Association of cytosine-adenine repeat polymorphism of the estrogen receptor-beta gene with rheumatoid arthritis symptoms.
The influence of the polymorphism of the estrogen receptor-beta gene, cytosine-adenine (CA) dinucleotide repeat in intron 6, in the occurrence of rheumatoid arthritis (RA) was investigated. Forty-seven RA patients and 36 control subjects with osteoarthritis (OA) were recruited. CA repeat polymorphism was examined using denaturing high-performance liquid chromatography (WAVE DNA Fragment Analysis System). The mean number of CA repeats was significantly higher in RA than in OA patients. Two groups were established: <or=21 repeats (short S), >or=22 repeats (long L); and 3 kinds of genotypes (SS, SL, LL) were found. In RA patients, the L allele frequency was higher (OR = 2.03; P <or= 0.05 Fisher's test) and more subjects carried the LL genotype (OR = 5.50; P <or= 0.05 Fisher's test) compared to OA patients. It was concluded that the presence of the L allele could be a risk factor for RA, although additional larger scale study is needed.


Spiral magnetic domain structure in cylindrically-shaped microwires.
Identification and characterisation of novel and unusual magnetization states remains a topic of research in modern magnetism. Recently, control of the magnetization state between the surface and volume in cylindrical microwires with the giant magneto-impedance effect has been demonstrated. Herein, the phenomenon of spatial migration of spiral magnetic domains inside a microwire is demonstrated using the magneto-optical Kerr effect. The main properties of the inclined spiral structure were determined, where the surface domain structure possessed a length limited only by actual sample length. Transformation of the structure from a spiral to an elliptical structure could be controlled by external torsion stress. Hysteresis and magnetic images were simulated based on a model assuming a spatial distribution of the internal stress inside the microwire, whose results were consistent with the experimental results. A consistent interpretation of the results in terms of the formation and transformation of the spiral magnetic domain structure is proposed.


Tuberculous hepatic abscess appearing after liver transplantation in a patient with human immunodeficiency virus and hepatitis B and C virus co-infection.
Tuberculosis infection occurs relatively frequently in solid organ transplant recipients, although the occurrence of tuberculous hepatic abscesses is uncommon. Anti-tuberculous therapy has several concerns in transplant recipients, including an increased risk of cellular rejection and potential hepatotoxicity. We present the case of a human immunodeficiency virus-infected liver transplant patient who developed multiple tuberculous liver abscesses. Treatment with isoniazid, ethambutol, pyrazinamide, and moxifloxacin was efficacious, well tolerated, and safe.


Child allergic symptoms and mental well-being: the role of maternal anxiety and depression.
To determine whether maternal mental health mediates the relationship between eczema or asthma symptoms and mental well-being in children. Analysis of 7250 children from the Avon Longitudinal Study of Parents and Children. Child mental well-being at 8 years was measured by the Strengths and Difficulties Questionnaire. Binary outcomes were high 'internalizing' (anxious/depressive) and 'externalizing' (oppositional/hyperactive) problems (high was >90th percentile). Child rash and wheeze categories were 'none'; 'early onset transient' (infancy/preschool only); 'persistent' (infancy/preschool and at school age); and 'late onset' (school age only). Maternal anxiety and depression were reported during pregnancy and when child was 8 years old. Persistent wheezing symptoms were associated with high externalizing (OR 1.74, 95% CI, 1.41-2.15) and internalizing (1.67, 1.35-2.06) problems compared with never wheeze. Maternal anxiety and depression, and disrupted child sleep, attenuated these associations. Persistent rash (externalizing: 1.74, 1.40-2.15; internalizing: 1.42, 1.16-1.74) and late onset rash (externalizing: 1.62, 1.17-2.25; internalizing: 1.46, 1.07-1.99) symptoms were associated with poorer mental well-being compared with no rash at any age. Maternal anxiety and depression, particularly when child was aged 8 years rather than during pregnancy, accounted for the association with internalizing symptoms and partly for externalizing symptoms. Sleep disruption did not mediate the association. Maternal anxiety and depression may mediate the association between child rash and wheeze and child mental well-being.


Outcome Analysis and Risk Factors for Perioperative Myocardial Ischemia After Elective Aortic Surgery.
Perioperative myocardial ischemia (PMI) after non-cardiac surgery remains a serious postoperative complication. This study analyzed the risk factors and outcomes of patients who suffered from PMI after elective aortic surgery. Data from 863 patients who underwent elective aortic surgery for aneurysms or Leriche syndrome were retrospectively analysed with regard to PMI. The diagnosis of PMI was based on a positive serum troponin diagnostic test. According to the clinical signs and symptoms, the patients with PMI were divided into two groups: symptomatic and asymptomatic patients. Comorbidities, preoperative medication, intraoperative parameters, postoperative complications, mortality, length of intensive care stay and hospitalisation, as well as the long-term follow-up, were compared in a matched-pair analysis (1:3) with patients without PMI. Logistic regression analyses were performed to identify independent risk factors for PMI. Thirty-two patients with PMI were identified. Cardiac comorbidities (previous myocardial ischemia, P = 0.0099; left ventricular systolic dysfunction, P = 0.0429), ASA score ≥III (P = 0.0114) and preoperative elevated creatinine (P = 0.0194) were more common in patients who suffered PMI. The regression analysis confirmed that peripheral artery disease and prolonged operative duration >180 min are significant predictors of PMI. Surgical complications (wound healing deficit, P = 0.0027; rate of secondary interventions during primary admission, P = 0.0057) and medical complications (pneumonia, P = 0.0002; renal dysfunction, P = 0.0041) were more common in patients with PMI compared to the control group. Patients who suffered PMI remained in intensive care for a significantly longer period (P = 0.0001) and were also hospitalized for longer (P = 0.0001) than the control group. The long-term survival of patients who suffered PMI after aortic surgery was significantly worse than the control group (P < 0.0001, median 53 vs. 84 months), independent of clinical ischemia-associated symptoms. PMI after aortic surgery not only affects long-term survival, but also correlates with worsening of surgical outcome. Thus, meticulous preoperative risk stratification is required for high-risk patients, together with routine postoperative monitoring of troponin levels after aortic surgery.


Induction of alkaline phosphatase in the inflamed intestine: a novel pharmacological target for inflammatory bowel disease.
This study demonstrates the upregulation of alkaline phosphatase and the mechanisms involved in experimental colitis. All models of ileal and colonic inflammation examined, which were characterized by significant oxidative stress and neutrophil infiltration, resulted in an increase in alkaline phosphatase activity which was attributable to both epithelial cells and cells of the lamina propria, mainly leukocytes. The increase in alkaline phosphatase sensitivity to the inhibitors levamisole and homoarginine, together with changes in the apparent molecular size and in the sialization of the enzyme, indicated a change in the isoform expressed. An increase in tissue non-specific alkaline phosphatase expression was observed by Western blotting. Treatment with the bone/kidney alkaline phosphatase inhibitor levamisole or a monoclonal antibody resulted in significant protection from colonic inflammation. Taken together, these results indicate that the kidney isoform is a marker of intestinal inflammation and that it might even constitute a target for pharmacological intervention.


PD-1 gene polymorphic variation is linked with first symptom of disease and severity of relapsing-remitting form of MS.
Multiple sclerosis (MS) is an inflammatory disease of the central nervous system (CNS), where inflammation, demyelination together with the axonopathy are the cardinal features on pathologic ground, with a combined genetic and environmental background. The associations of PD-1 single nucleotide polymorphisms (SNPs): PD-1.3 (in intron 4), PD-1.5 and PD-1.9 (both in exon 5) with clinical manifestation of MS in 479 south Polish individuals including 203 MS patients were analyzed. Presence of PD-1.5T allele was linked with the first manifestations of disease: diplopia and pyramidal signs - favored pyramidal signs but protected against of diplopia development. Farther, PD-1.3G/PD-1.5C/PD-1.9C haplotype significantly favored whereas GTC protected against diplopia. Besides, GTT haplotype strongly favored non-severe RRMS outcome and ATC haplotype was specific only for these MS patients. Our population-based case-control study, investigating selected three PD-1 SNPs: PD-1.3, PD-1.5 and PD-1.9, revealed that polymorphic variation may be rather disease-modifying than MS risk factor.


Effects of D-penicillamine on some oxidative enzymes of rat organs in vivo.
In the liver of neonate rats and in tissues of adult rats a study was made of the effect of D-penicillamine treatment in vivo on the enzymes of peroxide metabolism, lipid peroxidation and other components. It was found that D-penicillamine primarily acts by decreasing lipid peroxidation, thus stabilizing the membrane.


Gill Histopathological Biomarkers in Fish Exposed to Trace Metals in the Todos os Santos Bay, Brazil.
Histopathologies are widely recognized as biomarkers of environmental pollution. In this sense, we evaluated the putative relationship of gill histopathologies and distinct ecological impacts in two regions of Todos os Santos Bay (BTS), Brazil, the largest bay in Northeastern Brazil, South Atlantic. We compared the presence and concentration of metals (Al, As, Ba, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb, V, and Zn) in water, sediments, and gills and gill histopathologies of a demersal fish (Diapterus rhombeus) and a benthic fish (Ogcocephalus vespertilio). As expected, fish and sediment samples from historically contaminated areas (Aratu) showed more remarkable traces of metals than apparently low-impact areas (Jaguaripe). Likewise, the DTC (degree of tissue change) index and the volume densities were higher in fish caught in Aratu. In addition, the Diapterus rhombeus species showed more potential than Ogcocephalus vespertilio for risk assessment as it showed more responses to the environment reflected on more histopathologies. These data support the effectiveness of incorporating functional gill morphology to monitor impacts on estuarine biota that can be used as a reference to improve the management of ecosystems and prevent harm to human health.


Paneth cell tumor. An additional case of duodenal adenoma with malignant change.
A papillary adenoma with malignant change consisting of only Paneth cells and their precursors is described. Neoplastic proliferation of Paneth cells is discussed from a point of view of cell biology.


Epikeratoplasty with nonlyophilized tissue in children with aphakia.
We studied 75 epikeratoplasty procedures using nonlyophilized tissue performed by eight ophthalmic surgeons in 70 eyes (47 patients) to correct for aphakia in children less than 8 years of age (mean age, 3.4 +/- 2.1 years). Of the 47 patients in the study, 24 were girls and 23 were boys; 23 patients had bilateral surgery. Seven of the epigrafts required removal; two were not replaced, and five underwent successful repeat epikeratoplasty. Overall, the success rate (that is, the percentage of epigrafts that remained optically and functionally clear throughout the course of this study) for the epikeratoplasty procedure was 89% (62 of 70 eyes) for initial surgery and 96% (67 of 70 eyes) for repeat surgery. The average spherical equivalent was +14.4 +/- 3.7 diopters preoperatively and +0.3 +/- 2.9 diopters one year after the operation. One year after the final surgical procedure, 42 of 56 eyes (75%) were within 3 diopters of emmetropia. In the 29 verbal patients, best-corrected visual acuity was 20/100 or better in 25 (86.2%) one year after the operation.


Hippo signalling in the G2/M cell cycle phase: lessons learned from the yeast MEN and SIN pathways.
Over the past decade Hippo kinase signalling has been established as an essential tumour suppressor pathway controlling tissue growth in flies and mammals. All members of the Hippo core signalling cassette are conserved from yeast to humans, whereby the yeast analogues of Hippo, Mats and Lats are central components of the mitotic exit network and septation initiation network in budding and fission yeast, respectively. Here, we discuss how far core Hippo signalling components in Drosophila melanogaster and mammals have reported similar mitotic functions as already established for their highly conserved yeast counterparts.


Visual cortical neurons influenced by the oculomotor input: characterization of their receptive field properties.
A class of cells in cat visual cortex (area 17 and 18) had a tight correlation with spontaneous and electrically evoked PGO waves under reserpine. They tended to have high maintained activity and a large receptive field which was preferentially excited by fast moving slits. They were also direction selective and influenced through both eyes. For these cells, the selectivity for visual stimuli was dramatically altered in the presence of PGO waves induced by pontine stimulation. They were complex cells. A second class of cortical cells showed a moderate correlation with spontaneous PGO waves. Visually evoked activity of these cells was either excited or inhibited by evoked PGO waves in response to pontine stimulation. A third class of cells, the majority, did not seem to have any correlation with PGO waves. The second and third classes of cells could be either one of the 3 main categories of visual cortical cells, predominantly simple cells in area 17 and complex cells in area 18. The present study provided further support to a previous proposal that PGO waves in the visual cortex, as neural correlates of saccadic eye movements, modulate specifically the ongoing activity of a selective type of cortical cell. A type of complex cell in the deeper layers is supposed to integrate visual and oculomotor inputs. It is hypothesized that the consequence of oculomotor inputs to the visual cortex is specified by the type of cells which receive oculomotor inputs, rather than by the nature of the inputs themselves.


Alterations in pancreatic islet phosphate content during secretory stimulation with glucose.
Isolated rat pancreatic islets were perifused and analyzed for phosphate content immediately following the transient increase in the efflux of orthophosphate which occurs when insulin secretion is stimulated by glucose. In some instances, islets were perifused directly following isolation to minimize preparative delay; in others, islets were prelabeled during incubation with [32P]orthophosphate for 90 min prior to perifusion. In both experimental situations, total islet phosphate content declined 40--50% following exposure to stimulating concentrations of glucose and initiation of enhanced insulin release. In the experiments with prelabeled islets, tissue content of [32P]orthophosphate fell to a similar extent so that the specific radioactivity of islet orthophosphate was unaffected. Inhibited of heightened insulin release with Ni2+ did not modify the decrements in total or radioactive tissue orthophosphate, thus indicating that these responses to islet stimulation reflect events which are proximal to activated exocytosis. Simultaneous analyses for tissue ATP and ADP demonstrated that the efflux in orthophosphate and reduction in tissue orthophosphate content were not mediated via net changes in islet adenine nucleotides. The observations represent the first documentation that a net reduction of tissue inorganic phosphate is one of the early components of stimulus-secretion coupling in isolated pancreatic islets.


Descending thoracic aorta aneurysmectomy: left-left centrifugal pump versus simple clamping technique.
Forty-six patients who had had an elective repair of a descending thoracic aortic aneurysm were reviewed, in order to investigate the efficacy of support by a centrifugal pump on distal organ perfusion and spinal cord protection during cross-clamping of the thoracic aorta. Two concurrent groups were analysed: 36 patients (78%) were supported by left atriofemoral arterial bypass with a centrifugal pump and 10 (22%) had no distal circulatory support. No patient was fully heparinized. The demographic data and preoperative characteristics of the groups, including location and type of aneurysm, were similar. The mean(s.d.) duration of cross-clamping was 37.8 (16) min in the centrifugal pump group and 42.3(21) min in the simple clamping group. Preoperative haemodynamic and laboratory data were similar in both groups. During cross-clamping, parameters of pH and blood urea varied but were better in the centrifugal pump group; changes from pre-intervention to early aortic cross-clamping time were not significant (pH, P < 0.0006; bases, P < 0.0003). Differences in creatinine values were caused mainly by the change from pre-intervention to the first postoperative day (P < 0.03); this continued throughout the hospital stay. The cerebrospinal fluid pressure measurement indicated a significant difference in time change (P < 0.0001) and mean level over time (P < 0.0002): levels were significantly lower in the centrifugal pump group throughout aortic cross-clamping. Three patients in the simple clamping group and none in the centrifugal pump group (P < 0.02) required cerebrospinal fluid drainage.(ABSTRACT TRUNCATED AT 250 WORDS)


Facile synthesis of magnetic sludge-based carbons by using Electro-Fenton activation and its performance in dye degradation.
Highly stable iron based magnetic carbon were prepared by sequential Electro-Fenton (EF) activation and pyrolysis of sewage sludge. The applied voltage exerts great influence on EF treated sludge flocs and thus poses significant effect on physiochemical properties of the as-prepared carbons. High insertion rate of iron into sludge from EF activation resulted in carbons with highly dispersed iron oxides, which had average size of iron nanoparticles being 4.77nm. The carbon also presented well developed porosity which had Brunauer-Emmett-Teller (BET) surface area attaining 341m2/g. Carbons prepared by traditional Iron Impregnation (IM) were used as comparison to gain further insight into their catalytic role as Fenton-like catalyst. Results showed that EF-activated sludge carbon could yield 96.1% of Methyl Orange (MO) removal in 60min together with only 1.4% of iron leaching. After three cycles, the MO removal can still reach 80% with EF-activated carbons.


[Attitude to the disease model and self-help groups as predictors of participation in after care and of therapeutic outcome in alcoholic patients].
The study was conducted to determine the predictive value of attitudes towards the goals derived from the disease model of alcoholism (life-long acceptance of alcoholic status and endorsement of abstinence) as well as towards self-help groups. The criteria to be predicted were the intention to participate and actual participation in self-help groups as well as four indicators of therapeutic success. Only in cross-sectional analysis were significant correlations found. Longitudinally, there were no relevant relationships between the attitudes expressed by the patients at the end of inpatient treatment (t1) and subsequent participation in self-help groups or indicators of therapeutic success (t2: follow-up, 9 month after discharge). The discrepancy between cross-sectional and longitudinal analysis can be attributed to the fact that attitudes become increasingly less stable with time. This indicates that it is only possible to a limited extent to predict from the attitudes expressed during inpatient treatment how patients will actually behave in daily life outside the clinic. The consequences of these findings for therapy are discussed.


Aqua-glutarato(2,4,6-tri-2-pyridyl-1,3,5-triazine)nickel(II) trihydrate.
In the title compound, [Ni(C(5)H(6)O(4))(C(18)H(12)N(6))(H(2)O)(2)]·3H(2)O, the Ni(II) atom shows a distorted octa-hedral coordination by three N atoms of the tridentate chelating ligand and three O atoms of two aqua ligands and an O atom of one carboxylate group of the glutarate anion. Mol-ecules are self-assembled via inter-molecular O-H⋯O and O-H⋯N hydrogen-bonding inter-actions and π-π stacking inter-actions [centroid-centroid distance = 3.836 (3) Å] into a supra-molecular network.


Cultivation and diversity analysis of novel marine thraustochytrids.
Thraustochytrids are a group of unicellular marine heterotrophic protists, and have long been known for their biotechnological potentials in producing squalene, polyunsaturated fatty acids (PUFAs) and other bioactive products. There are less than a hundred known strains from diverse marine habitats. Therefore, the discovery of new strains from natural environments is still one of the major limitations for fully exploring this interesting group of marine protists. At present, numerous attempts have been made to study thraustochytrids, mainly focusing on isolating new strains, analyzing the diversity in specific marine habitats, and increasing the yield of bioactive substances. There is a lack of a systematic study of the culturable diversity, and cultivation strategies. This paper reviews the distribution and diversity of culturable thraustochytrids from a range of marine environments, and describes in detail the most commonly used isolation methods and the control of culture parameters. Furthermore, the perspective approaches of isolation and cultivation for the discovery of new strains are discussed. Finally, the future directions of novel marine thraustochytrid research are proposed. The ultimate goal is to promote the awareness of biotechnological potentials of culturable thraustochytrid strains in industrial and biomedical applications.


Regulation and pathophysiological role of epithelial turnover in the gut.
Cell death in the intestinal epithelium has to be tightly controlled. Excessive or misplaced epithelial cell death can result in barrier dysfunction and, as a consequence thereof, uncontrolled translocation of components of the microbial flora from the lumen into the bowel wall. Susceptibility to gastrointestinal infections or chronic inflammation of the gut, as observed in patients with inflammatory bowel disease, can be the result of such dysregulation. Conversely, defects in cell death initiation might lead to an irregular accumulation of epithelial cells and cause intestinal cancer development. Until recently, activation of caspases in the intestinal epithelium was considered as a potential contributor to barrier dysfunction and as a pathogenic factor in the development of intestinal inflammation. Thus blocking of caspases appeared to be a potential therapeutic option for patients with inflammatory bowel disease. Recent studies on necroptosis however demonstrated that also inhibition of caspases can cause barrier dysfunction and intestinal inflammation. Caspase-8 on top of its functions in the extrinsic apoptosis pathway also controls necroptosis and turns out to be an essential molecule in regulating tissue homeostasis in the gut. Epithelial caspase-8 therefore emerges as a checkpoint not only of cell survival and cell death, but also as a regulator of the mode of cell death. According to this model, both excessive activity as well as a lack of activity of caspase-8 results in epithelial cell death and intestinal inflammation and caspase-8 needs to be tightly controlled to warrant tissue homeostasis in the gut.


Transcription factor Oct-2A contains functionally redundant activating domains and works selectively from a promoter but not from a remote enhancer position in non-lymphoid (HeLa) cells.
In non-lymphoid cells such as HeLa cells, ectopic expression of the lymphocyte-specific transcription factor Oct-2A can activate reporter genes whose promoters consist of a single octamer sequence (ATTTGCAT) upstream of a TATA box. While the factor is strongly active in a promoter position, it tails as an enhancer factor: an enhancer consisting of multiple copies of the octamer sequence placed downstream of the reporter gene is not active in HeLa cells, even at high concentration of Oct-2A. In B lymphoid cells, however, the same enhancer is highly active. This could mean that an additional factor is required for enhancer activation in B cells. Furthermore, we have tested the transcriptional activation potential of Oct-2A with a series of N-terminal and C-terminal deletions. We show that a glutamine-rich domain near the N-terminus is required for full activity. Otherwise, large segments of the N-terminal half or the entire C-terminal region are dispensable in our assay, as long as the deletions do not impinge on the conserved POU domain which is sufficient for DNA binding. While N-terminal and C-terminal regions can functionally compensate for each other, a combined deletion that only retains the POU domain is a strong down mutation. We also find that activity depends on the promoter structure of the reporter gene: the POU domain by itself shows some activity with a promoter where the octamer sequence is located very close to the TATA box, but no activity with another promoter construction where the octamer sequence is located further upstream. The two promoters also respond differently to the deletion of the glutamine-rich stretch important for transcriptional activation. From these experiments we consider it likely that the natural octamer factor variants can selectively activate the different naturally occurring octamer-containing promoters.


Analysis of vertebrate genomes suggests a new model for clade B serpin evolution.
The human genome contains 13 clade B serpin genes at two loci, 6p25 and 18q21. The three genes at 6p25 all conform to a 7-exon gene structure with conserved intron positioning and phasing, however, at 18q21 there are two 7-exon genes and eight genes with an additional exon yielding an 8-exon structure. Currently, it is not known how these two loci evolved, nor which gene structure arose first--did the 8-exon genes gain an exon, or did the 7-exon genes lose one? Here we use the genomes of diverse vertebrate species to plot the emergence of clade B serpin genes and to identify the point at which the two genomic structures arose. Analysis of the chicken genome indicated the presence of a single clade B serpin gene locus, containing orthologues of both human loci and both genomic structures. The frog genome and the genomes of three fish species presented progressively simpler loci, although only the 7-exon structure could be identified. The Serpinb12 gene contains seven exons in the frog genome, but eight exons in chickens and humans, indicating that the additional exon evolved in this gene. We propose a new model for clade B serpin evolution from a single 7-exon gene (either Serpinb1 or Serpinb6). An additional exon was gained in the Serpinb12 gene between the tetrapoda and amniota radiations to produce the 8-exon structure. Both structures were then duplicated at a single locus until a chromosomal breakage occurred at some point along the mammalian lineage resulting in the two modern loci.


The role of reconstructive surgery in patients with end-stage interstitial cystitis/bladder pain syndrome: is cystectomy necessary?
Interstitial cystitis is a debilitating condition that has a profound effect on quality of life. Although many approaches to treatment have been explored, no consistently effective treatment has been identified. Reconstructive surgery is offered to patients with refractory IC/BPS; however, expert opinion is divided as to whether simultaneous cystectomy is necessary to achieve symptomatic cure. The aim of this study was to report our experience in the surgical management of IC/BPS in a university teaching hospital. The hospital inpatient enquiry (HIPE) system was used to identify patients with IC/BPS who underwent surgery between 1997 and 2013. Medical records were examined and patients were invited to complete three symptom-based questionnaires. Twelve patients were identified (8 female, 4 male). Reconstructive procedures included urinary diversion without cystectomy (9) and augmentation ileocystoplasty (4). One patient failed to have a sustained improvement in symptoms following ileocystoplasty and later underwent successful urinary diversion. All other patients noted a "marked improvement" in overall symptoms on global response assessment (GRA) and the resolution of bladder pain on a visual analogue sale (VAS). There were no persistent symptoms or complications related to the retained bladder following diversion. Our study adds to the existing evidence that cystectomy is not necessary to provide symptomatic cure in patients with end-stage IC/BPS. Urinary diversion without cystectomy is a highly effective operation and a successful outcome was achieved in all patients. Ileocystoplasty may be offered in carefully selected cases.


Sex-related differences in endogenous pituitary adenylate cyclase-activating polypeptide (PACAP) in the thalamic paraventricular nucleus: Implications for addiction neuroscience.
Males and females exhibit differences in motivated and affective behavior; however, the neural substrates underlying these differences remain poorly understood. In the paraventricular nucleus of the thalamus (PVT), sex-related differences in neuronal activity have been identified in response to motivated behavior tasks and affective challenges. Within the PVT, the neuropeptide, pituitary adenylate cyclase-activating polypeptide (PACAP), is highly expressed and is also involved in motivated and affective behavior. The purpose of this study was to compare the expression of PACAP mRNA and peptide in the PVT of males and females. Analysis with quantitative real-time PCR in mice revealed that females had significantly higher levels of PACAP mRNA than males in the whole PVT, but no differences in the neuropeptides enkephalin or corticotropin releasing factor (CRF) in this brain region. While in rats, females demonstrated a trend for greater gene expression than males in the anterior/middle and middle/posterior PVT, they again showed no differences in enkephalin or CRF. Analysis with immunofluorescent histochemistry revealed that female mice had significantly more PACAP-containing cells than males as a function of area throughout the PVT, and that female rats had significantly more PACAP-27 and PACAP-38-containing cells than males, both as a percentage of total cells and as a function of PVT area. For PACAP-27, this specifically occurred in the anterior PVT, and for PACAP-38, it occurred throughout the anterior, middle, and posterior PVT. These results suggest that sex-related differences in PVT PACAP may underly some of the established sex-related differences in motivated and affective behavior.


Intracoronary shunts reduce transient intraoperative myocardial dysfunction during off-pump coronary operations.
This study investigated the hemodynamic changes in patients undergoing multiple vessel beating heart coronary revascularization in the presence or absence of an intracoronary shunt. Forty patients were randomized to off-pump with a shunt (n = 20) or with the proximal coronary artery occluded by a soft snare (n = 20). Hemodynamic measurements were recorded at base line, during construction, and after completion of each distal anastomosis. Grafting of the left anterior descending coronary artery anastomosis was associated with a significant decrease in stroke volume, cardiac index, and mean arterial pressure, and an increase in pulmonary capillary wedge pressure and systemic vascular resistance in the snare but not in the shunt group. During grafting of the posterior descending coronary artery there was a marked decrease in stroke volume and cardiac index, and an increase in central venous pressure in both groups, and an increase in heart rate, mean pulmonary arterial pressure, pulmonary capillary wedge pressure, and systemic vascular resistance only in the snare group. The most extensive changes were observed during the circumflex coronary artery anastomosis with a reduction in stroke volume, cardiac index, and mean arterial pressure, and an increase in central venous pressure, pulmonary capillary wedge pressure, pulmonary arterial pressure, and systemic vascular resistance in both groups. In all settings, these changes were transient and recovered after the heart was returned to its anatomical position in the shunt group, whereas stroke volume and cardiac index remained reduced, and systemic vascular resistance was elevated in all settings in the snare group. Stabilization of the left anterior descending coronary artery to perform the anastomosis resulted in temporary hemodynamic changes, which are prevented by the use of an intracoronary shunt. The hemodynamic deterioration during the construction of the posterior descending coronary artery and circumflex coronary artery anastomoses is transient in the shunt group, whereas the snaring technique is associated with an impairment of early functional recovery.


Correlations between concentrations of interleukin-12 and interleukin-13 and lymphocyte subsets in the follicular fluid of women with and without polycystic ovary syndrome.
To investigate a possible correlation between interleukin-12 (IL-12) and IL-13 levels and lymphocyte subsets in the preovulatory follicles of patients with and without polycystic ovarian syndrome (PCOS). Controlled clinical study. University hospital. Seventy-eight infertile women undergoing IVF-embryo transfer. The subjects underwent blood sampling, ovum retrieval, and embryo transfer. Follicular fluid levels of T, androstenedione (A); IL-12, IL-13, activated T cells, T helper, and T-suppressor lymphocytes. The level of IL-12 detected in follicular fluid (FF) was significantly lower in patients with PCOS than in normally ovulating women (mean: 1.47 +/- 0.3 pg/mL vs. 2.25 +/- 0.7 pg/mL, respectively); in contrast, FF IL-13 concentrations were significantly higher in the patients with PCOS than in the normally ovulating women (mean: 32.5 +/- 3.7 pg/mL vs. 19.6 +/- 2.5 pg/mL, respectively), as was the total number of activated T lymphocytes (11.5% +/- 1.5% vs. 4.8% +/- 0.4%). A significant correlation was observed between FF activated T-cell concentrations and FF IL-12, IL-13, T, and A levels. No significant differences were observed when these data were compared with embryological parameters. The present study shows significant differences in the correlation between FF IL-12 and IL-13 levels and T lymphocyte numbers in the subset of patients with PCOS as compared to normally ovulating women.


Cyclosporine treatment of inflammatory bowel disease.
Numerous uncontrolled studies and one controlled trial of cyclosporine therapy suggest that this selective inhibitor of cell-mediated immunity may be of benefit in patients with chronically active Crohn's disease. In addition, uncontrolled studies indicate that cyclosporine may be useful in the following settings: fistulous Crohn's disease; corticosteroid sparing in Crohn's disease; severe ulcerative colitis; and refractory proctosigmoiditis. The major advantages of cyclosporine therapy are apparent efficacy in patients with refractory disease and a rapid onset of response. Nevertheless, the incidence of relapse is high after cyclosporine therapy is discontinued, and this outcome is not prevented by low-dose maintenance therapy. Furthermore, the frequency of occurrence of cyclosporine-related side effects during treatment of patients with inflammatory bowel disease is high; paresthesias and hypertrichosis are the two most common adverse effects reported in the literature. Although the potential for permanent renal damage is of concern, serious, irreversible toxicity seldom occurs. Ongoing clinical trials will provide additional information about the efficacy and safety of cyclosporine for the aforementioned indications.


Selective uptake of noradrenaline, dopa, and 5-hydroxytryptamine by the nervous system of Phocanema decipiens (Nematoda): a light autoradiographic and ultrastructural study.
As determined by light microscopic autoradiography, parts of the nervous system of Phocanema decipiens have selective and high affinity mechanisms for the uptake of tritiated noradrenaline, dihydroxyphenylalanine (dopa) and 5-hydroxytryptamine. In the nervous system, noradrenaline is accumulated only by the four papillary nerves and two fibers in the nerve ring. The precursor dopa is also taken up by these neurons and, in addition, by the lateral nerves. 5-Hydroxytryptamime is accumulated by the three pharyngeal nerves, two cells in each lateral ganglion, and two other fibers in the nerve ring. With adjacent ultrathin sections, the labelled papillary nerve and lateral ganglion were examined ultrastructurally and found to contain various dense core vesicles which are similar to those in other aminergic neurons. The adjoining unlabelled cells of the same neurons are found, on the other hand, to contain dense agranular vesicles. With these results, the noradrenaline accumulating neurons are suggested to be noradrenergic and to contain the amine synthesizing enzymes. The lateral nerves are regarded, for the present, as dopaminergic neurons. These suggestions are in agreement with the previous demonstration of catecholaminergic neurons in this nematode. The 5-hydroxytryptamine accumulating neurons are tentatively identified as tryptaminergic.


Simple two-dimensional ultrasound technique to assess intrapartum cervical dilatation: a pilot study.
To describe a two-dimensional (2D) ultrasound technique to measure cervical dilatation in labor, and to compare ultrasound with digital measurements. 2D transperineal ultrasound was performed in 21 nulliparous women in labor with a singleton fetus in cephalic presentation and cervical dilatation measured before or after a digital vaginal examination. The absolute difference was calculated and Bland-Altman analysis was used to assess the mean difference between digital vaginal examination and ultrasound examination of cervical dilatation. Pearson analysis was used to determine the correlation between digital and ultrasound measurements. Intraclass correlation coefficients (ICCs) with 95% CI were used to evaluate the reliability of the two methods. Satisfactory quality images of the cervix were obtained in 19 of 21 cases. There was positive correlation between 2D ultrasound measurement of cervical dilatation and digital vaginal examination (Pearson coefficient r = 0.821, n = 19, P < 0.001). Bland-Altman analysis showed a mean difference between digital and ultrasound measurements of 0.08 cm (95% limits of agreement: -1.83 to 2.00) and the mean absolute difference was 1.24 cm. The ICC between the two methods was 0.81 (95% CI, 0.58-0.92). Assessment and measurement of cervical dilatation by 2D transperineal ultrasound is feasible, with close agreement shown between the technique and digital vaginal examination. The technique that we describe could represent an important component of a 'sonopartogram' for ultrasound assessment of labor progress.


[Orthopedic aspects in interdisciplinary multimodal therapy of chronic back pain].
The effect of interdisciplinary multimodal therapy of chronic back pain is well documented. With elapsing time changing diagnostic focuses, therapeutic strategies and objectives have to be considered. The chronicity leads to a modification of the relevance of structure-related diagnosis and therapy and changes the significance of the classic orthopedic instruments. The requirement of a rational causal therapy in chronic back pain still remains but the focal points shift to the consideration of somatic, psychological and social disposing and supporting factors.The aim of this paper is to reflect the necessary orthopedic expertise in the context of the pathomechanics of chronic back pain and the interdisciplinary teamwork.


LINC00665 promotes the viability, migration and invasion of T cell acute lymphoblastic leukemia cells by targeting miR-101 via modulating PI3K/Akt pathway.
T cell acute lymphoblastic leukemia (T-ALL) is a high-risk malignancy. The effects of cancer growth-related Long Intergenic Non-Protein Coding RNA 665 (LINC00665) in T-ALL remained obscure, and therefore further exploration was conducted on that in this study. The expression of LINC00665 in acute myeloid leukemia (LAML) tissues and myeloid tissues was analyzed using Gene Expression Profiling Interactive Analysis (GEPIA) 2. The target microRNA (miR) of LINC00665 was predicted by LncBase Predicted v.2 and verified using dual-luciferase reporter assay. After LINC00665 and miR-101 in T-ALL cells were overexpressed or silenced, the viability, migration and invasion of cell were detected using cell counting kit-8 and Transwell assays. The expressions of LINC00665, miR-101, Cyclin D1, Matrix metalloproteinases (MMP)-2, MMP-9, phosphorylated (p)-phosphatidylinositol 3-kinase (PI3K), PI3K, p-Akt, Akt were detected by quantitative real-time Polymerase Chain Reaction (qRT-PCR) and western blot. High expression of LINC00665 was presented in LAML tissues, the peripheral blood samples from patients with T-ALL and T-ALL cells. Overexpression of LINC00665 promoted the viability, migration and invasion of T-ALL cells and downregulated miR-101 expression, whereas silencing of LINC00665 did oppositely. MiR-101 could competitively bind to LINC00665, and was low-expressed in T-ALL. MiR-101 mimic inhibited viability, migration and invasion of T-ALL cells, and reversed effects of overexpressed LINC00665, whilst miR-101 inhibitor reversed the effects of LINC00665 silencing. Besides, overexpressed LINC00665 upregulated the expressions of Cyclin D1 MMP-2, and MMP-9 and the ratios of p-PI3K/PI3K and p-Akt/Akt, which were reversed by miR-101 mimic. LINC00665 could enhance the viability, migration and invasion abilities of T-ALL cells by targeting miR-101 via activating PI3K/Akt pathway.


Cold-induced amnesia blocks escape deficits in the cockroach (Periplaneta americana).
For three consecutive days, four groups of adult cockroaches (Periplaneta americana) were yoked to cockroaches receiving escapable shock. The four groups of subjects received either inescapable shock followed by exposure to 12 minutes of cold (2 degrees C), inescapable shock followed by no exposure to cold, no-shock followed by exposure to cold, or no-shock followed by no exposure to cold. 24 hr. later all five groups were run on a different escape task. The cockroaches exposed to inescapable shock followed by no exposure to cold showed the usual escape deficit. The cockroaches exposed to inescapable shock followed by exposure to cold did not alter from the escapable shock cockroaches or the controls.


Toxicant-mediated redox control of proteostasis in neurodegeneration.
Disruption in redox signaling and control of cellular processes has emerged as a key player in many pathologies including neurodegeneration. As protein aggregations are a common hallmark of several neuronal pathologies, a firm understanding of the interplay between redox signaling, oxidative and free radical stress, and proteinopathies is required to sort out the complex mechanisms in these diseases. Fortunately, models of toxicant-induced neurodegeneration can be utilized to evaluate and report mechanistic alterations in the proteostasis network (PN). The epidemiological links between environmental toxicants and neurological disease gives further credence into characterizing the toxicant-mediated PN disruptions observed in these conditions. Reviewed here are examples of mechanistic interaction between oxidative or free radical stress and PN alterations. Additionally, investigations into toxicant-mediated PN disruptions, specifically focusing on environmental metals and pesticides, are discussed. Finally, we emphasize the need to distinguish whether the presence of protein aggregations are contributory to phenotypes related to neurodegeneration, or if they are a byproduct of PN deficiencies.


Forearm Interosseous Membrane Maintains the Stability of Proximal Radioulnar Joint.
To evaluate the effect of the proximal and central bundles of the interosseous membrane on the stability of proximal radioulnar joint. Twenty fresh samples of human forearm provided by the anatomy room of the Department of Human Anatomy of Nanjing Medical University were included in this study. They were used to explore the effect of proximal interosseous membrane bundle on the stability of proximal radioulnar joint. The proximal bundle was reconstructed along the original attachment point. The reconstructions of central bundle were divided into the reconstruction of original attachment point on radius-midpoint of the ulnar original attachment point (reconstruction A) and original attachment point reconstruction (reconstruction B). The loads of the proximal radioulnar joint in different positions were measured. The load of the proximal radioulnar joint was analyzed in neutral, pronation, and supination positions. After resection of proximal and central fascicles, the loads of proximal radioulnar joint in neutral, pronation, and supination positions were significantly lower than those before resection (P < 0.05). After reconstruction, the loads of proximal radioulnar joint in neutral and supination positions were higher than those after resection (P < 0.05). After reconstruction, the loads of proximal radioulnar joint in neutral and supination positions were higher than those after resection (P < 0.05), and that after reconstruction B in pronation position was higher than that after resection (P < 0.05), while there was no significant difference between reconstruction A and after resection (P > 0.05). In supination position, the load of reconstruction B was higher than that of reconstruction A (P < 0.05). After reconstruction of the proximal and central bundles, the proximal radioulnar joint could not reached the same load as it could before resection (P < 0.05). The stability of proximal radioulnar joint is affected by central bundle and proximal bundle. Reconstruction can increase the stability of proximal radioulnar joint.


Evaluation of resistance mechanisms and serotype and genotype distributions of macrolide-resistant strains in clinical isolates of Streptococcus pneumoniae [corrected] in Aydın, Turkey.
Macrolide resistance mechanisms in 89 Streptococcus pneumoniae strains isolated from several clinical samples between February 2007 and May 2009 were investigated. Erythromycin resistance was noted in 35 (40%) S. pneumoniae strains. In these strains, the most frequent resistance phenotype was cMLS(B) (74%), and the most frequent resistance genotype was ermB (82%). Both ermB and mefA genes were positive in 20% of macrolide-resistant strains. While no resistance to vancomycin, linezolid and telithromycin was noted in 89 S. pneumoniae strains, 12 (13%) strains were penicillin resistant, 26 (30%) strains were clindamycin resistant, 35 (40%) were azithromycin resistant, 32 (36%) strains were tetracycline resistant, and 1 (1%) strain was levofloxacin resistant. The serotype distribution of 35 macrolide-resistant S. pneumoniae strains revealed that the most frequent serotype was serogroup 19 (45%). Multidrug resistance was present in 19 (86%) of 22 strains carrying only the ermB resistance gene. No clonal dissemination was noted in the macrolide-resistant pneumococcal strains. These findings suggest that macrolide resistance rates, resistance phenotype and genotype, as well as resistant serotypes of S. pneumoniae strains should be continuously monitored in our country.


Cytoplasmic localization of IRF5 induces Wnt5a/E-cadherin degradation and promotes gastric cancer cells metastasis.
IRF5, a nucleoplasm shuttling protein, is a pivotal transcription factor regulating immune system activity. It's well known that immunosuppression is involved in the development of gastric cancer. However, no data exist for the expression and function of IRF5 in gastric cancer. This study demonstrated that IRF5 was cytoplasm-enriched in gastric cancer cells. IRF5 promoted gastric cancer cell migration, which involved the inhibition of Wnt5a and E-cadherin proteins expression. IRF5 (LA) localized in nucleus had no significant effect on Wnt5a and E-cadherin expressions, while mutation of IRF5 (ΔNLS), which prevents IRF5 nuclear translocation, had more impact on these inhibitory effects. In addition, degradation rates of both Wnt5a and E-cadherin were enhanced by resiquimod, an IRF5 agonist. Further in vivo experiments indicated that IRF5 knockout of gastric cancer cells repressed their pulmonary metastasis in nude mice. Finally, the expression and clinical significance of IRF5 were analyzed using gastric cancer tissue microarrays, which suggested that the expression of IRF5 varied procedurally in different progressive stages of gastric cancer. Our data revealed that IRF5 cytoplasmic localization were associated with Wnt5a and E-cadherin degradation and gastric cancer cell metastasis. Inhibiting IRF5 expression and/or its cytoplasmic localization may provide a novel target for gastric cancer therapy.


Effects of oxygen availability on maximum aerobic performance in Mus musculus selected for basal metabolic rate or aerobic capacity.
Maximum aerobic metabolism cannot increase indefinitely in response to demands for ATP production and, therefore, must be constrained by one (or many) of the steps of the oxygen transport and utilization pathways. To elucidate those constraints we compared peak metabolic rate elicited by running (V(.)(O₂,run)) in hypoxia (14% O₂), normoxia (21% O₂) and hyperoxia (30% O₂) of laboratory mice divergently selected for low and high basal metabolic rate (L-BMR and H-BMR, respectively), mice selected for maximum metabolic rate elicited by swimming (V(.)(O₂,swim)) and mice from unselected lines. In all line types (V(.)(O₂,run)) was lowest in hypoxia, intermediate in normoxia and highest in hyperoxia, which suggests a 'central' limitation of oxygen uptake or delivery instead of a limit set by cellular oxidative capacity. However, the existence of a common central limitation is not in agreement with our earlier studies showing that selection on high V(.)(O₂,swim) (in contrast to selection on high BMR) resulted in considerably higher oxygen consumption during cold exposure in a He-O₂ atmosphere than V(.)(O₂,run). Likewise, between-line-type differences in heart mass and blood parameters are inconsistent with the notion of central limitation. Although responses of V(.)(O₂,run) to hypoxia were similar across different selection regimens, the selection lines showed contrasting responses under hyperoxic conditions. V(.)(O₂,run) in the H-BMR line type was highest, suggesting that selection on high BMR led to increased cellular oxidative capacity. Overall, between-line-type differences in the effect of the oxygen partial pressure on V(.)(O₂,run) and in the components of O₂ flux pathways are incompatible with the notion of symmorphosis. Our results suggest that constraints on V(.)(O₂,max) are context dependent and determined by interactions between the central and peripheral organs and tissues involved in O₂ delivery.


Direct measurement of nitrous oxide MAC and neurologic monitoring in rats during anesthesia under hyperbaric conditions.
The minimum alveolar concentration (MAC) of nitrous oxide necessary to prevent purposeful movement in rats has not been directly measured; rather, it has been extrapolated because the required partial pressure exceeds 760 mm Hg, or 1 atm absolute pressure (ATA). Values reported have ranged from 1.36 to 2.20 ATA (136-220 vol%, or 1034-1672 mm Hg). By maintaining general anesthesia at 2.25 ATA (1710 mm Hg), we directly measured the nitrous oxide MAC in 17 Long-Evans rats during mechanical ventilation and monitoring of two-channel electroencephalogram, compressed spectral array and cortical evoked potentials, electrocardiograph, and respiratory and anesthetic gases by mass spectrometry. After a minimal stabilization period of 30 min during ventilation by 1.8 ATA nitrous oxide and 0.45 ATA oxygen, MAC measurements were begun. Each rat was given up to three noxious electrical stimulations of 50 V by 10-ms-duration pulses at 50/s for 45 s. The partial pressure of nitrous oxide was decreased by approximately 10% after each negative response. The MAC was taken as the nitrous oxide concentration midway between that at which there was no response and that at which the rat moved purposefully. The nitrous oxide MAC in Long-Evans rats was determined to be 1.55 +/- 0.16 ATA (mean +/- SD). Hyperbaric nitrous oxide decreased electroencephalogram wave frequency to a predominantly theta rhythm of increased amplitude. Cortical evoked potentials had decreased wave amplitudes and increased latencies with increasing partial pressures > 0.75 ATA.(ABSTRACT TRUNCATED AT 250 WORDS)


MiR-98 suppresses the effects of tumor-associated macrophages on promoting migration and invasion of hepatocellular carcinoma cells by regulating IL-10.
We aim to investigate the role of miR-98-mediated macrophage polarization in hepatocellular carcinoma (HCC) progression and to explore the underlying mechanism. A total of 25 paired HCC and matched adjacent normal tissues (ANTs) were collected. We incubated human blood monocytes isolated from healthy male donors with culture medium collected from HepG2 cells for 7 days. The mRNA and protein expression was detected by qRT-PCR and Western blot, respectively. Levels of cytokines secreted in culture medium were measured using the specific ELISA kits. The miR-98 mimic/inhibitor was transfected to explore the role of miR-98 in HCC-conditioned tumor-associated macrophages (TAMs). HepG2 cells were then cultured with condition medium from HCC-conditioned TAMs pretreated with miR-98 mimic/inhibitor, and cell migration and invasion assays were performed. Luciferase reporter assay was performed to analyze the interaction between miR-98 and interleukin (IL)-10. Our results showed that miR-98 was downregulated and IL-10 was upregulated in HCC tissues and HCC-conditioned TAMs. Further studies identified that IL-10 was a direct target gene of miR-98 in HCC-conditioned TAMs. Moreover, miR-98 regulated the levels of inflammatory cytokines in HCC-conditioned TAMs. HCC-conditioned TAMs pretreated with miR-98 regulated migration and invasion of HepG2 cells in vitro, and the effects were significantly reversed by IL-10. In conclusion, miR-98 not only regulated expression of inflammatory cytokines in HCC-conditioned TAMs, but also modulated the capacity of HCC-conditioned TAMs to regulate HepG2 cell migration and invasion, at least in part, by targeting IL-10. As a result, miR-98 may suppress the progress of HCC.


The many roles of myosins in filopodia, microvilli and stereocilia.
Filopodia, microvilli and stereocilia represent an important group of plasma membrane protrusions. These specialized projections are supported by parallel bundles of actin filaments and have critical roles in sensing the external environment, increasing cell surface area, and acting as mechanosensors. While actin-associated proteins are essential for actin-filament elongation and bundling in these protrusions, myosin motors have a surprising role in the formation and extension of filopodia and stereocilia and in the organization of microvilli. Actin regulators and specific myosins collaborate in controlling the length of these structures. Myosins can transport cargoes along the length of these protrusions, and, in the case of stereocilia and microvilli, interactions with adaptors and cargoes can also serve to anchor adhesion receptors to the actin-rich core via functionally conserved motor-adaptor complexes. This review highlights recent progress in understanding the diverse roles myosins play in filopodia, microvilli and stereocilia.


Quality management of pharmacology and safety pharmacology studies.
Pharmacology has traditionally been excluded from the mandatory application of good laboratory practice (GLP) principles. Consensus has been reached through the process of the International Conference on Harmonisation (ICH, Topic S7A) with regard to the definitions of the different types of pharmacology studies (ICH S7A): primary pharmacodynamic, secondary pharmacodynamic and safety pharmacology studies, and guidance on the quality standards (expectations for GLP conformity) for these study types have been provided. Primary pharmacodynamic studies are the only study types that are fully exempt from GLP requirements. Secondary pharmacodynamic and safety pharmacology studies are expected to be conducted to GLP quality standards -- preferably to full, formal GLP compliance, when results are used for human safety assessment. At the present time, regulatory authorities will most likely be prepared to exercise flexibility in their requirement for GLP compliance, however, if non-clinical studies used in human safety assessment are not formally in compliance with the principles of GLP, regulatory acceptance may not be guaranteed. Historically, the application of formal GLP standards in safety pharmacology studies appears to vary between test facilities, and a number of study components in safety pharmacology studies, e.g. ECG monitoring, may not always conform to formal GLP standards. Apparently, however, formal GLP standards for these study components can be implemented for a relatively low additional cost. Based on the guidance given in the ICH S7A guideline, it thus appears logical to recommend that test facilities and sponsors consider their organisation of safety pharmacology studies in view of sound study management and formal implementation of GLP, where needed. Organisation of study management should facilitate collaboration across scientific disciplines because a plethora of data, originating from basic pharmacodynamics, toxicology, kinetics, and metabolism, as well as from clinical investigations, are involved in a safety pharmacology assessment. Applying formal GLP standards to safety pharmacology studies, and, when indicated, to secondary pharmacodynamic studies, does not influence the scientific standards of studies. However, applying formal GLP standards will ensure the quality, reliability and integrity of studies, which reflect sound study management. It is important to encourage a positive attitude among researchers and academics towards these lines, whenever possible. GLP principles applied to the management of non-clinical safety studies are appropriate quality standards when studies are used in the context of protecting public health, and these quality standards are therefore obviously pivotal for regulatory acceptance of non-clinical safety studies in general, and of safety pharmacology studies in particular.


Thrombin induces ACSL4-dependent ferroptosis during cerebral ischemia/reperfusion.
Ischemic stroke represents a significant danger to human beings, especially the elderly. Interventions are only available to remove the clot, and the mechanism of neuronal death during ischemic stroke is still in debate. Ferroptosis is increasingly appreciated as a mechanism of cell death after ischemia in various organs. Here we report that the serine protease, thrombin, instigates ferroptotic signaling by promoting arachidonic acid mobilization and subsequent esterification by the ferroptotic gene, acyl-CoA synthetase long-chain family member 4 (ACSL4). An unbiased multi-omics approach identified thrombin and ACSL4 genes/proteins, and their pro-ferroptotic phosphatidylethanolamine lipid products, as prominently altered upon the middle cerebral artery occlusion in rodents. Genetically or pharmacologically inhibiting multiple points in this pathway attenuated outcomes of models of ischemia in vitro and in vivo. Therefore, the thrombin-ACSL4 axis may be a key therapeutic target to ameliorate ferroptotic neuronal injury during ischemic stroke.


Translation of Taenia crassiceps mRNA in cell-free heterologous systems.
1. mRNA isolated from larval Taenia crassiceps directs efficiently the synthesis of proteins in cell-free heterologous systems. 2. Part of the newly synthesized proteins in a reticulocyte system are precipitable by a rabbit antiserum against T. crassiceps proteins. 3. Analysis of the antiserum-protein dissociated complex by sodium dodecyl sulfate polyacrylamide gel electrophoresis indicates that most of the proteins synthesized are of low molecular weight (13,000-22,000) although a protein of mol. wt. 260,000 is also produced. 4. Whether the newly synthesized proteins which are precipitable by specific antisera correspond to parasite antigens or to proteins with closely antigenic similarities remains to be established.


Molecular identification and safety assessment of the potential probiotic strain Bacillus paralicheniformis HMPM220325 isolated from artisanal fruit dairy products.
Bacillus probiotics exhibit considerable economic potential owing to their heightened resilience to external stressors and relatively lower costs related to production and preservation. Although Bacillus paralicheniformis has been acknowledged as a plant-promoting bacterium for a long time, understanding its potential as a probiotic is still in its nascent stages. In this study, the safety and probiotic characteristics of a strain of HMPM220325, isolated from artisanal fruit dairy products, were examined through whole-genome sequencing and phenotypic analysis. The whole genome of HMPM220325 was analyzed for antimicrobial resistance genes, pathogenicity factors, and genes associated with probiotic traits including stress resistance, spore formation, gut adhesion, competitive exclusion of pathogens, bacteriocin expression, and carbohydrate metabolism related to prebiotic utilization. Also, wet lab experiments were conducted for the characterization of probiotics. The identification of the organism as B. paralicheniformis was verified. Its safety was assessed through in silico analysis, the haemolytic activity test, and the acute oral toxicity test. B. paralicheniformis HMPM220325 demonstrated its ability to survive in the pH range of 4-10 and bile salt concentrations of 0-0.9% (w/v), tolerate temperatures between 20 and 60 °C, and exhibit a robust antioxidant capacity. Moreover, B. paralicheniformis HMPM220325 demonstrated a moderate level of hydrophobicity, had the ability to form biofilms, achieved a self-aggregation rate of 51.77 ± 1.01% within 6 hours, and successfully colonized the mouse intestine for a duration of up to 17 days. Additionally, the genome of B. paralicheniformis HMPM220325 contains three gene clusters associated with the biosynthesis of bacteriocins and exhibits co-aggregation with Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella enterica serovar Typhimurium. The findings of the genomic analysis align with those obtained from the experimental investigation, thereby substantiating the potential of B. paralicheniformis HMPM220325 as a probiotic suitable for incorporation in dairy functional foods and feed applications.


Geranyl diphosphate synthase: an important regulation point in balancing a recombinant monoterpene pathway in Escherichia coli.
The expression level of geranyl diphosphate synthase (GPPS) was suspected to play a key role for geraniol production in recombinant Escherichia coli harboring an entire mevalonate pathway operon and a geraniol synthesis operon. The expression of GPPS was optimized by using ribosomal binding sites (RBSs) designed to have different translation initiation rates (TIRs). The RBS strength in TIR window of 500 arbitrary unit (au)-1400 au for GPPS appears to be suitable for balancing the geraniol biosynthesis pathway in this study. With the TIR of 500 au, the highest production titer of geraniol was obtained at a level of 1119mg/L, which represented a 6-fold increase in comparison with the previous titer of 183mg/L. The TIRs of GPPS locating out of range of the optimal window (500-1400 au) caused significant decreases of cell growth and geraniol production. It was suspected to result from metabolic imbalance and plasmid instability in geraniol production by inappropriate expression level of GPP synthase. Our results collectively indicated GPPS as an important regulation point in balancing a recombinant geraniol synthesis pathway. The GPPS-based regulation approach could be applicable for optimizing microbial production of other monoterpenes.


Wnt/β-catenin signal pathway stabilizes APP intracellular domain (AICD) and promotes its transcriptional activity.
Amyloid precursor protein (APP), a key protein in pathogenesis of Alzheimer's disease (AD), is a type I transmembrane protein which can be cleaved by β- and γ-secretase to release the amyloidogenic β-amyloid peptides (Aβ) and the APP intracellular domain (AICD). While Aβ has been widely believed to initiate pathogenic cascades culminating AD, the physiological functions and regulations of AICD remain elusive. In present study, endogenous AICD was demonstrated to be increased by canonical Wnt signal. Instead of due to γ-secretase activity, enhanced AICD expression was found due to the increased protein stability by Wnt/β-catenin. β-Catenin was demonstrated to be an associating partner of AICD, capable of promoting AICD mediated transcriptional activity. Investigation by AICD mutants proved that Fe65, a previously identified AICD binding partner, is not involved in this regulation. Taken together, our results suggest that AICD is stabilized and the AICD mediated transcriptional activity is promoted by canonical Wnt/β-catenin signaling independent of Fe65.


Phosphorylation-dependent conformational transition of the cardiac specific N-extension of troponin I in cardiac troponin.
We present here the solution structure for the bisphosphorylated form of the cardiac N-extension of troponin I (cTnI(1-32)), a region for which there are no previous high-resolution data. Using this structure, the X-ray crystal structure of the cardiac troponin core, and uniform density models of the troponin components derived from neutron contrast variation data, we built atomic models for troponin that show the conformational transition in cardiac troponin induced by bisphosphorylation. In the absence of phosphorylation, our NMR data and sequence analyses indicate a less structured cardiac N-extension with a propensity for a helical region surrounding the phosphorylation motif, followed by a helical C-terminal region (residues 25-30). In this conformation, TnI(1-32) interacts with the N-lobe of cardiac troponin C (cTnC) and thus is positioned to modulate myofilament Ca2+-sensitivity. Bisphosphorylation at Ser23/24 extends the C-terminal helix (residues 21-30) which results in weakening interactions with the N-lobe of cTnC and a re-positioning of the acidic amino terminus of cTnI(1-32) for favorable interactions with basic regions, likely the inhibitory region of cTnI. An extended poly(L-proline)II helix between residues 11 and 19 serves as the rigid linker that aids in re-positioning the amino terminus of cTnI(1-32) upon bisphosphorylation at Ser23/24. We propose that it is these electrostatic interactions between the acidic amino terminus of cTnI(1-32) and the basic inhibitory region of troponin I that induces a bending of cTnI at the end that interacts with cTnC. This model provides a molecular mechanism for the observed changes in cross-bridge kinetics upon TnI phosphorylation.


Capillary zone electrophoresis-mass spectrometry for top-down proteomics.
Mass spectrometry (MS)-based top-down proteomics characterizes complex proteomes at the intact proteoform level and provides an accurate picture of protein isoforms and protein post-translational modifications in the cell. The progress of top-down proteomics requires novel analytical tools with high peak capacity for proteoform separation and high sensitivity for proteoform detection. The requirements have made capillary zone electrophoresis (CZE)-MS an attractive approach for advancing large-scale top-down proteomics. CZE has achieved a peak capacity of 300 for separation of complex proteoform mixtures. CZE-MS has shown drastically better sensitivity than commonly used reversed-phase liquid chromatography (RPLC)-MS for proteoform detection. The advanced CZE-MS identified 6,000 proteoforms of nearly 1,000 proteoform families from a complex proteome sample, which represents one of the largest top-down proteomic datasets so far. In this review, we focus on the recent progress in CZE-MS-based top-down proteomics and provide our perspectives about its future directions.


Induction and disappearance of thymine dimers in human skin exposed to UVB radiation: flow cytometric measurements in replicating and nonreplicating epidermal cells.
We have earlier reported on determining UV-induced DNA damage in murine epidermal cell suspensions by flow cytometric analysis of the fluorescence from a fluorescein isothiocyanate-labeled antibody (H3) directed against thymine dimers (T < > T). Here we present an optimization of the technique for analysis of epidermal cell suspensions from 4 mm biopsies from human skin. Cells with different DNA contents can easily be distinguished in flow cytometry by the intensity of DNA-specific 7-amino-actinomycin D fluorescence. Genuine G2-M-phase cells can further be distinguished from cell doublets by pulse-shape discrimination. Thus, T < > T levels in individual cells with different DNA contents (i.e. G0-G1, S or G2-M phases) can be determined after in vivo exposure of human skin to environmentally relevant UVB (280-315 nm) doses. The method was applied to measure the decrease of T < > T in nonreplicating cells (G0-G1 phase) and replicating cells (S phase or G2-M phase) from seven volunteers exposed to twice their minimal erythema dose. The reduction in the average T < > T-specific fluorescence at 24 h after exposure was 46% (ranging between 16% and 66%) for the G0-G1 cells and 70% (ranging between 37% and 100%) for the S + G2-M cells. The difference was statistically highly significant. Determination of individual DNA repair capacities with this method can become a convenient diagnostic tool for patients with DNA repair disorders, or it may even be used to identify individuals with low repair proficiencies and increased risk of developing skin cancers.


Performance evaluation of an electrostatic precipitator with a copper plate using an aerosolized SARS-CoV-2 surrogate (bacteriophage phi 6).
The global spread of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has reminded us of the importance of developing technologies to reduce and control bioaerosols in built environments. For bioaerosol control, the interaction between researchers and biomaterials is essential, and considering the characteristics of target pathogens is strongly required. Herein, we used enveloped viral aerosols, bacteriophage phi 6, for evaluating the performance of an electrostatic precipitator (ESP) with a copper-collecting plate (Cu-plate). In particular, bacteriophage phi 6 is an accessible enveloped virus that can be operated in biosafety level (BSL)-1 as a promising surrogate for SARS-CoV-2 with structural and morphological similarities. ESP with Cu-plate showed >91% of particle removal efficiency for viral aerosols at 77 cm/s of airflow face velocity. Moreover, the Cu-plate presented a potent antiviral performance of 5.4-relative log reduction within <15 min of contact. We believe that the evaluation of ESP performance using an aerosolized enveloped virus and plaque assay is invaluable. Our results provide essential information for the development of bioaerosol control technologies that will lead the post-corona era.


Mucormycosis of Paranasal Sinuses of Odontogenic Origin Post COVID19 Infection: A Case Series.
The Severe acute respiratory syndrome and the Middle East respiratory syndromes emerged in 2002 and 2012 respectively. Currently the world is witnessing a global pandemic caused by a novel coronavirus, the severe acute respiratory syndrome coronavirus 2 (SARS CoV- 2) causing the Coronavirus 2019 (COVID-19). Mucormycosis is a fungal infection primarily affecting individuals with an immunocompromised state like diabetes mellitus, malignancies etc. Patients who have or have had COVID-19 infection with pre-existing uncontrolled Type 2 Diabetes mellitus are presumably more vulnerable for emergence of fungal infections cases. This article presents a report of 6 cases with histopathological proven mucormycosis associated with COVID-19 and uncontrolled Diabetes mellitus.


Fluid intake and risk of stress, urgency, and mixed urinary incontinence.
We investigated the relation between total fluid intake and incident urinary incontinence in the Nurses' Health Study cohorts. We measured daily fluid intake using food frequency questionnaires among 65,167 women, who were 37-79 years old, without urinary incontinence at study baseline (2000-2001). Women reported incontinence incidence on questionnaires during 4 years of follow-up evaluation. Multivariable-adjusted hazard ratios and 95% confidence intervals were calculated with Cox proportional hazards models. We found no association between total fluid intake and risk of incident incontinence (hazard ratio, 1.04; 95% confidence interval, 0.98-1.10; comparing top vs bottom quintile of fluid intake). In analyses of incontinence type, total fluid intake was not associated with risks of incident stress, urgency, or mixed incontinence. No significant risk of incident urinary incontinence was found with higher fluid intake in women. These findings suggest that women should not restrict their fluid intake to prevent incontinence development.


Expression, crystallization and preliminary crystallographic study of the C-terminal half of nsp2 from SARS coronavirus.
SARS coronavirus (SARS-CoV) is the aetiological agent of the highly infectious severe acute respiratory syndrome (SARS). To gain a better understanding of SARS-CoV replication and transcription proteins, a preliminary X-ray crystallographic study of the C-terminal domain of SARS-CoV nonstructural protein 2 (nsp2) is reported here. The C-terminal domain of SARS-CoV nsp2 was cloned, overexpressed, purified and crystallized using polyethylene glycol 5000 monomethyl ether as the precipitant; the crystals diffracted to 2.5 Å resolution. The crystals belonged to space group P6(5), with unit-cell parameters a=b=112.8, c=91.1 Å, α=β=90, γ=120°. One molecule is assumed to be present per asymmetric unit, which gives a Matthews coefficient of 2.89 Å3 Da(-1) and a solvent content of 56.2%.


C-reactive protein 2 days after laparoscopic gastric bypass surgery reliably indicates leaks and moderately predicts morbidity.
The aim of the present study was to evaluate whether serum C-reactive protein (CRP) is a useful predictor of early post-operative complications, particularly of intestinal leaks after laparoscopic Roux-en-Y gastric bypass (LRYGB) surgery. The present study was a retrospective analysis of a prospectively maintained database with 809 patients who underwent LRYGB from 2002 until 2011. For 410 of these patients, at least one CRP measurement within the first seven post-operative days was available. The diagnostic value was determined by the area under the curve (AUC) of the receiver operating characteristic (ROC) curve. Forty-nine of 410 patients (12.0 %; 95 % confidence intervals [95 % CI], 9.2-15.5 %) developed surgery-related complications. Leaks occurred in 17 patients (4.1 %; 95 % CI, 2.6-6.5 %) at a median of 5 days after surgery. CRP levels 2 days after surgery showed the highest diagnostic value for post-operative complications (AUC, 0.74; 95 % CI, 0.60-0.89). Sensitivity was 0.53 (95 % CI, 0.31-0.74) and specificity was 0.91 (95 % CI, 0.79-0.96) on day 2 (cutoff level, 229 mg/l). The sensitivity for intestinal leaks was 1.00 (95 % CI, 0.51-1.00). CRP on post-operative day 2 is a valuable predictor of post-operative complications, in particular intestinal leaks. Radiological imaging studies for intestinal leaks could be restricted to patients with CRP values exceeding 229 mg/l.


Phosphoantigen sensing combines TCR-dependent recognition of the BTN3A IgV domain and germline interaction with BTN2A1.
Vγ9Vδ2 T cells play critical roles in microbial immunity by detecting target cells exposed to pathogen-derived phosphoantigens (P-Ags). Target cell expression of BTN3A1, the "P-Ag sensor," and BTN2A1, a direct ligand for T cell receptor (TCR) Vγ9, is essential for this process; however, the molecular mechanisms involved are unclear. Here, we characterize BTN2A1 interactions with Vγ9Vδ2 TCR and BTN3A1. Nuclear magnetic resonance (NMR), modeling, and mutagenesis establish a BTN2A1-immunoglobulin V (IgV)/BTN3A1-IgV structural model compatible with their cell-surface association in cis. However, TCR and BTN3A1-IgV binding to BTN2A1-IgV is mutually exclusive, owing to binding site proximity and overlap. Moreover, mutagenesis indicates that the BTN2A1-IgV/BTN3A1-IgV interaction is non-essential for recognition but instead identifies a molecular surface on BTN3A1-IgV essential to P-Ag sensing. These results establish a critical role for BTN3A-IgV in P-Ag sensing, in mediating direct or indirect interactions with the γδ-TCR. They support a composite-ligand model whereby intracellular P-Ag detection coordinates weak extracellular germline TCR/BTN2A1 and clonotypically influenced TCR/BTN3A-mediated interactions to initiate Vγ9Vδ2 TCR triggering.


The Neurological Hand Deformity Classification: Construct validity, test-retest, and inter-rater reliability.
The Neurological Hand Deformity Classification (NHDC) is an impairment-based tool that classifies hand deformity into one of two ordinal scales: flexion or extension deformities. Classification is made from live observation or from recorded video footage. Differentiation between the levels is determined by wrist position and wrist and finger movement. To examine aspects of validity and reliability of the NHDC. A measurement study design. Data from a convenience sample of 127 children with cerebral palsy, 66 males: 61 females, ranging in age from 8 months to 15 years, across all Manual Ability Classification System levels I to V, were analyzed. Construct validity was assessed by testing predetermined hypotheses of relationships between the NHDC and measures of body function and activity measures with observed performance using the Chi Squared Test of Independence and Spearman Correlation Coefficient. Test-retest and inter-rater reliability were assessed by calculating agreement between repeated measures and paired raters using weighted kappa and Cohen's kappa with 95% confidence intervals. Predicted hypotheses for the NHDC were met in nine of 10 Spearman's rho correlations with body structure measures and in 2 of 7 correlations with activity measures. Test-retest for flexion deformities: κw = 0.84; 95% CI 0.70-0.98; and extension deformities: κ = 1.0; 95% CI 1.0-1.0 was good to excellent; inter-rater reliability for flexion deformities: κw = 0.76; 95% CI 0.67-0.85; and extension deformities κ = 0.75; 95% CI 0.43-1.0 was moderate to excellent. Expected relationships between the NHDC and other measures, stability between repeated measures and acceptable between-rater agreement supports confidence classifying hand deformity in children with cerebral palsy with the NHDC.


Two-dimensional speckle-tracking echocardiography of the left atrium: feasibility and regional contraction and relaxation differences in normal subjects.
Increased left atrial (LA) size and reduced global contractility are related to adverse cardiac events. The potential incremental value of assessing regional LA contractility is unknown. To assess the feasibility of measuring this variable angle, independent 2-dimensional speckle-tracking strain echocardiography (2D-SpTr) was used to measure regional LA strain (epsilon) and strain rate (SR) in normal individuals of various ages. From standard apical views, 2D-SpTr was used on 84 normal subjects to measure longitudinal velocity, epsilon, and SR in 13 LA segmental regions. The values obtained from the different atrial regions were compared with each other and corresponding LA volumes before and after LA contraction. Regional LA epsilon and SR could be measured in 77 of 84 normal subjects (94%). A consistent pattern of differences in LA regional function was noted with the annular regions, and particularly the inferior wall having a larger average peak velocity and epsilon and SR values in comparison with the mid and superior LA segments. Peak epsilon and SR during LA contraction had only a modest correlation with LA volumes. The angle-independent technique of 2D-SpTr tracking can analyze regional LA epsilon and SR in 94% of normal subjects. Regional differences in LA contractility are consistently present. The annular regions, and especially the inferior wall have the highest values for LA epsilon and SR. The significance of these findings and their possible use in identifying disease states will require further study.


Characteristics of men who aggress sexually and of men who imagine aggressing: risk and moderating variables.
The authors showed that the extent to which men's personalities were self-centered rather than sensitive to others' needs moderated the connection between risk factors and sexually aggressive behavior. Men who were at risk for committing aggression but who were also sensitive to others' feelings aggressed less than the corresponding group, who had relatively self-centered personalities. However, both groups showed high levels of imagined sexual aggression. The authors suggest that imagined sexual aggression may reveal information about the presence of underlying risk factors even when actual aggression is inhibited by personality characteristics such as those studied here. The implications for therapeutic interventions of the finding of aggression attenuation are also discussed.


Consecutive administration of paraquat to rats induces enhanced cholesterol peroxidation and lung injury.
It is our hypothesis that as a consequence of increased oxidative stress, rats develop lung injury with increased cholesterol-derived hydroperoxides and oxysterols in lung after consecutive exposure of the rats to paraquat. To test this we administered 10 mg/kg of paraquat i.p. once or seven times (once a day) to Wistar rats. Rats were killed, and lung tissue was collected 24 h after the last paraquat injection. We found that in response to consecutive paraquat doses, there were significant increases in 7alpha- and 7beta-hydroperoxycholest-5-en-3beta-ol (7alpha-OOH and 7beta-OOH; P=0.01) as well as 7alpha- and 7beta-hydroxycholesterol (7alpha-OH and 7beta-OH; P=0.01), and 7-ketocholesterol (7-keto; P=0.03). In addition, pulmonary hemorrhage, thickening of alveolar septum, and inflammatory cell infiltration of macrophages were observed. This is the first report showing enhanced cholesterol peroxidation and lung injury of rats due to consecutive doses of paraquat.


Switching stable patients with schizophrenia from depot and oral antipsychotics to long-acting injectable risperidone: efficacy, quality of life and functional outcome.
This international, non-randomised study evaluated maintained efficacy and safety of Risperidone Long-Acting Injectable (RLAI) compared to previous medications. To investigate the possible effect of differences in national health care systems across Europe, the UK subset efficacy, health-related quality of life (HRQoL) and functioning data are reported here. Patients with schizophrenia or other psychotic disorders, symptomatically stable on antipsychotic medication, received intramuscular injections of RLAI 25 mg (to a maximum of 50 mg) every 2 weeks for 6 months. Of 182 UK patients recruited; 79% had schizophrenia (87% paranoid), 21% other psychotic disorders; 96% had been hospitalised at sometime. Improvement in mean Positive and Negative Syndrome Scale (PANSS) total score was significant at 1 month through to endpoint (p = 0.0001). There were significant improvements in PANSS subscales (p <or= 0.001), PANSS-Marder subscores (p <or= 0.0391), and in Clinical Global Impression-Severity (p = 0.0001) at endpoint. Patients reported improvements in: HRQoL (p < 0.05), Global Assessment of Functioning (p = 0.0001) and patient satisfaction (p = 0.0001) at endpoint. The proportion of patients rating RLAI treatment as 'very good' increased from 10% to 40%. 55% (22/40) of patients hospitalised at baseline were discharged by endpoint. Sustained, significant improvement in symptom control, HRQoL and functioning was seen in stable patients treated with RLAI over 6 months following a switch from previous antipsychotic medications.


Palmitate and thapsigargin have contrasting effects on ER membrane lipid composition and ER proteostasis in neuronal cells.
The endoplasmic reticulum (ER) is an organelle that performs several key functions such as protein synthesis and folding, lipid metabolism and calcium homeostasis. When these functions are disrupted, such as upon protein misfolding, ER stress occurs. ER stress can trigger adaptive responses to restore proper functioning such as activation of the unfolded protein response (UPR). In certain cells, the free fatty acid palmitate has been shown to induce the UPR. Here, we examined the effects of palmitate on UPR gene expression in a human neuronal cell line and compared it with thapsigargin, a known depletor of ER calcium and trigger of the UPR. We used a Gaussia luciferase-based reporter to assess how palmitate treatment affects ER proteostasis and calcium homeostasis in the cells. We also investigated how ER calcium depletion by thapsigargin affects lipid membrane composition by performing mass spectrometry on subcellular fractions and compared this to palmitate. Surprisingly, palmitate treatment did not activate UPR despite prominent changes to membrane phospholipids. Conversely, thapsigargin induced a strong UPR, but did not significantly change the membrane lipid composition in subcellular fractions. In summary, our data demonstrate that changes in membrane lipid composition and disturbances in ER calcium homeostasis have a minimal influence on each other in neuronal cells. These data provide new insight into the adaptive interplay of lipid homeostasis and proteostasis in the cell.


Somatic mutation theory, DNA repair rates, and the molecular epidemiology of p53 mutations.
The theory of somatic mutagenesis predicts that the frequency pattern of induced selectable mutations along a gene is the product of the probability patterns of the several sequential steps of mutagenesis, e.g., damage, repair, polymerase misreading, and selection. Together, the variance of these component steps is propagated to generate a mutagen's induced mutational spectrum along a gene. The step with the greatest component of variance will drive most of the variability of the mutation frequency along a gene. This most variable step, for UV-induced mutations, is the cyclobutyl pyrimidine dimer repair rate. The repair rate of cyclopyrimidine dimers is quite variable from nucleotide position to nucleotide position and we show that this variation along the p53 gene drives the C-->T transition frequency of non-melanocytic skin tumors. On showing that the kinetics of cyclopyrimidine dimer repair at any one nucleotide position are first order, we use this kinetic and the somatic mutation theory to derive Leq, the adduct frequency along a gene as presented to a DNA polymerase after a cell population reaches damage-repair equilibrium from a chronic dose of mutagen. Leq is the product of the first two sequential steps of mutagenesis, damage and repair, and the frequency of this product is experimentally mapped using ligation-mediated PCR. The concept of Leq is applied to mutagenesis theory, chronic dose genetic toxicology, genome evolution, and the practical problems of molecular epidemiology.


Spanish implantable cardioverter-defibrillator registry. 18th official report of the Heart Rhythm Association of the Spanish Society of Cardiology (2021).
This article presents the data corresponding to implantable cardioverter-defibrillator (ICD) implantations in Spain in 2021. The data were drawn from implanting centers, which voluntarily completed a data collection sheet during the procedure. In 2021, 7496 implant data sheets were received, compared with 7743 reported by Eucomed (European Confederation of Medical Suppliers Associations), indicating that data were collected from 96.8% of the devices implanted in Spain. Data completion ranged from 99.9% for "name of implanting hospital" to 8.9% for "implanting hospital". In 2021, 199 hospitals participated in the registry, exceeding the figures of previous years, with around 170 participating hospitals. The total rate of registered implants was 158/million inhabitants (163 according to Eucomed), making 2021 the year with the highest activity. However, the registry continues to show significant differences among the various autonomous communities and the lowest implantation rate of all the European countries participating in Eucomed. The Spanish implantable cardioverter-defibrillator registry for 2021 recorded an increase in the number of ICD implantations, reflecting the recovery of hospital activity after the initial impact of the COVID-19 pandemic in 2020. Although the total number of implants has increased in Spain, figures are still much lower than the European Union average, with differences persisting among Spanish autonomous communities.


[Transformation of malignant non-Hodgkin's lymphomas from low- to high-grade malignancy].
Clinical and histological findings of six patients with low grade non-Hodgkin lymphoma (NHL) which transformed to high grade tumours are reported: 5 out of the 6 cases showed transformation from the centroblastic-centrocytic type to the centroblastic-polymorphic type (Kiel classification). Clinical aggressivity, large tumour mass, high frequency of extranodal sites of disease, systemic symptoms and resistance to treatment were observed. This type of transformation is not rare, estimated to be about 20 p. 100 of cases in the literature, and is more common than Richter's syndrome in chronic lymphocytic leukaemia.


Childhood neurodevelopmental disorders and maternal hypertensive disorder of pregnancy.
To examine the association of maternal chronic hypertension and pregnancy-induced hypertension (PIH)/preeclampsia with childhood neurodevelopmental disorders (NDDs) in a large-scale population-based cohort. We conducted a linked Taiwan National Health Insurance Research Database cohort study of children born between 2004 and 2008 (n=877 233). Diagnoses of autism spectrum disorder (ASD), attention-deficit/hyperactivity disorder (ADHD), developmental delay, intellectual disability, cerebral palsy (CP), and epilepsy/infantile spasms were identified from birth to the end of 2015. Cox proportional hazards models were fitted with adjustment for potential confounders to estimate the effect of maternal hypertensive disorder of pregnancy on childhood outcomes. Compared with the offspring of unexposed mothers, offspring of mothers with chronic hypertension or PIH/preeclampsia exhibited increased risk of developing a wide spectrum of NDDs. Chronic hypertension was associated with increased risks of ADHD (hazard ratio 1.22, 95% confidence interval [CI] 1.13-1.31), developmental delay (1.29, 1.21-1.38), intellectual disability (1.67, 1.43-1.95), CP (1.45, 1.14-1.85), and epilepsy/infantile spasms (1.31, 1.10-1.56) in the offspring, whereas PIH/preeclampsia was associated with increased risks of ASD (1.27, 1.12-1.43), ADHD (1.23, 1.17-1.29), developmental delay (1.29, 1.24-1.35), intellectual disability (1.53, 1.37-1.71), CP (1.44, 1.22-1.70), and epilepsy/infantile spasms (1.36, 1.22-1.52) in the offspring after adjustment for potential confounders. The co-occurrence of maternal diabetes, preterm deliveries, or fetal growth restriction further increased the risk. Chronic hypertension or PIH/preeclampsia seems to be sufficient to increase the risk of childhood NDDs. What this paper adds Children exposed to maternal hypertensive disorders have a higher cumulative incidence of neurodevelopmental disorders (NDDs) than unexposed children. Chronic hypertension or pregnancy-induced hypertension/preeclampsia seems to be sufficient to increase the risk of childhood NDDs. Co-occurrence of maternal diabetes, preterm deliveries, or fetal growth restriction further increases the risk.


Baseline functioning among individuals with posttraumatic stress disorder and alcohol dependence.
Comorbid posttraumatic stress disorder (PTSD) and alcohol dependence (AD) may lead to a complicated and potentially severe treatment profile. Our study examined 167 individuals with both PTSD and AD compared with 105 individuals with PTSD without an alcohol use disorder (AUD) and 240 individuals with AD without PTSD on baseline psychosocial functioning. We hypothesized that individuals with PTSD/AD would be more socially and functionally impaired than individuals with only one disorder. Results indicated that participants with PTSD/AD were more likely to be unemployed, have less education, and report less income and were less likely to live with a partner than the participants with only a single disorder. However, they did not differ on symptom severity within these disorders (drinking frequency/quantity, PTSD, and anxiety symptoms) with the exception of depression and alcohol craving. This contradicts clinical lore that comorbid patients are more impaired at treatment initiation and adds support for concurrent treatment as not only feasible but also possibly ideal for these patients.


Recordkeeping, logistics, and translation: a study of homeless services systems as infrastructure.
Homeless services systems provide unhoused individuals access to emergency shelter, subsidized housing, and other life-sustaining resources. In this paper, we present a qualitative study that draws on the experiences of fifteen social service workers to examine how recordkeeping practices sustain homeless services systems and unite a tangled web of institutions and actors, including public housing systems, nonprofit agencies, and local governments. We address the following research questions: How is the infrastructure of homeless services sustained by recordkeeping? How are social service workers affected by increasing recordkeeping demands? In what ways do social service workers work against or 'find the play' in this system? To address these questions, we collected interviews and conducted artifact walkthroughs with our study participants. We analyzed the data using an infrastructural lens and found that current recordkeeping practices within homeless services systems comprise an "infrastructure of last resort" that functions logistically, prioritizing efficiency and speed. We also found that social service workers "speak back" to logistification by making the homeless services infrastructure more legible to their unhoused clients through mediation and acts of translation that help to produce better resource outcomes. Our findings show how structuring recordkeeping in ways that privilege efficiency and speed disrupts social service work and interferes with social service workers' ability to provide care for vulnerable individuals facing life-altering and life-threatening hardships.


Lesions of the accessory parotid gland in children.
The accessory parotid gland, a little-known and seldom-mentioned anatomical variant, is a nodule of normal salivary tissue separate from the main parotid gland, located on the masseter muscle and connected to the Stensen duct at that level. It can be the site of both congenital and acquired lesions.


Square microchannel enables to focus and orient ellipsoidal Euglena gracilis cells by two-dimensional acoustic standing wave.
Flow cytometry has become an indispensable tool for counting, analyzing, and sorting large cell populations in biological research and medical practice. Unfortunately, it has limitations in the analysis of non-spherically shaped cells due to the variation of their alignment with respect to the flow direction and, hence, the optical interrogation axis, resulting in unreliable cell analysis. Here, we present a simple on-chip acoustofluidic method to fix the orientation of ellipsoidal cells and focus them into a single, aligned stream. Specifically, by generating acoustic standing waves inside a 100 ⋅ 100 µm square-shaped microchannel, we successfully aligned and focused up to 97.7% of a population of Euglena gracilis (an ellipsoidal shaped microalgal species) cells in the center of the microchannel with high precision at a volume rate of 25 to 200 µL min-1. Uniform positioning of ellipsoidal cells is essential for making flow cytometry applicable to the investigation of a greater variety of cell populations and is expected to be beneficial for ecological studies and aquaculture.


Characterization of T-cell receptors and immunoglobulin heavy chains loci and identification of T/B cell clusters in teleost.
Bacterial disease is one of the important factors leading to economic losses in the turbot (Scophthalmus maximus) cultivation industry. T lymphocytes are major components of cellular immunity, whereas B lymphocytes produce immunoglobulins (Ig) that are key elements of humoral immune responses against infection. However, the genomic organization of genes encoding T-cell receptors (TCR) and immunoglobulin heavy chains (IgHs) in turbot remains largely unknown. In this study, abundant full-length transcripts of TCRs and IgHs were sequenced by Isoform-sequencing (Iso-seq), and we investigated and annotated the V, D, J and C gene loci of TCRα, TCRβ, IgT, IgM and IgD in turbot. Furthermore, through single-cell RNA sequencing (scRNA-seq) of blood leukocytes, we confirmed that these identified TCRs and IgHs were highly expressed in T/B cell clusters, respectively. Meanwhile, we also identified the IgM+IgD+ B and IgT+ B cells with differential gene expression profiles and potential functions. Taken together, our results provide a comprehensive understanding of TCRs and IgHs loci in turbot, which will contribute to evolutionary and functional characterization of T and B lymphocytes in teleost.