Quantitative Monitoring Spatiotemporal Activation of Ras and PKD1 Using Confocal Fluorescent Microscopy.
Receptor activation upon ligand binding induces activation of multiple signaling pathways. To fully understand how these signaling pathways coordinate, it is essential to determine the dynamic nature of the spatiotemporal activation profile of signaling components at the level of single living cells. Here, we outline a detailed methodology for visualizing and quantitatively measuring the spatiotemporal activation of Ras and PKD1 by applying advanced fluorescence imaging techniques, including multichannel, simultaneous imaging and Förster resonance energy transfer (FRET).


Anticipation of affective image modulates visual evoked magnetic fields (VEF).
We investigated the interaction between anticipation of positive and negative affective images and visual evoked magnetic fields (VEF). Participants (n = 13) were presented emotionally positive or negative images under different anticipatory conditions, and their subsequent brain responses were recorded by magnetoencephalography (MEG). In the Affective Cue conditions, the cue stimulus indicated the emotional valence of the image, which followed 2 s later. In the Null Cue conditions, the cue stimulus did not include any information about the valence of the image. In the No Cue conditions, the affective image was suddenly presented, without a cue stimulus. The VEF amplitude for the negative image in the Affective Cue condition was smaller than that of the positive image in the Affective Cue condition and that of the negative image in the Null Cue condition. This result suggests that anticipation of the valence of affective images modulates the processes of the visual cortex.


Delayed seroconversion and rapid onset of lymphoproliferative disease after transmission of human T-cell lymphotropic virus type 1 from a multiorgan donor.
Human T-cell lymphotropic virus type 1 (HTLV-1) screening of blood and organ donors is not mandatory in Germany because of its low prevalence (about 7/100 000). An HTLV-1 transmission event caused by a multiple organ donor was investigated. Validity of diagnostic procedures and HTLV-1 disease association in immunosuppressed organ recipients were analyzed. Two screening immunoassays and an immunoblot (confirmatory assay) were used for detection of HLTV-1/2 antibodies. Proviral DNA was quantified in blood and biopsies of organ recipients by HTLV-1 real-time polymerase chain reaction (PCR). Proviral HTLV-1-DNA was detected in all blood samples of 3 organ recipients (1-100 copies/10(2) cells), but seroconversion was delayed for up to 2 years in screening assays and >6 years in the confirmatory assay. In 2 of 3 organ recipients, a cutaneous T-cell lymphoma was diagnosed 2 and 3 years after infection, respectively. Proviral HTLV-1 DNA concentration was almost 100 copies/10(2) cells in cutaneous lymphoma biopsies whereas in biopsies of other tissues ≤3.0 copies/10(2) cells were found. The third organ recipient did not suffer from lymphoma, but detailed clinical data on this patient were not available to us. Biopsy results support an etiological role for HTLV-1 in these cases of primary cutaneous T-cell lymphoma after solid organ transplant. HTLV-1-associated lymphoma can arise quickly in immunocompromised transplant recipients. The diagnosis of potentially HTLV-1-associated disease in organ recipients may require PCR because of delayed seroconversion.


Efficient self-organizing multilayer neural network for nonlinear system modeling.
It has been shown extensively that the dynamic behaviors of a neural system are strongly influenced by the network architecture and learning process. To establish an artificial neural network (ANN) with self-organizing architecture and suitable learning algorithm for nonlinear system modeling, an automatic axon-neural network (AANN) is investigated in the following respects. First, the network architecture is constructed automatically to change both the number of hidden neurons and topologies of the neural network during the training process. The approach introduced in adaptive connecting-and-pruning algorithm (ACP) is a type of mixed mode operation, which is equivalent to pruning or adding the connecting of the neurons, as well as inserting some required neurons directly. Secondly, the weights are adjusted, using a feedforward computation (FC) to obtain the information for the gradient during learning computation. Unlike most of the previous studies, AANN is able to self-organize the architecture and weights, and to improve the network performances. Also, the proposed AANN has been tested on a number of benchmark problems, ranging from nonlinear function approximating to nonlinear systems modeling. The experimental results show that AANN can have better performances than that of some existing neural networks.


Radiotherapy for cancers of the oesophagus, cardia and stomach.
We present the updated recommendations of the French society for radiation oncology on radiotherapy of oesophageal cancer. Oesophageal cancer still remains a malignant tumour with a poor prognosis. Surgery remains the standard treatment for localized cancers, regardless of histology. For locally advanced stages, surgery remains a standard for adenocarcinomas after neoadjuvant treatment with chemotherapy or chemoradiotherapy. However, it is a therapeutic option after initial chemoradiotherapy for stage III squamous cell carcinomas, given the increased morbidity and mortality with a multimodal treatment, which results in an equivalent overall survival with or without surgery. Preoperative or exclusive chemoradiotherapy should be delivered according to validated regimens with an effective total dose (50Gy), if surgery is not planned or if the tumour is deemed resectable before chemoradiotherapy. Intensity-modulated radiotherapy significantly reduces irradiation of the lungs and heart and may reduce the morbidity of this treatment, especially in combination with surgery. In case of exclusive chemoradiotherapy, dose escalation beyond 50Gy is not currently recommended. Some technical considerations still remain questionable, such as the place of prophylactic lymph node irradiation, adaptive radiotherapy, evaluation of response during and after chemoradiotherapy and the value of proton therapy.


Seasonal variations in hair zinc concentrations in Canadian and African children.
Proximal occipital scalp hair samples were collected during three survey periods from 30 omniverous Canadian boys aged 75.6 +/- 10.4 months (mean +/- SD) and 66 rural Malawian children (37 M; 29 F) aged 62 +/- 10 months (mean +/- SD) consuming predominantly plant-based diets. Hair samples were washed and then analyzed for zinc and copper using instrumental neutron activation analysis. Selected anthropometric measurements were also taken and dietary intakes calculated from weighed dietary records. A marked seasonal variation in hair zinc concentrations occurred in both groups; levels were highest in the season with shortest daylight hours (i.e. December/January in Canada vs July/August in Malawi). A similar trend was not observed for hair copper concentrations. Changes in hair zinc were not associated with changes in growth indices, age, socio-economic group, method of infant feeding, in either group, but were related to intakes of protein (per 1000 kcal) (p = 0.02, r = -0.36), zinc (mg day-1) (p = 0.01, r = -0.39), and dietary fiber (g day-1) (p = 0.04, r = 0.28), in the Malawian children during the second survey period. The seasonal effect on hair zinc concentrations must always be considered when interpreting hair zinc concentrations in children; its etiology remains unclear.


[Double-blind study on the influence of difenoxin hydrochloride on intestinal passage time in healthy volunteers (author's transl)].
The effect on the intestinal passage of 3 X 1 mg difenoxin hydrochloride (Lyspafen) was tested in a randomised double-blind study; the trial included 10 healthy volunteers and lasted one week. The time until elimination of both the first and twentieth of the total of 30 applied radiopaque stool markers was measured. The total weekly weight and number of stools served as further test criteria. The treatment with difenoxin and that without placebo were compared with the standard treatment using placebo. The extensive statistical evaluation failed to reveal significant differences in the criteria observed.


Development and validation of a stability indicating UPLC method for determination of ticlopidine hydrochloride in its tablet formulation.
The objective of the current study was the development of a simple, precise and accurate isocratic reversed-phase stability indicating Ultra Performance Liquid Chromatography [UPLC] assay method and validated for determination of ticlopidine hydrochloride in solid pharmaceutical dosage forms. Isocratic separation was achieved on a Zorbax SB-C18 (50 mm × 4.6 mm, 1.8 μm) column using mobile phase of methanol-0.01 M ammonium acetate buffer, pH 5.0 (80:20, v/v) at a flow rate of 0.8 ml min(-1), the injection volume was 4.0 μl and the detection was carried out at 235 nm by using photo-diode array detector. The drug was subjected to oxidation, hydrolysis, photolysis and heat to apply stress condition. The method was validated for specificity, linearity, precision, accuracy, robustness and solution stability. The method was linear in the drug concentration range of 62.5-375 μg ml(-1) with a correlation coefficient of 0.9999. The precision (relative standard deviation - RSD) of six samples was 1.31% for repeatability and the intermediate precision [RSD] among six-sample preparation was 0.77%. The accuracy (recovery) was between 98.80% and 101.50%. Degradation products produced as a result of stress studies did not interfere with detection of ticlopidine hydrochloride and the assay can thus be considered stability indicating.


Immunostimulant effect of a mixed herbal extract on infectious bursal disease virus (IBDV) vaccinated chickens in the context of a co-infection model of avian influenza virus H9N2 and IBDV.
This study was conducted to assess the comparative effects of a mixed herbal extract (MHE) containing Ocimum sanctum, Withania somnifera, Emblica officinalis, Tinospora cordifolia, Mangifera indica, and Asphaltum (shilajit) on infectious bursal disease virus (IBDV)-vaccinated (VAC) chickens infected with IBDV and avian influenza virus (AIV) H9N2. The experiment included three groups (G1-G3): G1, the negative control group; G2, the VAC + challenged (Ch) group; and G3, the VAC + Ch + MHE group. MHE was orally administered continuously for 5 weeks post-vaccination (PV) with IBDV at 12 days of age, and the chicks were simultaneously challenged with virulent IBDV (intraocularly) and AIV H9N2 (intranasally) at 21 days PV. Blood and tissue samples as well as tracheal and cloacal swabs were gathered at different times PV and post-challenge. Immunological and haematological parameters, histopathological lesions, relative organ weights and final live weights revealed significant differences (P ≤ 0.05) between G2 and G3 groups. Furthermore, in the G3 group, the protection rates, ELISA and HI titers and CD4+/CD8+ ratio were significantly increased, whereas viral shedding titers and the heterophil/lymphocyte ratio were decreased. In conclusion, the oral administration of the mixed herbal extract for 5 weeks can stimulate the immune response to IBDV vaccination and relieves the pathogenicity of an AIV H9N2 and IBDV co-infection in chickens.


Assessing Knowledge of Pharmacokinetics in an Integrated Medical Curriculum.
Pharmacokinetics is the branch of pharmacology that describes how the body processes drugs. As most physicians will prescribe drugs during their career, knowledge of pharmacokinetics is indispensable for medical students. Students, however, experience pharmacokinetics as difficult, probably due to its abstract and mathematical nature. In many medical curricula, pharmacokinetic topics are taught and examined as a part of integrated medical courses. As pharmacokinetics is a relatively small subject, unit examinations contain only few questions on the topic. The combination of a difficult subject and a few questions has raised concerns that students could perform poorly in pharmacokinetics and still pass the examinations and, hence, end up with insufficient knowledge of pharmacokinetics. In this study, we investigate this issue by contrasting students' performance on pharmacokinetics questions with their performance on the rest of the examinations (all non-pharmacokinetics questions lumped together). The results expressed as pass-fail scores showed that students failed more often on the pharmacokinetics part of the test than on the other questions, in two consecutive academic years. Despite the suboptimal knowledge in pharmacokinetics, students can still acquire their bachelor's degree. These results show that poor knowledge in pharmacokinetics could be a side effect of curricular integration. Attention should therefore be paid to provide insight into one's own performance in individual disciplines. This would avoid knowledge deficiency and incompetence in the future.


Effect of endosulfan pretreatment on organ weights and on pentobarbital hypnosis in rats.
The effects of endosulfan on the weights of the liver, adrenal and ovary, on pentobarbital blood and brain levels and on sleeping time (ST) have been investigated in female rats after daily oral doses of 0, 1.0, 2.5 and 5.0 mg/kg for a period of 7 or 15 days. No significant change in body weight was observed. With higher doses (2.5--5.0 mg/kg) the liver weight was significantly increased, but ovary and adrenal weights did not increase. Endosulfan treatment shortened sleeping time, while induction time was significantly increased. The concentration of pentobarbital in the blood and brain of rats after 30 min and upon awakening indicated that there was a significant decrease at 30 min. No change at awakening was observed in endosulfan-treated rats as compared to controls. It is suggested tha endosulfan may shorten the duration of pentobarbital-induced sleep, perhaps by induction of hepatic microsomal enzyme activity.


Antioxidant activity of phenylethanoid glycosides from Brandisia hancei.
Brandisia hancei is a medicinal herb in China. The ethanol extract of this plant and four phenylethanoid glycosides, acteoside (1), 2'-acetylacteoside (2), poliumoside (3) and brandioside (4), isolated from it were shown to have inhibitory effects on free radical-induced hemolysis of red blood cells and free radical scavenging activities in vitro. Brandioside (4) and poliumoside (3) showed stronger antioxidant effect than acteoside (1), 2'-acetylacteoside (2) and trolox.


Evolutionary and functional relationships between the basic and acidic beta-crystallins.
beta-Crystallins are complex oligomers composed of many related subunits. In order to understand their interactions we have built molecular models of several bovine beta-crystallins, based on their sequence similarity to the well-defined gamma-II crystallin structure, using interactive computer graphics techniques. Their common origin with gamma-crystallin is displayed in both the retention of four-fold sequence repeats of critical residues involved with stabilizing a folded beta-hairpin and the conservation of core-filling hydrophobic side-chains. The beta-crystallins have been built as bilobal molecules with each domain composed of two 'Greek key' motifs which associate about an approximate two-fold axis to form beta-sheets. The beta-crystallin sequences have previously been shown to comprise two families, the basic and acidic subunits, which have extensions of sequence. The three-dimensional models show how the two families appear to stabilize the folded beta-hairpin in the N- and C-terminal domains in ways which suggest that they have diverged from a common ancestor in different ways. Acidic beta-crystallins, like gamma-crystallins, have a regular array of charges on their N-terminal domain which has been interrupted in basic beta-crystallins by hydrophobic residues which may be related to the presence of a C-terminal extension. beta-Crystallins are more highly charged than gamma-crystallins although their charge density is higher in certain regions of the N-terminal domain, particularly in beta B1-crystallin. beta-crystallins also differ from gamma-crystallins in the virtual absence of core-filling sulphydryl groups whereas they have numerous sulphur-containing side-chains together with tryptophan and histidine rings protruding from the globular domains, particularly in the acidic subunits. The burial of these residues in subunit contacts is consistent with their spectroscopic and electrostatic properties. Protein subunit aggregation commonly occurs through hydrophobic interaction or beta-sheet extension. Analysis of the subunit surfaces has identified an N-terminal hydrophobic region common to beta B1 and beta B2 whereas a C-terminal hydrophobic loop region is common to beta B1 and beta A1 and may be correlated with their association properties. It is suggested that the polar C-terminal domain of beta B2 contributes towards the solubility of higher aggregates by interactions involving beta-sheet structure.


A generic method for intact and subunit level characterization of mAb charge variants by native mass spectrometry.
Monoclonal antibodies (mAbs) are heterogeneous macromolecules that display a complex isoform profile as a result of the large series of modifications they can undergo. Product-related charge variants that are associated with a loss of biological activity or affected half-life and immunogenicity are especially important. Consequently, they are often considered critical quality attributes such that acceptance criteria and controls should be established. The characterization of mAbs charge variants has long been a time and resource consuming task. Recent successes in the use of salt mediated pH gradient ion exchange chromatography with volatile mobile phases have shown there to be significant promise in using online mass spectrometric (MS) detection to facilitate peak detection. In this study, a newly developed 3 µm non-porous cation exchange column technology was investigated for its capability to be hyphenated to MS for the purpose of characterizing mAb charge variants. A 2 mm ID format was selected for the ease of configuring it to classical MS ESI ion sources. A monoclonal antibody reference material from NIST (RM 8671; NISTmAb) was used in its intact and IdeS/IgdE-digested forms to test for column performance and MS sensitivity. Furthermore, three different mAbs with highly basic isoelectric points (pI) were analyzed in their native and proteolyzed forms to demonstrate the straightforward application of the developed technique even with mAbs having strong retention on cation exchange media. The MS detection of low-abundance charge variant species (<0.1%) demonstrated there to be acceptable sensitivity and dynamic range even from routine analyses. The capability of the column to separate different mAbs having high basic pI was demonstrated, and it was found that slight adjustment of ammonium acetate concentration in the eluent can be a convenient way to rapidly optimize a separation if necessary. Linearity was shown to exist between protein mass loads of 2.5 and 50 µg while an optimal balance between chromatographic resolution and MS sensitivity was observed between 5 and 10 µg. Excellent run-to-run and column-to-column repeatability was achieved in terms of retention times, resolution and recovery.


The Comprehensive Complication Index for Advanced Monitoring of Complications Following Endoscopic Surgery of the Lower Urinary Tract.
Purpose: To evaluate the Comprehensive Complication Index (CCI) for reporting complications in lower urinary tract transurethral procedures and compare it with the Clavien-Dindo classification (CDC). Materials and Methods: A total of 450 consecutive patients were included into the analyses [150 each of transurethral resection of bladder tumors (TURBT), transurethral resection of the prostate (TURP), and transurethral enucleation of the prostate using Tm:YAG, (ThuLEP)]. Complications were assessed according to the modified CDC. The CCI was calculated using a freely accessible online tool. Descriptive statistics and correlation analyses were applied to quantify operational differences and length of stay (LOS) between CDC and CCI. Sample size calculations for hypothetical clinical trials were contrasted for CDC and CCI application. Results: Overall n = 150 patients with complications (33.3%) within the first 60 days after operation were identified. Of these, n = 125 (83.4%) were minor complications up to CDC grade IIIa. Of patients with complications, n = 57 patients (12.6%) experienced more than one complication. Here, the cumulative CCI led to an upgrade of at least one CDC grade in 33 patients. Hence, in 22.0% of cases, the highest CDC grade underestimated the degree of complications. CCI showed higher correlation with LOS compared with CDC (all r > 0.2, all p-values ≤0.0001). Using CCI instead of CDC for sample calculation resulted in a strong reduction of the required number of patients for all three interventions (percentage of patient number decrease: -93.2% for TURBT, -71.8% for TURP, and -81.1% for ThuLEP). Conclusion: The CCI gives a more precise interpretation of the postinterventional complications of TURBT, TURP, and ThuLEP. CCI application may reduce the required sample size for clinical trials and will relieve their recruitment in the future.


Down-regulation of natural killer cell activity by autologous polymorphonuclear leucocytes. Role of indomethacin.
It has been recently reported that neutrophils are involved in the regulation of NK cell activity. However, the mechanism of such regulation is unclear. The present study was designed to investigate the mechanisms involved in the regulation of NK cytotoxicity by human neutrophils. The role of indomethacin, an anti-inflammatory drug, in this interaction was studied. NK cells were purified from peripheral blood obtained from normal individuals. NK cell cytotoxicity was tested on K 562 cell line by Cr release assay. Autologous neutrophils obtained from peripheral blood were stimulated by opsonized zymosan either in the presence or absence of indomethacin. The role of neutrophil supernatant containing oxygen radicals and prostaglandins on NK cytotoxicity was examined. It was shown that supernatants from stimulated neutrophils significantly inhibited (P less than 0.05) the autologous NK cell cytotoxicity. The presence of indomethacin in the in vitro reaction mixture, or given orally to donors, partially or completely abolished the inhibitory effect of neutrophil supernatant. Indomethacin inhibited prostaglandin E2 release, and luminol-enhanced, myeloperoxidase-mediated chemiluminescence of activated PMN. Diafiltration of neutrophil supernatant showed that the inhibitory activity was present in the fraction containing molecules lower than 5,000 daltons. In conclusion, our findings indicate that down-regulation of NK cytotoxicity is mediated by prostaglandins produced by stimulated neutrophils and possibly by oxygen radicals.


Ultrasound-facilitated epidurals and spinals in obstetrics.
Regional anesthesia is currently the gold standard of practice for pain control in obstetrics. Failures and complications of regional anesthesia can be related to many causes, one of the most important being the blind nature of such techniques. The practice of epidurals and spinals relies primarily on the palpation of anatomic landmarks that are not always easy to find. Ultrasound has recently been introduced into clinical anesthesia to facilitate lumbar spinals and epidurals. The use of preprocedure ultrasound imaging or, eventually, real-time ultrasound guidance should improve not only clinical practice, but also teaching. This article describes the techniques, challenges, and benefits related to the use of ultrasound in guiding lumbar spinals and epidurals.


Giardia lamblia: the effects of extracts and fractions from Mentha x piperita Lin. (Lamiaceae) on trophozoites.
Giardia lamblia is a parasite that causes giardiasis in humans and other mammals. The common treatment includes different classes of drugs, which were described to produce unpleasant side effects. Mentha x piperita, popularly known as peppermint, is a plant that is frequently used in the popular medicine to treat gastrointestinal symptoms. We examined the effects of crude extracts and fractions from peppermint against G. lamblia (ATCC 30888) on the basis of trophozoite growth, morphology and adherence studies. The methanolic, dichloromethane and hexanic extracts presented IC(50) values of 0.8, 2.5 and 9.0microg/ml after 48h of incubation, respectively. The aqueous extract showed no effect against the trophozoites with an IC(50)>100microg/ml. The aqueous fraction presented a moderate activity with an IC(50) of 45.5microg/ml. The dichloromethane fraction showed the best antigiardial activity, with an IC(50) of 0.75microg/ml after 48h of incubation. The morphological and adhesion assays showed that this fraction caused several alterations on plasma membrane surface of the parasite and inhibited the adhesion of G. lamblia trophozoites. Cytotoxic assays showed that Mentha x piperita presented no toxic effects on the intestinal cell line IEC-6. Our results demonstrated antigiardial activity of Mentha x piperita, indicating its potential value as therapeutic agent against G. lamblia infections.


Rapid-onset obesity with hypothalamic dysfunction, hypoventilation, and autonomic dysregulation (ROHHAD): a collaborative review of the current understanding.
To provide an overview of the discovery, presentation, and management of Rapid-onset Obesity with Hypothalamic dysfunction, Hypoventilation, and Autonomic Dysregulation (ROHHAD). To discuss a search for causative etiology spanning multiple disciplines and continents. The literature (1965-2022) on the diagnosis, management, pathophysiology, and potential etiology of ROHHAD was methodically reviewed. The experience of several academic centers with expertise in ROHHAD is presented, along with a detailed discussion of scientific discovery in the search for a cause. ROHHAD is an ultra-rare syndrome with fewer than 200 known cases. Although variations occur, the acronym ROHHAD is intended to alert physicians to the usual sequence or unfolding of the phenotypic presentation, including the full phenotype. Nearly 60 years after its first description, more is known about the pathophysiology of ROHHAD, but the etiology remains enigmatic. The search for a genetic mutation common to patients with ROHHAD has not, to date, demonstrated a disease-defining gene. Similarly, a search for the autoimmune basis of ROHHAD has not resulted in a definitive answer. This review summarizes current knowledge and potential future directions. ROHHAD is a poorly understood, complex, and potentially devastating disorder. The search for its cause intertwines with the search for causes of obesity and autonomic dysregulation. The care for the patient with ROHHAD necessitates collaborative international efforts to advance our knowledge and, thereby, treatment, to decrease the disease burden and eventually to stop, and/or reverse the unfolding of the phenotype.


Identification and characterization of an alternatively spliced variant of the MHC class I-related porcine neonatal Fc receptor for IgG.
The neonatal Fc receptor for immunoglobulin G (IgG) (FcRn) functions to transport maternal IgG to the fetal/neonatal animals and protects IgG from catabolism. The present study identified two pFcRn cDNAs (1.071 and 0.795kb) from intestinal epithelial cells. The corresponding mRNA transcripts were detected in porcine kidney cell line LLC-PK1, peripheral blood mononuclear cells, and porcine tissues by reverse transcription-PCR (RT-PCR) and Northern blot. Sequence analysis showed that the 1.071kb cDNA encodes the full-length pFcRn (pFcRn-L), whereas the 0.795kb cDNA codes for a truncated pFcRn (pFcRn-S) with deletion of 92 amino acids matching the alpha2 domain of pFcRn-L. pFcRn-L was constitutively expressed by epithelial cells; however, pFcRn-S was not detectable in porcine tissues and cell lines although its transcript was abundant. Despite the lack of native pFcRn-S, pFcRn-S was readily detected in transfected cells. Recombinant pFcRn-L was confirmed to bind IgG at pH 6.0, but not at pH 7.5; however, pFcRn-S failed to bind IgG at both pH 5.0-6.0 and 7.5. pFcRn-L was expressed on the cell surface and mainly localized in early endosomes. In contrast, pFcRn-S was absent from cell surface and primarily localized in the lysosome and pFcRn-S trafficking to lysosomes was independent of beta(2)-microglobulin (beta(2)m). The accumulation of pFcRn-S in the lysosome may explain the absence of native pFcRn-S protein expression. In addition, the trafficking of pFcRn-S to the lysosomal compartment suggests that in addition to sorting signals in its cytoplasmic tail, the FcRn structural integrity may be important for proper intracellular trafficking and function.


Phosphoproteomics of Acute Cell Stressors Targeting Exercise Signaling Networks Reveal Drug Interactions Regulating Protein Secretion.
Exercise engages signaling networks to control the release of circulating factors beneficial to health. However, the nature of these networks remains undefined. Using high-throughput phosphoproteomics, we quantify 20,249 phosphorylation sites in skeletal muscle-like myotube cells and monitor their responses to a panel of cell stressors targeting aspects of exercise signaling in vivo. Integrating these in-depth phosphoproteomes with the phosphoproteome of acute aerobic exercise in human skeletal muscle suggests that co-administration of β-adrenergic and calcium agonists would activate complementary signaling relevant to this exercise context. The phosphoproteome of cells treated with this combination reveals a surprising divergence in signaling from the individual treatments. Remarkably, only the combination treatment promotes multisite phosphorylation of SERBP1, a regulator of Serpine1 mRNA stability, a pro-fibrotic secreted protein. Secretome analysis reveals that the combined treatments decrease secretion of SERPINE1 and other deleterious factors. This study provides a framework for dissecting phosphorylation-based signaling relevant to acute exercise.


Astrocytic phosphatase and tensin homolog deleted on chromosome 10 regulates neuropathic pain by facilitating 3-hydroxy-3-methylglutaryl-CoA reductase-dependent cholesterol biosynthesis.
Recent studies have noted the role of the phosphatase and tensin homolog deleted on chromosome 10 (PTEN) in developing neuropathic pain, but the underlying mechanisms are obscure. We found that PTEN was mainly expressed in astrocytes in the rat spinal cord and dramatically downregulated after chronic constriction injury (CCI). Intrathecal injection of a PTEN inhibitor induced pain-related behaviors in naive rats. By contrast, administration of a PTEN protector effectively mitigated CCI-induced pain. Adeno-associated virus-mediated overexpression of astrocytic PTEN in the spinal cord reduced glial activation and neuroinflammation and subsequently alleviated pain-related behaviors. Importantly, astrocyte-specific PTEN knockout ( Pten conditional knockout , Pten CKO) mice showed nociceptive sensitization and glial activation. Proteomic analysis revealed that PTEN overexpression upregulated at least 7 enzymes in the cholesterol biosynthesis pathway and the total cholesterol level in the spinal cord of CCI rats. Furthermore, PTEN directly interacted with enzymes, including 3-hydroxy-3-methylglutaryl-CoA reductase, in the cholesterol biosynthesis pathway. Astrocytic 3-hydroxy-3-methylglutaryl-CoA reductase overexpression alleviated both CCI-induced pain and mechanical allodynia in Pten CKO mice. Finally, cholesterol replenishment attenuated CCI-induced pain and suppressed spinal glial activation. Taken together, these findings imply that spinal astrocytic PTEN plays a beneficial role in CCI-induced pain by regulating cholesterol biosynthesis, and an increased level of PTEN may accelerate cholesterol biosynthesis and reduce glial activation, thereby alleviating neuropathic pain. Recovery of PTEN or cholesterol might be an effective therapeutic strategy for neuropathic pain.


SPECT/CT imaging for tracking subendothelial retention of electronegative low-density lipoprotein in vivo.
The fifth subfraction of low-density lipoprotein (L5 LDL) can be separated from human LDL using fast-protein liquid chromatography with an anion exchange column. L5 LDL induces vascular endothelial injury both in vitro and in vivo through the lectin-like oxidized LDL receptor-1 (LOX-1). However, no in vivo evidence shows the tendency of L5 LDL deposition on vascular endothelium and links to dysfunction. This study aimed to investigate L5 LDL retention in vivo using SPECT/CT imaging, with Iodine-131 (131I)-labeled and injected into six-month-old apolipoprotein E knockout (apoE-/-) mice through tail veins. Besides, we examined the biodistribution of L5 LDL in tissues and analyzed the intracellular trafficking in human aortic endothelial cells (HAoECs) by confocal microscopy. The impacts of L5 LDL on HAoECs were analyzed using electron microscopy for mitochondrial morphology and western blotting for signaling. Results showed 131I-labeled-L5 was preferentially deposited in the heart and vessels compared to L1 LDL. Furthermore, L5 LDL was co-localized with the mitochondria and associated with mitofusin (MFN1/2) and optic atrophy protein 1 (OPA1) downregulation, leading to mitochondrial fission. In summary, L5 LDL exhibits a propensity for subendothelial retention, thereby promoting endothelial dysfunction and the formation of atherosclerotic lesions.


Treatment of testicular cancer: influence on pituitary-gonadal axis and sexual function.
To investigate the influence of treatment for testicular cancer on the pituitary-gonadal axis and sexual function in long-time survivors after unilateral orchiectomy. Blood was drawn from patients treated for testicular cancer during routine oncologic follow-up for measurement of luteinizing hormone, follicle-stimulating hormone, sexual hormone-binding globulin, testosterone, and bioavailable testosterone. Sexual function was evaluated using the International Index of Erectile Function 15-item (IIEF-15) questionnaire. Patients were grouped according to treatment: group 1 followed a surveillance strategy, group 2 received two cycles of carboplatin monotherapy, and group 3 underwent cisplatin, etoposide, and bleomycin chemotherapy. No statistically significant difference was found in the serum hormonal levels among the three groups, and all hormonal levels were within the 95% confidence range, except for follicle-stimulating hormone. The median serum testosterone level was 3.5 ng/mL in group 1, 3.9 ng/mL in group 2, and 4.2 ng/mL in group 3. In group 1, the median IIEF-15 score was 64.0, and the median Erectile Function (EF) domain score was 28. The median scores in groups 2 and 3 were 62.5 for IIEF-15 and 27.5 for EF and 65.0 for IIEF-15 and 30.0 for EF, respectively. No correlation was found between testosterone level and IIEF-15 or EF score. None of the treatments investigated had a significant influence on the serum hormonal levels in long-time survivors of testicular cancer. Patients undergoing chemotherapy have no greater risk of developing a hormonal disorder than those following a surveillance strategy, and therapy for testicular cancer is not a risk factor for erectile dysfunction.


3D dense connectivity network with atrous convolutional feature pyramid for brain tumor segmentation in magnetic resonance imaging of human heads.
The existing deep convolutional neural networks (DCNNs) based methods have achieved significant progress regarding automatic glioma segmentation in magnetic resonance imaging (MRI) data. However, there are two main problems affecting the performance of traditional DCNNs constructed by simply stacking convolutional layers, namely, exploding/vanishing gradients and limitations to the feature computations. To address these challenges, we propose a novel framework to automatically segment brain tumors. First, a three-dimensional (3D) dense connectivity architecture is used to build the backbone for feature reuse. Second, we design a new feature pyramid module using 3D atrous convolutional layers and add this module to the end of the backbone to fuse multiscale contexts. Finally, a 3D deep supervision mechanism is equipped with the network to promote training. On the multimodal brain tumor image segmentation benchmark (BRATS) datasets, our method achieves Dice similarity coefficient values of 0.87, 0.72, and 0.70 on the BRATS 2013 Challenge, 0.84, 0.70, and 0.61 on the BRATS 2013 LeaderBoard, 0.83, 0.70, and 0.62 on the BRATS 2015 Testing, 0.8642, 0.7738, and 0.7525 on the BRATS 2018 Validation in terms of whole tumors, tumor cores, and enhancing cores, respectively. Compared to the published state-of-the-art methods, the proposed method achieves promising accuracy and fast processing, demonstrating good potential for clinical medicine.


Effect of long retention time in the settler on phosphorus removal from communal wastewater.
The objective of the paper is to illustrate the solution of a chronic problem occurring in a wastewater treatment facility with a capacity of 12,000 inhabitant equivalent. A drastic phosphorus concentration increase (two or three times higher than the Hungarian limit of 1 mg/l) was observed in the wastewater treatment facility for a long time (12-24 h) with changing time periods indicating malfunctioning in the operation of the facility. Computer-aided simulation technique was used to develop a solution for the treatment of the problem using a software developed by the Department of Environmental and Chemical Engineering, University of Veszprém, Hungary. The simulation studies show that if the nightly inflow is less than 200 m3/h at least for two hours, the system doesn't then receive enough fresh nutrients which can cause a deficiency in the nutrient uptake of the PAOs in the anaerobic zones. This can result in the fact that the PAOs accumulate less phosphorus into the aerobic zones. Long retention time (10 h) is the reason for the problem, namely--under special conditions--the phosphorus in the sludge of the settling tank of the wastewater treatment plant. The problem was caused by phosphorus dissolution from the sludge of the settling tank during the undesirably long retention time.


Restless legs syndrome in adolescents: relationship with sleep quality, cardiorespiratory fitness and body fat.
The aim of the current study was to investigate the relationship between restless legs syndrome (RLS) and cardiorespiratory fitness, body composition and sleep quality in a sample of adolescents. One hundred fifty seven volunteer adolescents (16.6 ± 0.7 yrs) participated in the study. Sleep quality was assessed by the Pittsburg sleep quality index. Cardiorespiratory fitness was assessed by the 20 m shuttle run test and body composition by bioelectrical impedance analysis. The prevalence of RLS was 5.1%. The adolescents with RLS were found to exhibit significantly higher body fat levels (p=0.019) and poorer sleep quality score (p=0.000) compared with their free-RLS counterparts. Adolescents with RLS are subjects of higher body fat and impaired sleep quality compared with adolescents without RLS. Early diagnosis and appropriate management of RLS is essential in the adolescents.


25KDa branched polyethylenimine increases interferon-γ production in natural killer cells via improving translation efficiency.
Ex vivo cultivation is a promising strategy for increasing the number of NK cells and enhancing their antitumor activity prior to clinical application. Recent studies show that stimulation with 25KDa branched polyethylenimine (25KbPEI) generates NK cells with enhanced antitumor activity. To better understand how 25KbPEI primes NK cells, we explored the mechanism underlying increase in production of IFN-γ. Chemical priming was performed on NK-92MI cells by incubating them with 5 μg/ml of 25KbPEI. The production of IFN-γ was evaluated by RT-qPCR, ELISA, and Flow cytometry. By evaluating the effect of pharmacological inhibition of ERK/mTOR-eIF4E signaling pathways on IFN-γ translation, the function of these signaling pathways in IFN-γ translation was examined. To comprehend the level of 25KbPEI activity on immune-related components in NK cells, RNA sequencing and proteomics analyses were conducted. 25KbPEI enhances the production of IFN-γ by NK cells without transcriptional activation. Activation of ERK and mTOR signaling pathways was found to be associated with 25KbPEI-mediated calcium influx in NK cells. The activation of ERK/mTOR signaling was linked to the phosphorylation of 4E-BP1, which resulted in the activation of translation initiation complex and subsequent IFN-γ translation. Analysis of RNA sequencing and proteomics data revealed that the activity of 25KbPEI to improve translation efficiency in NK cells could be extended to additional immune-related molecules. This study provides substantial insight into the process by which 25KbPEI primes NK cells. Our data demonstrated that the 25KbPEI mediated activation of ERK/mTOR signaling and subsequent stimulation of eIF4E is the primary mechanism by which the chemical stimulates translation of IFN-γ in NK cells. Video abstract.


Salade malade: malignant ventricular arrhythmias due to an accidental intoxication with Aconitum napellus.
Intoxication with Aconitum napellus is rare in our regions. Aconite alkaloids can cause ventricular arrhythmia by a prolonged activation of sodium channels. Because the margin of safety is low between the analgesic and toxic dose, intoxication is not rare when Aconite is used in herbal medicine. We present a case in which a 39-year-old male was accidentally intoxicated with Aconite. Even though no antidote or adequate therapy is available he was successfully resuscitated. (Neth Heart J 2008;16:96-9.).


Hypoconnectivity within the cingulo-opercular network in patients with mild cognitive impairment in Chinese communities.
At rest, the brain's higher cognitive systems engage in correlated activity patterns, forming networks. With mild cognitive impairment (MCI), it is essential to understand how functional connectivity within and between resting-state networks changes. This study used resting-state functional connectivity to identify significant differences within and between the cingulo-opercular network (CON) and default mode network (DMN). We assessed cognitive function in patients using the Chinese version of the Alzheimer's disease assessment scale-Cognitive subscale (ADAS-Cog). A group of MCI subjects (ages 60-83 years, n = 45) was compared to age-matched healthy controls (n = 70). Resting-state functional connectivity was used to determine functional connectivity strength within and between the CON and DMN. Compared to healthy controls, the MCI group showed significantly lower functional connectivity within the CON (F = 10.76, df = 1, p = 0.001, FDR adjusted p = 0.003). Additionally, the MCI group displayed no distinct differences in functional connectivity within DMN (F = 0.162, df = 1, p = 0.688, FDR adjusted p = 0.688) and between CON and DMN (F = 2.270, df = 1, p = 0.135, FDR adjusted p = 0.262). Moreover, we found no correlation between ADAS-Cog and within- or between-connectivity metrics among subjects with MCI. Our findings indicate that specific patterns of hypoconnectivity within CON circuitry may characterize MCI relative to healthy controls. This work improves our understanding of network dysfunction underlying MCI and could inform more targeted treatment.


The effect of dextransulfate 500 on the pathogenesis of herpes simplex virus infections in weanling mice.
Intraperitoneal (i.p.) injection of Dextran Sulfate (D.S.) 500 during a limited period of time influences the course of herpes simplex-virus-infections. D.S.500 was found to reduce the resistance of mice for some herpes simplex-virus strains (Len, L3--2s, Haase) if given between 16 hours and 2 hours after i.p. infection. The decrease of resistance could be correlated with an increase of the virus content of liver, spleen, brain and spinal cord. Injection of herpes simplex-virus-specific immune serum counteracted the effect of D.S.500 on the course of infections. Conversely, D.S.500 increased the resistance of mice to another group of herpes simplex-viruses (strains D-316, Thea, DD), if given 3 to 8 hours before infection. These effects are ascribed to a special interaction of D.S.500 with macrophages and probably other virus-susceptible cells of the peritoneal cavity and elsewhere with a resulting counteraction to the virus infection.


Evaluating Acalabrutinib In The Treatment Of Mantle Cell Lymphoma: Design, Development, And Place In Therapy.
Mantle cell lymphoma (MCL) is an incurable intermediate-grade lymphoma representing 5-6% of non-Hodgkin's lymphomas diagnosed in the United States. The introduction of inhibitors of Bruton's tyrosine kinase (BTK) into targeted therapy for MCL has significantly improved outcomes in patients with relapsed/refractory (R/R) disease. Since the initial approval of the first-generation inhibitor, ibrutinib, several second-generation inhibitors have been explored. Acalabrutinib, a second-generation BTK inhibitor, has demonstrated impressive efficacy in clinical trials along with a safety profile that thus far appears improved compared to ibrutinib. The results of a Phase II trial in patients with R/R MCL led to the approval of acalabrutinib in this patient population while fueling further exploration of acalabrutinib in several ongoing clinical trials.


Strand-preferred base editing of organellar and nuclear genomes using CyDENT.
Transcription-activator-like effector (TALE)-based tools for base editing of nuclear and organellar DNA rely on double-stranded DNA deaminases, which edit substrate bases on both strands of DNA, reducing editing precision. Here, we present CyDENT base editing, a CRISPR-free, strand-selective, modular base editor. CyDENT comprises a pair of TALEs fused with a FokI nickase, a single-strand-specific cytidine deaminase and an exonuclease to generate a single-stranded DNA substrate for deamination. We demonstrate effective base editing in nuclear, mitochondrial and chloroplast genomes. At certain mitochondrial sites, we show editing efficiencies of 14% and strand specificity of 95%. Furthermore, by exchanging the CyDENT deaminase with one that prefers editing GC motifs, we demonstrate up to 20% mitochondrial base editing at sites that are otherwise inaccessible to editing by other methods. The modular nature of CyDENT enables a suite of bespoke base editors for various applications.


Surgery for sight: outcomes of congenital and developmental cataracts operated in Durban, South Africa.
To study the visual outcomes of congenital and developmental cataract surgery and determine variables for presentation for pediatric cataract surgery in KwaZulu Natal province of South Africa. Care-givers of children presenting with cataract to a quaternary centre were asked when they first detected the condition. The reasons for delay between detection and surgery were studied. The children underwent a comprehensive eye examination and then appropriate surgery. They were prospectively followed up for 3 months and visual acuity and stereopsis were noted. Delay in presentation for surgery and visual outcomes were co-related with demographic and clinical factors. Eighty-three non-traumatic cataract surgeries in 50 children were studied. Twenty-six (52%) were males, mean age was 3 years 10 months (SD 3yrs 4 months). The mean delay between identification and surgery was 20.7 months (SD 18 months). Twenty-six (52%) children had >15 months interval between diagnosis and surgery. Only mother's occupation was significantly associated with delay (P=0.017). Post-surgery 17/69 (24.7%) had visual acuity ≥6/18, 20/69 (29.0%) had vision between 6/24-6/60, whereas 32/69 (46.3%) had visual acuity ≤6/60. The final vision was associated with age (P=0.031), delay between diagnosis and surgery (P<0.001), type of surgery (P=0.046) and preoperative vision (P<0.001). Although the children's vision improved substantially, a longer follow-up and amblyopia treatment would be necessary to optimize the visual outcome, which depended on age and preoperative vision. Health promotion activities aimed at mothers are important in improving visual outcomes.


Navigating the bone marrow niche: translational insights and cancer-driven dysfunction.
The bone marrow niche consists of stem and progenitor cells destined to become mature cells such as haematopoietic elements, osteoblasts or adipocytes. Marrow cells, influenced by endocrine, paracrine and autocrine factors, ultimately function as a unit to regulate bone remodelling and haematopoiesis. Current evidence highlights that the bone marrow niche is not merely an anatomic compartment; rather, it integrates the physiology of two distinct organ systems, the skeleton and the marrow. The niche has a hypoxic microenvironment that maintains quiescent haematopoietic stem cells (HSCs) and supports glycolytic metabolism. In response to biochemical cues and under the influence of neural, hormonal, and biochemical factors, marrow stromal elements, such as mesenchymal stromal cells (MSCs), differentiate into mature, functioning cells. However, disruption of the niche can affect cellular differentiation, resulting in disorders ranging from osteoporosis to malignancy. In this Review, we propose that the niche reflects the vitality of two tissues - bone and blood - by providing a unique environment for stem and stromal cells to flourish while simultaneously preventing disproportionate proliferation, malignant transformation or loss of the multipotent progenitors required for healing, functional immunity and growth throughout an organism's lifetime. Through a fuller understanding of the complexity of the niche in physiologic and pathologic states, the successful development of more-effective therapeutic approaches to target the niche and its cellular components for the treatment of rheumatic, endocrine, neoplastic and metabolic diseases becomes achievable.


Clinical Trial of Human Umbilical Cord Blood-Derived Stem Cells for the Treatment of Moderate-to-Severe Atopic Dermatitis: Phase I/IIa Studies.
Mesenchymal stem cells (MSCs) have been proven to be therapeutically effective against atopic dermatitis (AD) in preclinical studies. However, the safety and efficacy of MSCs against AD have not yet been investigated in a clinical study. To establish the safety and efficacy of human umbilical cord blood-derived MSCs (hUCB-MSCs) in AD, 34 adult patients with moderate-to-severe AD were enrolled in two phase trials with a follow-up for 1 month and 3 months, respectively. Patients were randomly allocated to receive low dose (2.5 × 107 ) or high dose (5.0 × 107 ) of hUCB-MSCs subcutaneously. An Eczema Area and Severity Index (EASI) score, Investigator's Global Assessment (IGA) score, Severity Scoring for Atopic Dermatitis (SCORAD) score, adverse effect assessments, and serum biomarker levels were evaluated as end points. A single treatment of hUCB-MSCs resulted in dose-dependent improvements in AD manifestation. Fifty-five percent of patients in high dose hUCB-MSC-treated group showed a 50% reduction in the EASI score. The IGA score and SCORAD score decreased by 33% and 50%, respectively, in high dose-treated group. Particularly, the administration of high dose hUCB-MSCs reduced the pruritus score by 58%. The serum IgE levels and number of blood eosinophils were downregulated by the treatment. No serious adverse events occurred, and none of the patients discontinued the trial due to adverse events. This is the first report to demonstrate a marked improvement of AD features with cell therapeutics. These data suggest that the infusion of hUCB-MSCs might be an effective therapy for patients with moderate-to-severe AD. Stem Cells 2017;35:248-255.


Synthesis and preliminary evaluation of some substituted coumarins as anticonvulsant agents.
Some new substituted coumarinylthiazolines, coumarinylthiazolidin-4-ones, and substituted chromenothiazoles were synthesized and evaluated for anticonvulsant activity. Some selected compounds were assayed against seizures induced by pentylenetetrazole (PTZ) and strychnine in mice. Compounds 3b, 6b, and 7b were the most active of the series against PTZ induced seizures. Compound 7b provided anticonvulsant activity (PD(50)=95mg/kg, ip) at a dose 200mg/kg compared to phenobarbital (PD(50)=16mg/kg, ip) at a dose 30mg/kg (90% protection). No clear correlation was observed between the antiepileptic activity and molecular lipophilicity descriptors of the tested compounds.


Spontaneous ureteral stent fragmentation: a case report and review of the literature.
Ureteral stents have been widely used for more than two decades with different indications. Due to the widespread usage of ureteral stents, the number of possible complications of ureteral stents has increased, including stent migration, encrustation, stone formation, and fragmentation. Among these complications, ureteral stent fragmentation is rare. Herein, we present a case of spontaneous ureteral stent fragmentation and review the relevant literature.


Non-random reconstitution of HMG1 and HMG2 in chromatin. Determination of the histone contacts.
We have studied how non-histone proteins HMG1 and HMG2 interact with rat liver chromatin using reconstitution and chemical cross-linking procedures. Both proteins were found to associate to chromatin only to some extent and always with a marked preference for short oligonucleosomes, mainly mono- and dinucleosomes. However, a slight reconstitution with the long polynucleosomal fraction can be observed in H1-depleted chromatin. Reconstitution is non-random and a clear preference for regions highly sensitive to staphylococcal nuclease (EC 188.8.131.52) is observed. Chemical cross-linking has allowed us to identify H1, H2A and H2B as the histones contacted by HMG1 and HMG2 upon reconstitution. Also, we present evidence that HMG1 and HMG2 interact with the nucleosomal particle without replacing H1 or any other histone.


Population tailored modification of tuberculosis specific interferon-gamma release assay.
Blood-based Interferon-Gamma Release Assays (IGRA) identify Mycobacterium tuberculosis (MTB) sensitisation with increased specificity, but sensitivity remains impaired in human immunodeficiency virus (HIV) infected persons. The QuantiFERON-TB Gold In-Tube test contains peptide 38-55 of Rv2654c, based on data indicating differential recognition between tuberculosis patients and BCG vaccinated controls in Europe. We aimed to fine map the T cell response to Rv2654c with the view of improving sensitivity. Interferon-gamma ELISpot assay was used in HIV uninfected persons with latent and active tuberculosis to map peptide epitopes of Rv2654c. A modified IGRA was tested in two further groups of 55 HIV uninfected and 44 HIV infected persons, recruited in South Africa. The most prominently recognised peptide was between amino acids 51-65. Using p51-65 to boost the QuantiFERON-TB Gold In-Tube assay, the quantitative performance of the modified IGRA increased from 1.83 IU/ml (IQR 0.30-7.35) to 2.83 (IQR 0.28-12.2; p = 0.002) in the HIV uninfected group. In the HIV infected cohort the percentage of positive responders increased from 57% to 64% but only after 3 months of ART (p = ns). Our data shows the potential to population tailor detection of MTB sensitization using specific synthetic peptides and interferon-gamma release in vitro.


Changes in lymph node size following systemic irradiation for malignant lymphoma.
Changes in surface area of lymph nodes visualised on abdominal radiographs following lymphography were measured in 18 patients treated by systemic irradiation for non-Hodgkin's lymphoma. Eleven radiologically normal nodes in testicular tumour patients receiving higher doses of external irradiation were measured for comparison. Following lymphography spontaneous shrinkage by up to 20% of the surface area of the node was observed. Since spontaneous and continual regression of abdominal nodes can occur exceptionally in nodular lymphoma, treatment was deferred until there was evidence of an increase in the size of nodes judged as being involved by lymphoma. Two forms of systemic therapeutic irradiation were employed, total body (TBI) and hemi-body (HBI). The rate of nodal regression with both was comparable but the amount of regression, time to nadir of node size and subsequent growth delay, was greater for HBI than TBI.


[Calcium pyrophosphate dihydrate-crystal induced arthropathy].
Calcium pyrophosphate dihydrate (CPPD) crystal induced arthropathy (CPPD-CA; systemic articular chondrocalcinosis) characterized by deposition of CPPD-crystals in fibro- and hyaline cartilage, joint capsule and periarticular tissues is associated with a variable clinical spectrum of inflammatory degenerative and occasionally destructive joint and vertebral manifestations including neurologic complications and rarely tophaceous-tumoral tissue calcifications. Microcrystal formation in the extracellular matrix of cartilage and tendons is based on genetic and acquired dysregulation of chondrocyte pyrophosphate metabolism and presumably linked to gene mutants on the short arm of chromosome-5 (gene locus 5p15.1 and ANKH gene). Idiopathic disease occurs rarely in hereditary-familial and frequently in sporadic manifestation with increasing prevalence due to aging; a secondary form is significantly related to endocrine and metabolic disorders (as to hyperparathyroidism, hemochromatosis etc.). The impact of the clinical syndrome in practice is important in differential diagnosis of age related conditions like acute mono- or oligoarthritis, systemic osteoarthritis and polymyalgia rheumatica. Actual treatment ignores established prevention of systemic cartilage calcification and is oriented to symptomatic relief.


Comparison of atopic and nonatopic children with chronic cough: bronchoalveolar lavage cell profile.
Chronic cough is a common complaint in children and its relationship with asthma is controversial. The aim of the present study was to determine the pattern of airway inflammation in atopic and nonatopic children with chronic cough, and to investigate whether atopy is a predictive factor for eosinophilic inflammation in cough. Bronchoalveolar lavage (BAL; three aliquots of 1 ml/kg saline) was performed in the right middle lobe of 24 (11 atopic and 13 nonatopic) children with persistent cough (8 females, 16 males), mean age 4.7 years (range: 1-11). Atopy was defined as an elevated total serum IgE or a positive RAST test. Both atopic and nonatopic children with persistent cough had an increase in total cells/ml in BAL (atopic: median 39 x 10(4), range: 20-123; nonatopic: median 22 x 10(4), range: 17-132) compared to nonatopic controls (median 11 x 10(4), range 9-30). The increases were mainly in neutrophils (atopic: median 17%, range 2.5-88.5%; nonatopic: median 6%, range 1.0-55.0%) compared to controls (median 1.55%, range 0.5-7.0%; atopics vs. controls, P < 0.005). There were no significant increases in eosinophils, lymphocytes, epithelial cells, or mast cells. Eosinophils were elevated in only 5/11 atopic and none of the nonatopic children. The increased percentage of neutrophils in the BAL fluid of atopic and nonatopic children with persistent cough could be due to an underlying inflammatory process driving the cough, or even conceivably, due to the effect of coughing itself. In this highly selected series, the absence of eosinophilic inflammation in the majority suggests that most would be predicted not to respond to inhaled corticosteroid therapy. This study underscores the need to be cautious about treating coughing children with inhaled corticosteroids, even in the context of a tertiary referral practice.


Scope and mechanism of formal S(N)2' substitution reactions of a monomeric imidozirconium complex with allylic electrophiles.
The zirconium imido complex Cp2(THF)Zr=NSi(t-Bu)Me2 (1) reacts with allylic ethers, chlorides, and bromides to give exclusively the products of the SN2' reaction; i.e., attack at the allylic position remote from the leaving group with migration of the double bond. The primary amine products can be isolated in excellent yields, after in situ Cbz protection, in the presence of variety of functional groups. Good diastereoselectivity and complete stereoselectivity allowed the formation of enantioenriched allylic amines from enantioenriched allylic ethers. Regiospecific substitution with 1 has also been achieved with allylic fluorides, which are notoriously poor substrates in other substitution reactions. On the basis of rate and kinetic isotope effect studies, we propose a general mechanism for the allylic substitution reactions with 1 which involves dissociation of THF and binding of the substrate, followed by the substitution step. In a DFT study of the substitution reaction, we identified a six-membered closed transition state for the substitution step and other relevant stationary points along the reaction coordinate. This study shows that the substitution reaction can be described as a concerted asynchronous [3,3]-sigmatropic rearrangement. This detailed knowledge of the reaction mechanism provides a rationale for the origins of the observed regio-, diastereo-, and stereoselectivity and of the unusual reactivity profile observed in the reaction.


Immunosuppressive agents in penetrating keratoplasty.
One of the problems with corneal grafting is the occurrence of rejection. Immune reaction is considered to be an important cause of graft failure. Early recognition of such graft reaction can frequently be treated successfully with intensive topical administration of corticosteroids. Generally, I prefer to add systemic corticosteroids and immunosuppressive agents such as azathioprine. In the present study two cases are described to illustrate the use of azathioprine and demonstrate that even severe and recurrent graft reactions can be successfully treated with immunosuppressive agents.


Interaction of interstitial trophoblast with placental bed capillaries and venules of normotensive and pre-eclamptic pregnancies.
While endovascular trophoblast invasion of the human placental bed spiral arteries has been studied extensively, no information is available on the interaction between interstitially invading trophoblast and uterine capillaries and venules. Placental bed biopsies of eight normotensive and 15 pre-eclamptic patients were double-immunostained for cytokeratin and the endothelial marker CD31, providing satisfactory staining results in six and 10 biopsies, respectively. Interstitial trophoblast tissue density did not differ between the two series of biopsies, implying that this pathway of invasion is not impaired in pre-eclampsia. Both groups showed a similar incidence of approach of non-arterial vascular structures by perivascular trophoblast. Differences in CD31 staining intensity were noticed in different vascular cross-sections. Lower staining intensity was related to the presence of perivascular trophoblast. Because of the identity of CD31 with the platelet-endothelial cell adhesion molecule (PECAM)-1, the trophoblast-dependent downregulation of CD31 may play a role in the control of leukocytic traffic within the placental bed. The phenomena described in this paper did not show any difference between the normotensive and pre-eclamptic patients, implying that interaction of interstitial trophoblast with venous and capillary structures is not related to the pathogenesis of pre-eclampsia.


Flow cytometric characterization of chronic lymphocyte leukaemias using orthogonal light scattering and quantitative immunofluorescence.
Light scattering properties and antigen distribution of lymphocytes labeled with the monoclonal antibodies CD 5 and CD 20 were determined for 19 patients with a chronic B-cell derived leukaemia. The density of the antigen detected by the monoclonal antibody CD 5 appeared to be considerably lower on malignant B-lymphocytes of the patients as compared with T lymphocytes. A large variation was observed in the amount of receptors for the monoclonal antibodies CD 5 and CD 20 on the malignant cells of the different patients. B-cell chronic lymphocytic leukaemia (B-CLL) patients were clearly distinguishable from leukaemic follicular non Hodgkin lymphoma patients (LF-NHL, formerly lymphosarcoma cell leukaemia) and from a patient with a prolymphocytoid transformation (PLT) of the B-CLL according to the amount of the antigens for CD 5 and CD 20. Within the B-CLL patient population, no relation of progression of the disease with distribution of these antigens could be observed. In one patient the extraordinary phenotype CD 20+, CD 11+, leu 8+, CD 5- of the malignant lymphocytes was observed. An experimentally simple method to differentiate between the various chronic lymphocytic leukaemias (CLL) appeared to be the determination of orthogonal light scattering properties of lymphocytes. In healthy donors one can always distinguish two populations of lymphocytes in the orthogonal light scatter histograms. Lymphocytes of B-CLL patients show one uniform population with a relatively small orthogonal light scattering signal, lymphocytes of our patients with PLT of B-CLL or with LF-NHL show one uniform population with a relatively large orthogonal light scattering signal.


Destabilizing single chain major histocompatibility complex class I protein for repurposed enterokinase proteolysis.
The lack of a high throughput assay for screening stabilizing peptides prior to building a library of peptide-major histocompatibility complex class I (pMHC-I) molecules has motivated the continual use of in silico tools without biophysical characterization. Here, based on de novo protein fragmentation, the EASY MHC-I (EZ MHC-I) assay favors peptide antigen screening to an unheralded hands-on time of seconds per peptide due to the empty single chain MHC-I protein instability. Unlike tedious traditional labeling- and antibody-based MHC-I assays, repurposed enterokinase directly fragments the unstable single MHC-I chain protein unless rescued by a stabilizing peptide under luminal condition. Herein, the principle behind EZ MHC-I assay not only characterizes the overlooked stability as a known better indicator of immunogenicity than classical affinity but also the novel use of enterokinase from the duodenum to target destabilized MHC-I protein not bearing the standard Asp-Asp-Asp-Asp-Lys motif, which may protend to other protein instability-based assays.


Pdgfrβ+ Mural Preadipocytes Contribute to Adipocyte Hyperplasia Induced by High-Fat-Diet Feeding and Prolonged Cold Exposure in Adult Mice.
The expansion of white adipose tissue (WAT) in obesity involves de novo differentiation of new adipocytes; however, the cellular origin of these cells remains unclear. Here, we utilize Zfp423(GFP) reporter mice to characterize adipose mural (Pdgfrβ(+)) cells with varying levels of the preadipocyte commitment factor Zfp423. We find that adipose tissue contains distinct mural populations, with levels of Zfp423 distinguishing adipogenic from inflammatory-like mural cells. Using our "MuralChaser" lineage tracking system, we uncover adipose perivascular cells as developmental precursors of adipocytes formed in obesity, with adipogenesis and precursor abundance regulated in a depot-dependent manner. Interestingly, Pdgfrβ(+) cells do not significantly contribute to the initial cold-induced recruitment of beige adipocytes in WAT; it is only after prolonged cold exposure that these cells differentiate into beige adipocytes. These results provide genetic evidence for a mural cell origin of white adipocytes in obesity and suggest that beige adipogenesis may originate from multiple sources.


[Cerebral lactate dehydrogenase isoenzymes in different hypoxic conditions].
Spectrum of LDH isozymes and their activity were compared in animal brain tissue under circulatory and hypoxic forms of hypoxia. Dynamics and manifestation of alterations observed correlated primarily with the type of oxygen deficiency as well as with its severity and length of action. Development of oxygen deficiency in brain tissue was accompanied by increase in total LDH activity and by elevation in content of M-subunits, typical for "anaerobic" spectrum of LDH isozymes, with simultaneous decrease in amount of H-subunits.


[Motor rehabilitation of a child after a stroke].
The rehabilitation of a child after a stroke is different from that of an adult. The child is in the process of growth, brain maturation and psychomotor development. A multidisciplinary rehabilitation team works to ensure the best possible neuromotor recovery. To be effective, the rehabilitation must be adapted to the child, his or her age, the age at which the stroke occurred and also take into account his or her environment.


A comparison of intrascleral bleb height by anterior segment OCT using three different implants in deep sclerectomy.
To compare intrascleral blebs characteristics after deep sclerectomy (DS) with three intrascleral implants using the Visante anterior segment optical coherence tomography. This is a cross-sectional study including 60 eyes of 51 patients that underwent DS with Sk-Gel, Esnoper, and Aquaflow implant. Intraocular pressure (IOP) measurement, slit-lamp examination, and Visante scans were performed the same day in all the patients. Visante scans were done through the intrascleral bleb at 45°, 90°, and 135° and the bleb height was measured. Sk-Gel was used in 19 eyes (31.66%), Esnoper in 22 eyes (36.66%), and Aquaflow in 19 eyes (31.66%). The median time lapsed from the surgery was 15.50 months 25th and 75th percentiles (p(25) 8.25; p(75) 20). The median IOP was 13 mm Hg (p(25) 10; p(75) 15), with no significant differences among implants (P = 0.232). Overall, the correlation between the scleral bleb height and the IOP was statistically significant at 45° (r=-0.359; P = 0.004), 90° (r = -0.410; P = 0.001), and 135° (r = -0.417; P = 0.001). However, Sk-Gel did not show any statistically significant correlation between the scleral height and IOP whereas the other two groups (Esnoper and Aquaflow) showed a significant correlation. There were no differences in the bleb height among implants. There was a moderate inverse correlation between the scleral bleb height and the IOP measurement after DS with Esnoper and Aquaflow implants. There were no differences in bleb height among the three implants.


Early onset of urea synthesis and ammonia detoxification pathways in three terrestrially developing frogs.
Frogs evolved terrestrial development multiple times, necessitating mechanisms to avoid ammonia toxicity at early stages. Urea synthesis from ammonia is a key adaptation that reduces water dependence after metamorphosis. We tested for early expression and plasticity of enzymatic mechanisms of ammonia detoxification in three terrestrial-breeding frogs: foam-nest-dwelling larvae of Leptodactylus fragilis (Lf) and arboreal embryos of Hyalinobatrachium fleischmanni (Hf) and Agalychnis callidryas (Ac). Activity of two ornithine-urea cycle (OUC) enzymes, arginase and CPSase, and levels of their products urea and CP in tissues were high in Lf regardless of nest hydration, but reduced in experimental low- vs. high-ammonia environments. High OUC activity in wet and dry nests, comparable to that under experimental high ammonia, suggests terrestrial Lf larvae maintain high capacity for urea excretion regardless of their immediate risk of ammonia toxicity. This may aid survival through unpredictably long waiting periods before rain enables their transition to water. Moderate levels of urea and CP were present in Hf and Ac tissues and enzymatic activities were lower than in Lf. In both species, embryos in drying clutches can hatch and enter the water early, behaviorally avoiding ammonia toxicity. Moreover, glutamine synthetase was active in early stages of all three species, condensing ammonia and glutamate to glutamine as another mechanism of detoxification. Enzyme activity appeared highest in Lf, although substrate and product levels were higher in Ac and Lf. Our results reveal that multiple biochemical mechanisms of ammonia detoxification occur in early life stages of anuran lineages that evolved terrestrial development.


Novel 2-oxo-2-phenylethoxy and benzyloxy diaryl urea hybrids as VEGFR-2 inhibitors: Design, synthesis, and anticancer evaluation.
Two series of diaryl urea derivatives, 6a-k and 7a-n, were synthesized. All the newly synthesized compounds were tested against the NCI (US) cancer cell lines via SRB assay. The p-chloro-m-trifluoromethyl phenyl derivatives 6e-g and 7e-g showed the most potent cytotoxic activity with a GI50 value range of 1.2-15.9 µM. Furthermore, the p-fluorobenzyloxy diaryl urea derivative 7g revealed the most potent cytotoxicity against eight cancer cell lines in the MTT assay with IC50 values below 5 µM. Compounds 6a-k and 7a-n were tested for their vascular endothelial growth factor receptor-2 (VEGFR-2) kinase inhibitory activities. The p-chloro-m-trifluoromethyl diaryl urea benzyloxy derivatives 7e-i and the p-methoxydiaryl urea benzyloxy derivatives 7k, 7l, and 7n were found to be the most active compounds as VEGFR-2 inhibitors in the benzyloxy series 7, with an IC50 range of 0.09-4.15 µM. In the 2-oxo-2-phenylethoxy series 6, compounds 6e-g and 6i were reported with IC50 values of 0.94, 0.54, 2.71, and 4.81 µM, respectively. Moreover, compounds 7e and 7g induced apoptosis, causing cell cycle arrest in the G2/M phase. In addition, 7g showed an antimigratory effect in A-375 cells and inhibited the VEGFR-2 expression in an immunohistofluorescence study. Molecular docking simulations on VEGFR-2 as well as ADME properties prediction were also performed.


Disturbed ovarian morphology, oestrous cycling and fertility of high fat fed rats are linked to alterations of incretin receptor expression.
Obesity is a major cause of infertility in females with a direct correlation between energy intake and reproductive dysfunction. To explore underlying mechanisms, disturbances in reproductive health and incretin/reproductive hormone receptor expression were studied in female Wistar rats fed a high-fat-diet for 20-weeks. Metabolic parameters and ovarian/adrenal gene expression were monitored along with estrous cycling and fertility upon mating. High-fat-feeding significantly increased body weight, plasma insulin and HOMA-IR, indicative of obesity and insulin resistance. Estrous cycles were prolonged compared to normal chow-fed rats, with 50 % having an average cycle length ≥ 7days. Reproductive outcomes revealed high-fat-diet reduced litter size by 48 %, with 16 % rats unable to achieve pregnancy. Furthermore, 80 % of the high-fat group took > 35 days to become pregnant compared to 33 % fed a normal-diet. Also, 35 % of pups born to high-fat-fed rats were eaten by mothers or born dead which was not observed with control rats. These changes were associated with downregulation of Amh, Npy2R and GcgR gene expression in ovaries with upregulation of InsR and Glp-1R genes. In adrenals, Glp-1R, GipR, Npy2R, InsR, GcgR, GshR and Esr-1 genes were upregulated. Histological analysis of high-fat-diet ovaries and adrenals revealed changes in morphology with significantly increased number of cysts and reduced adrenal capsule thickness. Circulating levels of insulin, testosterone and progesterone was significantly higher in high-fat group with reduced FSH levels in plasma. These data demonstrate that high-fat feeding disrupts female reproductive function and suggest important interactions between gut and reproductive hormones in ovaries and adrenals which merit further investigation.


Atherosclerotic Renal Artery Stenosis Prevalence and Correlations in Acute Myocardial Infarction Patients Undergoing Primary Percutaneous Coronary Interventions: Data From Nonrandomized Single-Center Study (REN-ACS)—A Single Center, Prospective, Observational Study.
We are the first to evaluate the prevalence of renal artery stenosis (RAS) in consecutive patients with acute myocardial infarction (AMI) referred for primary percutaneous coronary intervention from a single tertiary center. As a novelty, we assessed hydration and metabolic status and measured arterial stiffness. We elaborated a predicting model for RAS in AMI. One hundred and eighty-one patients with AMI underwent concomitantly primary percutaneous coronary intervention and renal angiography. We obtained data on demographics, medical history, cardiovascular risk factors, echocardiography, Killip class, and blood tests. In the first 24 hours post-primary percutaneous coronary intervention, we assessed bioimpedance through Body Composition Monitoring(®) and arterial stiffness through pulsed-wave velocity, SphygmoCor(®). Significant RAS (>50% lumen narrowing, RAS+) was present in 16.6% patients. In the RAS+ group we recorded significantly higher stiffness, CRUSADE score and dehydration, and more women with higher prevalence of multivascular coronary artery disease and heart failure. In our multivariate models, variables independently associated with RAS+ were previous percutaneous coronary intervention, low estimated glomerular filtration rate, multivascular coronary artery disease, and total/extracellular body water. These models had good specificity and low sensitivity. We observed that RAS+ AMI patients have a particular hydration, metabolic, and endothelial profile that could generate more future major adverse cardiac events. Hence, renal angiography in AMI should be considered in specific subsets of patients. URL: https://www.clinicaltrials.gov/. Unique identifier: NCT02388139.


Intra-rater Reliability of a Qualitative Landing Scale for the Single-Hop Test: A Pilot Study.
The test battery classically used for return-to-sport (RTS) decision-making after anterior cruciate ligament (ACL) reconstruction (ACLR) may not be sufficient, as it does not include a qualitative analysis of movement. Therefore, the Landing Error Scoring System (LESS) scale was adapted to a primary functional test in the typical RTS test battery: the single leg hop for distance (SHD). The aim of this study was to determine the intra-rater reliability of the LESS scale adapted to the SHD (SHD-LESS scale) in healthy young athletes. Reliability analysis. Nineteen healthy individuals (14 men, 5 women; mean age: 22.4 years) participated in the study. Participants performed the SHD tasks on both limbs (dominant and non-dominant) using a standardized protocol in two sessions that were one week apart (single reviewer; 2-dimensional video). Intra-class correlation coefficients (ICC2,1) were used to measure the reproducibility of the scale in the dominant (dom) and non-dominant (nondom) limbs. Additionally, limb data (dom and nondom) were pooled and evaluated collectively with intra-class correlation coefficients. The Kappa coefficient was used to assess the reproducibility of each individual item of SHD-LESS scale. The intra-rater reliability was good (ICCdom = 0.77; ICCnondom = 0.87; ICCpooled = 0.87) for the overall SHD-LESS scale scores. Agreement of SHD-LESS individual items ranged from 62% to 100%. Dorsiflexion at initial contact (97% agreement; kappa value=0.79) and knee valgus after landing (88% agreement; kappa value=0.65) had excellent agreement and kappa values. The newly-adapted SHD-LESS scale showed good intra-rater reliability overall. Further studies should evaluate the impact of using the SHD-LESS scale within the RTS test battery on outcomes in patients after ACLR. 3.


A day in the EMG laboratory: case studies of 10 patients with different clinical problems.
This article reviews 10 cases that have been evaluated in the electromyography laboratory, which represent a variety of clinical problems that may be referred for electrodiagnostic evaluation. The clinical and electrodiagnostic findings are presented, and practical points related to the approach taken in the evaluation are discussed.


Ultrasound-Triggered Delivery of Iproplatin from Microbubble-Conjugated Liposomes.
The PtIV prodrug iproplatin has been actively loaded into liposomes using a calcium acetate gradient, achieving a 3-fold enhancement in drug concentration compared to passive loading strategies. A strain-promoted cycloaddition reaction (azide- dibenzocyclooctyne) was used to attach iproplatin-loaded liposomes L(Pt) to gas-filled microbubbles (M), forming an ultrasound-responsive drug delivery vehicle [M-L(Pt)]. Ultrasound-triggered release of iproplatin from the microbubble-liposome construct was evaluated in cellulo. Breast cancer (MCF-7) cells treated with both free iproplatin and iproplatin-loaded liposome-microbubbles [M-L(Pt)] demonstrated an increase in platinum concentration when exposed to ultrasound. No appreciable platinum uptake was observed in MCF-7 cells following treatment with L(Pt) only or L(Pt)+ultrasound, suggesting that microbubble-mediated ultrasonic release of platinum-based drugs from liposomal carriers enables greater control over drug delivery.


[Effects of low dose mifepristone on population and subsets of natural killer cells in human endometrium during receptive phase].
To investigate the immunologic mechanism of low dose mifepristone as an anti-implantation contraceptive drug. Endometrial biopsies were obtained from fourteen normally cycling women during the receptive phase. Each endometrial tissue, divided into three equal parts, was assigned to three groups: control group, 65 nmol/L mifepristone group (group A) and 200 nmol/L mifepristone group (group B). Endometrial explants were cultured in vitro. The numbers of natural killer cells and the percentages of CD3- CD56+ CD16- subset and CD3- CD56+ CD16+ subset were analysed by immunohistochemistry and flow cytometry. (1) The mean number of CD56+ NK cells in group A (148 +/- 11) and group B (150 +/- 12) were significantly higher when compared to the control group (121 +/- 7, P < 0.05). There was no significant difference between the two mifepristone-treated groups (P > 0.05). (2) The percentage of CD3- CD56+ Nk cells in group A (44 +/- 5)% and in group B (48 +/- 4)% were significantly higher when compared to the control group (35 +/- 3)% (P < 0.05), there was no significantly difference between the two mifepristone-treated groups (P > 0.05); The percentage of CD3- CD56+ CD16- subset in group A (42 +/- 5)% and in group B (45 +/- 5)% were significantly higher when compared to the control group (33 +/- 3)%, (P < 0.05), there was no significantly difference between the two mifepristone-treated groups (P > 0.05); the percentages of CD3- CD56+ CD16+ subset in three groups [(2.70 +/- 0.24)% (3.26 +/- 0.37)% and (2.33 +/- 0.29)%] were not significantly different (P > 0.05). Low dose mifepristones increase the number of CD56+ NK cells and the percentage of CD3- CD56+ CD16- NK subset, which might result in the disturbance of human endometrial immuno-microenvironment during receptive phase and lead to implantation failure.


Natural course of myopic traction maculopathy and factors associated with progression or resolution.
To investigate the natural course of myopic traction maculopathy and to identify the factors associated with its progression or resolution in 207 highly myopic eyes. Retrospective, consecutive case series. We investigated 207 highly myopic eyes with myopic traction maculopathy by optical coherence tomography (OCT) and followed the patients for at least 24 months. The size of the macular retinoschisis was categorized according to its extent and location relative to the fovea. The clinical data, status of the myopic traction maculopathy, and changes in the posterior vitreous and inner limiting membrane (ILM) were analyzed during the natural course of the disease process. During a mean follow-up of 36.2 ± 6.2 months, 8 of 207 eyes (3.9%) showed a decrease or complete resolution of the macular retinoschisis. Of these 8 eyes, 6 had release of retinal traction in the OCT images, and 4 of the 6 eyes developed posterior vitreous detachment. A spontaneous disruption of the ILM occurred in 2 eyes before the improvement of the myopic traction maculopathy. Of the 207 eyes, 24 (11.6%) experienced progression of the myopic traction maculopathy. The eyes with more extensive macular retinoschisis showed progression significantly more commonly (42.9%) than the eyes having less extensive macular retinoschisis areas (6.7%). The natural course of myopic traction maculopathy varies, and eyes with more extensive macular retinoschisis tend to progress more commonly. A complete resolution of macular retinoschisis can occur after a release of traction on the retina. These findings can be used to determine whether surgery should be performed.


Betamethasone for Preterm Birth: Auckland Steroid Trial Full Results and New Insights 50 Years on.
The objective of this study was to use modern analysis and reporting methods to present the full results of the first randomized trial of antenatal corticosteroids, performed 50 years ago. In this single-center trial, women at risk of preterm birth at 24 to less than 37 weeks of gestation were randomized to receive 2 doses of betamethasone or placebo, 24 hours apart. Women and their caregivers were blinded to treatment allocation. The primary outcome was respiratory distress syndrome. Secondary outcomes included measures of neonatal mortality and morbidity, mode of birth, and maternal infection. Between 1969 and 1974, 1115 women (1142 pregnancies) were randomized, 560 pregnancies (601 infants) to betamethasone and 582 (617 infants) to placebo. The risk of respiratory distress syndrome was significantly reduced in the betamethasone group compared with placebo (8.8% vs 14.4%, adjusted relative risk 0.62, 95% CI 0.45-0.86, P = .004). Subgroup analyses indicated greater efficacy in male than female infants but no effect of tocolytic therapy or doubling of betamethasone dose. Fetal or neonatal death, neonatal or maternal infection, neonatal hypoglycaemia, cesarean delivery, and lactation status at discharge were not different between the groups. Antenatal betamethasone administered to women at risk of preterm birth between 24 and less than 37 weeks of gestation reduces the incidence of respiratory distress syndrome, with greater effect in male than in female infants. Doubling the dose of betamethasone does not provide additional benefit.


Profile of territrem metabolism and cytochrome P-450 3A expression in liver microsomes from Wistar rats of both genders as a function of age.
This study determined territrem metabolites after incubation of territrem A, B, or C with NADPH and liver microsomes from Wistar rat of both genders aged 2 to 76 wk. The liver microsomal cytochrome P-450 content, NADPH-cytochrome P-450 reductase activity, and CYP3A1 and CYP3A2 protein and mRNA levels were also analyzed. Male rats had significantly higher liver microsomal cytochrome P-450 content and NADPH-cytochrome P-450 reductase activities than females at 14 to 26 wk. Microsomal cytochrome P-450 content was decreased in senescence in both genders compared with postpubertal and adulthood stages. The activity of 6beta-testosterone hydroxylase in male rats, which was significantly higher than those in females at all ages, decreased after 52 wk. After 26 wk, the levels of CYP3A1 protein markely declined in both genders, which resulted in a large gender difference (male greater than female). The protein levels and mRNA of CYP3A2 were constitutively expressed in 2- to 52-wk-old male rats, but they decreased after 76 wk, and decreased in females after 6 wk. The expression of CYP3A1 or CYP3A2 in males are generally higher than in females. The metabolites of territrems MA1, MAX, MA2, MB2, MB4, and MC were measured by high-performance chromatography (HPLC). Formation of MA1, MAX, and MA2 decreased after 52 wk in males, and MAX and MA2 were not formed after 6 wk in females. The amount of MB2 formed in females was less than in males, but the amount of MC (TRC metabolites) formed in females was higher than in males. The gender differences in metabolism of TRA were related to the protein and mRNA expression of CYP3A2. The protein levels and mRNA expression of CYP3A2 and efficiency of territrems metabolism were decreased after 76 wk. The results suggested that the effects of age and gender on territrem metabolism are due to differences in CYP3A1 and CYP3A2 expression in the liver microsomes.


Structural and cytochemical features of perineuronal glial nets in the rat brain.
Perineuronal nets formed by glial cells could be visualized in different regions of the rat brain with Golgi methods. Although mostly their cellular origin was not detectable, in some cases the nets were found to originate from glial cell processes possessing characteristic features of microglia. Using the colloidal iron hydroxide binding method and carbohydrate binding lectins the perineuronal nets were shown to be enriched with anionic carbohydrate components. Some functional aspects of these structures have been discussed.


2022 ISPNE Bruce McEwen Lifetime Achievement award: Stress, from molecules to societies.
The International Society for Psychoneuroendocrinology meeting in Chicago in 2022 was thrilled to recognize Dr. Robert Sapolsky with the Bruce McEwen Lifetime Achievement award. This is the second year for the award to be named to honor Bruce McEwen and it marks the completion of a special issue edited by Blazej Miziak and Robert Paul Juster in the journal Psychoneuroendocrinology dedicated to Bruce's legacy and the unfathomable contribution of Allostatic Load to the stress field. Yet, as our award winner writes, Bruce's legacy is more than scientific as he was well known for mentorship and being an exemplary person, theorist, and scientist. Perhaps understandably for a career favored by humble introverts and shy reclusives, the science shines in the spotlight and personal reflections are cut to accommodate word count limits. For scholars entering the field, stargazing at larger than life luminaries in the field is thrilling yet intimidating as it feels impossible that these experts have the same doubts and distractions as the rest of us primates. Thus, Psychoneuroendocrinology is thrilled to kick off the first perspectives piece in the Cell to Selves series with Dr. Robert Sapolsky sharing that, like his Baboon troops in Kenya, he too sometimes has a bad-hair day. This paper is a written version of a lecture I gave on September 8th, 2022, when receiving the first Bruce McEwen Lifetime Achievement Award from the ISPNE. This was a bittersweet honor; Bruce was my graduate advisor at Rockefeller University and over the next forty years, he was my mentor, teacher and father figure. His death in 2020 left a hole in my life.


Use of PCR in detection of Mycobacterium avium complex (MAC) bacteremia: sensitivity of the assay and effect of treatment for MAC infection on concentrations of human immunodeficiency virus in plasma.
We evaluated the sensitivity and specificity of a PCR-based qualitative test for the rapid diagnosis of Mycobacterium avium-M. intracellulare complex (MAC) bacteremia in patients with AIDS disease. Eleven subjects with newly culture-proven MAC bacteremia had the following tests performed at biweekly intervals during the first 8 weeks of therapy: blood culture, Mycobacterium-specific PCR, and quantitative human immunodeficiency virus (HIV) viral-load testing. Mycobacterium genus-specific biotinylated primers were used to amplify a sequence of approximately 582 nucleotides within the 16S rRNA genes of M. avium and M. intracellulare. Detection of the amplified product was performed with an oligonucleotide probe-coated microwell plate combined with an avidin-horseradish peroxidase-tetramethylbenzidine conjugate-substrate system. While not as sensitive as BACTEC culture, PCR detected 17 of 18 specimens which grew >/=40 organisms/ml (94.4% sensitivity) and 9 of 16 specimens which grew </=40 organisms/ml (56.3% sensitivity). No clear change in HIV viremia occurred in response to successful treatment of patients' MAC bacteremia. Use of the PCR test allowed detection of MAC bacteremia in 1 day, with a sensitivity similar to those of quantitative blood culture techniques, and it may prove useful for rapid screening of suspected cases. HIV viremia was unaffected by 8 weeks of MAC therapy.


Behavior of field-applied triadimefon, malathion, dichlorvos, and their main metabolites during barley storage and beer processing.
The behavior and fate of 3 pesticides (triadimefon, malathion, and dichlorvos) and the main metabolites (triadimenol and malaoxon) during barley storage or beer processing were assessed using a pilot-plant equipment. The residues of all products were determined using liquid chromatography coupled with tandem mass spectrometry. Field investigation of the dissipation rate kinetics for triadimefon and malathion during storage indicated that their half-life was twice as high when 5 times the recommended dosage was used. Milling had little effect on the removal of dichlorvos and malathion residues, whereas these were substantially removed when the spent grains were filtered after mashing. The calculated processing factors were all <1, indicating the residual ratios of dichlorvos and malathion were reduced during the entire process. In conclusion, storage and processing considerably reduced pesticide residue levels in barley and beer; however, greater focus needs to be paid to the toxicity of their metabolites in commercial by-products.


Serologic evidence of arboviral infections in white-tailed deer from central Wisconsin.
A survey conducted during 1979-1980 on white-tailed deer (Odocoileus virginianus) in central Wisconsin revealed serological evidence of infection by selected arboviruses. Among sera from 41 deer, antibody was detected for Jamestown Canyon virus (56%) and Bunyamwera group virus (80%), demonstrating their continuing endemic activity. Antibody for La Crosse virus, not found previously in sera from deer in central Wisconsin, also was detected (5%) in this study.


2-(4-Methoxyphenyl)ethyl-2-acetamido-2-deoxy-β-d-pyranoside confers neuroprotection in cell and animal models of ischemic stroke through calpain1/PKA/CREB-mediated induction of neuronal glucose transporter 3.
Salidroside is proven to be a neuroprotective agent of natural origin, and its analog, 2-(4-Methoxyphenyl)ethyl-2-acetamido-2-deoxy-β-d-pyranoside (named SalA-4g), has been synthesized in our lab. In this study, we showed that SalA-4g promoted neuronal survival and inhibited neuronal apoptosis in primary hippocampal neurons exposed to oxygen and glucose deprivation (OGD) and in rats subjected to ischemia by transient middle cerebral artery occlusion (MCAO), respectively, and that SalA-4g was more neuroprotective than salidroside. We further found that SalA-4g elevated glucose uptake in OGD-injured primary hippocampal neurons and increased the expression and recruitment of glucose transporter 3 (GLUT3) in ischemic brain. Signaling analysis revealed that SalA-4g triggered the phosphorylation of CREB, and increased the expression of PKA RII in primary hippocampal neurons exposed to OGD injury, while inhibition of PKA/CREB by H-89 alleviated the elevation in glucose uptake and GLUT3 expression, and blocked the protective effects of SalA-4g. Moreover, SalA-4g was noted to inhibit intracellular Ca(2+) influx and calpain1 activation in OGD-injured primary hippocampal neurons. Our results suggest that SalA-4g neuroprotection might be mediated by increased glucose uptake and elevated GLUT3 expression through calpain1/PKA/CREB pathway.


Phycotoxins in bivalves from the western Canadian Arctic: The first evidence of toxigenicity.
This study presents the first evidence that a diverse suite of phycotoxins is not only being actively produced by the toxigenic algal communities in the Canadian Arctic waters, but is also entering the marine food web. We detected measurable amounts of Amnesic Shellfish Toxins (ASTs) and Paralytic Shellfish Toxins (PSTs), as well as trace amounts of other lipophilic toxin groups including pectenotoxins, yessotoxins, and cyclic imines, in bivalves collected from the Canadian Beaufort Sea in 2014 and 2018. There appear to be species-specific differences in accumulation and retention of AST by Arctic bivalves, with significantly higher concentrations recorded in Nuculanidae than Propeamussiidae, likely reflecting physiological and allometric differences. We further confirm the omnipresence of potentially toxic taxonomically-versatile phytoplankton communities in the western Canadian Arctic comprising Pseudo-nitzschia delicatissima group, P. obtusa, Dinophysis acuminata, Prorocentrum minimum, Alexandrium tamarense, and Gymnodinium spp. Although measurements of actual toxicity levels and profiles of these species at the time of sampling fall outside of the scope of this study, we show that high abundance and competitive success of known AST-producers, Pseudo-nitzschia spp., are possible in Canadian Arctic waters. In 2014, a strong dominance of Pseudo-nitzschia spp. was observed at a few shallow coastal stations, representing nearly 40% of the total phytoplankton cell abundances with > 106 cells/L at the depth of maximum chlorophyll a. We further describe oceanographic conditions conducive to high abundances of toxin-producing algae, indicating that temperature is likely a key factor. Even though measured AST and PST concentrations in bivalve tissue remained well below the Health Canada's levels at which monitored fisheries would close, i.e., 5% and 4%, respectively, their presence demonstrate that phycotoxin accumulation is occurring in food webs of the Canadian Beaufort Sea. Yet, the phycotoxin production controls and trophic transfer mechanisms remain unknown. Canadian Arctic marine ecosystems are rapidly changing and temperatures are expected to continue to increase. Given that these changes simultaneously affect multiple, and often co-occurring, species of primary producers, adaptive capacity is likely to play an important role in the structure of phytoplankton communities in the Canadian Arctic.


Addition of [CH(CN)2]- and [TCNE]˙- to Ru(VI)≡N bearing 8-quinolinolato ligands.
Reaction of Ru(VI)≡N complexes bearing 8-quinolinolato ligands with NCCH(2)CN/piperidine and NaTCNE afford novel ruthenium(ii) dicyanoimine and diimine/imino-oxazolone complexes, respectively.