in p38alpha-deficient cells. Akt was also present in caveolae and incubation of wild-type cells with the PP2A inhibitor okadaic acid increases the levels of Akt activity. Thus, p38alpha-induced re-localization of PP2A to caveolae can lead to dephosphorylation and inhibition of Akt, which in turn would contribute to the decreased survival observed in wild type cells. However, cell detachment impairs the formation of the PP2A/caveolin-1 complex and, as a consequence, phospho-Akt levels and survival are no longer regulated by p38alpha in detached wild type cardiomyocytes. Our results suggest that p38alpha can negatively modulate Akt activity, independently of PI3K, by regulating the interaction between caveolin-1 and PP2A through a mechanism dependent on cell attachment.


High-resolution paraventricular nucleus serial section model constructed within a traditional rat brain atlas.
As a starting point for constructing a high-resolution, resliceable computer graphics model for the extraction, quantitative analysis, display, and modeling of neuroanatomical data the outer border and the boundaries of inner divisions and parts of the paraventricular nucleus have been drawn for all 39 serial histological sections prepared for a published reference atlas of the rat brain. This careful parceling revealed three new features of paraventricular nucleus topography: the full rostral extent of the anterior parvicellular part, the caudal end of the medial magnocellular part, and a thin rostrolateral extension of the dorsal medial parvicellular part composed at least in part of neurons expressing corticotropin-releasing hormone. The vector graphics drawings were aligned using the already established alignment of nine consecutive, relevant Atlas Levels, and then contours were smoothed to eliminate nonlinear distortions associated with histological mounting. This dataset was then used to create three-dimensional contour and surface models of the paraventricular nucleus, as well as two-dimensional horizontal and sagittal projections of its outer border. The computer graphics files containing raw and smoothed drawings for all 39 serial sections are supplied for use by researchers interested in developing new or better computer graphics analysis tools involving the paraventricular nucleus. This work may also stimulate the long range goal of creating a high-resolution, resliceable, computer graphics model of the whole brain, and eventually the whole nervous system, that is useful for quantitative analysis and topological transformation.


Discovery of natural product ellagic acid as a potent CD73 and CD39 dual inhibitor.
The ATP-adenosine pathway has been recently identified as an attractive immune-oncology target and several drug candidates have been entered clinic trials. Inspired by the report of the first small-molecule CD73inhibitor AB680, we describe the discovery of natural product ellagic acid as a dual CD73 and CD39 inhibitor with an IC50 value of 1.85 ± 0.21 μM and 0.50 ± 0.22 μM, respectively. The result of cytotoxicity assays indicated that ellagic acid is a valuable lead compound with low cytotoxicity effect for immune therapy.


Studies Exploring the Interaction of the Organophosphorus Compound Paraoxon with Fullerenes.
In vitro experiments previously published demonstrated the ability of fullerenes to decrease the capability of organophosphorus (OP) compounds to inhibit acetylcholinesterase. Experiments described herein demonstrate molecular level affinity interactions between fullerenes and the OP test compound paraoxon with NMR spectroscopy. The calculated binding constant of 19 M-1 indicates that this binding was not covalent.


Generation of human embryonic stem cell line chHES-472 from abnormal embryos diagnosed with Spinocerebellar ataxia type 3.
Spinocerebellar ataxia type3 (SCA3) is an autosomal dominant neurodegenerative disorder. Human embryonic stem cell line chHES-472 was derived from abnormal embryo donated by SCA3 patient after preimplantation genetic diagnosis (PGD) treatment. This cell line had a normal karyotype and retained the disease-causing mutant in ATXN3 gene. Characteristic tests proved that the embryonic stem cell line presented typical markers of pluripotency and had the capability to form the three germlayers in vivo.


Fate of the food mutagen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) in Sprague-Dawley rats. I. Mutagens in the urine.
2-Amino-3-methylimidazo[4,5-f]quinoline (IQ) is a potent bacterial mutagen formed during cooking of beef. IQ was administered intravenously to Sprague-Dawley rats at concentrations ranging from 7.5-50 mg/kg body weight. Urine was collected and analyzed for mutagenicity. Urinary mutagens were found which required activation by S9 mix, and reverted Ames test strains TA98 and TA100, but not TA1535 or TA1537. The amount of urinary mutagen(s) were related to IQ dose administered and were excreted within 48 h. Additional mutagenic activity was not released after incubation with beta-glucuronidase or aryl sulfatase. Analysis of urinary mutagens by HPLC indicates that the majority of mutagenic activity is due to unchanged IQ, but a small peak of mutagenic activity may correspond to N-acetyl or 3-N-demethylated metabolite. Since only 1% of the administered mutagenic activity is recovered in the urine, IQ may be readily detoxified in vivo.


Alleviating effects of selenium on fluoride-induced testosterone synthesis disorder and reproduction toxicity in rats.
Fluoride (F) exists widely in food, water and other natural resources, and can cause damage to the reproductive system of human and animals. Studies have shown that selenium (Se) is a necessary trace element to maintain the normal male reproductive system. However, it is not clear whether it can alleviate the damage of reproductive system induced by F. Hence, sodium fluoride (NaF) was administered singly in drinking water at 100 mg L-1 alone and co-administered by drinking with sodium selenite (Na2SeO3) at 0.5, 1.0, 2.0 mg L-1 for 10 consecutive weeks. The results demonstrated that the sperm deformity rate were increased significantly by F, however, it was improved significantly after the addition of 2.0 mg L-1 Na2SeO3. The contents of glutathione peroxidase 4 (GPX-4), selenoprotein P (SePP), pregnenolone (PREG), androstenedione (ASD), and testosterone (T) were reduced obviously in the F group, however, it was increased significantly after adding 0.5, 1.0 and 2.0 mg L-1 Na2SeO3. F decreased noticeably the mRNA and protein expression levels of steroidogenic acute regulatory protein (StAR), cytochrome P450 cholesterol side chain lyase (P450scc), 3β-hydroxysteroid dehydrogenase (3β-HSD), cytochrome P450 17α-hydroxylase (P450c17) and 17β-hydroxysteroid dehydrogenase (17β-HSD), which was increased obviously after the addition of 1.0 and 2.0 mg L-1 Na2SeO3. In summary, 2.0 mg L-1 Na2SeO3 can alleviate testosterone synthesis disorder induced by F via reducing oxidative stress, increasing the level of selenoprotein in testis and regulating the content of related hormones and enzyme activity during testosterone synthesis pathway.


Histamine metabolism in diabetic rats.
Streptozotocin-induced diabetes in rats has been used to study the effect of insulin deficiency on histamine metabolism. There were significant increases in the amine content in the pancreas and intestine, and a significant drop in intestinal diamine oxidase (DAO) activity. The reduced DAO activity may be of clinical relevance.


Evaluation of on-line solid-phase extraction capillary electrophoresis-mass spectrometry with a nanoliter valve for the analysis of peptide biomarkers.
On-line solid-phase extraction capillary electrophoresis-mass spectrometry (SPE-CE-MS) is a powerful technique for high throughput sample clean-up and analyte preconcentration, separation, detection, and characterization. The most typical design due to its simplicity and low cost is unidirectional SPE-CE-MS. However, in this configuration, the sample volumes introduced by pressure depend on the dimensions of the separation capillary and some matrix components could be irreversibly adsorbed in its inner walls. Furthermore, in many cases, the requirements of on-line preconcentration are incompatible with the background electrolyte necessary for an efficient separation and sensitive MS detection. Here, we present SPE-CE-MS with a nanoliter valve (nvSPE-CE-MS) to overcome these drawbacks while keeping the design simple. The nvSPE-CE-MS system is operated with a single CE instrument and two capillaries for independent and orthogonal SPE preconcentration and CE separation, which are interfaced through an external and electrically isolated valve with a 20 nL sample loop. The instrumental setup is proved for the analysis of opioid and amyloid beta peptide biomarkers in standards and plasma samples. NvSPE-CE-MS allowed decreasing the limits of detection (LODs) 200 times with regard to CE-MS. Compared to unidirectional SPE-CE-MS, peak efficiencies were better and repeatabilities similar, but total analysis times longer and LODs for standards slightly higher due to the heart-cut operation and the limited volume of the valve loop. This small difference on the LODs for standards was compensated for plasma samples by the improved tolerance of nvSPE-CE-MS to complex sample matrices. In view of these results, the presented setup can be regarded as a promising versatile alternative to avoid complicated matrix samples entering the separation capillary in SPE-CE-MS.


Gas-liquid chromatography-frequency pulse-modulated electron-capture detection in the diagnosis of infectious diseases.
The extremely sensitive and selective gas-liquid chromatography-frequency pulsed-modulated electron-capture detection (GLC-FP-ECD) procedure has been applied to the diagnosis of bacterial, fungal, viral, rickettsial, and parasitic diseases by the examination of various body fluids, effusions and exudates, and excretion products. Carboxylic acid and alcohol, hydroxy acid, and amine product profiles of microbial or host-response origin, have been used to establish specific etiologies, these profiles are reproducible, and can be used to aid in the diagnosis of infections. In addition, we have used the GLC-FP-ECD procedure to analyze microbial metabolic products in vitro and to provide data for identification and classification. We also explored computer time-sharing for data analysis, profile library comparison, and eventual profile matching for diagnosis.


Pharmacological reversal of post-transcriptional alterations implicated in alcohol-induced alveolar macrophage dysfunction.
Alcohol use disorders (AUD) cause alveolar macrophage (AM) immune dysfunction and increase risk of lung infections. Excessive alcohol use causes AM oxidative stress, which impairs AM phagocytosis and pathogen clearance from the alveolar space. Alcohol induces expression of NADPH oxidases (Noxes), primary sources of oxidative stress in AM. In contrast, alcohol decreases AM peroxisome proliferator-activated receptor gamma (PPARγ), a critical regulator of AM immune function. To explore the underlying molecular mechanisms for these effects of alcohol, we hypothesized that ethanol promotes CCAAT/enhancer-binding protein beta (C/EBPβ)-mediated suppression of Nox-related microRNAs (miRs), in turn enhancing AM Nox expression, oxidative stress, and phagocytic dysfunction. We also hypothesized that PPARγ activation with pioglitazone (PIO) would reverse alcohol-induced C/EBPβ expression and attenuate AM oxidative stress and phagocytic dysfunction. Cells from the mouse AM cell line (MH-S) were exposed to ethanol in vitro or primary AM were isolated from mice fed ethanol in vivo. Ethanol enhanced C/EBPβ expression, decreased Nox 1-related miR-1264 and Nox 2-related miR-107 levels, and increased Nox1, Nox2, and Nox 4 expression in MH-S cells in vitro and mouse AM in vivo. These alcohol-induced AM derangements were abrogated by loss of C/EBPβ, overexpression of miRs-1264 or -107, or PIO treatment. These findings identify C/EBPβ and Nox-related miRs as novel therapeutic targets for PPARγ ligands, which could provide a translatable strategy to mitigate susceptibility to lung infections in people with a history of AUD. These studies further clarify the molecular underpinnings for a previous clinical trial using short-term PIO treatment to improve AM immunity in AUD individuals.


Management of an extruded footplate of an anterior chamber lens with the Nd:YAG laser.
A footplate of an anterior chamber lens that extruded through clear cornea was treated by severing the protruding haptic with the Nd:YAG laser and removing it surgically. During the procedure, the remainder of the implant was repositioned and vitreous traction was relieved by pars plana vitrectomy. This technique appeared less injurious to the eye than attempting to cut all four fibrosed haptics and remove the entire implant.


The effect of prophylactic lithium treatment on mortality and suicidal behavior: a review for clinicians.
Prophylactic lithium treatment lowers the frequency of recurrences in manic depressive illness. Does it also reduce mortality and suicidal behavior? In this paper all relevant publications were reviewed. The mortality of lithium-treated patients was compared with the mortality in the general population, and the patients' suicidal behavior when they were on lithium was compared with their behavior when they were not. The mortality of manic depressive patients is two to three times higher than that of the general population, a highly significant difference. In the studies reviewed here the patients' mortality during lithium treatment was not significantly higher or only moderately higher than in the general population. After discontinuation of lithium the mortality was again significantly higher. The number of patients attempting suicide was 6-15 times lower and the number of patients completing suicide was 3-17 times lower when they were on lithium than they were not. Similar observations have not been reported for prophylactic treatment with other mood stabilizers. For methodological reasons explained in the introduction, these observations cannot prove that lithium treatment has a mortality-lowering, antisuicidal effect. But they are compatible with such an assumption. It must be the duty of psychiatrists to keep the possibility of an antisuicidal action of lithium in mind when they choose prophylactic treatment for patients with severe depressions or suicidal thoughts or suicide attempts in the past or combinations of these.


Experimental animal models of chronic inflammation.
Inflammation is a general term for a wide variety of both physiological and pathophysiological processes in the body which primarily prevents the body from diseases and helps to remove dead tissues. It has a crucial part in the body immune system. Tissue damage can recruit inflammatory cells and cytokines and induce inflammation. Inflammation can be classified as acute, sub-acute, and chronic. If it remained unresolved and lasted for prolonged periods, it would be considered as chronic inflammation (CI), which consequently exacerbates tissue damage in different organs. CI is the main pathophysiological cause of many disorders such as obesity, diabetes, arthritis, myocardial infarction, and cancer. Thus, it is critical to investigate different mechanisms involved in CI to understand its processes and to find proper anti-inflammatory therapeutic approaches for it. Animal models are one of the most useful tools for study about different diseases and mechanisms in the body, and are important in pharmacological studies to find proper treatments. In this study, we discussed the various experimental animal models that have been used to recreate CI which can help us to enhance the understanding of CI mechanisms in human and contribute to the development of potent new therapies.


Enriched environment exposure alters the input-output dynamics of synaptic transmission in area CA1 of freely moving rats.
We examined in freely moving animals whether EE exposure directly alters synaptic transmission in CA1 of the hippocampus. Adult male Sprague-Dawley rats were chronically implanted with recording and stimulating electrodes in CA1 and housed singly in standard cages. After stable recordings were established for field excitatory postsynaptic potentials (fEPSPs), the effects of three levels of environmental enrichment for 19 days were assessed. A change in fEPSPs was observed only after repeated overnight enriched environment exposure, such that response potentiation occurred at high stimulus intensities but a reduced response was evident at low stimulus intensities. Behavioural tests confirmed that EE exposure was sufficient to facilitate performance on spatial working memory tasks. These results suggest that overnight EE exposure refines the dynamics of CA1 synaptic transmission in a way that may contribute to augmented information processing and thus performance on hippocampus-dependent tasks.


Lumbar puncture needle length determination.
Appropriate lumbar puncture (LP) needle length selection may be more difficult for less experienced practitioners or for patients who are of unusually large or small body habitus. The purpose of this study is to determine if there is a relationship between body height and weight, and the percutaneous depth to the spinal canal, which can more reliably assist in selecting an LP needle length. This is a retrospective cohort study of patients who received an abdominal computed tomographic scan (for any reason) from July 1999 to December 2000. Lumbar puncture depth was measured on the computed tomographic scan and was used to derive a formula. The final data pool consisted of 175 patients, aged 25 days to 80 years, with height of 48 to 181.5 cm, weight of 3.0 to 127.3 kg, and body mass index of 11.7 to 49.7 kg/m2. Using this data set, the formula for predicting the required LP depth is (weight in kilograms, height in centimeters): LP depth (cm) = 1 + 17 x weight/height. Using linear regression comparing the skin to mid-spinal canal depth measurements with the calculated LP depths, R2 was 0.81 (P < .001). This formula selected a needle that was too short in 6% (less than that of 4 other previously published LP needle length selection methods) and a needle that was too long in 31%. Compared with other formulas, this formula might be a more reliable predictor for estimating the required LP needle length, but this must be validated by further studies. It should be noted that none of the formulas were perfect.


Effect of pH on production of cytokinin-like substances by bacteria isolated from soil, rhizosphere and mycorrhizosphere of pine (Pinus silvestris L.).
Cytokinin-like substances were produced in higher amounts by soil isolates than by those isolated from the rhizosphere and mycorrhizosphere. However more bacterial strains isolated from the rhizosphere and mycorrhizosphere were capable of synthesizing cytokinin-like compounds. A distinct effect of pH on the production of these substances was found. It is suggested that zeatin and zeatin riboside are likely to be among the cytokinin-like substances detected.


Indications for contrast medium enhancement in computed tomography of the brain.
The use of contrast medium enhancement in computed tomography almost doubles the duration of each examination. As the clinical demand is far greater than the scanning time available, it is desirable to use contrast medium with discrimination. The following indications are suggested for extended examinations with contrast: 1. Lesions shown by alternative neuroradiological investigations. 2. Local abnormality shown on plain scan. 3. Suspected posterior fossa lesions. 4. Suspected suprasellar lesions. In a retrospective review of 3000 patient examinations, 430 patients falling outside the above criteria had been given contrast medium. In only one patient in this latter group was an abnormality shown after contrast enhancement. In the other 429 patients, no additional information was obtained. These figures support the indications suggested for the use of contrast enhancement.


Viscoelastic shear properties of in vivo thigh muscles measured by MR elastography.
To measure the viscoelastic properties of passive thigh muscles using multifrequency magnetic resonance elastography (MMRE) and rheological models. Four muscles in five volunteers underwent MMRE tests set up inside a 1.5T magnetic resonance imaging (MRI) scanner. Compression excitation was generated with a driver attached around the thigh, and waves were generated at 70, 90, and 110 Hz. In vivo experimental viscoelastic parameters (G(ω) = G' + i G″) were extracted from the wavelength and attenuation measurements along a local profile in the direction of the wave's displacement. The data-processing method was validated on a phantom using MMRE and RheoSpectris tests. The complex modulus (G(ω)) related to elasticity (μ) and viscosity (η) was then determined using four rheological models. Zener was the best-fit model (χ ∼0.35 kPa) for the rheological parameters of all muscles. Similar behaviors for the elastic components for each muscle were found for the Zener and springpot models. The gracilis muscle showed higher elastic values (about 2 kPa) in both models compared to other muscles. The α-values for each muscle was equivalent to the ratio G″/G' at 90 Hz. MMRE tests associated with data processing demonstrated that the complex shear modulus G(ω) of passive muscles could be analyzed using two rheological models. The viscoelastic data can be used as a reference for future assessment of muscular dysfunction. J. Magn. Reson. Imaging 2015. J. Magn. Reson. Imaging 2016;43:1423-1433.


An Uncommon Case Report: Pathologic Fracture in a Proximal Femur Aneurysmal Bone Cyst in a Child.
Aneurysmal bone cysts (ABCs) are a group of benign, expansile, locally aggressive lesion characterized by fluid-filled cysts usually in the metaphyseal end of long bones. They usually affect children and young adults, with an atypical etiology and uncommon presentation. Treatment modalities include en bloc resection and curettage with or without bone graft or bone substitute augmentation with instrumentation, sclerosing agents, arterial embolization, and adjuvant radiotherapy. We report a rare case of ABC with pathological fracture in the proximal femur of a 13-year-old male patient, who presented to the emergency department with severe pain in the right hip and inability to walk following trivial fall while playing. Curettage with open biopsy was performed followed by implantation with modified hydroxyapatite granules and internal fixation for the subtrochanteric fracture with pediatric dynamic hip screw and four hole plate, with a favorable outcome. There is a lack of a standard guideline for the management on account of uniqueness of these cases; curettage with bone graft or bone substitutes in conjunction with internal fixation of associated pathologic fracture yields bony union with adequate clinical results.


Status of different anthropogenic organic pollutants accumulated in sediments from Olt River Basin, Romania: From distribution and sources to risk assessment.
Some organic pollutants including organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) have been banned from production worldwide, but due to their toxicity and persistence are still of concern. Also, unintentional by-products of combustion and industrial processes such as polycyclic aromatic hydrocarbons (PAHs), represent a permanent threat to the safety of the environment and the population. In this study, surface sediment samples from the middle and lower Olt River Basin (ORB), Romania, including dams, the main tributaries and the confluence with Danube River were collected during seasonal sampling campaigns in 2019 and analyzed for 13 OCPs, 12 PCBs and 15 PAHs in order to evaluate the impact of the main anthropogenic activities in the area (industrial activities and agriculture) and the ecological status of the ORB. The registered levels of OCPs, PCBs and PAHs in surface sediments varied from low to significantly polluted environments, indicating a clear spatial distribution between sites based on concentrations and congener profiles correlated with the influence of anthropogenic activities in the surrounding area. Based on some molecular diagnostic ratio and multivariate statistical analysis, both non-point sources and point sources deposition by surface runoff or atmospheric deposition were identified. Overall, the contamination profile of the study area reveals significant amounts of organochlorine compounds, resulting from the industrial production of chlorinated products, including lindane, but also from the long-term agricultural use of both HCHs and DDTs, more than half of the sites having levels that pose a potential risk for benthic organisms. Therefore, levels of POPs in the hot-spots sampling locations raise numerous concerns about the safety of the environment and the population in the region, requiring immediate actions.


Spontaneous thrombosis in a coronary artery fistula with aneurysmal dilatation of the sinus of Valsalva.
This case report describes spontaneous thrombosis of a congenital coronary artery fistula with associated aneurysmal dilatation of the right coronary sinus of Valsalva. The grossly dilated sinus of Valsalva was reconstructed with a Dacron patch. Coronary revascularization was performed with a sequential saphenous vein graft anastomosed to the right posterior descending and posterior ventricular arteries distally and to the aorta proximally.


Structural characteristics of thermostable immunogenic outer membrane protein from Salmonella enterica serovar Typhi.
In this work, we explored the acid-induced unfolding pathway of non-porin outer membrane protein (OMP), an immunogenic protein from Salmonella Typhi, by monitoring the conformational changes over a pH range of 1.0-7.0 by circular dichroism, intrinsic fluorescence, ANS binding, acrylamide quenching, and dynamic light scattering. The spectroscopic measurements showed that OMP in its native state at pH 7.0 exists in more stable and compact conformation. In contrast, at pH 2.0, OMP retains substantial amount of secondary structure, disrupted side chain interactions, increased hydrodynamic radii, and nearly four-fold increase in ANS fluorescence with respect to the native state, indicating that MG state exists at pH 2.0. Quenching of tryptophan fluorescence by acrylamide further confirmed the accumulation of a partially unfolded state between native and unfolded state. The effect of pH on the conformation and thermostability of OMP points towards its heat resistance at neutral pH (T m ~ 69 °C at pH 7.0, monitored by change in MRE222 nm). Acid unfolded state was also characterized by the lack of a cooperative thermal transition. All these results suggested that acid-induced unfolded state of OMP at pH 2.0 represented the molten globule state. The chemical denaturation studies with GuHCl and urea as denaturants showed dissimilar results. The chemical unfolding experiments showed that in both far-UV CD and fluorescence measurements, GuHCl is more efficient than urea. GuHCl is characterized by low C m (~1 M), while urea is characterized by high C m (~3 M). The fully unfolded states were reached at 2 M GuHCl and 4 M urea concentration, respectively. This study adds to several key considerations of importance in the development of therapeutic agents against typhoid fever for clinical purposes.


Infrared spectroscopy of serum as a potential diagnostic screening approach for naturally occurring canine osteoarthritis associated with cranial cruciate ligament rupture.
To evaluate infrared (IR) spectroscopy of serum as a screening tool to differentiate dogs affected by naturally occurring osteoarthritis (OA) associated with cranial cruciate ligament rupture (CrCLR) and controls. 104 adult dogs with CrCLR (affected group) and 50 adult control dogs were recruited for this prospective observational study. Serum samples were collected preoperatively from CrCLR dogs and from a subset of these dogs at 4-, and 12-week post-surgical intervention to stabilize the affected stifles. Serum was collected once from control dogs. Dry films were made from serum samples, and IR absorbance spectra acquired. Data preprocessing, principal component analysis and multivariate analysis of covariance were performed to separate samples from the two groups, and to evaluate temporal differences. Weighted logistic regression with L1 regularization method was used to develop a predictive model. Model performance based on an independent test set was evaluated. Spectral data analysis revealed significant separation between the sera of CrCLR and control dogs (P < 0.0001), but not amongst different time points in the OA group. The sensitivity, specificity, AUC and accuracy of the test set were 84.62%, 96.15%, 93.20% and 92.31% respectively. These findings confirm the potential of IR-spectroscopy of serum with chemometrics methods to differentiate controls from dogs with OA associated with CrCLR. This is the first step in development of an economic, and comparatively simple IR-based screening serum test for OA. Utility of this tool as a clinical screening and diagnostic test requires further investigation and validation.


Corrigendum: Validation of the GALAD model and establishment of a new model for HCC detection in Chinese patients.
[This corrects the article DOI: 10.3389/fonc.2022.1037742.].


Space-age operations.
Robotic surgery at Kent and Canterbury Hospital has enabled a nurse specialist to extend her role and assist a surgeon with operations.


The obscuration by dust of most of the growth of supermassive black holes.
Supermassive black holes underwent periods of exponential growth during which we see them as quasars in the distant Universe. The summed emission from these quasars generates the cosmic X-ray background, the spectrum of which has been used to argue that most black-hole growth is obscured. There are clear examples of obscured black-hole growth in the form of 'type-2' quasars, but their numbers are fewer than expected from modelling of the X-ray background. Here we report the direct detection of a population of distant type-2 quasars, which is at least comparable in size to the well-known unobscured type-1 population. We selected objects that have mid-infrared and radio emissions characteristic of quasars, but which are faint at near-infrared and optical wavelengths. We conclude that this population is responsible for most of the black-hole growth in the young Universe and that, throughout cosmic history, black-hole growth occurs in the dusty, gas-rich centres of active galaxies.


Rat bone marrow cells undergo thymopoiesis in mouse fetal thymic organ culture.
We have developed an in vitro differentiation assay allowing the study of thymopoiesis from rat bone marrow cells. In this assay, Wistar rat bone marrow cells repopulated fetal Swiss mouse thymic lobes depleted in endogeneous lymphoid cells by deoxyguanosine treatment. Due to the xenogeneic situation, repopulating rat cells from any hemopoietic lineage could be easily recognized by anti-rat monoclonal antibodies such as anti-Thy-1.1 that did not react with Swiss mouse thymocytes. After 15 days in vitro, 80% of the developing rat cells were Thy-1.1+ lymphoid cells and about 70% of the Thy-1.1+ cells expressed CD5, CD2 and leukosialin. The percentages of cells expressing pre-B cell, B cell and myeloid determinants were less than 20%. The developing thymocytes comprised CD4-CD8- T cell receptor (TcR) alpha/beta-, CD4-CD8+TcR alpha/beta low and CD4+CD8+TcR alpha/beta low cells, indicating that the early stages of rat thymopoiesis occurred within mouse thymic lobes. Limiting dilution assays showed that 50% of positive assays were obtained with 3000 nucleated bone marrow cells, which is in good agreement with recent estimates derived from in vivo reconstitution after intrathymical transfer. Moreover the limiting dilution assays proved to be sensitive enough to evidence a tenfold enrichment of pre-T cell activity in the low-density fraction of rat bone marrow. This xenogeneic system might greatly facilitate studies on prethymic and intrathymic stages of rat T cell development and permit new in vitro approaches of the colonizing bone marrow T cell precursor properties.


Paneth cell α-defensins HD-5 and HD-6 display differential degradation into active antimicrobial fragments.
Antimicrobial peptides, in particular α-defensins expressed by Paneth cells, control microbiota composition and play a key role in intestinal barrier function and homeostasis. Dynamic conditions in the local microenvironment, such as pH and redox potential, significantly affect the antimicrobial spectrum. In contrast to oxidized peptides, some reduced defensins exhibit increased vulnerability to proteolytic degradation. In this report, we investigated the susceptibility of Paneth-cell-specific human α-defensin 5 (HD-5) and -6 (HD-6) to intestinal proteases using natural human duodenal fluid. We systematically assessed proteolytic degradation using liquid chromatography-mass spectrometry and identified several active defensin fragments capable of impacting bacterial growth of both commensal and pathogenic origins. Of note, incubation of mucus with HD-5 resulted in 255-8,000 new antimicrobial combinations. In contrast, HD-6 remained stable with consistent preserved nanonet formation. In vivo studies demonstrated proof of concept that a HD-5 fragment shifted microbiota composition (e.g., increases of Akkermansia sp.) without decreasing diversity. Our data support the concept that secretion of host peptides results in an environmentally dependent increase of antimicrobial defense by clustering in active peptide fragments. This complex clustering mechanism dramatically increases the host's ability to control pathogens and commensals. These findings broaden our understanding of host modulation of the microbiome as well as the complexity of human mucosal defense mechanisms, thus providing promising avenues to explore for drug development.


Ubiquitin-dependent sperm quality control mechanism recognizes spermatozoa with DNA defects as revealed by dual ubiquitin-TUNEL assay.
Defective mammalian spermatozoa become ubiquitinated during epididymal passage, a mechanism that may mark the abnormal spermatozoa for proteolytic destruction (Sutovsky et al., 2001a: J Cell Sci 114:1665-1675). It is not known how such spermatozoa are recognized by the epididymal ubiquitination pathway and whether there is a selection against certain types of sperm defects. We examined the relationship between sperm ubiqutination, lifelong sperm morphology and sperm DNA defects using a single chanel, ubiquitin-activated flow cytometric assay, and a dual, ubiquitin-TUNEL assay. Semen samples from nine service sires of good-to-average fertility were screened. A positive correlation was found between sperm ubiquitination and the average frequency of morphological semen abnormalities from field evaluations performed throughout the reproductive life of individual sires. Sample correlation coefficients were r=0.65 for primary (head and tail) and r=0.60 for total semen abnormalities in the single channel assay. In a dual assay, we found a high, positive correlation (r=0.93) between the ubiquitin-positive sperm and the TUNEL positive sperm. Substantial correlations (r=0.47-0.64) were observed when the measurements from these two respective assays were compared for individual sires. While anti-ubiquitin antibodies recognized most of the TUNEL-positive sperm cells, the TUNEL-positive spermatozoa represented only a subset (approximately 20-40%) of all ubiquitin-positive cells. It appears that the ubiquitin-dependent sperm quality control, residing in the epididymal epithelium, has the ability to detect spermatozoa with apoptotic or necrotic DNA, while spermatozoa with defects other than DNA fragmentation are also recognized and ubiquitinated.


Discrimination between RRR- and all-racemic-alpha-tocopherols labeled with deuterium by patients with abetalipoproteinemia.
The ability to discriminate between stereoisomers of alpha-tocopherol was studied in five patients with abetalipoproteinemia (ABL) because an impairment in secretion of apolipoprotein B-containing lipoproteins might impede the normally enhanced plasma transport of RRR-alpha-tocopherol. An oral dose containing 3.7 g of each 2R, 4'R,8'R-alpha-[5-C2H3]tocopheryl acetate (d3RRR-alpha-tocopheryl acetate) and 2RS,4'RS,8'RS-alpha-[5,7-(C2H3)2]tocopheryl acetate (d6 all rac-alpha-tocopheryl acetate) was administered, then the labeled and unlabeled alpha-tocopherol contents of plasma and red blood cells from multiple blood samples obtained at selected times up to 72 h following the dose were quantitated. ABL plasma contained about 1%-10% of the d3-RRR-alpha-tocopherol concentrations of normal subjects given only 150 mg of each isotope. Three of the patients discriminated between forms of alpha-tocopherol with ratios of RRR-/allrac-alpha-tocopherol > or = 1.8, similar to normals. These data suggest that the hepatic tocopherol binding protein is present and functional in ABL patients. Although two of the patients did not discriminate between stereoisomers of alpha-tocopherol, it is likely that this resulted from nearly a complete block in very low density lipoprotein (VLDL) secretion. Thus, the ability of ABL patients to absorb and transport orally administered vitamin E is markedly impaired and variable among patients.


Yeast two-hybrid analysis of jasmonate signaling proteins.
Protein-protein interaction studies are crucial to unravel how jasmonate (JA) signals are transduced. Among the different techniques available, yeast two-hybrid (Y2H) is commonly used within the JA research community to identify proteins belonging to the core JA signaling module. The technique is based on the reconstitution of a transcriptional activator that drives the reporter gene expression upon protein-protein interactions. The method is sensitive and straightforward and can be adapted for different approaches. In this chapter, we provide a detailed protocol to perform targeted Y2H assays to test known proteins and/or protein domains for direct interaction in a pairwise manner and present the possibility to study ternary protein complexes through Y3H.


Mechanism of the glucocorticoid regulation of growth of the androgen-sensitive prostate-derived R3327H-G8-A1 tumor cell line.
The R3327H-G8-A1 cell line derived from the Dunning rat prostate adenocarcinoma contains both androgen and glucocorticoid receptors. Following steroid deprivation, androgens specifically increase the concentration of their receptors in these cells by approximately 2-fold within 6 h and 3-4-fold in 24 h. In the presence of potent glucocorticoids, androgen receptor augmentation is reduced by 40-50% in the first 6 h and completely inhibited during the subsequent 24 h. This event, which is specific for glucocorticoids, appears to be due to an inhibition of androgen receptor synthesis. Furthermore, glucocorticoids inhibit proliferation of these cells by inhibiting the release of growth factors and arresting them in the G0 or A state of the cell cycle. This inhibition can be overcome by addition of low concentrations of either epidermal growth factor or platelet-derived growth factor; however, the inhibitory effect of the glucocorticoid on androgen receptor augmentation is not released. These results suggest that glucocorticoids arrest cellular proliferation by altering the autoregulation of growth and that this event is not dependent upon inhibition of androgen receptor augmentation.


8-Aminoquinolines as anticoccidials--II.
During a chemistry program aimed at finding a novel analogue of pentaquine with improved in vivo activity, a number of hypotheses concerning the way this drug acts in the chicken were investigated. Consideration of the products of monoamine oxidase metabolism of pentaquine suggested that pentaquine aldehyde is the likely active metabolite. Although isolation of this unstable compound was not possible, oxime and cyclic acetal and ketal derivatives were obtained and shown to possess in vitro anticoccidial activity.


Correlations between the trigeminal nerve microstructural changes and the trigeminal-pontine angle features.
Morphological and microstructural changes of the trigeminal nerve due to neurovascular compression (NVC) have been reported in primary trigeminal neuralgia (PTN) patients. This investigation was to examine the relationship between the trigeminal-pontine angle and nerve microstructural changes. Twenty-five patients underwent microvascular decompression (MVD) for trigeminal neuralgia, and 25 age- and sex-matched controls were studied. The two groups underwent high-resolution three-dimensional MRI and diffusion tensor imaging (DTI). Bilateral trigeminal-pontine angle, cross-sectional area of cerebellopontine angle (CPA) cistern, and the length of trigeminal nerve were evaluated. The mean values of fractional anisotropy and apparent diffusion coefficient at the site of NVC were also measured. Correlation analyses were performed for the trigeminal-pontine angle and the diffusion metrics (FA and ADC) in PTN patients. The mean trigeminal-pontine angle and FA value on the affected side was significantly smaller than the unaffected side and the control group (p < 0.001), while the mean ADC value was significantly increased (p < 0.01). When taking the conflicting vessel types into consideration, the angle affected by the superior cerebellar artery (SCA) was statistically sharper than when affected by other vessels (p < 0.01). However, there were no significant changes in the area of the CPA cistern or the length of the trigeminal nerve between the groups. Correlation analyses showed that the trigeminal-pontine angle was positively correlated with FA and negatively correlated with ADC. A sharp trigeminal-pontine angle may increase the chance of NVC and exacerbate nerve degeneration, which may be one of the supplementary factors that contribute to the pathogenesis of trigeminal neuralgia.


A critical review of enzymes immobilized on chitosan composites: characterization and applications.
Enzymes with industrial significance are typically used in biological processes. However, instability, high sensitivity, and impractical recovery are the major drawbacks of enzymes in practical applications. In recent years, the immobilization technology has attracted wide attention to overcoming these restrictions and improving the efficiency of enzyme applications. Chitosan (CS) is a unique functional substance with biocompatibility, biodegradability, non-toxicity, and antibacterial properties. Chitosan composites are anticipated to be widely used in the near future for a variety of purposes, including as supports for enzyme immobilization, because of their advantages. Therefor this review explores the effects of the chitosan's structure, molecular weight, degree of deacetylation on the enzyme immobilized, effect of key factors, and the enzymes immobilized on chitosan based composites for numerous applications, including the fields of biosensor, biomedical science, food industry, environmental protection, and industrial production. Moreover, this study carefully investigates the advantages and disadvantages of using these composites as well as their potential in the future.


Droxinostat, a Histone Deacetylase Inhibitor, Induces Apoptosis in Hepatocellular Carcinoma Cell Lines via Activation of the Mitochondrial Pathway and Downregulation of FLIP.
The current chemotherapeutic outcomes for hepatocellular carcinoma (HCC) are not encouraging, and long-term survival of this patient group remains poor. Recent studies have demonstrated the utility of histone deacetylase inhibitors that can disrupt cell proliferation and survival in HCC management. However, the effects of droxinostat, a type of histone deacetylase inhibitor, on HCC remain to be established. The effects of droxinostat on HCC cell lines SMMC-7721 and HepG2 were investigated. Histone acetylation and apoptosis-modulating proteins were assessed via Western blot. Proliferation was examined with 3-(4, 5 dimetyl-2-thiazolyl)-2, 5-diphenyl 2H-tetrazolium bromide, cell proliferation, and real-time cell viability assays, and apoptosis with flow cytometry. Droxinostat inhibited proliferation and colony formation of the HCC cell lines examined. Hepatoma cell death was induced through activation of the mitochondrial apoptotic pathway and downregulation of FLIP expression. Droxinostat suppressed histone deacetylase (HDAC) 3 expression and promoted acetylation of histones H3 and H4. Knockdown of HDAC3 induced hepatoma cell apoptosis and histone H3 and H4 acetylation. Droxinostat suppresses HDAC3 expression and induces histone acetylation and HCC cell death through activation of the mitochondrial apoptotic pathway and downregulation of FLIP, supporting its potential application in the treatment of HCC.


Spontaneous rupture of the extensor carpi radialis brevis in a 51-year-old man: case report.
Dorsal hand osteophytes are common findings in the general population, frequently presenting with dorsal pain and treated with surgical excision. We report the spontaneous rupture of the extensor carpi radialis brevis in association with a previously asymptomatic dorsal scaphoid spur. Following conservative management, surgical excision of dorsal hand osteophytes should be considered for both resolution of pain and prevention of attritional tendon rupture.


Reprogrammed Functional Brown Adipocytes Ameliorate Insulin Resistance and Dyslipidemia in Diet-Induced Obesity and Type 2 Diabetes.
Brown adipocytes (BAs) play important roles in body temperature regulation, energy balance, and carbohydrate and lipid metabolism. Activities of BAs are remarkably diminished in obese and diabetic patients, providing possibilities of transplanting functional BAs resulting in therapeutic benefit. Here, we show generation of functional BAs by cellular reprogramming procedures. Transduction of the PRDM16 gene into iPSC-derived embryoid bodies induced BA phenotypes (iBAs). Moreover, normal human fibroblasts were directly converted into BAs (dBAs) by C/EBP-β and C-MYC gene transduction. Approximately 90% of the fibroblasts were successfully converted within 12 days. The dBAs were highly active in mitochondrial biogenesis and oxidative metabolism. Mouse dBAs were induced by Prdm16, C/ebp-β, and L-myc genes, and after transplantation, they significantly reduced diet-induced obesity and insulin resistance in an UCP1-dependent manner. Thus, highly functional BAs can be generated by cellular reprogramming, suggesting a promising tailor-made cell therapy against metabolic disorders including type 2 diabetes mellitus.


BK Virus Nephropathy: Prevalence, Impact and Management Strategies.
BK virus reactivation as a result of therapeutic immunosuppression following renal transplant can result in BK polyomavirus nephropathy and renal allograft loss. This is a complex and challenging clinical problem with a range of management options and practices reported in literature. The current standard for early diagnosis and treatment is surveillance by measuring viral DNA in blood using qPCR. Immunosuppression reduction is the cornerstone of effective management but is associated with a risk of acute rejection following treatment.


Cow's milk-induced pulmonary hemosiderosis.
Cow's milk protein-induced pulmonary disease is a serious condition which, if recognized, can be treated effectively by avoidance of milk protein; untreated it may lead to early death. It should be considered in any child who suffers recurrent episodes of pneumonia and anemia.


Genetic diversity and evolution of Hantaan virus in China and its neighbors.
Hantaan virus (HTNV; family Hantaviridae, order Bunyavirales) causes hemorrhagic fever with renal syndrome (HFRS), which has raised serious concerns in Eurasia, especially in China, Russia, and South Korea. Previous studies reported genetic diversity and phylogenetic features of HTNV in different parts of China, but the analyses from the holistic perspective are rare. To better understand HTNV genetic diversity and gene evolution, we analyzed all available complete sequences derived from the small (S) and medium (M) segments with bioinformatic tools. Eleven phylogenetic groups were defined and showed geographic clustering; 42 significant amino acid variant sites were found, and 19 of them were located in immune epitopes; nine recombinant events and eight reassortments with highly divergent sequences were found and analyzed. We found that sequences from Guizhou showed high genetic divergence, contributing to multiple lineages of the phylogenetic tree and also to the recombination and reassortment events. Bayesian stochastic search variable selection analysis revealed that Heilongjiang, Shaanxi, and Guizhou played important roles in HTNV evolution and migration; the virus may originate from Zhejiang Province in the eastern part of China; and the virus population size expanded from the 1980s to 1990s. These findings revealed the original and evolutionary features of HTNV, which will help to illustrate hantavirus epidemic trends, thus aiding in disease control and prevention.


Complete sequence of the Sendai virus NP gene from a cloned insert.
A DNA molecule representing all but the three terminal bases of the Sendai virus nucleoprotein (NP) gene, copied from viral mRNA, was inserted into pBR322. The NP insert comprised 1673 bases. The first AUG protein initiation codon, at position 65, began an open reading frame of 1551 bases, encoding a protein of 517 amino acids with an amino acid composition corresponding to previously published data. The NP gene sequence determined in the present work is similar to that described by Shioda et al. [ Nucl . Acids Res. 11, 7317 (1983)], but there are 14 amino acid differences that probably reflect differences in virus strains. The predicted secondary structure of the NP molecule and the locations within that structure of potential protease cleavage sites are in accord with structural domains previously defined by controlled protease digestion.


Comparison and Evaluation of Force Fields for Intrinsically Disordered Proteins.
Molecular dynamics (MD) simulations of six upgraded empirical force fields were compared and evaluated with short peptides, intrinsically disordered proteins, and folded proteins using trajectories of 1, 1.5, 5, or 10 μs (five replicates of 200 ns, 300 ns, 1 μs, or 2 μs) for each system. Previous studies have shown that different force fields, water models, simulation methods, and parameters can affect simulation outcomes. Here, the MD simulations were done in an explicit solvent with RS-peptide, HEWL19, HIV-rev, β amyloid (Aβ)-40, Aβ-42, phosphodiesterase-γ, CspTm, and ubiquitin using ff99IDPs, ff14IDPs, ff14IDPSFF, ff03w, CHARMM36m, and CHARMM22* force fields. The IDP ensembles generated by six all-atom empirical force fields were compared against NMR data. Despite using identical starting structures and simulation parameters, ensembles obtained with different force fields exhibit significant differences in NMR RMDs, secondary structure contents, and global properties such as the radius of gyration. The intrinsically disordered protein (IDP)-specific force fields could substantially reproduce the experimental observables in force field comparison: they have the lowest error in chemical shifts and J-couplings for short peptides/proteins, reasonably well for large IDPs and reasonably well with the radius of gyration. A high population of disorderness was observed in the IDP-specific force field for the IDP ensemble with a fraction of β sheets for β-amyloids. CHARMM22* performs better for many observables; however, it still has a preference toward the helicity for short peptides. The results of β-amyloid 42 starting from two different initial structures (Aβ421Z0Q and Aβ42model) were also compared with DSSP and NMR data. The results obtained with IDP-specific force fields within 2 μs simulation time are similar, even though starting from different structures. The current force fields perform equally well for folded proteins. The results of currently developed or modified force fields for IDPs are capable of enlightening the overall performance of the force field for disordered as well as folded proteins, thereby contributing to force field development.


A pericentric inversion of chromosome 9 and a rearrangement involving chromosomes 9 and 10, observed in two generations. Clinical description of chromosome 9 (p12-p21) deletion syndrome.
The clinical picture associated with a deletion of a central part of the short arm of chromosome no. 9 is described in two siblings. The clinical signs differ from those described in deletion of the terminal part of the short arm. Pericentric inversion of chromosome no. 9, combined with a rearrangement involving chromosomes 9 and 10, was found in the mother and the maternal grandmother of the propositus.


Attenuation of auditory N1 results from identity-specific action-effect prediction.
The auditory N1 event-related potential has previously been observed to be attenuated for tones that are triggered by human actions. This attenuation is thought to be generated by motor prediction mechanisms and is considered to be important for agency attribution. The present study was designed to rigorously test the notion of action prediction-based sensory attenuation. Participants performed one of four voluntary actions on each trial, with each button associated with either predictable or unpredictable action effects. In addition, actions with each hand could result in action effects that were either congruent or incongruent with hand-specific prediction. We observed no significant differences in N1 amplitude between predictable and unpredictable tones. When contrasting action effects that were congruent or incongruent with hand-specific prediction, we observed significant attenuation for prediction-congruent compared to prediction-incongruent action-effects. These novel findings suggest that accurate action-effect prediction drives sensory attenuation of auditory stimuli. These findings have important implications for understanding the mechanisms of action-effect prediction and sensory attenuation, and may have clinical implications for studies investigating action awareness and agency in schizophrenia.


Topological mapping of 13 epitopes on a subunit of Androctonus australis hemocyanin.
A topological localization of epitopes on the surface of the Aa6 subunit of Androctonus australis hemocyanin has been carried out. First, immunocomplex strings composed of native hemocyanin and monoclonal antibodies were examined in the electron microscope and submitted to an image processing by correspondence analysis. The average images were then compared to a three-dimensional model of the 24-mer suggesting that 11 of the 13 epitopes are located in three zones of the subunit surface. Second, the overlaps between the epitopes were then studied by polyacrylamide gel electrophoresis, competitive binding inhibition, and immunoelectron microscopy. Four groups of epitopes were identified. One group was capable of binding exclusively to the free subunit. The other three groups were identical to those found in immunoelectron microscopy. The data are consistent with the existence of a small number of immunodominant regions on the surface of the Aa6 subunit.


A new perspective on volatile halogenated hydrocarbons in Chinese agricultural soils.
Soil contamination by volatile organic compounds has been greatly studied. However, there is still limited information regarding the occurrence, distribution and health effects of typical volatile halogenated hydrocarbons (VHCs) in soils on a national scale. In this study, headspace-based extraction with gas chromatography/mass spectrometry (HS-GC/MS) detection was optimized for the simultaneous analysis of 18 VHCs (haloalkanes, haloalkenes and halogenated aromatics) in 112 surface agricultural soil samples across China in 2016. The results show that 100% of the soil samples were contaminated by VHCs, and 13 of the 18 VHCs investigated were detected. The haloalkanes were the dominant group. Five VHC components had detection rates greater than 35%, including dichloromethane (DCM), chloroform (CF), 1,2-dichloroethane (1,2-DCA), chlorobenzene (CB) and 1,4-dichlorobenzene (1,4-DCB). For total VHCs, North and Southwest China were the hotspots for contamination. The highest concentrations were found in Shanxi Province and Chongqing municipality, which could be the result of long-term aerial deposition and wet deposition from various accidental industrial leakages or natural sources. In addition, the applications of sewage sludge and pesticides in agricultural activities may also contribute to soil VHC pollution. By using the health risk-assessing models recommended by the U.S. Environment Protection Agency (U.S. EPA), the estimated hazard indices (HIs) of all VHCs were below 1, and the carcinogenic risk (CR) values were all at acceptable levels (<1 × 10-6). These findings indicated that the agricultural soils may not trigger serious long-term health impacts on public health nationwide. The results from this study can initially grasp the agricultural soil VHC pollution level and provide an understanding to avoid potential ecological and human health risks.


The effect of 4 days methionine sulfoximine administration on net muscle protein breakdown in rats.
It has been suggested that the amino acid glutamine is essential for the regulation of protein turnover in muscle, but in vivo data are lacking. Therefore, administration of methionine sulfoximine (MSO) was used to inhibit glutamine synthetase activity and to induce in vivo muscle glutamine depletion. Glutamine metabolism of the hindquarter was measured by determining fluxes and intracellular concentrations after an overnight fast in ether anaesthetized normal rats, MSO treated rats and their pairfed controls. Fluxes and intracellular concentrations of several other amino-acids including 3-methylhistidine and ammonia were also determined. MSO treatment resulted in a 50% decrease in arterial glutamine concentration, a 55% reduction in intracellular muscle glutamine and a 50% increase in muscle ammonia. Four day MSO treatment significantly increased hindquarter muscle plasma flow. Precursors of muscle glutamine synthesis seem to be preferentially used for increased production of alanine. Ammonia was found to be released in increased amounts. The increased efflux of amino-acids including phenylalanine and tyrosine indicate that MSO treatment induces net muscle protein catabolism. These results suggest that, in vivo, reduced intramuscular glutamine concentrations correlate with increased net muscle protein breakdown.


Better prediction by use of co-data: adaptive group-regularized ridge regression.
For many high-dimensional studies, additional information on the variables, like (genomic) annotation or external p-values, is available. In the context of binary and continuous prediction, we develop a method for adaptive group-regularized (logistic) ridge regression, which makes structural use of such 'co-data'. Here, 'groups' refer to a partition of the variables according to the co-data. We derive empirical Bayes estimates of group-specific penalties, which possess several nice properties: (i) They are analytical. (ii) They adapt to the informativeness of the co-data for the data at hand. (iii) Only one global penalty parameter requires tuning by cross-validation. In addition, the method allows use of multiple types of co-data at little extra computational effort. We show that the group-specific penalties may lead to a larger distinction between 'near-zero' and relatively large regression parameters, which facilitates post hoc variable selection. The method, termed GRridge, is implemented in an easy-to-use R-package. It is demonstrated on two cancer genomics studies, which both concern the discrimination of precancerous cervical lesions from normal cervix tissues using methylation microarray data. For both examples, GRridge clearly improves the predictive performances of ordinary logistic ridge regression and the group lasso. In addition, we show that for the second study, the relatively good predictive performance is maintained when selecting only 42 variables.


Vitrectomy in infants and children with retinal detachments caused by cicatricial retrolental fibroplasia.
Four infants and one child with retinal detachments from cicatricial retrolental fibroplasia involving traction judged too severe to be relieved by scleral buckling alone underwent lensectomy and vitrectomy. Three retinas were successfully reattached. Because of the incomplete development of the pars plana in infants and the manner in which peripheral retina is pulled anteriorly to a position near or in contact with the posterior lens capsule, entry through the pars plana can create iatrogenic tears in the peripheral retina or introduction of irrigation fluid under the retina. Our experience has shown that a small contact lens makes it possible to work in the posterior vitreous cavity through the limbal approach in these infants. The circumferential traction produced by the ring of fibrovascular proliferation behind the lens can be relieved and tractional bands running forward from the posterior pole can be cut. Fibrovascular proliferative tissue lying on the posterior pole, however, proved resistant to membrane peeling or dissection, and its severity seemed to determine the success or failure of the surgery.


Effect of sodium molybdate on carbohydrate metabolizing enzymes in alloxan-induced diabetic rats.
We evaluated the effect of sodium molybdate on carbohydrate metabolizing enzymes and mitochondrial enzymes in diabetic rats. Diabetic rats showed a significant reduction in the activities of glucose metabolising enzymes like hexokinase, glucose-6-phosphate dehydrogenase, glycogen synthase and in the level of glycogen. An elevation in the activities of aldolase, glucose-6-phosphatase, fructose 1,6- bisphosphatase, glycogen phosphorylase and in the level of blood glucose were also observed in diabetic rats when compared to control rats. The activities of mitochondrial enzymes isocitrate dehydrogenase, alpha-ketoglutarate dehydrogenase, succinate dehydrogenase, malate dehydrogenase, NADH-dehydrogenase and cytochrome-C-oxidase were also significantly lowered in diabetic rats. Molybdate administration to diabetic rats reversed the above changes in a significant manner. From our observations, we conclude that administration of sodium molybdate regulated the blood sugar levels in alloxan-induced diabetic rats. Sodium molybdate therapy not only maintained the blood glucose homeostasis but also altered the activities of carbohydrate metabolising enzymes. Molybdate therapy also considerably improved the activities of mitochondrial enzymes, thereby suggesting its role in mitochondrial energy production.


When the clock ticks wrong with COVID-19.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a member of the coronavirus family that causes the novel coronavirus disease first diagnosed in 2019 (COVID-19). Although many studies have been carried out in recent months to determine why the disease clinical presentations and outcomes can vary significantly from asymptomatic to severe or lethal, the underlying mechanisms are not fully understood. It is likely that unique individual characteristics can strongly influence the broad disease variability; thus, tailored diagnostic and therapeutic approaches are needed to improve clinical outcomes. The circadian clock is a critical regulatory mechanism orchestrating major physiological and pathological processes. It is generally accepted that more than half of the cell-specific genes in any given organ are under circadian control. Although it is known that a specific role of the circadian clock is to coordinate the immune system's steady-state function and response to infectious threats, the links between the circadian clock and SARS-CoV-2 infection are only now emerging. How inter-individual variability of the circadian profile and its dysregulation may play a role in the differences noted in the COVID-19-related disease presentations, and outcome remains largely underinvestigated. This review summarizes the current evidence on the potential links between circadian clock dysregulation and SARS-CoV-2 infection susceptibility, disease presentation and progression, and clinical outcomes. Further research in this area may contribute towards novel circadian-centred prognostic, diagnostic and therapeutic approaches for COVID-19 in the era of precision health.


Selective enrichment of plasma cell-free messenger RNA in cancer-associated extracellular vesicles.
Extracellular vesicles (EVs) have been shown as key mediators of extracellular small RNA transport. However, carriers of cell-free messenger RNA (cf-mRNA) in human biofluids and their association with cancer remain poorly understood. Here, we performed a transcriptomic analysis of size-fractionated plasma from lung cancer, liver cancer, multiple myeloma, and healthy donors. Morphology and size distribution analysis showed the successful separation of large and medium particles from other soluble plasma protein fractions. We developed a strategy to purify and sequence ultra-low amounts of cf-mRNA from particle and protein enriched subpopulations with the implementation of RNA spike-ins to control for technical variability and to normalize for intrinsic drastic differences in cf-mRNA amount carried in each plasma fraction. We found that the majority of cf-mRNA was enriched and protected in EVs with remarkable stability in RNase-rich environments. We observed specific enrichment patterns of cancer-associated cf-mRNA in each particle and protein enriched subpopulation. The EV-enriched differentiating genes were associated with specific biological pathways, such as immune systems, liver function, and toxic substance regulation in lung cancer, liver cancer, and multiple myeloma, respectively. Our results suggest that dissecting the complexity of EV subpopulations illuminates their biological significance and offers a promising liquid biopsy approach.


In Vivo Antitumor and Antimetastatic Efficacy of a Polyacetal-Based Paclitaxel Conjugate for Prostate Cancer Therapy.
Prostate cancer (PCa), one of the leading causes of cancer-related deaths, currently lacks effective treatment for advanced-stage disease. Paclitaxel (PTX) is a highly active chemotherapeutic drug and the first-line treatment for PCa; however, conventional PTX formulation causes severe hypersensitivity reactions and limits PTX use at high concentrations. In the pursuit of high molecular weight, biodegradable, and pH-responsive polymeric carriers, one conjugates PTX to a polyacetal-based nanocarrier to yield a tert-Ser-PTX polyacetal conjugate. tert-Ser-PTX conjugate provides sustained release of PTX over 2 weeks in a pH-responsive manner while also obtaining a degree of epimerization of PTX to 7-epi-PTX. Serum proteins stabilize tert-Ser-PTX, with enhanced stability in human serum versus PBS (pH 7.4). In vitro efficacy assessments in PCa cells demonstrate IC50 values above those for the free form of PTX due to the differential cell trafficking modes; however, in vivo tolerability assays demonstrate that tert-Ser-PTX significantly reduces the systemic toxicities associated with free PTX treatment. tert-Ser-PTX also effectively inhibits primary tumor growth and hematologic, lymphatic, and coelomic dissemination, as confirmed by in vivo and ex vivo bioluminescence imaging and histopathological evaluations in mice carrying orthotopic LNCaP tumors. Overall, the results suggest the application of tert-Ser-PTX as a robust antitumor/antimetastatic treatment for PCa.


Imaging in children with hearing loss.
Since the introduction of hearing screening in Germany in 2009, pediatric hearing disorders are detected at an early stage. Early therapy is essential for language development. Imaging plays a central role in diagnosis and therapy planning. Imaging findings of the most relevant causes of pediatric hearing disorders are presented. Specific attention is given to the method used in each case - CT or MRI. While CT is the method of choice for conductive hearing loss, a combination of CT and MRI with high-resolution T2-3D sequences has been established as the best diagnostic method for sensorineural hearing loss. The most common causes of conductive hearing loss in childhood are chronic inflammation and cholesteatoma. Congenital malformations of the outer or middle ear are less frequent. In the case of sensorineural hearing loss, the cause is located in the inner ear and/or the cochlear nerve or the cerebrum. In these cases, congenital malformations are the most common cause. · CT and MRI are necessary to identify morphological causes of hearing disorders and to clarify the possibility of hearing-improving ear surgery or cochlear implantation.. · Contraindications for surgical procedures must be excluded.. · Anatomical variants that may be risk factors for surgery must be described.. · Sorge I, Hirsch F, Fuchs M et al. Imaging diagnostics for childhood hearing loss. Fortschr Röntgenstr 2023; 195: 896 - 904.


Distribution of endocrine disruptors in the Mondego River estuary, Portugal.
Recent studies in the Mondego River estuary demonstrated signs of pollution in the area, but the nature of the contamination remains unexplored. Because there seems to be any studies in that zone checking for the simultaneous presence of xenoestrogenic endocrine disrupting compounds, either of animal (estradiol and estrone), vegetal (daidzein, genistein and biochanin A), pharmaceutical (17alpha-ethynylestradiol) or industrial (bisphenol A, 4-octylphenol, 4-nonylphenol) origins, the main objective of this study was to investigate their presence, in every year season and at eight points along the estuarine gradient. For this propose, water samples (2 L) were collected in high and low tides, preconcentrated in the Oasis HLB cartridges and cleaned in silica cartridges before their analysis by HPLC-DAD and GC-MS. The current data showed the absence of contamination by animal or pharmaceutical estrogens and by alkylphenols, but demonstrated the presence of high levels of phytoestrogens (up to 1.1 mug/L) and of bisphenol A (up to 880.0 ng/L). Because these two chemicals existed in high environmental concentrations in the most upstream sampling station, and taking in account that these pollutants may additively contribute to endocrine disruption, namely on fish, it was concluded that at least this particularly area of the Mondego estuary deserves continuous monitoring programmes.


Insulin-like growth factor I (IGF-I) measurements in growth hormone (GH) therapy of idiopathic short stature (ISS).
Growth hormone (GH) therapy has evolved rapidly over the past decade. Ongoing research has demonstrated a clear role for therapeutic GH in a wide spectrum of pediatric disorders involving both poor growth and abnormal body composition. Although guidelines for GH dosing are not fully established, a series of key studies has delineated the range of dosages that are useful in the treatment of children with growth disorders. The recent approval of idiopathic short stature (ISS) as an indication for GH therapy presents further challenges in optimizing the care of GH-treated patients. ISS is now recognized as a diverse collection of environmental and molecular abnormalities, some of which involve the GH-IGF axis. Emerging data indicate that serum IGF-I measurements are not only useful in the diagnosis of growth abnormalities but, in conjunction with auxological measurements, are also a powerful tool for assessing GH efficacy. While it is clear that many ISS patients respond to GH, some individuals will not show a satisfactory response. Monitoring IGF-I levels and change in height SDS during treatment can assist the physician in distinguishing those patients in whom GH successfully and safely induces statural growth from those with partial or complete GH insensitivity who might benefit from modified GH treatment protocols or alternate therapies. In addition, serum IGF-I measurements are increasingly used as part of a rational monitoring strategy to ensure safety of GH dosing in light of cumulative data associating high IGF-I levels with potential malignancy risk, and low IGF-I levels with cardiovascular disease risk.


Taming the hydra of specialized metabolism: how systems biology and comparative approaches are revolutionizing plant biochemistry.
Specialized (traditionally called 'secondary') metabolism can be thought of as a hydra with hundreds of thousands of compounds produced by thousands of enzymes across the entire plant kingdom. Until recently, plants that produce the most interesting and valuable metabolites were recalcitrant to modern molecular biology approaches for gene and pathway discovery. Recent advances in technologies for genomic, transcriptomic, proteomic, and metabolomic methods now allow for deployment of 'systems biology' approaches to help elucidate unknown steps in specialized metabolite pathways, for example through co-expression analyses. Inexpensive transcriptome and whole genome sequencing (WGS) promises to provide direct access to metabolic pathways in plants not currently used as reference organisms. For example, WGS has uncovered cases of physical proximity of genes of specialized metabolism. Further integration of multiple 'omics' datasets through advances in bioinformatics tools will increase our knowledge of pathway architecture and regulation at an ever-increasing rate. As such the era of systems biology is rapidly providing a broader and deeper understanding of plant specialized metabolism.


Aspergillus peritonitis in a lupus patient on chronic peritoneal dialysis.
A woman on continuous ambulatory peritoneal dialysis (CAPD) due to renal failure in systemic lupus erythematosus (SLE) developed fungal peritonitis and survived following treatment with amphotericin B and removal of the dialysis catheter. The causative organism, Aspergillus fumigatus is very rare in fungal peritonitis and may be related in this case to the combination of SLE, end-stage renal disease (ESRD) and their treatment.


Bowel obstruction in the postoperative period of laparoscopic inguinal hernia repair (TAPP): review of the literature.
A case is presented of a male patient that presented with intestinal obstruction in the early postoperative period of a transabdominal preperitoneal inguinal repair (TAPP) that was diagnosed and repaired successfully using laparoscopy. Whenever a total extraperitoneal procedure (TEP) cannot be performed, the peritoneal closure of the TAPP should be done water-tight using interrupted stitches of absorbable monofilament sutures.


Artificial double vagina in association with Rokitansky syndrome.
A case of Rokitansky syndrome is described where the typical anomalies of the entity were associated with cribriform hymen. The appearance of the external genitalia led to an erroneous diagnosis of complete vaginal agenesis and to surgical creation of an artificial vagina by the McIndoe technique at the patient's age of 19. She had a satisfactory sexual life subsequently. Ten years following the surgery, routine examination revealed that the patient had an essentially normal vagina with a blind upper ending. Prior to this, she and her husband had been unaware of the fact that their marital relations were conducted through her natural vagina during the years of their sexual activities.


Genetic determinants of normal variation in coagulation factor (F) IX levels: genome-wide scan and examination of the FIX structural gene.
High-normal and elevated plasma FIX activity (FIX:C) levels are associated with increased risk for venous- and possibly arterial-thrombosis. Because the broad normal range for FIX:C involves a substantial unknown genetic component, we sought to identify quantitative-trait loci (QTLs) for this medically important hemostasis trait. We performed a genome-wide screen and a resequencing-based variation scan of the known functional regions of every distinct FIX gene (F9) in the genetic analysis of idiopathic thrombophilia project (GAIT), a collection of 398 Spanish-Caucasians from 21 pedigrees. We found no evidence for linkage (LOD scores <1.5) despite genotyping more than 540 uniformly-spaced microsatellites. We identified 27 candidate F9 polymorphisms, including three in cis-elements responsible for the increase in FIX:C that occurs with aging, but found no significant genotype-specific differences in mean FIX:C levels (P-values > or = 0.11) despite evaluating every polymorphism in GAIT by marginal multicovariate measured-genotype association analysis. The heritable component of interindividual FIX:C variability likely involves a collection of QTLs with modest effects that may reside in genes other than F9. Nevertheless, because the alleles of these 27 polymorphisms exhibited a low overall degree of linkage disequilibrium, we are currently defining their haplotypes to interrogate several highly-conserved non-exonic sequences and other F9 segments not examined here.


Oncolysis by Clostridium oncolyticum M55 and subsequent enzymatic determination of sialic acid in serum.
Since the discovery of the "Clostridium tetani phenomenon", various apathogenic clostridia have been used for tumour lysis. Experiments have been conducted to achieve a tumour diagnosis using radiolabelled antibodies to clostridia. In addition, a method has been described that distinguishes, with variable success, between healthy and tumour-carrying animals by means of hemagglutination. The method outlined here uses the fact that malignant cells produce a multitude of sialic acid compounds which lie on the cell membrane and are also connected to the lipid layer of the tumour cell membrane. The apathogenic Clostridium oncolyticum M55 only germinates and multiplies in the malignant tumour tissue. Thus; bacterial hydrolases can enter the tumour tissue and lead to oncolysis. Subsequently the glycocompounds which can be detected by means of an enzymatic determination of the concentration of neuraminic acid (one of the sialic acids) in the serum are washed out into the peripheral blood. We observed these processes in mice in the Ehrlich ascites solid carcinoma and in the Lewis lung carcinoma. Using this method it was possible to detect tumour growth at an early stage with impressive accuracy. The Lewis lung carcinoma which secretes only small amounts of sialic acid glycocompounds cannot be distinguished from the control group by determination of sialic acid concentration. It was possible to detect a 52% increase in the amount of sialic acid after administration of spores of clostridia. This method makes it possible to increase the tumour marker sialic acid through manipulation of the tumour, using apathogenic clostridia, and to measure of sialic acid concentration as an indicator of the metabolic products of the tumour.


Deoxynivalenol triggers porcine intestinal tight junction disorder: Insights from mitochondrial dynamics and mitophagy.
Deoxynivalenol (DON) is universally detected trichothecene in most cereal commodities, which is considered as a major hazardous material for human and animal health. Intestine is the most vulnerable organ with higher concentration of DON than other organs, owing to the first defense barrier function to exogenous substances. However, the underling mechanisms about DON-induced intestinal toxicity remain poorly understood. Here, DON poisoning models of IPEC-J2 cells was established to explore adverse effect and the potential mechanism of DON-induced enterotoxicity. Results showed that DON exposure destroyed IPEC-J2 cells morphology. Results showed that DON exposure destroyed IPEC-J2 cells morphology. Intestinal epithelial barrier injury was caused by DON with increasing LDH release, decreasing cell viability as well decreasing tight junction protein expressions (Occludin, N-Cad, ZO-1, Claudin-1 and Claudin-3). Moreover, DON caused mitochondrial dysfunction by opening mitochondrial permeability transition pore and eliminating mitochondrial membrane potential. DON exposure upregulated protein and mRNA expression of mitochondrial fission factors (Drp1, Fis1, MIEF1 and MFF) and mitophagy factors (PINK1, Parkin and LC3), downregulated mitochondrial fusion factors (Mfn1, Mfn2, except OPA1), resulting in mitochondrial dynamics imbalance and mitophagy. Overall, these findings suggested that DON induced tight junction dysfunction in IPEC-J2 cells was related to mitochondrial dynamics-mediated mitophagy.


General and selective inhibition of pancreatic enzyme discharge using a proteinase inhibitor (FOY-305).
The guanidino acid esters (FOY, FOY-305) represent a new class of potent proteinase inhibitors and are thought to have a beneficial effect on the course of acute pancreatitis. Because of their structure and low molecular size they might enter cells and interfere with cellular processes. To test this possibility in the case of the exocrine pancreas a series of in vivo and in vitro studies was carried out to analyse intracellular transport and discharge of pancreatic enzymes in the presence of FOY-305. The infusion of FOY-305 to conscious rats led to a transient inhibition of protein and enzyme discharge from the cannulated pancreas accompanied by lower serum enzyme levels and increased enzyme content in the pancreas. An identical inhibition of discharge of newly synthesized proteins was observed in vitro in the presence of 1 microM FOY-305. The analysis of the release of individual enzymes using separation on two-dimensional gels showed a pronounced inhibition of mainly the release of acidic proteins. FOY-305 not only interfered with discharge of serine proteinases (trypsinogen, chymotrypsinogen, proelastase) but also with procarboxypeptidases and lipase. It was concluded that FOY-305 enters the acinar cell and due to an unspecific binding to acidic proteins interferes with the intracellular transport of individual enzyme proteins during their passage through the membrane-bound cellular compartments. This charge-dependent effect is independent of the inhibitory effect on enzymatic activity of serine proteinases.


Malignant transformation of recurrent meningioma with pulmonary metastases.
A patient presented with a histologically benign intracranial meningioma which, after multiple recurrences, underwent malignant transformation. The patient survived 18 years following initial presentation. Pulmonary metastases were present over the final 8 years. Eleven subtotal resections of the meningioma were performed, including exenteration of the right eye and a thoracotomy. Radiation therapy and chemotherapy were relatively ineffective. The problem and treatment of recurrent meningiomas is briefly reviewed.


Interaction of DPI 201-106 with cardiac muscarinic receptors.
The effects of the new cardiotonic compound, DPI 201-106, on muscarinic responses and muscarinic receptor binding were studied in the guinea pig heart. DPI 201-106 exerted a pronounced anticholinergic action in isolated auricles and a moderate one in papillary muscles, which resembled the pattern of a functional antagonism. However, in competition binding experiments, DPI 201-106 inhibited binding of the specific muscarinic antagonist [3H]NMS with equal potency in atrial and ventricular homogenates (apparent KI = 0.7 mumol/l in atria and 1.2 mumol/l in ventricles). At higher concentrations (greater than 3 mumol/l), DPI 201-106 slowed the dissociation of [3H]NMS from cardiac muscarinic receptors, indicating that DPI 201-106 affects in addition a site allosteric to the muscarinic receptor. It is concluded that DPI 201-106 is able to inhibit cholinergic responses in the heart, not only by a functional antagonism but also by direct interaction with muscarinic receptors.


Tm3+/Ho3+ profiled co-doped core area optical fiber for emission in the range of 1.6-2.1 µm.
Double-clad optical fiber with a multi-ring core profile doped with thulium and holmium fabricated by Modified Chemical Vapor Deposition Chelate Doping Technology (MCVD-CDT) is presented. The measured Tm2O3 and Ho2O3 complexes' weight concentrations were 0.5% and 0.2% respectively. Numerical analyses show weakly guiding conditions and 42.2 µm of MFD LP01 at 2000 nm. The low NA numerical aperture (NA = 0.054) was obtained for the 20/250 µm core/cladding ratio optical fiber construction. The emission spectra in the range of 1.6-2.1 µm vs. the fiber length are presented. The full width at half maximum (FWHM) decreases from 318 to 270 nm for fiber lengths from 2 to 10 m. The presented fiber design is of interest for the development of new construction of optical fibers operating in the eye-safe spectral range.


Chemotherapy of human head and neck cancer xenografts with three clinically active drugs: cis-platinum, bleomycin and methotrexate.
Human head and neck tumours were successfully transplanted in athymic nude mice. In 14 xenograft lines the effect of 1 to 3 clinically active agents could be tested. Maximum tolerated doses were given daily for 3-7 days. Growth delay was estimated in terms of the number of volume doubling times gained by the treatment. Cis-platinum and bleomycin appeared to be effective agents. In all 6 lines in which cis-platinum was examined, growth delay sometimes followed by complete regression was achieved. In 6/7 lines a response to bleomycin was observed. There was wide variation in sensitivity to cis-platinum and bleomycin among the different lines. Methotrexate, effective in 40-60% of patients with head and neck cancer, essentially showed no activity. Methotrexate produced a minimal growth delay in 1/11 lines treated. Two of the patients from whom xenografts were obtained responded to methotrexate treatment. The observed lack of activity of methotrexate against these tumour xenografts indicates that this model has limitations in the screening of new anticancer agents.


Continuing education needs assessment: a brief report.
A needs assessment was conducted to quantify the need for, and the capacity to provide, public health continuing education in San Diego County, California. The study was conducted with the objective of developing public health continuing education courses to be offered by the San Diego State University Graduate School of Public Health (SDSU-GSPH) to address the unmet demands of local practitioners. Survey instruments were developed to separately assess the continuing education needs of public health practitioners and the capacity of SDSU-GSPH faculty to provide courses in San Diego County. The faculty survey was distributed to all full-time SDSU-GSPH faculty. Practitioner surveys were distributed to a systematic sample of divisions and subdivisions in the County of San Diego Department of Health Services. The overall survey response rates were 71% for faculty (22/31) and 40% for practitioners (190/470). Practitioners indicated a strong interest in several of the 64 specific course topics they were asked to rate. Reported faculty interest and capacity to teach continuing education courses appears to be limited. The analysis confirms a need to improve and increase SDSU-GSPH public health continuing education offerings. Further, our results suggest that external instructional resources will be needed to meet this educational demand.


Could Mycobacterium leprae Infection Be an Occupational Disease? A Survey in Healthcare Workers From an Endemic Area in the Amazonian Region.
A serologic survey was conducted to evaluate the prevalence of Mycobacterium leprae infection among healthcare workers and associated factors. Of 280 workers, 26 (9.3%) were positive using immunoglobulin M serology for PGL-I M. leprae antigen. Exposure to leprosy patients in the workplace was significantly associated with seropositivity (P=.044).


Surgical Treatment of Nontuberculous Mycobacterial Pulmonary Disease and a Combination of Nontuberculous Mycobacterium Pulmonary Disease and Pulmonary Tuberculosis.
Nontuberculous mycobacterial pulmonary disease (NTMPD) is an important health system problem worldwide. Surgical treatment experience is limited, and the safety of such operations is still unclarified. Therefore, improving our knowledge of NTMPD is important. In this retrospective observational study, we analyzed the medical records of 2,432 patients operated on from January 2011 through December 2017 in our hospital. There were 20 patients with NTMPD in group 1 and 23 patients with combined NTMPD and pulmonary tuberculosis in group 2. Patients received antibiotic treatment before and after surgery as per Russian Federal clinical guidelines and individual drug susceptibility. Postoperative complications occurred in 2 (10%) cases in group 1 and 4 (17.4%) in group 2, and intraoperative complications occurred in 2 (8.7%) cases in group 2. There was no 30-day mortality in both groups. After 1 year, all 40 traced patients had neither bacterial excretion nor cavities in the lungs. After 3 years and 5 years, the efficacy among 32 and 13 traced patients was 100%, respectively. Modern surgery for small NTMPD forms is safe - helping improve outcomes for ineffective antibiotic treatment. Combined surgery and antibiotic treatment are acceptable in both the short and long term.


Morphological and chemical analyses of Eriocauli Flos sold in Taiwan markets.
Eriocauli Flos (Gujingcao; EF), the dried capitulum with the peduncle of Eriocaulon buergerianum Koern. (Eriocaulaceae), is a Chinese herbal medicine for treating eye diseases and inflammation. However, several species of the Eriocaulon genus are used as substitutes in different areas. To examine the species of EF used in Taiwan and to establish the quality control platform, morphological and chemical analyses have been performed. Ten major compounds, including apigenin (7) and its 7-O-β-D-glucopyranoside (1) and 7-O-(6-O-E-coumaroyl)-β-D-glucopyranoside (6), hispidulin (8) and its 7-O-β-D-glucopyranoside (2) and 7-O-(6-O-E-coumaroyl)-β-D-glucopyranoside (5), jaceosidin (9) and its 7-O-β-D-glucopyranoside (3), and toralactone (10) and its 9-O-β-D-glucopyranosyl(1→6)-β-D-glucopyranoside (4), were isolated and identified from commercially available EF. Morphological investigation showed that two kinds of EFs and most of the EFs sold in Taiwan herbal markets are capitulum without the peduncle. A simultaneous high performance liquid chromatography and ultra performance liquid chromatography analyses of multiple components (1-10) in commercially available EFs, collected from different areas of Taiwan, was conducted. Results showed wide variations in morphology and chemical profiles between capitulum with and without the peduncle. In comparison with an authentic E. buergerianum, we found not only the morphology but also the chemical profile was different from both collected samples. In terms of the morphological examination, the samples without peduncle are closer to the authentic one. To ensure the correct EF materia medica is used in Taiwan so as to guarantee their therapeutic efficacy in clinical practice, further monitoring is necessary.


Associations between SARS-CoV-2 variants and risk of COVID-19 hospitalization among confirmed cases in Washington State: a retrospective cohort study.
The COVID-19 pandemic is dominated by variant viruses; the resulting impact on disease severity remains unclear. Using a retrospective cohort study, we assessed the hospitalization risk following infection with seven SARS-CoV-2 variants. Our study includes individuals with positive SARS-CoV-2 RT-PCR in the Washington Disease Reporting System with available viral genome data, from December 1, 2020 to January 14, 2022. The analysis was restricted to cases with specimens collected through sentinel surveillance. Using a Cox proportional hazards model with mixed effects, we estimated hazard ratios (HR) for hospitalization risk following infection with a variant, adjusting for age, sex, calendar week, and vaccination. 58,848 cases were sequenced through sentinel surveillance, of which 1705 (2.9%) were hospitalized due to COVID-19. Higher hospitalization risk was found for infections with Gamma (HR 3.20, 95%CI 2.40-4.26), Beta (HR 2.85, 95%CI 1.56-5.23), Delta (HR 2.28 95%CI 1.56-3.34) or Alpha (HR 1.64, 95%CI 1.29-2.07) compared to infections with ancestral lineages; Omicron (HR 0.92, 95%CI 0.56-1.52) showed no significant difference in risk. Following Alpha, Gamma, or Delta infection, unvaccinated patients show higher hospitalization risk, while vaccinated patients show no significant difference in risk, both compared to unvaccinated, ancestral lineage cases. Hospitalization risk following Omicron infection is lower with vaccination. Infection with Alpha, Gamma, or Delta results in a higher hospitalization risk, with vaccination attenuating that risk. Our findings support hospital preparedness, vaccination, and genomic surveillance. Hospitalization risk following infection with SARS-CoV-2 variant remains unclear. We find a higher hospitalization risk in cases infected with Alpha, Beta, Gamma, and Delta, but not Omicron, with vaccination lowering risk. Our findings support hospital preparedness, vaccination, and genomic surveillance.


Kinetochores on chromosomes enclosed within the nuclear envelope.
The kinetochore plate which develops after nuclear envelope breakdown in normal cells can be seen to be formed on condensed chromosomes still enclosed in the nuclear envelope in fused multinucleate cells where some nuclei show delayed envelope breakdown caused by nuclear interaction. This suggests that neither nuclear envelope breakdown nor assembly of microtubules is directly related to the formation of the kinetochore plate. Furthermore, it can be clearly observed in these cells that the kinetochores do not have any special association with the nuclear envelope.


Photoinhibition of the CAM succulent Opuntia basilaris growing in Death Valley: evidence from 77K fluorescence and quantum yield.
Diurnal measurements of low temperature (77K) fluorescence at 690 nm (PS II) from north, south, east, and west facing cladode surfaces of Opuntia basilaris in Death Valley, California were made on six occasions during 1985. The absolute levels of F o(instantaneous fluorescence) and F m(maximum fluorescence), as well as the ratio F v/F m(variable fluorescence, F m-F o, over maximum fluorescence), were greater in the north face relative to the other faces. Diurnal decreases in F o, F mand F v/F mwere found concomitant with increases in incident photon flux area density (PFD). F v/F mwas fairly low throughout the year, indicative of photoinhibition, but became somewhat elevated after a spring rain. In early fall the quantum yield of the south face was considerably depressed relative to that of the north face, and corresponding differences were observed in F v/F m. A decrease in PFD during growth of glasshouse plants led to an increase in chlorophyll concentration, F oand F m, but not F v/F m. Although there was some variability in the quantum yield of well watered glasshouse cladodes, a correlation was found between quantum yield and the light and CO2 saturated rate of photosynthesis. When O. basilaris was water stressed under glasshouse conditions, reductions in quantum yield, F m, and F v/F mwere observed. Reductions in F v/F malways indicated a reduced quantum yield, although the converse was not necessarily so in well watered glasshouse plants. The results of this study indicate that O. basilaris is likely to experience photoinhibition throughout much of its life in Death Valley.


Pargyline increases 6-hydroxydopamine levels in the neostriatum of methamphetamine-treated rats.
Neostriatal 6-hydroxydopamine (6-OHDA) was detected in 6 of 13 rats pretreated 2 or 4 hr earlier with methamphetamine (MA; 100 mg/kg, SC) and pargyline (25 mg/kg, IP, 30 min before MA injection). Neostriatal 6-OHDA was detected in 2 of 16 rats treated 2 or 4 hr earlier with MA. These results suggest that pargyline pretreatment may enhance formation of 6-OHDA from endogenous stores of dopamine (DA) following MA administration. Alternatively, these results suggest that pargyline pretreatment may protect endogenously formed 6-OHDA from oxidative deamination by monoamine oxidase. Enhancement of MA-induced neostriatal 6-OHDA levels may be the mechanism by which pargyline enhances the long-term neurotoxic effects of MA upon dopaminergic nerve terminals. These observations support the hypothesis that MA toxicity to DA-containing fibers is caused by the conversion of released DA into 6-OHDA.