Kinase activity of the type V transforming growth factor beta receptor.
The type V TGF-beta receptor purified from bovine liver plasma membranes catalyzed the phosphorylation of casein using [gamma-32P]ATP as co-substrate (Km,app approximately 10 microM). TGF-beta stimulated the casein phosphorylation by the type V receptor with a half-optimal concentration of approximately 0.3 nM. Both TGF-beta-stimulated and -unstimulated phosphorylations occurred at serine residues. Amino acid sequences of 32P-labeled peptides from reverse phase HPLC of the tryptic digests of casein 32P-phosphorylated by the type V receptor were analyzed by automated Edman degradation. Alignment of phosphorylation site amino acid sequences of the 32P-labeled peptides revealed a recognition motif (S-X-E/S(P)) for the kinase activity of the type V receptor. The type V receptor catalyzed the phosphorylation of an octadecapeptide designated peptide SESTE (SKDIGS*ESITEDQAMEDKK) (the asterisk indicates the phosphorylated residue) containing the major phosphorylation site sequence of casein. The Km,app and Vmax for peptide SESTE were determined to be 0.3 microM and 2.2 nmol of 32P incorporation/min/mg of enzyme, respectively. The phosphorylation of peptide SESTE by the type V receptor was stimulated by TGF-beta or polylysine but inhibited by heparin. Like intact casein, peptide SESTE was phosphorylated at 2 close serine residues (-S*-E-S*-T-E-) by the type V receptor. The type V receptor also phosphorylated a variant of peptide SESTE, peptide AESTE (SKDIGAES*TEDQAMEDKK). However, the type V receptor appeared not to phosphorylate peptide SEATE (SKDIGSEATEDQAMEDKKK) in which an alanine residue replaced 1 of the 2 close serine residues (at the C-terminal side) of peptide SESTE. These results suggest that the type V receptor is capable of exerting interdependent phosphorylation.


Haplotypes and effects on growth traits of bovine Wnt7a gene in Chinese Qinchuan cattle.
Wnt7a is a member of the WNT gene family, which encodes secreted signaling proteins and responds to many biological processes. Specifically Wnt7a influences satellite stem cells and regulates the regenerative potential of the muscle. However, similar researches about the bovine Wnt7a gene are lacking. Therefore, in this study, polymorphisms of the bovine Wnt7a gene were detected in 488 individuals from Chinese Qinchuan cattle by DNA pooling, forced PCR-RFLP, and DNA sequencing methods. 3 novel SNPs were identified, two SNPs (g.T4926C and g.A21943G) were in the intron and the last one (g.C63777T) was in the exon. Five haplotypes involved in these three variant sites in the Wnt7a gene were identified and their effects on growth traits were analyzed. The results revealed that haplotype 1 had the highest haplotype frequencies and was highly significantly associated with body height (P<0.01), body weight (P<0.05), chest width (P<0.05) and height at hip cross (P<0.01) respectively.


Influence of steroids on complement and cytokine generation after cardiopulmonary bypass.
It is recognized that the inflammatory mediators complement and cytokines are generated during cardiopulmonary bypass as an endogenous response to extracorporeal circulation. Nineteen randomized patients (10 steroid/9 nonsteroid) entered an institutional review board-approved protocol to measure complement and interleukin level generation before and after elective coronary revascularization. The steroid regimen involved 1 g of methylprednisolone sodium succinate intravenously before bypass and 4 mg of dexamethasone every 6 hours for four doses during the first 24 hours of recovery. Complement and interleukin levels were measured before bypass, immediately after bypass, and at 24, 48 and 72 hours of recovery. In the nonsteroid group, there was a significant elevation in all inflammatory mediators relative to the steroid group. The predominant changes occurred at 24 hours after operation. Steroids produced a dramatic reduction in complement and interleukin levels. The number of patients was clearly too small to document a clinical consequence of steroid administration.


High-performance liquid chromatographic determination of amantadine in urine after micelle-mediated pre-column derivatization with 1-fluoro-2,4-dinitrobenzene.
Cationic micelles have been used for the derivatization of the anti-Parkinson drug amantadine with the chromophore 1-fluoro-2,4-dinitrobenzene in urine. In the presence of 90 mM cetyltrimethylammonium bromide (CTAB), the conversion of amantadine into its derivative is complete within 4 min at 60 degrees C and pH 11. Such a short reaction time allows a fully automated pre-column derivatization of amantadine in an on-line combination with reversed-phase high-performance liquid chromatography. This cannot be attained when using purely aqueous derivatization mixtures because then the reaction takes some 20 min at the same temperature. Without the use of an internal standard, the repeatability of the automated determination at the 0.5 microgram ml-1 level is ca. 6%, whilst the detection limit is 75 ng ml-1 (S/N = 3). The present study clearly demonstrates that micellar systems can be beneficially used for the on-line precolumn derivatization of amines in urine.


Age-related morphometrics of normal adrenal glands based on deep learning-aided segmentation.
This study aims to evaluate the morphometrics of normal adrenal glands in adult patients semiautomatically using a deep learning-based segmentation model. A total of 520 abdominal CT image series with normal findings, from January 1, 2016, to March 14, 2019, were retrospectively collected for the training of the adrenal segmentation model. Then, 1043 portal venous phase image series of inpatient contrast-enhanced abdominal CT examinations with normal adrenal glands were included for analysis and grouped by every 10-year gap. A 3D U-Net-based segmentation model was used to predict bilateral adrenal labels followed by manual modification of labels as appropriate. Quantitative parameters (volume, CT value, and diameters) of the bilateral adrenal glands were then analyzed. In the study cohort aged 18-77 years old (554 males and 489 females), the left adrenal gland was significantly larger than the right adrenal gland [all patients, 2867.79 (2317.11-3499.89) mm3 vs. 2452.84 (1983.50-2935.18) mm3, P < 0.001]. Male patients showed a greater volume of bilateral adrenal glands than females in all age groups (all patients, left: 3237.83 ± 930.21 mm3 vs. 2646.49 ± 766.42 mm3, P < 0.001; right: 2731.69 ± 789.19 mm3 vs. 2266.18 ± 632.97 mm3, P = 0.001). Bilateral adrenal volume in male patients showed an increasing then decreasing trend as age increased that peaked at 38-47 years old (left: 3416.01 ± 886.21 mm3, right: 2855.04 ± 774.57 mm3). The semiautomated measurement revealed that the adrenal volume differs as age increases. Male patients aged 38-47 years old have a peaked adrenal volume.


Intravenous injection of high and medium concentrations of computed tomography contrast media and related heat sensation, local pain, and adverse reactions.
To evaluate the incidence of acute adverse reactions and degrees of heat sensation and local pain after intravenous injection of high and medium concentrations of iodinated contrast medium for computed tomography (CT). A prospective study was performed involving 729 patients who underwent contrast CT scans. High-concentration (370 mgI/mL) and medium-concentration (300 mgI/mL) iodinated contrast medium was assigned to 342 patients (group H, aged 20-90 years, mean = 59.8 years) and to 387 patients (group M, aged 20-95 years, mean = 61.7 years), respectively. An injection rate of contrast medium (1, 2, or 4 mL/s) at a base volume of 2 mL/kg of body weight was selected according to the protocols for the evaluated diseases and regions. Each patient was assessed for heat sensation and local pain at the injection site using a visual analog scale (ranging from none for 0 to severe for 10). Acute adverse reactions were recorded when they occurred. There were no significant differences in patient background factors, including age, sex, history of prior adverse reactions, and allergies, between the 2 groups. The score for heat sensation was significantly higher in group H than in group M (4.46 +/- 2.44 vs. 3.44 +/- 2.45; P < 0.0001 for heat sensation). The data did not show a higher incidence of adverse reactions in group H than in group M (5 [1.46%] of 342 patients vs. 2 [0.52%] of 387 patients; P = 0.26) or a higher score for local pain in group H than in group M (0.98 +/- 1.70 vs. 0.88 +/- 1.49; P = 0.66), respectively. High and medium concentrations of iodinated contrast medium can be used for CT study with comparable safety profiles even though heat sensation produced by the high-concentration CM is greater than that produced by the medium-concentration CM.


Effects of actinomycin D on brain RNA synthesis and discrimination learning in the goldfish (Carassius auratus).
Intracranial injection of actinomycin D 2 microgram inhibited about 70% of the brain RNA synthesis from 3 hr to 4 days after injection in the goldfish. Under these conditions, fish were given 4-day-training of visual discrimination between a card with vertical stripes and one with horizontal stripes. Fish injected intracranially with actinomycin D showed deficits in between-day retention (long-term memory) but not interruption of within-day acquisition (short-term memory). It is suggested that brain RNA synthesis is necessary only for the formation of long-term memory but not short-term memory.


[Total cavopulmonary connection for grown diminutive pulmonary artery after staged Blalock-Taussig shunt].
We report a two-year-old girl with asplenia, [A, L, L] DORV, pulmonary atresia, common AV valve, PDA, and TAPVC, who successfully underwent total cavo pulmonary connection (TCPC). Deep cyanosis was pointed out since birth. Cardiac catheterization performed on the sixth day after birth revealed a diminutive pulmonary artery tree of which PA index was 41 mm2/m2. Left modified Blalock-Taussig shunt was created at 27 days of age. The PA index increased to 282 mm2/mm2, but disparity in diameter between the left and the right pulmonary artery was yielded by PDA subsidence. Therefore additional contralateral B-T shunt was made at one year of age. Follow-up cardiac catheterization at 28 months of age showed well developed pulmonary artery; PA index of 460 mm2/m2, right pulmonary resistance (Rp) of 3.49 units, left Rp of 2.33 units, and estimated total Rp was 1.39. According to study, bidirectional Glenn procedure or TCPC was indicated. Considering necessity of urgent repair of common pulmonary vein obstruction, regurgitation of the common atrio-ventricular valve and pulmonary artery stenosis, TCPC was performed with concomitant repair of the associated lesions. Severe butterfly-figure stenosis of the central PA was augmented by anastomosing both the left SVC and the left-sided atrium. In conclusion, diminutive pulmonary artery could be adequately grown by phase-in Blalock-Taussig shunts. Pulmonary blood flow scintigraphy was thought to be useful for estimation of pulmonary resistance in such cases with different pulmonary resistance between right and left PA.


The effects of aerobic exercise on eGFR, blood pressure and VO2peak in patients with chronic kidney disease stages 3-4: A systematic review and meta-analysis.
CKD is associated with several comorbidities, cardiovascular disease being the most significant. Aerobic training has a beneficial effect on cardiovascular health in healthy and some well-defined non-healthy populations. However, the effect of aerobic training on glomerular filtration rate in patients with CKD stages 3-4 is unclear. To review the effects of aerobic exercise training on kidney and cardiovascular function in patients with chronic kidney disease (CKD) stages 3-4. A random-effects meta-analysis was performed to analyse published randomized controlled trials through February 2018 on the effect of aerobic training on estimated glomerular filtration rate, blood pressure and exercise tolerance in patients with CKD stages 3-4. Web of Science, PubMed and Embase databases were searched for eligible studies. 11 randomized controlled trials were selected including 362 participants in total. Favourable effects were observed on estimated glomerular filtration rate (+2.16 ml/min per 1.73m2; [0.18; 4.13]) and exercise tolerance (+2.39 ml/kg/min; [0.99; 3.79]) following an on average 35-week aerobic training program when compared to standard care. No difference in change in blood pressure was found. There is a small beneficial effect of aerobic training on estimated glomerular filtration rate and exercise tolerance, but not on blood pressure, in patients with CKD stages 3-4. However, data are limited and pooled findings were rated as of low to moderate quality.


Humoral immunity and antibody-mediated rejection in solid organ transplantation.
The humoral arm of the immune system provides robust protection against extracellular pathogens via the production of antibody molecules that neutralize or facilitate the destruction of microorganisms. However, the humoral immune system also provides a significant barrier to solid organ transplantation due to the antibody-mediated recognition of non-self proteins and carbohydrates expressed on transplanted organs. Historically, the presence of donor-specific antibodies (DSA) that recognize donor HLA molecules, incompatible ABO blood group antigens and other endothelial or xenogeneic antigens was considered a contraindication to transplantation. However, recent advances in antibody testing and immunosuppressive therapies have made it possible to cross certain antibody barriers successfully. In this article, we review our current understanding of antibody-mediated processes in solid organ transplantation and discuss the clinically available treatment options for preventing and treating antibody-mediated rejection.


How chronically administered valproate increases chlordiazepoxide transfer through the blood-brain barrier.
Sodium valproate (VPA) is a drug widely used in the treatment of epileptics often in association with benzodiazepines. Recent animal studies have shown that the addition of valproate increases diazepam levels in the cortex and the cerebellum (Hariton et al, 1985). The aim of our study was to determine the effect of VPA on the transfer of benzodiazepines through the blood-brain barrier. They were investigated using the intracarotid injection technique in rats as described by Oldendorf (1971). Our results show that the 14C-chlordiazepoxide brain extraction is significantly higher in rats on prolonged valproate treatment than in controls. With regard to plasma protein binding effects on chlordiazepoxide transport, our data indicate that a fraction of the protein-bound chlordiazepoxide could transfer from the intracapillary space to the brain tissue space because of enhanced drug dissociation from albumin in the brain microcirculation (Kd in vitro = 74.1 microM; Kd in vivo = 793.7 microM). Two distinct mechanisms can be deduced from this study: 1) chlordiazepoxide is displaced from HSA by valproate, 2) in addition, this fatty acid could increase drug permeation through the blood brain barrier (PS/F (chlordiazepoxide) = 0.60 in controls, PS/F (chlordiazepoxide) = 0.97 in treated rats). On the contrary, the washout of the benzodiazepine from the rat brain does not seem to be modified by the addition of valproate.


Sepiapterin reductase gene-disrupted mice suffer from hypertension with fluctuation and bradycardia.
(6R)-l-erythro-5,6,7,8-Tetrahydrobiopterin (BH4) is an essential cofactor for monoamine and nitric oxide (NO) production. Sepiapterin reductase (SPR) catalyzes the final step in BH4 biosynthesis. We analyzed the cardiovascular function of adult Spr gene-disrupted (Spr-/-) mice for the first time. After weaning, Spr-/- mice suffered from hypertension with fluctuation and bradycardia, while the monoamine contents in these mice were less than 10% of those in the wild-type mice as a result of BH4 depletion. Heart rate variability analysis indicated the sympathetic dominant state in Spr-/- mice. The endothelium-dependent vascular relaxation in response to acetylcholine was significantly impaired in Spr-/- mice after sexual maturation (above 4 months old). Protein amounts of α1 adrenergic receptor and eNOS in the aorta were not altered. Spr-/- mice exhibited hypoglycemia and elevation of plasma renin activity. Our results suggest that the hypertension with fluctuation and bradycardia of Spr-/- mice would be caused by an imbalance of sympathetic and parasympathetic input and impaired nitric oxide production in endothelial cells. We suggest an important role of BH4 and SPR in age-related hypertension and a possible relationship with the cardiovascular instabilities in autonomic diseases, including Parkinson's disease and spinal cord injury.


Reversal learning after prenatal or early postnatal alcohol exposure in juvenile and adult rats.
Learning tasks that require the reversal of a previously learned contingency are disrupted in animals and humans exposed to alcohol during the perinatal period. The current experiments examined how varying the time of alcohol exposure and the age at which subjects were tested would affect the expression of reversal deficits in a T-maze task. Groups of rats were exposed to alcohol from gestational day (GD) 8 to GD20 or from postnatal day (PN) 4 to PN9, and then tested in a spatial reversal task at either PN28 or PN63. Results indicate that exposure to alcohol during the prenatal period did not lead to substantial dose-dependent reversal learning deficits in males or females at either age tested. However, exposure to alcohol during the early postnatal period, a period that corresponds to the third trimester in human neural development, selectively disrupted reversal learning performance in male rats at PN28 but not PN63. Statistically significant sex differences were seen when subjects were tested at PN63. These results demonstrate how the timing of alcohol exposure leads to variability in the age-dependent expression of learning deficits associated with fetal alcohol effects.


An unusual excitatory action of adenosine on the ventricular muscle of the South African clawed toad (Xenopus laevis).
The effects of adenyl compounds and some of their analogues were examined on atrial and ventricular muscle of Xenopus laevis. In contrast to all heart muscle preparations studied previously, adenosine produced excitation of the ventricle of Xenopus. 2-Chloroadenosine, ATP, beta, gamma-methylene ATP (APPCP) and alpha, beta-methylene ATP (APCPP) also elicited excitation of Xenopus ventricles; the order of potency being 2-chloroadenosine greater than ATP greater than or equal to adenosine greater than APPCP = APCPP. The excitatory effects of 2-chloroadenosine, adenosine, ATP and APPCP were antagonised by 8-phenylthe-ophylline (8-PT). A small excitatoty response to ATP persisted in the presence of 8-PT. The effects of adenosine were not potentiated by dipyridamole. Propranolol antagonised the excitatory responses to adrenaline, phenylephrine, 5-hydroxytryptamine (5-HT) and dopamine. High concentrations of histamine were needed to produce even small excitatory responses. Neither propranolol nor phentolamine affected the responses to adenosine and ATP. Adenosine, ATP and APPCP elicited inhibition of Xenopus atria which was antagonised by 8-PT but was not potentiated by dipyridamole. APCPP, which is resistant to degradation, did not produce a similar inhibitory response. It is concluded that excitation, mediated largely by excitatory P1- and possibly by some P2-purinoceptors, occurs in the Xenopus ventricle and that there are inhibitory P1-purinoceptors in the Xenopus atrium.


Prophylaxis of graft-versus-host disease by lentiviral-mediated expression of herpes simplex virus-thymidine kinase and ganciclovir treatment.
Our aim was to study whether herpes simplex virus-thymidine kinase/ganciclovir (HSV-TK/GCV)-mediated lentiviral gene transfer diminished graft-versus-host disease (GVHD) in a mouse model of allogeneic bone marrow transplantation (BMT). Donor splenic lymphocytes infected with lentiviral vectors carrying HSV-TK were mixed with donor bone marrow cells before cotransplantation into recipient mice irradiated with 60Co gamma-ray. GCV (25 mg/kg/d) was intraperitoneally administered beginning on days 0, 7, and 12 after transplantation for 7 days. The survival time, severity and incidence of GVHD, T-lymphocyte immune reconstitution, and percentage of allogeneic chimeras were observed after allo-BMT. The average survival times of mice in the TK/GCV 0 day, TK/GCV 7 day, and TK/GCV 12 day groups were 30.10 +/- 5.21, 36.40 +/- 5.28, and 28.20 +/- 4.82 days, respectively, all of which were longer than that of the control group (P < .05). The effect at 7 days was best: it was significantly different from the 0 and 12 day groups. The incidence of grade III to IV GVHD after allo-BMT in the control group was 100%, whereas there were pathological changes of grade II to III GVHD in the experimental groups. After allo-BMT, CD4+ and CD4+/CD8+ T lymphocytes in the experimental groups were all higher than in the control group (P < .05), but CD8+ T lymphocyte percentages were all lower than the latter. HSV-TK/GCV expression mediated by lentiviral transduction plays a role to prevent and treat GVHD after allo-BMT. The effect to control GVHD was most pronounced when GCV was administered on day 7 after transplantation.


Progesterone receptor concentration differences in the chick oviduct cells and apparent down-regulation by ligand. A semiquantitative immunohistochemical study.
A semiquantitative immunohistochemical technique was developed for identification of chick progesterone receptor (PR). The mouse monoclonal antibody PR6 was used. The nuclear PR concentration was analyzed with Leitz Orthoplan MPV-3 light microscope. The target tissue was chick oviduct, with epithelial, glandular, mesenchymal, smooth muscle and peritoneal cells analyzed separately. PR concentration varied between different cell types and also from cell to cell within a single cell type. A significant decrease of PR concentration, as noted by decrease in staining, was also observed in all studied cell types, 6 h after a single injection of progesterone. This technique allows for histological identification of biochemical events that should help lead to the understanding of the role of PR changes in a variety of experimental situations.


OFER GON, YUN-CHIH LIAO & SHAO KWANG-TSAO (2015) A new species of the cardinalfish genus Jaydia (Teleostei: Apogonidae) from the Philippines. Zootaxa, 3980 (2): 286-292.
Table 1, catalogue number of holotype: replace ASIZP 68202 with PNM 15192. Table 1, catalogue number of 34.9 mm paratype: replace CAS 236506 with USNM 435707. Table 1, catalogue number of 46.5 mm paratype: replace CAS 236506 with SAIAB 200712


Biological signal generators: integrating synthetic biology tools and in silico control.
Biological networks sense extracellular stimuli and generate appropriate outputs within the cell that determine cellular response. Biological signal generators are becoming an important tool for understanding how information is transmitted in these networks and controlling network behavior. Signal generators produce well-defined, dynamic, intracellular signals of important network components, such as kinase activity or the concentration of a specific transcription factor. Synthetic biology tools coupled with in silico control have enabled the construction of these sophisticated biological signal generators. Here we review recent advances in biological signal generator construction and their use in systems biology studies. Challenges for constructing signal generators for a wider range of biological networks and generalizing their use are discussed.


Dysfunctional Networks in Functional Dystonia.
Functional dystonia, the second most common functional movement disorder, is characterized by acute or subacute onset of fixed limb, truncal, or facial posturing, incongruent with the action-induced, position-sensitive, and task-specific manifestations of dystonia. We review neurophysiological and neuroimaging data as the basis for a dysfunctional networks in functional dystonia. Reduced intracortical and spinal inhibition contributes to abnormal muscle activation, which may be perpetuated by abnormal sensorimotor processing, impaired selection of movements, and hypoactive sense of agency in the setting of normal movement preparation but abnormal connectivity between the limbic and motor networks. Phenotypic variability may be related to as-yet undefined interactions between abnormal top-down motor regulation and overactivation of areas implicated in self-awareness, self-monitoring, and active motor inhibition such as the cingulate and insular cortices. While there remain many gaps in knowledge, further combined neurophysiological and neuroimaging assessments stand to inform the neurobiological subtypes of functional dystonia and the potential therapeutic applications.


Fracture risk assessment in patients with chronic kidney disease.
Fractures are common in patients with chronic kidney disease (CKD) and associated with substantially high morbidity and mortality. Bone mass measurements are commonly used to assess fracture risk in the general population, but the utility of these measurements in patients with CKD, and specifically among those on hemodialysis, is unclear. This review will outline the epidemiology and etiology of fractures in patients with CKD with a particular emphasis on men and women on hemodialysis. As well, we will summarize the published data, which describes the association between risk factors for fracture (including bone mass measurements, biochemical markers of mineral metabolism, and muscle strength) and fractures in patients with CKD. Patients with CKD suffer from fractures due to impairments in bone quantity, bone quality, and abnormalities of neuromuscular function. There is a paucity of evidence on the associations between bone quality, bone turnover markers, neuromuscular function, and fractures in patients with CKD. Furthermore, the complex etiology of fractures combined with the technical limitations of bone mineral density testing, both by dual energy X-ray absorptiometry (DXA) and by peripheral quantitative tomography (pQCT), limits the clinical utility of bone mass measurements for fracture prediction in CKD; this is particularly true among patients with stages 4 and 5 CKD. Further prospective studies to identify noninvasive measures of bone strength that can be used for fracture risk assessment are needed.


Further analysis of the antinociceptive action caused by p-methoxyl-diphenyl diselenide in mice.
The objective of this study was to extend our previous findings by investigating in greater detail the mechanisms that might be involved in the antinociceptive action of p-methoxyl-diphenyl diselenide, (MeOPhSe)(2), in mice. The pretreatment with nitric oxide precursor, l-arginine (600 mg/kg, intraperitoneal, i.p.), reversed antinociception caused by (MeOPhSe)(2) (10 mg/kg, p.o.) or N(G)-nitro-l-arginine (l-NOARG, 75 mg/kg, i.p.) in the glutamate test. Ondansetron (0.5 mg/kg, i.p., a 5-HT(3) receptor antagonist) and SCH23390 (0.05 mg/kg, i.p.., a D(1) receptor antagonist) blocked the antinociceptive effect caused by (MeOPhSe)(2). Conversely, pindolol (1 mg/kg, i.p., a 5-HT(1A)/(1B) receptor/beta adrenoceptor antagonist), WAY 100635 (0.7 mg/kg, i.p., a selective 5-HT(1A) receptor antagonist), ketanserin (0.3 mg/kg, i.p., a selective 5-HT(2A) receptor antagonist), prazosin (0.15 mg/kg, i.p., an alpha(1)-adrenoreceptor antagonist), yohimbine (1.0 mg/kg, i.p., an alpha(2)-adrenoreceptor antagonist), sulpiride (5 mg/kg, i.p., a D(2) receptor antagonist), naloxone (1 mg/kg, i.p., a non-selective opioid receptor antagonist) and caffeine (3 mg/kg, i.p., a non-selective adenosine receptor antagonist) did not change the antinociceptive effect of (MeOPhSe)(2). (MeOPhSe)(2) significantly inhibited nociception induced by intraplantar (i.pl.) injection of bradykinin (10 nmol/paw) and Des-Arg(9)-bradykinin (10 nmol/paw, a B(1) receptor agonist). (MeOPhSe)(2) significantly inhibited phorbol myristate acetate (PMA, 0.03 mug/paw, a protein kinase C (PKC) activator)-induced licking response. These results indicate that (MeOPhSe)(2) produced antinociception in mice through mechanisms that involve an interaction with nitrergic system, 5-HT(3) and D(1) receptors. The antinociceptive effect is related to (MeOPhSe)(2) ability to interact with kinin B(1) and B(2) receptors and PKC pathway mediated mechanisms.


A single-agent extension of the SIR model describes the impact of mobility restrictions on the COVID-19 epidemic.
Mobility restrictions are successfully used to contain the diffusion of epidemics. In this work we explore their effect on the epidemic growth by investigating an extension of the Susceptible-Infected-Removed (SIR) model in which individual mobility is taken into account. In the model individual agents move on a chessboard with a Lévy walk and, within each square, epidemic spreading follows the standard SIR model. These simple rules allow to reproduce the sub-exponential growth of the epidemic evolution observed during the Covid-19 epidemic waves in several countries and which cannot be captured by the standard SIR model. We show that we can tune the slowing-down of the epidemic spreading by changing the dynamics of the agents from Lévy to Brownian and we investigate how the interplay among different containment strategies mitigate the epidemic spreading. Finally we demonstrate that we can reproduce the epidemic evolution of the first and second COVID-19 waves in Italy using only 3 parameters, i.e , the infection rate, the removing rate, and the mobility in the country. We provide an estimate of the peak reduction due to imposed mobility restrictions, i. e., the so-called flattening the curve effect. Although based on few ingredients, the model captures the kinetic of the epidemic waves, returning mobility values that are consistent with a lock-down intervention during the first wave and milder limitations, associated to a weaker peak reduction, during the second wave.


A monoclonal antibody against a neuron-specific 65-kDa protein with laminar expression in the developing cerebral cortex.
The extracellular matrix component chondroitin sulfate supports the survival of neocortical neurons and influences their differentiation in culture. During development of the rat forebrain expression of chondroitin sulfate peaks at around birth. To elucidate functional partners of this important player of neurogenesis, a monoclonal antibody, termed MAb-9, was generated after immunization with chondroitin sulfate-binding proteins from neonatal rat brain. In western blots of neonatal tissue, MAb-9 recognized a major brain-specific 65-kDa protein band. While most of the 65-kDa protein was present in the soluble compartment, a significant fraction was membrane-associated. Prolonged extraction of brain membranes with CHAPS revealed three additional minor protein bands of approximately 48, 50, and 58 kDa. Of these, the 50-kDa protein specifically bound to chondroitin sulfate C-Sepharose. Immunocytochemical studies and western blot analyses of cultures of neocortical neurons and astrocytes demonstrated that MAb-9 recognizes a neuron-specific cytosolic protein. In the developing cerebral cortex the protein was detectable by immunohistochemistry in the preplate and ventricular zones as early as embryonic day 15. On embryonic day 18, the protein was found in the marginal zone and the subplate, but it was not present in the emerging cortical plate. At the neonatal stage the immunoreactivity was distributed throughout the cerebral cortex with prominent staining of the marginal zone. A similar pattern was observed in the adult animal. Notably, the laminar distribution of MAb-9 immunoreactivity in the cerebral cortex during the prenatal period closely resembled the expression pattern reported for chondroitin sulfate. Thus, MAb-9 recognizes a neuronal cytosolic protein which might participate in neurotrophic signaling events triggered by chondroitin sulfate.


Tosylhydrazine-promoted self-conjugate reduction-Michael/aldol reaction of 3-phenacylideneoxindoles towards dispirocyclopentanebisoxindole derivatives.
An efficient tosylhydrazine-mediated conjugate reduction of 3-phenacylideneoxindole and sequential Michael/intramolecular aldol reaction is reported under base-catalyzed conditions towards the formation of densely substituted dispirocyclopentanebisoxindole derivatives. The reaction proceeded in a diastereoselective manner to afford four chiral stereocenters. The method also has advantages of wide substrate scope, readily available starting materials and operational simplicity through one pot reaction.


[Bilateral carotid artery dissection in a severe preeclamptic setting: an unusual cause of postpartum headache].
A 30-year-old woman with severe preeclampsia presented at 27 weeks of amenorrhea with left headache, neck pain, blurred vision and numbness of left hemiface that resolved spontaneously within 2 hours. A week later, hypertension remained poorly controlled despite combination of nicardipine and labetalol intravenous therapy; an urgent caesarean section was eventually performed due to onset of HELLP syndrome. At day 5 postpartum, the patient had a Horner syndrome with right ipsilateral disabling tinnitus. A CT-angiography of supra-aortic trunks was performed urgently; it showed a bilateral carotid arterial dissection without stroke, which was subsequently confirmed by MRI angiography. The patient was transferred in neurovascular intensive care unit. Anticoagulant therapy was implemented to prevent cerebral and retinal ischemic lesions. Symptoms resolved quickly and the patient was discharged at day 7 postpartum. MR-angiography performed 4 months later showed a full resolution of the bilateral carotid dissection. Anticoagulant therapy was therefore discontinued.


[Is distal femoral replacement an adequate therapeutic option after complex fractures of the distal femur?].
Modular distal femur replacements originally were developed for reconstructing a full weight-bearing and functional extremity after resection of primary bone tumours with large bony defects located in proximity to joints. The aim of this study was to examine the use of the modular distal femur replacements for complex fractures and severe post-traumatic sequelae of the distal femur in context to comparable studies. Fourteen patients with complex fractures or post-traumatic sequelae and extensive bone defects requiring distal femur replacement were analysed retrospectively. Median age of the patients at the time of distal femoral replacement was 77 years and median follow-up interval was 27 months. Median follow-up was 27.0 (IQB 13.5-37.5) months (range 10-49 months). The indication for distal femur replacement was a periprosthetic fracture in three cases. Three further periprosthetic fractures were treated with a megaendoprosthesis after failure of osteosynthesis. In eight patients a megaendoprosthesis was implanted due to complications following ostheosynthesis for distal femoral fractures. Distal femoral arthroplasty was performed as a two-stage procedure in five patients with implant-associated infections. A lateral approach was used in six patients and a medial arthrotomy was conducted in eight patients. The median Knee Society score (KSS) improved significantly from 20.0 (IQB 7.5-30.0) points preoperatively to 80.0 (IQB 62.3-89.0) points at follow-up (p < 0.001). Complications requiring surgical intervention were documented in seven of 14 patients (50 %). In two patients wound-healing disorders and superficial infections necessitated surgery. In one patient a rupture of the patellar tendon was diagnosed. This patient subsequently also sustained a periprosthetic fracture. Another patient developed early aseptic loosening of the femoral component. The most common complication was a periprosthetic fracture in four patients. Three patients died for reasons not related to distal femoral replacement. Distal femoral replacement is an important option in reconstituting a full weight-bearing and functional lower extremity after complex fractures and post-traumatic sequelae with massive bone destruction. Particularly elderly patients regain ambulatory ability in the vast majority of cases. The relatively high complication rate demands very thorough preoperative planning as well as prompt allocation of extensive surgical procedures in the case of an adverse event.


Photoaffinity labeling of procaine-binding sites in normal and sickle cell membranes.
A photoaffinity probe, procaine azide, was employed to determine the sites of interaction of procaine in normal and sickle cell erythrocytes. Studies show that the number of binding sites and affinity of procaine to membranes derived from normal and sickled cell erythrocytes were similar, although procaine retards the in vitro formation of irreversibly sickled cells from cells, The results show that procaine azide, a photoaffinity analogue of procaine, is covalently incorporated into both protein (60--70%) and lipid (40--30%) components of the membrane. Sodium dodecyl sulfate-gel electrophoresis of the labeled ghosts show that procaine binds specifically to band 3 and periodic acid Schiff staining bands in membranes derived from labeled erythrocytes. Binding of procaine or covalent incorporation of procaine azide into membrane proteins does not affect the phosphate transport. Moreover, pre treatment of intact erythrocytes with 4,4-'diisothiocyano-2,2'-stilbene disulfonate, an anion transport inhibitor, did not affect either the binding or covalent incorporationof procaine azide into erythrocytes. These results indicate that the binding of procaine azide to Band 3 protein occurs at a locus different than that involved in anion translocation process.


Retention of mild asthmatics in the navy (REMAIN): a low-risk approach to giving mild asthmatics an opportunity for military service.
Rising U.S. asthma prevalence will be reflected in military applicants. We studied retaining mild asthmatics on active duty. A cohort study at Great Lakes Naval Training Center from 2000 to 2002 compared recruits diagnosed during basic training with mild asthma to matched comparison recruits on outpatient visits, hospitalizations, and discharge through August 2003. A total of 136 asthmatic and 404 control subjects were enrolled. Overall attrition was greater among the asthma cohort (p < 0.01), largely during training. Asthmatics used more health care than controls during training (0.1 vs. 0.004 per person-month). No asthma-related hospitalizations or deaths occurred during the study. Although attrition during recruit training was higher in mild asthmatics, nearly 40% of recruits were retained on active duty without significant risk of hospitalization or excessive outpatient treatment after recruit training. These findings argue for consideration of a trial on active duty for recruits with mild asthma.


In vivo drug resistance during treatment for typhoid fever: a case report.
A case of a 27 year old male who developed resistance to some previously sensitive antibiotics while still on treatment for salmonella typhi infection is presented. The possible mechanism for such a drug resistance is discussed.


Subacute steroid-induced paraparesis: surgical treatment of a devastating "invisible" side effect.
To emphasize an underestimated side effect following long-term use of steroids. We report on surgical treatment of two patients with serious neurologic deficits caused by epidural spinal lipoma following long-term intake of cortisone. Early decompression of the spinal cord by removal of epidural lipoma was the most effective treatment in these patients with progressive symptoms. Diagnostic work-up of such patients should include early spinal MRI resulting in surgical intervention, if indicated. Decompression of the spinal cord eventually combined with fusion is necessary.


Intrahepatic IL-8 producing Foxp3⁺CD4⁺ regulatory T cells and fibrogenesis in chronic hepatitis C.
Regulatory CD4(+) T cells (Tregs) are considered to affect outcomes of HCV infection, because they increase in number during chronic hepatitis C and can suppress T-cell functions. Using microarray analysis, in situ immunofluorescence, ELISA, and flowcytometry, we characterised functional differentiation and localisation of adaptive Tregs in patients with chronic hepatitis C. We found substantial upregulation of IL-8 in Foxp3(+)CD4(+) Tregs from chronic hepatitis C. Activated GARP-positive IL-8(+) Tregs were particularly enriched in livers of patients with chronic hepatitis C in close proximity to areas of fibrosis and their numbers were correlated with the stage of fibrosis. Moreover, Tregs induced upregulation of profibrogenic markers TIMP1, MMP2, TGF-beta1, alpha-SMA, collagen, and CCL2 in primary human hepatic stellate cells (HSC). HSC activation, but not Treg suppressor function, was blocked by adding a neutralizing IL-8 antibody. Our studies identified Foxp3(+)CD4(+) Tregs as an additional intrahepatic source of IL-8 in chronic hepatitis C acting on HSC. Thus, Foxp3(+)CD4(+) Tregs in chronic hepatitis C have acquired differentiation as regulators of fibrogenesis in addition to suppressing local immune responses.


Simplified gonadotropin-releasing hormone (GnRH) stimulation test.
Provocative gonadotropin-releasing hormone (GnRH) stimulation testing indirectly assesses testicular function with more sensitivity than determination of basal gonadotropin levels alone. Unfortunately, the drawbacks of multiple blood sampling and high cost have limited the clinical usefulness of this test. We herein present a simplified, two-point, thirty-minute GnRH stimulation test. Statistical analysis of data from 55 men with normal baseline gonadotropin levels, reveal that this simplified test is just as accurate as the traditional test (p less than 0.0001) without the latter's attendant difficulties. In addition, we found that normal basal gonadotropin levels had little correlation to the actual responses obtained from GnRH stimulation testing (r = 0.20 and r = 0.39 for luteinizing hormone and follicle-stimulating hormone, respectively).


Adaptation to cyclic stance perturbations in Parkinson's disease depends on postural demands.
The postural reactions of 10 moderate PD patients and 10 age matched controls were studied during stance on a sinusoidally back and forth moving platform during 5 blocks of 10, 1-min trials. Free stance was followed by weight bearing, lowering or raising the center of gravity (COG) by 10%. Next, a book was balanced on the head and finally, forward inclination was constrained. Normal stance strategy, including predictive muscle responses was found during free stance. Modifying COG height caused little response changes. Stance strategy became abnormal when head trajectory or forward inclination was restricted, indicating that ability to generate a nonstandard multi-joint stance strategy, rather than production of adequate stabilizing forces, is impaired in moderate, medicated PD patients.


Regional comparison of the lumped constants of deoxyglucose and fluorodeoxyglucose.
We determined the regional relationships of the lumped constants for deoxyglucose (DG) and fluorodeoxyglucose (FDG). Awake male rats were given simultaneous intravenous injections of [14C]DG and [18F]FDG, and sacrificed after 45 min. The brains were removed and small pieces of cortical tissue were sampled by dissection. The remainder of the brains were frozen and sliced into 20-microns-thick sections that were placed on coverslips and dried on a hot plate. Two sets of brain autoradiograms were prepared from the brain sections, one representing predominantly [14C] and the other [18F]. The autoradiograms were digitized, and after correcting for cross-contamination, tracer-concentration images of the DG and FDG were generated. Based on relative tracer concentrations in the tissue samples, the lumped constant of FDG was found to be 1.25 times that of DG, or 0.6. Using this value, images of glucose metabolism for the DG and FDG were generated from the tracer concentration images and compared. No significant regional differences were found, indicating that the relationship between the lumped constant of DG and that of FDG was stable throughout the normal brain.


Exercise-induced bronchoconstriction.
To review the literature regarding the pathophysiology of exercise-induced bronchoconstriction (EIB). The databases of PubMed, Ovid MEDLINE, and Scopus were searched for articles using the subject headings and/or keywords asthma, exercise-induced/etiology, exercise, mechanism, pathogenesis, and bronchoconstriction. Articles were selected based on their relevance to the focus of this review, with emphasis on the specific pathophysiologic mechanisms of EIB. EIB occurs in response to the loss of water from the lower airways that results from heating and humidifying large volumes of air in a short period. The resulting hyperosmolar environment activates various cellular mechanisms to release mediators from mast cells, eosinophils, epithelial cells, and sensory nerves. These mediators, in turn, lead to airway smooth muscle contraction and bronchoconstriction. Airway hyperresponsiveness in elite athletes may develop from a process of airway injury and changes in the contractile properties of airway smooth muscle. EIB commonly affects individuals with and without clinically recognized asthma, especially those who participate in competitive athletics. Through years of research, the pathophysiology of EIB is now better understood and involves a complex interaction between several different cell types and mediators. Continued research to improve the knowledge regarding the mechanisms of EIB should aid the identification, diagnosis, and treatment of this common condition.


Individual-level trait diversity predicts phytoplankton community properties better than species richness or evenness.
Understanding how microbial diversity influences ecosystem properties is of paramount importance. Cellular traits-which determine responses to the abiotic and biotic environment-may help us rigorously link them. However, our capacity to measure traits in natural communities has thus far been limited. Here we compared the predictive power of trait richness (trait space coverage), evenness (regularity in trait distribution) and divergence (prevalence of extreme phenotypes) derived from individual-based measurements with two species-level metrics (taxonomic richness and evenness) when modelling the productivity of natural phytoplankton communities. Using phytoplankton data obtained from 28 lakes sampled at different spatial and temporal scales, we found that the diversity in individual-level morphophysiological traits strongly improved our ability to predict community resource-use and biomass yield. Trait evenness-the regularity in distribution of individual cells/colonies within the trait space-was the strongest predictor, exhibiting a robust negative relationship across scales. Our study suggests that quantifying individual microbial phenotypes in trait space may help us understand how to link physiology to ecosystem-scale processes. Elucidating the mechanisms scaling individual-level trait variation to microbial community dynamics could there improve our ability to forecast changes in ecosystem properties across environmental gradients.


Calcium dependency of the endothelium-dependent hyperpolarization in smooth muscle cells of the rabbit carotid artery.
1. In smooth muscle cells of the rabbit carotid artery, ACh (greater than 10(-8) M) generated a hyperpolarization with two components (transient followed by sustained), only in the tissues with an intact endothelium. There were no detectable changes in the membrane potential, as elicited by ACh (up to 10(-5) M) in tissues with no endothelium or in the presence of atropine (10(-6) M). 2. Reduction of [Ca2+]o inhibited the sustained component which was not apparent in [Ca2+]o below 0.16 mM. In Ca2+-free (EGTA-containing) solution, the generation of the transient component of the hyperpolarization remained sustained but with a substantially reduced amplitude. 3. Procaine (greater than 10(-6) M) inhibited the ACh-induced hyperpolarization in a concentration-dependent manner, and at a concentration of procaine (10(-3) M) which caused substantial depolarization of the membrane, no detectable change was elicited by ACh. 4. Caffeine (10(-6)-10(-3) M) produced a transient hyperpolarization, independent of the presence or absence of the endothelium, and inhibited the sustained component of the ACh-induced hyperpolarization more so than the initial component. 5. A23187 (greater than 10(-8) M) hyperpolarized the smooth muscle membrane in a concentration-dependent manner, and this hyperpolarization was not generated in Ca2+-free solution or in the absence of endothelial cells. 6. In intact tissues, pre-treatment with A23187 resulted in a reduction of the subsequently generated ACh-induced hyperpolarization, in an irreversible manner. 7. It would thus appear that in the rabbit carotid artery, the endothelium-dependent hyperpolarization induced by ACh has Ca2+-dependent and Ca2+-independent components, and each may be related to the increase in endothelial [Ca2+]i by release from the intracellular store and by influx from the extracellular medium, respectively. The increased [Ca2+]i would trigger a release of an endothelium-derived hyperpolarizing factor (EDHF) from the endothelial cells.


Bile acid profiles in neonatal intrahepatic cholestasis caused by citrin deficiency.
Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is characterized by conjugated hyperbilirubinemia and increased plasma bile acid concentrations. However, the underlying mechanisms remain unclear. We established a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for simultaneously quantifying plasma bile acids and examined bile acid profiles in NICCD infants. We measured 15 bile acids within 15min and found a wide linear range for individual bile acids. The within-run and run-to-run CV of all bile acids was 1.2-10.9% and 3.1-10.8%, respectively, with a mean recovery of 90.5-112.6%. Compared to infants with citrullinemia without mutations in SLC25A13 (non-NICCD), NICCD infants showed increased plasma total bile acid concentrations (mean: 201 vs. 42μM, p<0.001), with a distinct bile acid profile characterized by increased conjugated primary bile acid concentrations. The calculated ratios, including primary/secondary bile acid (714 vs. 235, p<0.05) and conjugated/free bile acid (371 vs. 125, p<0.05) ratios, were higher in NICCD infants. The area under receiver operating characteristic curve for conjugated/free bile acid ratio to identify infants with NICCD was 0.871 (95% confidence interval, 0.713-1.0). Together, our findings indicated plasma bile acid profile as a potential noninvasive diagnostic biomarker for NICCD.


Vasovasostomy and vas occlusion: preliminary observations using artificial devices in guinea pigs.
Intravas devices designed to reverse a vasectomy or to occlude the vasa instead of transecting them were evaluated in guinea pigs. Absorbable intravas devices (AID) were inserted into the vasa at the time of vasovasostomy and produced a return of normal semen quality in 73% of the experimental animals as compared with only 40% in the control group. Seventy per cent of the AID group sired litters. In a second group of guinea pigs, vasectomy was replaced by the bilateral insertion of reversible intravas plugs (RIP) which occluded the lumina. After a period of azoospermia, the central, occlusive pins were removed from the devices. Sperm-positive animals were bred and three of five sired normal offspring. Histologic examination of the vasa revealed morphologic changes in some AID animals, these being attributed to the vasectomy procedure. A similar evaluation of the functional RIP insertion site revealed the desired occlusive tissue-device interfaces.


Evaluation of the hemagglutination test for epidemiologic studies of leptospiral antibodies in wild mammals.
Sera from 153 wild animals of 18 species were tested for antibodies against 12 serovars of Leptospira by the microscopic agglutination (MA) test. Seventy-five percent of the animals tested were seropositive against one or more of the 12 serovars used. The most commonly found serovars were pomona, autumnalis, pyrogenes, icterohaemorrhagiae, australis, and canicola. Of 62 carnivores representing 7 species, 55 (89%) were seropositive, as were 46 (60%) of 77 rodents from 9 species. Leptospira of the serovar copenhageni serogroup icterohaemorrhagiae were recovered from kidney tissues of a Norway rat (Rattus norvegicus). Of 443 wildlife sera tested by the indirect hemagglutination (IHA) test using cells sensitized with L. illini antigen, 47 (11%), mainly carnivores and deer, gave a heterophile reaction. Of the remaining 396 sera, 164 (41%) were seropositive for leptospirosis by the IHA test. To compare the IHA test with the MA test, 143 serum samples were tested by both methods. There was 84% concordance between the two tests.


How the comparison of age-related macular degeneration treatments trial results will impact clinical care.
To provide a perspective on the impact of the Comparison of Age-related Macular Degeneration Treatments Trial (CATT) on future clinical practices. Interpretation of trial outcomes relative to clinical use of neovascular age-related macular degeneration (AMD) treatments, assessment of the influence of study design and execution on results, and review of unanalyzed safety data in the online supplement. Expert opinion. The CATT study supports the selection of either ranibizumab or bevacizumab for treatment of AMD based on factors other than efficacy, such as cost, because monthly administration of bevacizumab was noninferior to the reference treatment of monthly ranibizumab in improving visual acuity at 1 year. Visual acuity results for bevacizumab as needed were inconclusive for noninferiority relative to monthly administration of either drug. The secondary outcome of decrease in thickness at the foveal center as measured by time-domain optical coherence tomography significantly favored the monthly ranibizumab group vs the bevacizumab-as-needed group but is more difficult to interpret as it did not correlate with visual acuity and is less appropriate for a noninferiority design. Bevacizumab groups had a statistically higher observed risk of serious adverse events; however, scrutiny of the online supplements shows similar numbers of cardiac and neurologic events in bevacizumab and ranibizumab users. Information regarding fellow eye treatment with anti-VEGF agents was not given. CATT provides the first level I evidence for bevacizumab in a large number of patients with neovascular AMD. The trial supports use of either drug as primary therapy and suggests that modification of monthly dosing regimens is feasible. A difference in cardiovascular safety between the 2 drugs was not apparent on inspection of the supplementary safety data.


Connecting macroscopic diffusion metrics of cardiac diffusion tensor imaging and microscopic myocardial structures based on simulation.
To investigate the relationship between microscopic myocardial structures and macroscopic measurements of diffusion tensor imaging (DTI), we proposed a cardiac DTI simulation method using the Bloch equation and the Monte Carlo random walk in a realistic myocardium model reconstructed from polarized light imaging (PLI) data of the entire human heart. To obtain a realistic simulation, with the constraints of prior knowledge pertaining to the maturational change of the myocardium structure, appropriate microstructure modeling parameters were iteratively determined by matching DTI simulations and real acquisitions of the same hearts in terms of helix angle, fractional anisotropy (FA) and mean diffusivity (MD) maps. Once a realistic simulation was obtained, we varied the extra-cellular volume (ECV) ratio, myocyte orientation heterogeneity and myocyte size, and explored the effects of microscopic changes in tissue structure on macroscopic diffusion metrics. The experimental results demonstrated the feasibility of simulating the DTI of the whole heart using PLI measurements. When varying ECV from 15% to 55%, mean FA decreased from 0.55 to 0.26, axial diffusivity increased by 0.6 μm2/ms, and radial diffusivity increased by 0.7 μm2/ms. When orientation heterogeneity was varied from 0 to 20∘, mean FA decreased from 0.4 to 0.3, axial diffusivity decreased by 0.08 μm2/ms, and radial diffusivity increased by 0.03 μm2/ms. When mean diameter of myocytes was varied from 6 μm to 10 μm, FA decreased from 0.67 to 0.46, axial and radial diffusivities increased by 0.05 and 0.2 μm2/ms, respectively.


Inhibition of hedgehog signalling attenuates UVB-induced skin photoageing.
The hedgehog (Hh) signalling pathway regulates normal development and cell proliferation in metazoan organisms, but its aberrant activation can promote tumorigenesis and progression of a variety of aggressive human cancers including skin cancer. Despite its importance, little is known about its role in photoageing, a type of UV-induced skin lesions. In this study, we investigated the involvement of Hh signalling in the photoageing process as well as the use of an Hh-regulating alkaloid compound as a novel therapeutic drug to regulate photoageing in keratinocytes. We found that UVB induced Hh signalling by the expression of Hh ligands and Hh-mediated transcription factors, respectively. Moreover, UVB-induced Hh activation relied on mitogen-activated protein kinase (p38, ERK and JNK) activity and inflammatory responses (upregulation of COX-2, IL-1β, IL-6 and TNF-α), resulting in premature senescence and photoageing in vitro and in vivo. Notably, a selected Hh inhibitor, evodiamine, mediated photoageing blockade in a mouse skin model. Taken together, our findings demonstrated that Hh signalling is associated with UVB-induced photoageing, while pharmacological inhibition of Hh signalling significantly reduced experimental photoageing, indicating its potential for use as a therapeutic target for this disease.


Chemical profiling of Justicia vahlii Roth. (Acanthaceae) using UPLC-QTOF-MS and GC-MS analysis and evaluation of acute oral toxicity, antineuropathic and antioxidant activities.
Justicia vahlii Roth. (Acanthaceae), also called as kodasoori and bhekkar is an annual therophyte erect or decumbent herb used traditionally in toothache, skin diseases (itching, topical inflammation) and for the treatment of various respiratory disorders. The current study aimed at exploring pain cessation potential of J. vahlii Roth. via murine model of neuropathic pain and its phytochemical, toxicological and antioxidant profiles. The hydro-alcoholic extract of J. vahlii (HAEJv) prepared by maceration technique was subjected to preliminary phytochemical screening, total bioactive content determination, UPLC-QTOF-MS and GC-MS analysis. Toxicity assessment was carried out by using brine shrimp lethality assay and acute oral toxicity test. Murine model of neuropathic pain was applied to assess the antineuropathic potential of the species. Furthermore effect of the extract on catalase, superoxide oxide dismutase (SOD), Glutathione (GSH), interleukin-1beta (IL-1β) and total necrosis factor-alpha (TNF-α) was also studied. In vitro antioxidant profile was explored by using four methods; 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis(3-ethylbenothiazoline)-6-sulfonic acid (ABTS), CUPric reducing antioxidant capacity (CUPRAC) and Ferric reducing antioxidant power (FRAP) assay. The phytochemical screening revealed the presence of phenols, flavonoids, coumarins, alkaloids and lignans as the major classes of secondary metabolites. The extract was found rich in total phenolics content (TPC) and total flavonoids content (TFC) with identification of total 59 bioactives in UPLC-QTOF-MS and 40 compounds in GC-MS analysis. The extract was found nontoxic up to 4000 mg/kg (p.o.) in mice and no mortality observed in brine shrimp lethality assay. The HAEJv significantly reduced number of acetic acid induced abdominal constrictions at 100 mg/kg (p < 0.01) and 200 mg/kg (p < 0.001) and increased paw withdrawal threshold p < 0.05 at 100 mg/kg and p < 0.001 at 200 mg/kg, and an increase in tail withdrawal latency time p < 0.001 at 200 mg/kg was observed. The extract significantly increased levels of catalase, SOD and GSH while decreased IL-1β and TNF-α levels in sciatic nerve tissue of mice. HAEJv showed highest antioxidant activity through CUPRAC method 121.32 ± 1.22 mg trolox equivalent per gram of dry extract (mg TE/g DE) followed by DPPH 81.334 ± 4.35 mg TE/g DE, FRAP 69.89 ± 3.05 mg TE/g DE and ABTS 38.17 ± 2.12 mg TE/g DE. The current study back the traditional use of J. vahlii in pain cessation through antioxidant based antineuropathic pain activity and revealed the extract non-toxic with number of functional phytoconstituents and warrants further research on isolation of the compounds and sub-acute toxicity studies.


HIV-1 drug resistance profiling using amino acid sequence space cartography.
Human immunodeficiency virus (HIV) drug resistance is a global healthcare issue. The emergence of drug resistance influenced the efficacy of treatment regimens, thus stressing the importance of treatment adaptation. Computational methods predicting the drug resistance profile from genomic data of HIV isolates are advantageous for monitoring drug resistance in patients. However, existing computational methods for drug resistance prediction are either not suitable for emerging HIV strains with complex mutational patterns or lack interpretability, which is of paramount importance in clinical practice. The approach reported here overcomes these limitations and combines high accuracy of predictions and interpretability of the models. In this work, a new methodology based on generative topographic mapping (GTM) for biological sequence space representation and quantitative genotype-phenotype relationships prediction purposes was introduced. The GTM-based resistance landscapes allowed us to predict the resistance of HIV strains based on sequencing and drug resistance data for three viral proteins [integrase (IN), protease (PR) and reverse transcriptase (RT)] from Stanford HIV drug resistance database. The average balanced accuracy for PR inhibitors was 0.89 ± 0.01, for IN inhibitors 0.85 ± 0.01, for non-nucleoside RT inhibitors 0.73 ± 0.01 and for nucleoside RT inhibitors 0.84 ± 0.01. We have demonstrated in several case studies that GTM-based resistance landscapes are useful for visualization and analysis of sequence space as well as for treatment optimization purposes. Here, GTMs were applied for the in-depth analysis of the relationships between mutation pattern and drug resistance using mutation landscapes. This allowed us to predict retrospectively the importance of the presence of particular mutations (e.g. V32I, L10F and L33F in HIV PR) for the resistance development. This study highlights some perspectives of GTM applications in clinical informatics and particularly in the field of sequence space exploration. https://github.com/karinapikalyova/ISIDASeq. Supplementary data are available at Bioinformatics online.


Preparation of N,O-carboxymethyl chitosan coated alginate microcapsules and their application to Bifidobacterium longum BIOMA 5920.
In order to greatly improve vitality of probiotic bacteria within the application, a novel biocompatible vehicle, N,O-carboxymethyl chitosan (NOCs) with appropriate degrees of substitution coat alginate (ALg) microparticles, was prepared by electrostatic droplet generation. The amount of chitosan (Cs) and N,O-carboxymethyl chitosan (NOCs) coated on the ALg microparticles was determined by differential scanning calorimetry. The surface morphology of ALg microparticles, Cs coated ALg microparticles and NOCs coated ALg microparticles was determined using scanning electron microscopy. The coating thickness of Cs coated ALg microparticles and that of NOCs coated ALg microparticles was directly observed with confocal laser scanning microscopy. In order to assess pH sensitivity of microparticles, the bovine serum albumin release from the microspheres was tested in acid solution (pH 2.0) for 2 h and subsequently in alkaline solution (pH 7.0) for 2 h. The survival of Bifidobacterium longum BIOMA 5920 loaded in NOCs coated with ALg microparticle was improved in simulated gastric juice (pH 2.0, for 2 h) compared to that of B. longum BIOMA 5920 loaded in ALg microparticles and Cs coated ALg microparticles. After incubation in simulated intestinal juices (pH 7.0, 2 h), the release of microencapsulated B. longum BIOMA 5920 was investigated.


The spectrum of endocrinocarcinomas of the larynx.
The authors discuss the terminology, classification, pathology, diagnosis, treatment and prognosis of the endocrinocarcinomas of the larynx.


Cyclin D1 gene polymorphism and familial prostate cancer: the AA genotype of A870G polymorphism is associated with prostate cancer risk in men aged 70 years or older and metastatic stage.
Cyclin D1 is a key protein involved in cell cycle regulation. A common A870G single nucleotide polymorphism in exon 4 of the cyclin D1 gene (CCND1) has an effect on the transcription of two different cyclin D1 mRNAs. Association of the genetic polymorphism of A870G of CCND1 with several cancer risks has been reported. We performed a case-control study consisting of 99 familial prostate cancer cases and 115 noncancer controls to examine the genotypic frequency of the CCND1 polymorphism. We did not observe a significant association of the genotypic frequency of CCND1 with overall cases and controls. However, the AA genotype of CCND1 showed a tendency to increase prostate cancer risk in subjects aged 70 years or older (odds ratio [OR] = 2.14, 95% confidence interval [CI] = 0.86-5.27, p = 0.096). Stratification of cases according to clinical stages showed that the AA genotype was significantly frequently observed in metastatic cancer in comparison with GG + AG genotypes (OR = 3.03, 95% CI = 1.11-8.23, p = 0.026). The present study suggested that the genetic polymorphism might be associated with familial prostate cancer risk in a Japanese population.


Analysis of cell-cycle checkpoint pathways in head and neck cancer cell lines: implications for therapeutic strategies.
To determine the mechanism of action of paclitaxel (Taxol) and carboplatin in cell lines of head and neck squamous cell carcinoma (HNSCC). Four HNSCC cell lines were treated with paclitaxel and carboplatin, alone or in combination, and evaluated for cell-cycle position by means of flow cytometry, for molecular determinants of cell cycle by means of Western blotting and kinase analysis, and for anchorage-independent growth by means of soft-agar assays. Paclitaxel was more effective at inducing apoptosis and inhibiting anchorage-independent cell growth, compared with carboplatin. The activity of paclitaxel was correlated with an elevation of cyclin B1/CDC2 activity, prolonged mitotic arrest, and Bcl-2 phosphorylation. In contrast, carboplatin arrested cells before mitosis. Combination treatment with both agents, simultaneously or sequentially, was more effective at inhibiting cell growth than either single agent. Cellular outcome was the same regardless of which drug was used first. The order of addition of these 2 drugs differentially affected cell-cycle position. Paclitaxel pretreatment arrested cells in mitosis, whereas carboplatin pretreatment or cotreatment resulted in premitotic arrest. To our knowledge, this study is the first to explore how paclitaxel and carboplatin, alone or in combination, differentially affect cell-cycle checkpoint response and HNSCC cell growth. These results provide molecular validation for the current clinical use of both drugs in combination and set the stage for analyses of patient tumor specimens.


Experiments, molecular dynamics simulations, and quantum chemistry calculations on the effect of gemini surfactants' headgroup on the oil-water interfacial tension.
The effect of the Gemini surfactant headgroup on the oil-water interfacial tension has yet to be systematically revealed. In this work, anionic Gemini surfactants with different hydrophilic headgroups (carboxylic, sulfuric, and sulfonic) were designed and synthesized. The oil-water interfacial tension was tested. The essential parameters for evaluating the interface characteristics, including the oil-water interfacial layer thickness, the coordination number, and the diffusion coefficient, were calculated employing molecular dynamics simulation. The surface electrostatic potential explained the quantitative mechanism of the hydrophobicity and lipophilicity of three types of Gemini surfactants through quantum chemical calculations. The oil-water interfacial tension difference of the Gemini surfactants was revealed at the electronic level. This paper will provide theoretical guidance for designing Gemini surfactants with a high-efficiency performance to enhance oil recovery.


In vitro activities of new quinolones and oxazolidinones against Actinomadura madurae.
In this work, we studied the in vitro sensitivity of 24 strains of Actinomadura madurae to a new oxazolidinone (DA-7867), gatifloxacin, moxifloxacin, and garenoxacin by using a broth microdilution method. We observed that the A. madurae strains had a high level of sensitivity to all the antimicrobials tested. The most active drug was DA-7867, with a MIC at which 90% of the strains are inhibited (MIC(90)) of 0.125 micro g/ml and a MIC(50) of 0.06 micro g/ml.


The plasma lncRNA acting as fingerprint in non-small-cell lung cancer.
Recent studies have indicated that long non-coding RNAs (lncRNAs) could act as non-invasive tumor markers in both diagnosis and predicting the prognosis. In this study, we focused to determine the expression of circulating lncRNAs in patients suffering from non-small-cell lung cancer (NSCLC), aiming to found the potential lncRNA as predictor. Twenty-one lncRNAs which previously identified were selected as candidate targets for subsequent circulating lncRNA assay. The candidate lncRNAs were validated by qRT-PCR arranged in the training and validation sets. Circulating SPRY4-IT1, ANRIL, and NEAT1 were significantly increased in plasma samples of NSCLC patients during training set and validation set. Receiver operating characteristic curve (ROC) analysis revealed that plasma ANRIL provided the highest diagnostic performance with an area under ROC curve value (AUC) of 0.798. Further combination with the three factors indicated a higher power (AUC, 0.876; sensitivity, 82.8 %; specificity, 92.3 %). The stableness detection of the three factors indicated that circulating SPRY4-IT1, ANRIL, and NEAT might serve as a predictor for the early warning of non-small-cell lung cancer.


Pharmaceutical subsidy policy in Iran: a qualitative stakeholder analysis.
Over the past three decades, allocation of foreign currency subsidies has been the primary strategy of various administrations in Iran to improve access to medicines. This strategy has resulted in several challenges, including stakeholder conflicts of interest. To identify the power, interest, and role of the stakeholders in allocating foreign currency subsidies to medicines in the Iranian health system. In this qualitative study, 39 semi-structured interviews were conducted. Key informants were recruited using a purposive sampling technique. The theoretical framework adopted by Varvasovszky and Brugha was employed. The data were analysed using directed content analysis. The foreign currency subsidy for medicines included 21 stakeholders in five main categories: governmental organizations, Iranian Parliament, general population, nongovernmental organizations (NGOs), and the pharmaceutical industry. Stakeholders varied in their level of participation and support in the policy-making process. Among them, the Iranian Government, Planning and Budget Organization, the Ministry of Health and Medical Education (MoHME), and Iran Food and Drug Administration (IFDA) were the most important stakeholders, with highly supportive positions, while domestic drug manufacturers were the strongest opponents of this policy. The Government of Iran is the most powerful institution with regard to the ability to allocate foreign currency subsidies to medicines, followed by the MoHME and the IFDA. This study demonstrated that identifying and analysing the stakeholders involved in allocating foreign currency subsidies to medicines can provide valuable information for policy-makers to enable a more comprehensive understanding and better capacity to determine whether or not to eliminate these subsidies. Moreover, decision-making in this process is a long-term issue that requires consensus among all stakeholders. Because of the political and social consequences of eliminating foreign currency subsidies, the necessary political will is not institutionalized. We recommend a step-by-step approach in eliminating foreign currency subsidies if the requirements are met (i.e., those related to the consequences of such interventions). Therefore, revision of the current policy along with these requirements, in addition to financial transparency and enhanced efficiency, will facilitate progress towards achieving the Sustainable Development Goals by improving access to medicines.


Apples or candy? Internal and external influences on children's food choices.
The goal of this concise narrative review is to examine the current literature regarding endogenous and exogenous influences on youth food choices. Specifically, we discuss internal factors such as interoception (self-awareness) of pain and hunger, and neural mechanisms (neurofunctional aspects) of food motivation. We also explore external factors such as early life feeding experiences (including parenting), social influences (peers), and food marketing (advertising). We conclude with a discussion of the overlap of these realms and future directions for the field of pediatric food decision science.


Effect of pelleting on the enzymatic digestibility of corn stover.
Pelleting of lignocellulosic biomass to improve its transportation, storage and handling impacts subsequent processing and conversion. This work reports the role of high moisture pelleting in the enzymatic digestibility of corn stover prior to pretreatment, together with associated substrate characteristics. Pelleting increases the digestibility of unpretreated corn stover, from 8.2 to 15.5% glucan conversion, at 5% solid loading using 1 FPU Cellic® CTec2 per g solids. Compositional analysis indicates that loose and pelleted corn stover have similar non-dissolvable compositions, although their extractives are different. Enzymatic hydrolysis of corn stover after size reduction to normalize particle sizes and removal of extractives confirms that pelleting improves corn stover digestibility. Such differences may be explained by the decreased particle size, improved substrate accessibility, and hydrolysis of cross-linking structures induced by pelleting. These findings are useful for the development of processing schemes for sustainable and efficient use of lignocellulose.


New zwitterionic butanesulfonic acids that extend the alkaline range of four families of Good buffers: evaluation for use in biological systems.
Four new zwitterionic butanesulfonic acid buffers that are structurally related to four families of Good buffers were evaluated for use in biological systems. These buffers, with pKa values from 7.6 to 10.7, were compared with a variety of other buffers from the same family and with unrelated buffers to determine their effect on enzyme activity and on microbial growth. The activity of four enzymes with optimum pH values in the alkaline range were tested: beta-galactosidase, esterase, phosphodiesterase and alkaline phosphatase. In general, all the Good buffers, including the new butanesulfonic acid buffers, gave good activity; however, there was variation in activity of certain enzymes with certain buffers. Tris, glycine, and phosphate buffers typically showed variation in activity compared to the family of Good buffers. beta-Galactosidase, in particular, showed greater activity with Good buffers than with phosphate or Tris buffers. Similarly, growth of seven bacterial strains was consistent, with a few exceptions, for all the Good family of buffers with Tris often inhibiting growth. Quantitation of alkaline phosphatase conjugated to antibodies is an important tool in many applications in molecular biology. Several Good buffers gave good signals when compared with Tris at pH 9.5 for detection of proteins using alkaline phosphatase-conjugated antibodies.


14-item resilience scale (RS-14): psychometric properties of the Brazilian version.
The Resilience Scale (RS) was developed to evaluate the levels of resilience in the general population. Its reduced version (RS-14) has presented reliable internal consistency and external validity. However, its psychometric properties have not been systematically evaluated. The objective of this study was to present the psychometric properties of the Brazilian RS-14. A total of 1,139 subjects selected by convenience (62.9% women) from 14 to 59 years old (M = 26.1, SD = 11.61) participated in the study. Exploratory factor analyses (EFAs) and parallel analysis were conducted in order to assess the factor structure of the scale. A 13-item single-factor solution was achieved. Confirmatory factor analyses (CFA) and multigroup CFA (MGCFA) corroborated the goodness of fit and measurement invariance of the obtained exploratory solution. The levels of resilience correlated negatively with depression and positively with meaning in life and self-efficacy.


Guidance of engineered tissue collagen orientation by large-scale scaffold microstructures.
The tensile strength and stiffness of load-bearing soft tissues are dominated by their collagen fiber orientation. While microgrooved substrates have demonstrated a capacity to orient cells and collagen in monolayer tissue culture, tissue engineering (TE) scaffolds are structurally distinct in that they consist of a three-dimensional (3-D) open pore network. It is thus unclear how the geometry of these open pores might influence cell and collagen orientation. In the current study we developed an in vitro model system for quantifying the capacity of large scale ( approximately 200 microm), geometrically well-defined open pores to guide cell and collagen orientation in engineered tissues. Non-degradable scaffolds exhibiting a grid of 200 microm wide rectangular pores (1:1, 2:1, 5:1, and 10:1 aspect ratios) were fabricated from a transparent epoxy resin via high-resolution stereolithography. The scaffolds (n=6 per aspect ratio) were surface modified to support cell adhesion by covalently grafting GRGDS peptides, sterilized, and seeded with neonatal rat skin fibroblasts. Following 4 weeks of static incubation, the resultant collagen orientation was assessed quantitatively by small angle light scattering (SALS), and cell orientation was evaluated by laser confocal and scanning electron microscopy. Cells adhered to the struts of the pores and proceeded to span the pores in a generally circumferential pattern. While the cell and collagen orientations within 1:1 aspect ratio pores were effectively random, higher aspect ratio rectangular pores exhibited a significant capacity to guide global cell and collagen orientation. Preferential alignment parallel to the long strut axis and decreased spatial variability were observed to occur with increasing pore aspect ratio. Intra-pore variability depended in part on the spatial uniformity of cell attachment around the perimeter of each pore achieved during seeding. Evaluation of diamond-shaped pores [Sacks, M.S. et al., 1997. J. Biomech. Eng. 119(1), 124-127] suggests that they are less sensitive to initial conditions of cell attachment than rectangular pores, and thus more effective in guiding engineered tissue cell and collagen orientation.


Do silicone breast implants affect breastfeeding?
One of my patients has silicone breast implants. She gave birth to a healthy baby boy but is afraid to breastfeed because she read in a magazine that the baby might be affected by silicone secreted in the milk. How should I advise her? Neither silicone nor its components could be measured in the breast milk of women with silicone implants. Your patient should be able to breastfeed safely.


Metabolic and anti-inflammatory benefits of eccentric endurance exercise - a pilot study.
Eccentric endurance exercise (e.g. hiking downwards) is less strenuous than concentric exercise (e.g. hiking upwards) but its potential to reduce cardiovascular risk is unknown. We randomly allocated 45 healthy sedentary individuals (16 men and 29 women, mean age 48 years) to one of two groups, one beginning with two months of hiking upwards, the other with two months of hiking downwards the same route, with a crossover for a further two months. For the opposite way, a cable car was used where compliance was recorded electronically. The difference in altitude was 540 metres; the distance was covered three to five times a week. Fasting and postprandial metabolic profiles were obtained at baseline and after the two month periods of eccentric and concentric exercise, respectively. Forty-two of the 45 participants completed the study; the compliance rate was therefore 93%. Compared with baseline, eccentric exercise lowered total cholesterol (by 4.1%; P = 0.026), low-density lipoprotein (LDL) cholesterol (by 8.4%, P = 0.001), Apolipoprotein B/Apolipoprotein A1 ratio (by 10.9%, P < 0.001), homeostasis model assessment of insulin resistance scores (by 26.2%, P = 0.017) and C-reactive protein (by 30.0%; P = 0.007); the magnitude of these changes was comparable to that of concentric exercise. Eccentric exercise improved glucose tolerance (by 6.2%, P = 0.023), whereas concentric exercise improved triglyceride tolerance (by 14.9%, P = 0.022). Eccentric endurance exercise is a promising new exercise modality with favourable metabolic and anti-inflammatory effects and is well applicable to sedentary individuals.


Prognostic Value of the Perioperative Systemic Inflammation Score for Patients With Curatively Resected Gastric Cancer.
The systemic inflammation score (SIS) is a promising tool for the evaluation of prognosis. The present study aimed to evaluate the clinical impact of the preoperative SIS status in gastric cancer (GC) patients who underwent curative resection. This study retrospectively analyzed 258 patients with primary gastric cancer who received curative treatment at Yokohama City University. The SIS was evaluated before surgery as determined by the lymphocyte-to-monocyte ratio (cut-off value=4.44) and serum albumin level (cut-off value=4.0 g/dl). A high SIS was identified as an independent predictor of overall survival [hazard ratio (HR)=1.784, p<0.05] and multivariate analysis showed marginal significance for recurrence-free survival (HR=1.710, p<0.05). The preoperative SIS score was correlated with both the OS and RFS of GC patients, as well as the clinical course of adjuvant chemotherapy. Thus, the SIS score is a promising prognostic factor for GC.


Use of MRI in diabetic lumbosacral radiculoplexus neuropathy: case report and review of the literature.
Diabetic lumbosacral radiculoplexus neuropathy is often confused with radiculopathy in the context of spinal degenerative disc disease including spinal stenosis. Accuracy in diagnosis may prevent unnecessary interventional procedures including selective nerve root blocks or epidural steroid injections or even surgery in selected cases. Our patient with known diabetes and lumbar disc disease presented with acute onset of pain in L5-S1 distribution of the left lower extremity. Initial MR imaging of the lumbar spine did not show sufficient structural changes to explain her symptomatology. An MR neurogram of the lumbosacral plexus revealed inflammation within the bilateral sciatic and femoral nerves; subsequent EMG demonstrated a generalized sensorimotor neuropathy but no evidence of plexopathy. To our knowledge, this is the first case report that utilized MR imaging of the pelvis to assist in the diagnosis of diabetic lumbosacral radiculoplexus neuropathy (DLRPN).


[Diagnostic value of myelography as an expanded clinical study method].
The paper investigates 54 myelograms and their influence on diagnostic thinking efficacy, on patient management efficacy, and on ultimate outcome efficacy, using statistical methods. Conventional myelography contributes to diagnostic thinking efficacy having a LLR- value of approximately 0.9. With an accuracy value of 0.95 it can be regarded as a reliable method of diagnosis. It alters diagnostic thinking efficacy in no less than 83% (94%) of the cases and patients management efficacy in 89%. Thus, by definition, myelography may be said to affect the state of health of the myelographed patients in 94.4%.


[Protection of myocardiocytes against anoxia-reoxygeneration injury by heat shock protein 70 gene transfection: experiment with rats].
To study the protective role of heat shock protein (HSP)70 expression in the myocardial cells undergoing anoxia/reoxygeneration. Myocardiocytes of neonatal SD rats were cultured and randomly divided into 4 groups: anoxia/reperfusion (A/R) group undergoing anoxia/reperfusion; A/R + HS group, undergoing transfection of HS and A/R, pCDNA HSP70 + A/R group, undergoing transfection of liposome-coated HSP70 pCDNA and A/R, and control group. RT-PCR and Western blotting were used to detect the mRNA and protein expression of HSP70 in the cells. The cell viability was examined by MTT method. The levels of lactic dehydrogenase (LDH), and serum creatine phosphokinase isoenzyme (CPK) were detected. Transmission electronic microscopy was used to observe the ultrastructure of the cells. Apoptosis rate was measured. The cell viability rates of the HS + A/R group and pCDNA HSP70 + A/R group were (71.8 +/- 10.3)% and (73.4 +/- 12.2)% respectively, both significantly higher than that of the A/R group [(35.2 +/- 6.8)%, both P < 0.01]. The LDH activity levels of the HS + A/R and pCDNA HSP70 + A/R groups were (14.3 +/- 2.6) and (14. 6 +/- 2.9) IU/L respectively, both significantly lower than that of the A/R group [22.9 +/- 4.0 IU/L, P < 0.01]. The cell ultrastructure was abnormal in the A/R group e, whereas nearly normal in the HS + A/R and pCDNA HSP70 + A/R groups. The HSP70 mRNA land protein were only slightly expressed in the myocardiocytes of the A/R group; However, the expressions of HSP70 mRNA and protein were of the HS + A/R and pCDNA HSP70 + A/R groups were significantly higher than those of the A/R group (both P < 0.01). The apoptosis rates of the A/R group was (12.84 +/- 2.17), significantly higher than that of the control group [(2.16 +/- 0.15), P < 0.01], and the apoptotic rates of the HS + A/R and pCDNA HSP70 + A/R groups were (5.23 +/- 1.04) and (4.42 +/- 0.93) respectively, both significantly lower than that of the A/R group (P < 0.01). Overexpression of HSP70 alone can provide protection, related to the anti-apoptosis effect, for cardiomyocytes against anoxia-reoxygeneration.


[Preparation and monitoring of sedation and analgosedation carried out by pediatricians and pediatric training assistants].
Sedation and analgosedation form part of the every-day work in the clinical care of pediatric patients. The American Academy of Pediatrics (AAP), the American Society of Anesthesiologists (ASA) and the German Society for Anesthesiology and Intensive Care (DGAI) have established guidelines outlining the general conditions for sedating/analgosedating measures. Since the provisions of the specialist associations diverge in some respects, we have developed an own standard. This takes into account the existing guidelines, aiming at maximum safety for our patients and maximum practicability. In line with the guidelines of the above specialist societies and the international literature, we have developed a standard referring to the information of patients, eating and drinking, monitoring before, during and after the sedation/analgosedation as well as documentation. Over a 12-month period, this standard was applied to all sedations/analgosedations in our hospital which were not performed by the intensive care department. All measures were recorded in a protocol specifically designed for this purpose. In the twelve months under review, a total of 103 sedations/analgosedations were performed (39 of which for bone marrow puncture or punch biopsies, 27 for gastroscopies or coloscopies and 24 for arthrocenteses). None of the sedations or analgosedations implied complications endangering any of our patients. The standard applied proved to be practicable. It was only the documentation, which is closely related to anaesthesiologist protocols, which required adjustment from our staff initially. The standard we have developed and applied provides a practicable and safe protocol to regulate the preparation and monitoring of sedations and analgosedations for pediatric patients. A specific guideline to be issued by the Deutsche Gesellschaft für Kinder- und Jugendmedizin (German Pediatric Society) is considered desirable.


Low utility of diabetic eye care services and perceived barriers to optimal diabetic retinopathy management in Indonesian adults with vision-threatening diabetic retinopathy.
Among 100 Indonesian adults with type 2 diabetes having vision-threatening diabetic retinopathy, less than 25% of individuals had appropriate treatment within 12 months from their initial screening due to lack of information about the treatment cost (88%), no permission from family member (77.3%) and having no eye complaint (77%).


How parents and health professionals experience prematurity in an Italian neonatal intensive care: A grounded theory study.
The aim of this study was to explore prematurity as a psycho-social process from the point of view of both parents and healthcare professionals. We conducted a Grounded Theory study through semi-structured interviews. Participants were from an Italian Neonatal Intensive Care Unit, including parents of premature children, and health professionals. Interviews were analyzed according to Grounded Theory coding strategies, to hypothesize a theoretical model. On one side, behaviors, emotions, and feelings of parental couples; on the other one, the activities and reactions of professionals have been interpreted in a four-phase negotiated process: 1) the initial breakup; 2) the floating family; 3) the event processing; 4) the reconstruction of a new family. Managing prematurity is a negotiated process which could be better managed through a family-centered approach as fundamental in neonatal and pediatric services. Being open in listening, coherent in the given answers, and knowing the entire process and story of being suddenly parents of a premature child can make the difference in the future definition of a new family. Perinatal care would benefit from the following suggested practical implications: i) reducing feelings of separation and solitude by integrating diverse professionals around the family systems; ii) expediting prematurity-related changes through peer-support during hospitalization and attention to rooms' organization; iii) enhancing information exchange between all professionals involved in the care of dyads/parents and new-borns, starting since the pregnancy and continuing after the child's birth by implementing multidisciplinary meetings or appropriate care pathways.


Selective impregnation of developing neurons with the Loyez method and with two kinds of Luxol Fast Blue.
In chick embryos, unlike in adults, many neurons can be impregnated selectively with the LOYEZ iron-haematoxylin method, and with the MBSN and ARN types of Luxol Fast Blue. The same neurons are selected with each of the 3 methods. Their distribution at a given age is roughly predictable, but it changes with age, as is described in some detail. Large neurons are impregnated more readily than small neurons; glial cells are not impregnated. With each of the 3 methods, the same conditions of fixation and embedding are needed for the selective impregnation to occur: fixation with a dilute, alkaline solution of formaldehyde (or, non-optimally, a closely related aldehyde), and nitrocellulose embedding. Numerous other stains fail to produce the selective impregnation. The parameters of the LOYEZ method have been varied systematically in order to optimize the neuronal impregnation, and preliminary efforts have been made to deduce the substrate of the staining, which probably involves proteins or peptides.


Effect of the venereal diseases epidemic on the incidence of ectopic pregnancy -- implications for the evaluation of contraceptives.
This paper identifies a basis for quantifying the risk of ectopic pregnancy in pregnancy seekers and in users of the major means of contraception, with and without prior tubal infection. The doubling, in U.S. in recent years, of both numbers of ectopic pregnancies and the prevalence of nonsurgical sterility is correlated with the epidemic of gonorrhea and other sexually transmitted, salpingitis-producing diseases. The risk of ectopic pregnancy in women who have once had salpingitis is shown, on the basis of Weström's landmark prospective study and many retrospective studies, to be approximately 10-fold that of normal women. Fertilization-preventing contraceptives reduce uterine and ectopic pregnancies in the same proportion and in both categories of users--that is, in normal-risk and high-risk (post-salpingitis) women. Intrauterine contraceptives, however, markedly reduce uterine pregnancies, but have little effect on the incidence of ectopic pregnancy. Among IUD users, therefore, the incidence of ectopic pregnancy will be disproportionately great in relation to the fraction of high-risk women in the population of users. This disproportionate influence of prior salpingitis may confuse the comparative evaluation of contraceptive safety because the proportions of normal-risk and high-risk women probably vary widely, since public health data show extremely wide geographic differences in the incidence of gonorrhea -- the best known but not the only sexually transmitted, salpingitis-producing disease.


Firing pattern and location of respiratory neurons in cat medullary raphe nuclei.
Extracellular spikes of single respiratory neurons were explored in the medullary raphe nuclei in spontaneously breathing decerebrated or Nembutal-anesthetized cats. A total of 26 respiratory neurons, whose spikes were confirmed to originate from the cell bodies, was recorded in the raphe obscurus and pallidus. They could be classified into six types based on the relation of their firing to the phases of respiration: (1) inspiratory (I) decrementing (I-DEC, n = 14), (2) I-frequency modulated (n = 3), (3) I-augmenting (I-AUG, n = 2), (4) late I (n = 2), (5) expiratory (E) augmenting (E-AUG, n = 3) and (6) I-E phase-spanning (I-E PS, n = 2). These results suggest the existence of neurons relating to the control of respiration in the medullary raphe nuclei.


Maquiroside A, a new cytotoxic cardiac glycoside from Maquira calophylla.
Maquiroside A, a new cardiac glycoside, was isolated from the Peruvian plant Maquira calophylla. The structure was established using spectroscopic methods, and the aglycone was found to be cannogenol. The sugar portion was identified as D-cymarose. Maquiroside A demonstrated activity against the KB cell culture.


The precarization of the Mexican nursing labor market: a repeated cross-sectional analysis for the period 2005-2018.
Precarization of labor conditions has been expanding over the last three decades as a consequence of global economic transformations. The health workforce labor market is exposed to these transformations as well. In Mexico, analyses of the nursing labor market have documented high levels of unemployment and underemployment; however, precarization has been not considered as a relevant indicator in these analyses. In this study, precarization is analyzed using a quantitative approach to show its prevalence and geographic distribution between 2005 and 2018. A repeated cross-sectional study was carried out with data from the population-based National Occupation and Employment Survey (ENOE in Spanish) which is administered quarterly to people 15 years or older in over 120 000 households. All individuals who reported having nursing training (technical or university level) were selected for this study. Our main variable was labor precariousness, which included three dimensions: (i) economic, (ii) regulatory, and (iii) occupational safety. We show the evolution of the relative weight of nursing activity between the years 2005 and 2018, the main socio-demographic characteristics of nurses as well as their main labor conditions, and the geographic distribution of precariousness for the 32 federal states in México. Four of the five indicators of labor precariousness increased among the group of nurses analyzed: (a) the percentage of people with no written contract, (b) the percentage of people with incomes lower than two times the minimum wage, (c) the percentage of nurses without social security, and (d) the percentage of nurses without social benefits. The percentage of nurses that work under some condition of work precariousness increased from 46% in 2005-2006 to 54% in 2018. Finally, the number of states with high precariousness level increased from seven in 2005-2006 to 17 in 2018. Throughout Mexico, nursing precariousness has expanded reaching 53% by 2018. The advancement of precarization of nursing jobs implies a reduction in the capacity of the Mexican health system to reach its coverage and care goals as nurses represent 52% of all available workers that provide direct services to the population.