Tissue substance P levels in acute experimental burns.
To determine the tissue concentrations of substance P (SP) in burns of different depths and to see whether the concentrations of SP relate to the previously reported late increase in tissue histamine concentrations. Experimental animal study with pigs. Superficial, partial thickness and full thickness burns were created and the microdialysis method used to collect samples for substance P analysis from burned and non-burned control tissue during a 24-h follow-up. Substance P concentrations increase after 4h in the partial and full thickness burns reaching the peak at 18 and 12h, respectively. The increase was later and more modest in the superficial and control sites. At 24h the SP median concentrations in the superficial, partial and full thickness burns were 28%, 85% and 140% higher than in the control site, respectively. There was a peak in the SP concentration in serum at 4h followed by a decrease and stabilization at a level about 15 pg/ml. The release of substance P in tissue is a possible cause of the late increase in tissue histamine in burns. Medical inhibition of SP is of clinical interest in preventing late histamine liberation in burns.


Skull base chondrosarcoma: a case treated by the CyberKnife system.
Skull base chondrosarcomas represent 2% of all chondrosarcomas. They usually have a low grade of malignity and a complete resection may be curative. However, often they are considered irresectable because of the difficult approach and another kind of treatment is needed. We present a 54-year-old male case who, after weeks suffering from diplopia, was diagnosed with skull base chondrosarcoma and treated with the CyberKnife system. A review of radiosurgery techniques for skull base neoplasms is carried out.


Phospholipase D2 activity suppresses hydrogen peroxide-induced apoptosis in PC12 cells.
Phospholipase D (PLD) plays an important role as an effector in the membrane lipid-mediated signal transduction. However, the precise physiological functions of PLD are not yet well understood. In this study, we examined the role of PLD activity in hydrogen peroxide (H(2)O(2))-induced apoptosis in rat pheochromocytoma (PC12) cells. Treatment of PC12 cells with H(2)O(2) resulted in induction of apoptosis in these cells, which is accompanied by the activation of PLD. This H(2)O(2)-induced apoptosis was enhanced remarkably when phosphatidic acid production by PLD was selectively inhibited by pretreating the PC12 cells with 1-butanol. Expression of PLD2, but not of PLD1, correlated with increased H(2)O(2)-induced PLD activity in a concentration- and time-dependent manner. Concomitant with PLD activation, the PLD2 activity suppressed H(2)O(2)-induced apoptosis in PC12 cells. Expression of PLD2 lipase-inactive mutant (K758R) had no effect on either PLD activity or apoptosis. PLD2 activity also suppressed H(2)O(2)-induced cleavage and activation of caspase-3. Taken together, the results suggest that PLD2 activity is specifically up-regulated by H(2)O(2) in PC12 cells and that it plays a suppressive role in H(2)O(2)-induced apoptosis.


Perhexilene: effects on hepatic lysosomal function in rats.
1. Perhexilene, a long-acting anti-anginal drug, can induce adverse effects on the liver which may be dose-dependent. At high concentrations, perhexilene causes marked morphological changes in hepatocyte lysosomes. The current study examined the effect of 'therapeutic' doses of perhexilene on hepatic lysosomal function, particularly the biliary release of lysosomal enzymes, using an isolated perfused rat liver (IPRL) model. 2. Pharmacokinetic studies demonstrated that clearance of single doses of perhexilene by the perfused rat liver was dose-dependent and established a 'therapeutic' dose of 0.6 mg using the IPRL. A 5 day pretreatment regimen of 20 mg/kg per day was shown to produce 'therapeutic' perhexilene concentrations of 150-210 ng/ml. 3. At perhexilene concentrations equating the 'therapeutic' range in man, the major effect of perhexilene was at the biliary pole of the hepatocyte. In 5 day pretreatment dose studies, lysosomal enzyme excretion into bile was markedly increased. In single dose studies, the increase in biliary lysosomal enzyme output partially reflected an increase in bile water production which was not seen with the 5 day pretreatment regimen. Hepatic and perfusate lysosomal enzyme activities were not affected. 4. This selective effect of perhexilene on hepatocyte-to-bile lysosomal excretion may reflect intracellular lysosomal drug localization.


Absorption, serum levels and urinary excretion of inorganic sulfate after oral administration of sodium sulfate in the conscious rat.
The absorption of inorganic sulfate after ingestion was investigated in rats. After oral administration of Na235SO4, 35S radioactivity was measurable in plasma already after 15 min and its plasma concentration reached a peak after about 1.5--2 h. The 35S-radioactivity excreted in urine during 24 h after ingestion of Na235SO4 together with varying amounts of unlabelled Na2SO4 (0.25--5.0 mmol Na2SO4 per rat) indicated an almost complete absorption of inorganic sulfate from the gastrointestinal tract. Determination of the inorganic sulfate concentration in rat serum 2 h after oral administration of 5.0 mmol Na2SO4 revealed a three-fold increase in serum sulfate concentration. The data suggest a rapid and almost complete absorption of inorganic sulfate after oral administration in the rat. Its importance in relation to the sulfate availability for sulfate conjugation of drugs is discussed.


Studies on the effects of glucose in vitro and of the glycaemic state in vivo on the binding characteristics of mu, delta and kappa opiate receptors in mouse brain.
The effect of glucose on the binding characteristics of opiate receptor subtypes was investigated in brain membranes from normoglycaemic lean Aston (C57BL/6J) mice using [3H][D-Ala2,MePhe4,Gly5-ol]enkephalin (DAMGO), [3H][D-Pen2,D-Pen5]enkephalin (DPDPE) and [3H]U69,593 as selective ligands for mu, delta and kappa opiate receptors respectively. The equilibrium dissociation constants (Kd) and maximal binding capacities (Bmax) of [3H]DAMGO and [3H]DPDPE were unaltered by 20mM glucose in vitro. Similarly, [3H]U69,593 binding was not modified by increasing the concentration of glucose from 0 to 20mM (P between 0.10 and 0.05), or by the presence of 20mM fructose and of 20mM 3-O-me-glucose, a non-metabolisable sugar, in the incubation medium. The nonselective opiate ligand, [3H]diprenorphine, bound with similar affinity and binding capacity to brain membranes prepared from control and streptozotocin-diabetic Swiss (CD1) mice. The addition of 20mM glucose or of 20mM fructose in vitro induced no changes in their binding parameters. The affinity and binding capacity of [3H]U69,593 to STZ-diabetic Swiss mouse brain membranes was not significantly different to that of normoglycaemic controls; 20mM glucose in vitro had no effect on ligand binding to kappa sites in STZ-diabetic mouse brain membranes. We conclude that glucose does not interact directly with the opiate receptor to modfy it in such as way as could explain the altered sensitivity to different opioid agonists seen in obese and hyperglycaemic animal models in vivo.


Lipid-binding hSH3 domains in immune cell adapter proteins.
SH3 domains represent versatile scaffolds within eukaryotic cells by targeting proline-rich sequences within intracellular proteins. More recently, binding of SH3 domains to unusual peptide motifs, folded proteins or lipids has been reported. Here we show that the newly defined hSH3 domains of immune cell adapter proteins bind lipid membranes with distinct affinities. The interaction of the hSH3 domains of adhesion and degranulation promoting adapter protein (ADAP) and PRAM-1 (Promyelocytic-Retinoic acid receptor alpha target gene encoding an Adaptor Molecule-1), with phosphatidylcholine-containing liposomes is observed upon incorporation of phosphatidylserine (PS) or phosphoinositides (PIs) into the membrane bilayer. Mechanistically we show that stable association of the N-terminal, amphipathic helix with the beta-sheet scaffold favours lipid binding and that the interaction with PI(4,5)P(2)-containing liposomes is consistent with a single-site, non-cooperative binding mechanism. Functional investigations indicate that deletion of both amphipathic helices of the hSH3 domains reduces the ability of ADAP to enhance adhesion and migration in stimulated T cells.


Impact of respiratory symptoms on lung cancer: 30-year follow-up of an urban population.
We investigated the relationship between respiratory symptoms reported at one time and incidence of lung cancer the subsequent 30 years in an urban Norwegian population. A cohort of 19,998 persons, aged 15-70 years living in Oslo, was randomly selected for a respiratory survey in 1972. The response-rate was 89% and 17,670 respondents were followed up. The relationship between respiratory symptoms and lung cancer incidence was investigated separately for each symptom group, symptom score and sex, with adjustment for age, smoking habits and occupational exposure. Lung cancer developed in 352 persons (228 men and 124 women) during follow up. We found a significant positive association between the incidence of lung cancer and cough symptoms in both sexes, asthma-like symptoms among women and dyspnoea when walking uphill among men. The relative risk for lung cancer increased with the number of symptoms reported at baseline and was strongest the first decade and decreased with duration of follow up. This association was more pronounced for non-small cell lung cancer than for small cell lung cancer.


Objective determination of dark adaptation: accuracy of the evoked potential method.
The psychophysical threshold and the appearance of the visually evoked potential (VEP) were compared in the dark adapted state. Nineteen normal subjects and 11 persons with defective dark adaptation were studied. The VEP threshold was an average of 0.2 log units (standard deviation 0.2) above the subjective threshold in the normal group, and 0.4 log units above it in the night-blind subjects. In 7 normal subjects, the alpha-activity of the electroencephalogram interfered with the VEP recording and rendered the evaluation more difficult. The results indicate that the VEP can be used as an accurate method of objectively measuring dark adaptation.


Interactions of vitamin E and penicillamine in the treatment of hereditary avian muscular dystrophy.
Our prior work demonstrated that penicillamine treatment of dystrophic chickens delayed the onset of symptoms, partially alleviated contractures, improved muscle function, and lowered serum creatine kinase. Penicillamine, a sulfhydryl compound with reducing properties, also prevented inactivation of glycolytic enzymes by protecting thiol groups. The present study shows that vitamin E enhances the therapeutic effects of penicillamine. Interaction of these two reductants is dose related. With vitamin E as adjunct therapy, the dosage level of penicillamine could be lowered by 50%, thereby minimizing side effects. The therapeutic rationale for two antioxidants is that penicillamine may act primarily in the cytoplasm to prevent oxidative damage, whereas the more hydrophobic vitamin E may protect membrane bilayers. Additionally, penicillamine may prevent collagen cross-linking and, deposition of insoluble collagen in muscle and thus decrease contracture formation. General applications of combined penicillamine and vitamin E therapy are discussed regarding prevention of free radical and oxidative damage in Duchenne dystrophy and a wide range of human diseases.


Body mass index and risk of mental disorders in the general population: results from the Singapore Mental Health Study.
The aims of the current study were to elucidate the association between body mass index (BMI) and mental disorders and to examine whether these associations are moderated by socio-demographic correlates and comorbid physical disorders. The Singapore Mental Health Study (SMHS) surveyed adult Singapore residents (Singapore citizens and permanent residents) aged 18 years and above. The survey was conducted from December 2009 to December 2010. The diagnoses of mental disorders were established using the World Mental Health Composite International Diagnostic Interview version 3.0 (CIDI 3.0). BMI was calculated using height and weight which were self-reported by respondents. The Euro-Qol-5Dimensions (EQ-5D) was used to measure the health related quality of life (HRQoL) in the sample. Six thousand and six hundred sixteen respondents completed the study (response rate of 75.9%) and constituted a representative sample of the adult resident population in Singapore. Being underweight was associated with both lifetime (adjusted odds ratio (OR): 2.3) and 12-month obsessive-compulsive disorder (adjusted OR: 4.4). Obesity was associated with 12-month alcohol dependence (adjusted OR: 8.4). There were no significant differences in the EQ-5D indices or the EQ-VAS scores among the four BMI groups in the population. Our findings are somewhat unique and different from those reported in research from Western countries. There is a need for further cross-cultural research to explore and identify genetic, metabolic and cultural differences that underlie the interaction between obesity and mental illnesses.


Different expression of HLA class I antigens in liver of children with chronic hepatitis B, evaluated by immunohistochemical method.
Studies on the quantitative expression of the Major Histocompatibility Complex (MCH) in hepatocytes chronically infected by Hepatitis B Virus (HBV) report that an increased expression of these antigens could be related to a good immunological response. In the present work we analyze the expression of the MCH antigens in cryostatic sections of liver biopsies taken from subjects (19 children) with various forms of HBsAg positive chronic hepatitis. A high expression of HLA class I antigens and a high degree of hepatocyte necrosis was evident in Chronic Active Hepatitis (CAH) and Chronic Lobular Hepatitis (CLH). On the contrary, subjects with histological diagnosis of Chronic Persistent Hepatitis (CPH) showed a low expression of such antigens. There was however, the difference that in subjects with high hepatic cytolysis and high expression of HLA class I antigens, serum HBV-DNA was clearly present in almost all the cases with CAH, but not detectable in all cases with CLH. The expression of HLA class II antigens and of Beta2 microglobulin was the same in all 19 cases. All cases with HBV-DNA positivity with high class I antigen expression had active hepatitis which seems to suggest that all attempts at viral clearance on the part of the immune system have been in vain. We hope our paper will be an additional parameter for evaluating the course of hepatitis during Interferon treatment.


Intracavernous internal carotid artery mycotic aneurysms: comprehensive review and evaluation of the role of endovascular treatment.
Mycotic aneurysms may arise in the setting of many local or systemic infections. Those of the intracranial circulation are especially worrisome due to their potential to compress vital neural structures and their propensity for rupture with consequent hemorrhage. Mycotic aneurysms of the intracavernous internal carotid artery (ICA) represent an exceedingly rare clinical entity, described in less than fifty published cases. Typically presenting as a cavernous sinus syndrome with signs and symptoms of the underlying infection, they are often missed initially, with diagnosis and treatment commencing for the triggering infection or confused with cavernous sinus thrombophlebitis, which may be additionally coexistent, confounding timely diagnosis of the aneurysmal disease. Compared to non-mycotic aneurysms of the intracavernous ICA, which typically have a benign course, the infectious etiology of the mycotic variety increases their tendency to rupture, precludes surgical clipping as a viable treatment option, and requires institution of prolonged antibiotic therapy prior to definitive intervention. Their critical location, friability, and propensity to occur bilaterally result in an unpredictable risk of rapid neurological decline and death, making the timing and specific nature of treatment a unique dilemma facing the treating physician. This review seeks to discuss the natural history of and management strategies for mycotic aneurysms of the intracavernous ICA with special emphasis on the role, safety, and efficacy of endovascular therapies.


Characterization of Trypanosoma (Trypanozoon) evansi from dromedary camels in Kuwait by isoenzyme electrophoresis.
Blood from 115 camels in Kuwait was examined for blood parasites. Two camels of a local herd (1.7%) were found to be infected with Trypanosoma (Trypanozoon) evansi and three camels (2.6%) with microfilarial nematodes. The Trypanosoma stocks isolated from these two camels were screened for isoenzyme patterns of 10 enzymes using thin-layer starch-gel electrophoresis. The results revealed that these two stocks were identical to camel stocks of T. evansi from certain countries in Africa, as well as to two stocks isolated from dogs in Kuwait. This is the first record of Trypanosoma (Trypanozoon) evansi isolates and microfilariae from camels in Kuwait.


A pangolin-origin SARS-CoV-2-related coronavirus: infectivity, pathogenicity, and cross-protection by preexisting immunity.
Virus spillover remains a major challenge to public health. A panel of SARS-CoV-2-related coronaviruses have been identified in pangolins, while the infectivity and pathogenicity of these pangolin-origin coronaviruses (pCoV) in humans remain largely unknown. Herein, we comprehensively characterized the infectivity and pathogenicity of a recent pCoV isolate (pCoV-GD01) in human cells and human tracheal epithelium organoids and established animal models in comparison with SARS-CoV-2. pCoV-GD01 showed similar infectivity to SARS-CoV-2 in human cells and organoids. Remarkably, intranasal inoculation of pCoV-GD01 caused severe lung pathological damage in hACE2 mice and could transmit among cocaged hamsters. Interestingly, in vitro neutralization assays and animal heterologous challenge experiments demonstrated that preexisting immunity induced by SARS-CoV-2 infection or vaccination was sufficient to provide at least partial cross-protection against pCoV-GD01 challenge. Our results provide direct evidence supporting pCoV-GD01 as a potential human pathogen and highlight the potential spillover risk.


Potential intermediate hosts of Angiostrongylus cantonensis in the European Mediterranean region (Mallorca, Spain).
Angiostrongylus cantonensis is the main etiological agent of eosinophilic meningitis, a disease that often leads to severe neurological manifestations in mammals and birds. In recent years, the prevalence of this zoonotic nematode has dramatically increased as it expands into new territories beyond its native range in Southeast Asia and the Pacific Basin.Its arrival to Europe, the last continent to be invaded, has raised many questions concerning the parasite's life cycle, particularly in the Mediterranean region, where it is now endemic. This study aims to collect evidence about intermediate hosts (snail and slug species) involved in the transmission of the A. cantonensis in Mallorca. We have conducted a systematic surveillance of gastropods within 2 km radius areas, surrounding the specific locations where infected hedgehogs were found. We employed a sequence-based detection approach that included a species-specific PCR amplification followed by DNA sequencing of the internal transcribed spacer I (ITS-1). This conservative approach was essential to avoid cross reactions with the numerous metastrongylid species known to be circulating in Mallorca. Overall, we collected and identified 398 gastropods belonging to 17 species, of which 11% tested positive for A. cantonensis. These gastropods were collected from touristic settlements to agricultural lands. Five gastropod species: Theba pisana, Cornu aspersum, Deroceras reticulatum, Milax gagates and Otala lactea, had been previously incriminated as A. cantonensis intermediate hosts, while 6 species: Papillifera papillaris, Cochlicella acuta, Eobania vermiculata, Ganula lanuginosa, Milax nigricans and Rumina decollata, are newly recorded hosts. The findings of this study have important epidemiological implications, and further measures are discussed to prevent neuroangiostrongylosis cases.


[Use of the light-dispersion spectroscopy for detection of the precipitation reaction].
The light beating spectroscopy technique was used to detect the precipitation test. The system consisting of human serum globulin--rabbit antiserum against human globulins was studied. A possibility of detecting the precipitation test for much more diluted systems of the antigen and antibody by means of the light beating technique than by the routine visual method was demonstrated.


Explaining attitudes toward minority groups with human values in Germany - What is the direction of causality?
This paper examines the reciprocal relations between values and attitudes toward minorities over a period of fourteen months in 2015 and 2016. A representative sample of the adult population in Germany completed four waves of a panel study in which attitudes and values were each measured two times. Reciprocal relations over time between Schwartz's (1992) higher-order value of conservation and the value of universalism as well as attitudes toward four different minorities (Muslims, refugees, foreigners, Sinti/Roma) were examined using a modified cross-lagged longitudinal design. The results showed that values and attitudes had reciprocal longitudinal effects on one another, meaning that values predicted changes in attitudes and attitudes predicted changes in values. The findings also revealed that (1) values were more stable over time than attitudes, and (2) the longitudinal effect of values on attitudes was not significantly stronger than the longitudinal effect of attitudes on values.


Experience with primary carcinoma of the male urethra.
We reviewed 23 cases of primary carcinoma of the male urethra and compared the clinical findings, treatment and results to the experiences previously reported. Patients with lesions of the pendulous urethra managed by appropriate amputation of the penis with ilio-inguinal node dissection for groin metastases had a good prognosis, whereas patients with lesions of the bulbo-membranous or prostatic urethra had a poor prognosis. Despite occasional successful results with conservative surgical excision and over-all poor results with radical excision the generally extensive nature of such tumors necessarily makes radical excision the treatment of choice.


Prediction of pulmonary hypertension in older adults based on vital capacity and systolic pulmonary artery pressure.
Right heart catheterization (RHC) is associated with a higher procedural risk in older adults, but non-invasive estimation of pulmonary hypertension (PH) is a challenge. We aimed to elaborate a non-invasive prediction model to estimate PH. We retrospectively analysed 134 older adults (70.0 years ±12.3; 44.9% males) who reported to our clinic with unclear dyspnea between 01/2015 and 01/2020 and had received RHC as a part of their diagnostic workup. Lung function testing, analysis of blood gas samples, 6 min walk distance and echocardiography were performed within 24 hours of RHC. In a stepwise statistical approach by using an in/exclusion algorithm (using the AIC criterion) we analysed non-invasive parameters to test their value in predicting PH (defined as mean pulmonary artery pressure, PAmean, >25mmHg). Discrimination capability of the final model was measured by the AUC (area under curve) from an ROC (receiver operating characteristics) analysis. We yielded a sensitivity of 87.2% and a specificity of 62.5% in a combinatorial logistical model with systolic pulmonary artery pressure (sPAP) and forced vital capacity (VCmax), the discrimination index was 86.7%. The odds ratios for an increase of 10 mmHg of sPAP were 2.99 (2.08-4.65) and 1.86 (1.11-3.21) for a 1 l decrease in VCmax. On their own, VCmax proved to be specific (83.3%), while sPAP was a sensitive (79.1%) predictor for PH. We provide a combinatorial model to predict PH from sPAP and VCmax in older adults, which may help to avoid invasive procedures.


Effect of methyl farnesoate, alone and in combination with other hormones, on ovarian growth of the red swamp crayfish, Procambarus clarkii, during vitellogenesis.
The effect of methyl farnesoate (MF) on the ovaries of female red swamp crayfish, Procambarus clarkii, undergoing vitellogenesis was determined both in vivo and in vitro. The in vivo assay showed a positive effect of MF on oocyte growth when injected alone and in combination with 17 beta-estradiol, but not in combination with JHIII or 17 alpha-hydroxyprogesterone. A higher level of incorporation of labeled leucine was induced by MF on isolated pieces of ovary. The same effect was seen when ovary and mandibular organ (MO) were coincubated. These results suggest that MF stimulated the synthesis of vitellin in the ovary of crayfish. In vitro, 17 alpha-hydroxyprogesterone completely suppressed the stimulatory action of the MO on the ovary, suggesting a competitive inhibition between 17 alpha-hydroxyprogesterone and MF on the ovary and/or a negative feedback by that steroid on the MO.


Prevalence of lactose intolerance and malabsorption among children of two ethnic groups from the urban areas of Malaysia and its relation to calcium intake and bone health status.
Lactase insufficiency could develop into lactose intolerance (LI) and disrupt the intake of calcium, which is a core nutrient for bone development in children. However, data regarding the prevalence of LI and bone health status (BHS) among Malaysian children are scarcely reported. This study aimed to determine the prevalence of LI and lactose malabsorption (LM) in Malay and Chinese children and examine its relationship with calcium intake (CI) and BHS. A total of 400 children participated in this study. The prevalence of lactose tolerance (LT) was assessed using hydrogen breath test, LT test, and visual analogue scales. Assessment of CI was performed using a 24-h dietary recall interview (24-h DR) and food frequency questionnaire (FFQ). Calcaneal broadband ultrasound attenuation (BUA) was measured using a quantitative ultrasonometer. The prevalence of LI among Chinese children (37%) was significantly higher (p = 0.002) than among Malay children (22.5%). However, 61.5% of Malay and 54.5% of Chinese children were found to have LM. CI of the children fulfilled 30.5% and 33.9% of the Malaysian recommended CI (1300 mg/day) for 24-h DR and FFQ, respectively. The BUA score of Malay children was significantly higher (p < 0.001) than that of Chinese children. However, LT and CI were not correlated with BHS (p > 0.05, respectively). LI was diagnosed among Malay and Chinese children. However, the higher prevalence of LM is rather worrying as it could develop to LI. The prevalence possibly has been worsened by insufficient CI. Thus, effective approaches to increase CI are highly needed as bone development occurs rapidly at this age and is important for the attainment of the optimum peak bone mass during late adolescence.


C8-amino purine nucleosides. A well-defined steric determinant of glycosyl conformational preferences.
100 MHz proton magnetic resonance measurements were performed on dilute solutions of adenosine and guanosine and their 8MH2, 8-NHCH3 8-N(CH3)2 and 8-bromo derivatives. The chemical shift of the ribose C2'-H and especially the difference in chemical shifts between the C1'-H and C2'-H resonances clearly indicated whether the nucleoside exists in a syn glycosyl conformation (the C8-dimethylamino derivatives) or as a flexible syn-anti mixture (the monomethylamino and amino derivatives). The temperature dependent behavior of these indicators can be employed to define qualitative shifs in syn-anti equilibrium with temperature. An increased C1'-H-C2'-H chemical shift separation implies shift to more anti, a decreased separation a shift to more syn conformers.


Repetitive reduction lead to significant elevated IL-6 and decreased IL-10 levels in femoral osteotomies: A quantitative analysis of a robot-assisted reduction process in a rat model.
The field of robot-assisted fracture reduction has been developed by several research groups over more than one decade by now, with the main goals of increasing the fracture reduction accuracy. However, the influence of different reduction paths to patients' physiology is not fully known yet. The aim of our study was to compare the impacts of a robot-assisted direct reduction path versus an artificially prolonged reduction path by measuring the cytokine responses in an in vivo rat model. Thirty-six male CD(©) rats were assigned into three groups with an external fixator and osteotomy on the left femur. Seven days later, the robot was attached and one group was reduced in a single attempt, while the other group underwent 10 attempts by the robot. The third group was the control group without reduction. Before, and as well as 6, 24 and 48h after the reduction process blood samples were collected. IL-1, IL-6, IL-10, IL-17, and MCP-1 concentrations where analysed via ELISA or cytometric bead assay. Muscle biopsies in the osteotomy area were collected 48h after the reduction process for histological analyses. Statistical significance was set at p≤0.05. Analysis of the cytokines showed that the pro-inflammatory cytokine IL-6 of the Ten-Attempts reduction group significantly increased 6h after reduction compared to the control group. IL-6 further showed markedly elevated levels 6h after surgery in the Ten-Attempts reduction group compared to the Single-Attempt reduction group. On the anti-inflammatory side, IL-10 showed a significant decrease in the Ten-Attempts reduction group 6h after reduction compared to the Single-Attempt reduction and control group. Muscle biopsies showed a significant increase of pathological changes in both reduction groups and an increase in the severity of bleedings of the Ten-Attempts reduction group compared to the Single-Attempt reduction and control group. A direct and gentle reduction procedure as feasible by the aid of a robot is preferable over a prolonged reduction in terms of cytokine response and tissue changes.


Drivers of year-to-year variation in exacerbation frequency of COPD: analysis of the AERIS cohort.
The association between exacerbation aetiology and exacerbation frequency is poorly understood. We analysed 2-year follow-up data from a prospective observational study of patients with chronic obstructive pulmonary disease (COPD) (www.clinicaltrials.gov identifier number NCT01360398) to evaluate year-to-year variation in exacerbation frequency and related aetiology. A total of 127 patients underwent blood and sputum sampling monthly and at exacerbation to detect respiratory infections and eosinophilic inflammation; 103 continued into year 2 and 88 completed both years. The most common bacterial species at stable state and exacerbation was Haemophilus influenzae. Among infrequent exacerbators (one exacerbation per year), the incidence of viral infection at exacerbation was high (60.0% (95% CI 35.1-81.7%) in year 1 and 78.6% (53.4-94.2%) in year 2). Those with more frequent exacerbations tended to have higher relative incidence of bacterial than viral infection. Patients with at least two additional exacerbations in year 2 versus year 1 had a higher risk of H. influenzae colonisation at stable state than those with at least two fewer exacerbations, as detected by culture (OR 1.43 (95% CI 0.71-2.91) versus 0.63 (0.40-1.01), p=0.06) and PCR (1.76 (95% CI 0.88-3.51) versus 0.56 (0.37-0.86), p<0.01). This was not seen with other infection types or eosinophilic inflammation. Analysis of the same cohort over 2 years showed, for the first time, that changes in yearly COPD exacerbation rate may be associated with variations in H. influenzae colonisation.


Dietary overlap of grasshoppers on sandhill rangeland in northeastern Colorado.
Overlap in the diets of 14 species of grasshoppers on mixed-grass prairie in northeastern Colorado was estimated by microscopic examination of crop contents. Food availability estimates, taken by a weight-estimate method, facilitated the determination of food preferences. Two cases of high overlap in diets and period of habitat occupancy occurred between species which consumed equal or almost equal numbers of different foods (foodniche breadth) and had almost identical foraging behavior. The first case occurred early in the season between Xanthippus corallipes and Arphia conspersa with foodniche breadths of 14 and 11, respectively. The second case occurred later in the season between Amphitornus coloradus and Trachyrhachys kiowa both of which have foodniche breadths of 6. The amount of movement between plants while feeding was equal or almost equal in both cases, thus we observed high dietary overlap between species with equal ecological efficiencies. Dietary overlap between the 14 grasshopper species decreased as foodniche dimensions increased and the grasshopper population as a whole utilized each food resource approximately proportional to its relative abundance. Factors influencing the interpretation and importance of dietary overlap in this study included 1. non-synchronous life cycles, 2. seasonal changes in diets and food preferences, 3. complexity of the food resources, 4. variations in foodniche dimensions among grasshopper species, and 5. variations in diets between males and females within a species. Our findings agree with MacArthur and Levins theory that the number of competing species which can coexist is proportional to the total range of the environment divided by the niche breadth of the species.


Assessment of neutrophil function in the elderly using antibody coated polyacrylamide gel (immunobeads) and nitroblue tetrazolium (NBT) reduction as a combined test.
Neutrophil function was assessed in 35 elderly individuals (age greater than 75) and 20 normal young individuals (age 20-45) by combining ingestion of antibody coated polyacrylamide gel and nitroblue tetrazolium reduction in a single test. This test evaluates phagocytosis and metabolic integrity simultaneously and appears to be a sensitive and reliable test of neutrophil function. No significant difference was found (by using this test) between neutrophils from healthy elderly people and the neutrophils from young controls, or between the sexes in either age group.


Postoperative changes in liver function tests: randomized comparison of low- and high-pressure laparoscopic cholecystectomy.
Pneumporeitoneum at 14 mmHg causes dangerous hemodynamic disturbances in some patients, leading to splanchnic ischemia. Laparoscopic cholecystectomy (LC) using low-pressure pneumoperitoneum (7 mmHg) minimizes adverse hemodynamic effects on hepatic portal blood flow and hepatic function. This study evaluated the changes in liver function tests after high-pressure LC (HPLC; 14 mmHg) and low-pressure LC (LPLC; 7 mmHg). For this study, 50 patients were randomly assigned to undergo either HPLC (n = 25) or LPLC (n = 25) Liver function tests including total bilirubin, gamma-glutamyltransferase (GGT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were obtained preoperatively, then 24 and 48 h postoperatively. All patients had normal values on the preoperative liver function tests. The anesthesiologic protocol was uniform. The findings showed that ALT after 24 h (LPLC: 1473.72 +/- 654.85; HPLC: 2233.74 +/- 1247.33; p = 0.0096) and 48 h (LPLC: 1322.99 +/- 601.51; HPLC 2007.80 +/- 747.55; p = 0.0008) and AST after 24 h (LPLC: 1189.96 +/- 404.79 i.j.; HPLC: 1679.40 +/- 766.13; p = 0.0069) were increased in the patients who underwent HPLC. The AST levels after 48 h were statistically unchanged from baseline in both groups. Total bilirubin, ALP, and GGT levels remained unchanged from baseline in both groups, without a significant difference between the two groups. Because LPLC minimizes adverse hemodynamic effects on hepatic function, a low-pressure pneumoperitoneum should be considered for patients with compromised liver function, particularly those undergoing prolonged laparoscopic surgery.


AKR leukemogenesis: identification and biological significance of thymic lymphoma receptors for AKR retroviruses.
We have previously demonstrated that in vitro cell lines of mouse thymic lymphomas express surface receptors specific for the retrovirus that induced them. This study extends these observations to an analysis of receptor-bearing cells in the preleukemic and leukemic phases of spontaneous AKR thymic lymphomagenesis. AKR mice regularly begin expressing N-tropic retroviruses (as assayed on NIH fibroblasts by the XC plaque assay) in several tissues early in life; thymic lymphocytes also express these viruses, but are not autonomously transformed. Later thymic lymphomas emerge which are capable of metastasizing in the host of origin or transplanting leukemias into syngeneic hosts. Just prior to the appearance of thymic lymphomas, these mice also begin producing xenotropic retroviruses [as assayed in xenogeneic (For example, mink) fibroblasts], and concomitant with the appearance of the leukemias is the appearance of "recombinant" retroviruses which cause mink fibroblast foci (MCF); these viruses express elements of both N- and X-tropic virus envelopes and N-tropic viral gene products in their cores. Spontaneous AKR leukemias also produce other retroviruses which do not cause XC plaques or mink fibroblast foci; these are called SL viruses. The subject of this study was to test whether in vivo thymocytes in the preleukemic and leukemic periods also bear receptors specific for N-tropic, recombinant MCF and SL AKR retroviruses. We demonstrated that each spontaneous thymic lymphoma does bear receptors that bind viruses produced by the lymphomas and MCF-247 to a high degree and that bind N-ecotropic AKR retroviruses less well. Thymic lymphocytes predominating in the preleukemic period do not express detectable levels of receptors for either of the viruses. In some mice, receptor-positive cells co-exist with receptor-negative cells; only the receptor-positive cells are capable of transplanting leukemia to syngeneic hosts. We conclude that the presence of specific cell surface receptors for lymphoma cell-produced and recombinant AKR retroviruses is a marker for leukemia in these hosts.


[Changes of biological clock protein in neonatal rats with hypoxic-ischemic brain damage].
To study the effects of biological clock protein on circadian disorders in hypoxic-ischemic brain damage (HIBD) by examining levels of CLOCK and BMAL1 proteins in the pineal gland of neonatal rats. Seventy-two 7-day-old Sprague-Dawley (SD) rats were randomly divided into sham-operated and HIBD groups. HIBD model was prepared according to the modified Levine method. Western blot analysis was used to measure the levels of CLOCK and BMAL1 in the pineal gland at 0, 2, 12, 24, 36 and 48 hours after operation. Both CLOCK and BMAL levels in the pineal gland increased significantly 48 hours after HIBD compared with the sham-operated group (P<0.05). There were no significant differences in levels of CLOCK and BMAL proteins between the two groups at 0, 2, 12, 24 and 36 hours after operation (P>0.05). Levels of CLOCK and BMAL1 proteins in the pineal gland of rats increase significantly 48 hours after HIBD, suggesting that both CLOCK and BMAL1 may be involved the regulatory mechanism of circadian disorders in rats with HIBD.


Model-driven spatial evaluation of nutrient recovery from livestock leachate for struvite production.
Nutrient pollution is one of the major worldwide water quality problems, resulting in environmental and public health issues. Agricultural activities are the main source of nutrient release emissions, and the livestock industry has been proven to be directly related to the presence of high concentrations of phosphorus in the soil, which potentially can reach waterbodies by runoff. To mitigate the phosphorus pollution of aquatic systems, the implementation of nutrient recovery processes allows the capture of phosphorus, preventing its release into the environment. Particularly, the use of struvite precipitation produces a phosphorus-based mineral that is easy to transport, enabling redistribution of phosphorus to deficient locations. However, livestock leachate presents some characteristics that hinder struvite precipitation, preventing extrapolation of the results obtained from wastewater studies to cattle waste. Consideration of these elements is essential to determine the optimal operating conditions for struvite formation, and for predicting the amount of struvite recovered. In this work, a detailed thermodynamic model for precipitates formation from cattle waste is used to develop surrogate models to predict the formation of struvite and calcium precipitates from cattle waste. The variability in the organic waste composition, and how it affects the production of struvite, is captured through a probability framework based on the Monte Carlo method embedded in the model. Consistent with the developed surrogate models, the potential of struvite production to reduce the phosphorus releases from the cattle industry to watersheds in the United States has been assessed. Also, the more vulnerable locations to nutrient pollution were determined using the techno-ecological synergy sustainability metric (TES) by evaluating the spatial distribution and balance of phosphorus from agricultural activities. Although only struvite formation from cattle operations is considered, reductions between 22% and 36% of the total phosphorus releases from the agricultural sector, including manure releases and fertilizer application, can be achieved.


New Perspectives In The Objective Evaluation Of Animal Welfare, With Focus On The Domestic Pig.
Animal welfare can be viewed as the result of integrating repeated affective evaluations of success in coping with environmental challenges, i.e., subjective challenge adequacy. The present work summarizes why established physiological and behavioral welfare parameters are inadequate to assess challenge adequacy. Behavioral tests based on the mood-congruent judgment effect and physiologic parameters based on components of the somatotropic axis are proposed as an alternative. Here, the judgment bias measures an animal's subjective confidence to cope successfully with a challenge, which is in turn modulated by the animal's previous experience. The somatotropic axis incorporates the insulin-like growth factor (IGF) and its binding proteins (IGFBP), which are involved in the regulation of metabolism and growth. First results indicate that in particular IGF-1 and IGFBP-3 react with higher latency and higher inertness to short-term stressful events than established physiological stress parameters. Before these indicators can be utilized for overall welfare assessment, further validation studies are necessary that provide more insights into how repeatable the measurements are under different conditions and which other factors may confound the measures.


Nutritional, chemical, and antioxidant screening of selected varieties of lentils (Lens culinaris spp.) from organic and conventional agriculture.
Lentils are an ancient legume established worldwide for direct consumption and with great potential for application in food processing. In addition, it is a sustainable crop owing to its ability to scavenge nitrogen and carbon, and it improves the nutrient status of the soil. A diet rich in lentils has been linked to significant health benefits. However, the composition of lentils can be influenced by both the lentil variety and the growing conditions. The aim of this work was to evaluate the nutritional profiles and antioxidant potential, as well as the impact that the type of cultivation (conventional or organic) and the variety could have on these parameters, in different lentil varieties. Overall, carbohydrates are the major macronutrients in all varieties, with notable amounts of fibre (11.62-27.36%) and starch (41.98-50.27%). High amounts of protein and ash were also identified, particularly in the Beluga variety, with 21.9-23.3 and 1.38-1.82 g 100 g-1 fresh weight, respectively. Fructose and sucrose were detected (high-performance liquid chromatography (HPLC) with refraction index detection), along with oxalic, quinic, malic, and shikimic acids (ultra-fast liquid chromatography with photodiode array detection), and α- and γ-tocopherol isoforms (HPLC with fluorescence detection). Fatty acid methyl ester assessment showed the prevalence of polyunsaturated fatty acids (33.5-46.3%). Good antioxidant capacity (thiobarbituric acid reactive substances and oxidative haemolysis inhibition assay) was also noted. The results obtained showed that all the varieties analysed are an excellent source of fibre and have a good antioxidant capacity. Lentil variety has a greater influence on its nutritional composition than the type of cultivation. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Lack of effect of probucol on atheroma formation in cholesterol-fed rabbits kept at comparable plasma cholesterol levels.
Rabbits were fed cholesterol for 14 weeks to study the effect of probucol on atheroma formation. Three groups of animals were investigated: group CHOL was fed 1% cholesterol and served as control for group P + CHOL. fed 1% cholesterol and 1% probucol from the onset till the end of the experiment: group CHOL + P received 1% cholesterol throughout the experiment and 1% probucol during the last 4 weeks only. Plasma cholesterol concentrations were monitored at frequent intervals and were modulated by dietary perturbations so that the areas under the curve expressing plasma cholesterol changes with time, were similar in probucol and non-treated rabbits. The efficacy of long-term probucol treatment was evidenced by a significant reduction in plasma apolipoprotein A-I throughout the experiment and lower plasma TBARs during the first 6 weeks, when the hypocholesterolemic effect of probucol was also seen. Two weeks prior to the termination of the experiment, the rabbits were injected with rabbit plasma labeled with [3H]cholesteryl linoleyl ether [( 3H]CLE). Aortic atheromatosis was quantified by determination of total and cholesteryl ester (CE). The aortic cholesterol content was related to the arch, thoracic and abdominal segments, to the surface area of each segment or its dry defatted weight. Total and esterified cholesterol were highest in the aortic arch in all 3 groups when related to any of the above mentioned parameters. No statistically significant difference in aortic total cholesterol and CE content was seen among the three groups studied. The [3H]CLE recovered in the aortic segment correlated with the CE content and the [3H]CLE (dpm)/mg CE in all segments was similar. No statistically significant difference in the [3H]CLE recovered in the aortic segments among the 3 groups was seen. We conclude that in cholesterol-fed rabbits, in which the plasma cholesterol levels were maintained at comparable levels, probucol treatment did not affect plasma CE influx into the aorta and did not attenuate development of aortic atherosclerosis.


NMR structure and dynamics of the agonist dynorphin peptide bound to the human kappa opioid receptor.
The structure of the dynorphin (1-13) peptide (dynorphin) bound to the human kappa opioid receptor (KOR) has been determined by liquid-state NMR spectroscopy. (1)H and (15)N chemical shift variations indicated that free and bound peptide is in fast exchange in solutions containing 1 mM dynorphin and 0.01 mM KOR. Radioligand binding indicated an intermediate-affinity interaction, with a Kd of ∼200 nM. Transferred nuclear Overhauser enhancement spectroscopy was used to determine the structure of bound dynorphin. The N-terminal opioid signature, YGGF, was observed to be flexibly disordered, the central part of the peptide from L5 to R9 to form a helical turn, and the C-terminal segment from P10 to K13 to be flexibly disordered in this intermediate-affinity bound state. Combining molecular modeling with NMR provided an initial framework for understanding multistep activation of a G protein-coupled receptor by its cognate peptide ligand.


Refractive outcome of cataract surgery using partial coherence interferometry and ultrasound biometry: clinical feasibility study of a commercial prototype II.
To evaluate the refractive outcome of cataract patients 3 months postoperatively using optical biometry obtained with a prototype version (axial length measurement, ALM, Carl Zeiss Jena) of the commercial partial coherence interferometry (PCI) instrument (IOLMaster, Carl Zeiss Jena). Department of Ophthalmology, Vienna General Hospital, and Institute of Medical Physics, University of Vienna, Austria. Forty-five patients with age-related cataract in both eyes were scheduled for bilateral cataract surgery. Axial length was measured preoperatively with a prototype (ALM) of the commercial PCI instrument as well as with immersion ultrasound (IUS). Immersion US was performed by a single experienced investigator. In each patient, the first eye was randomly assigned to receive an intraocular lens (IOL) using the Holladay IOL power formula based on ALM or IUS biometry. The other biometric technique was used in the contralateral eye. Subjective refractive outcome was assessed 3 months postoperatively. Refractive outcomes with the 2 techniques did not differ significantly (P = .28). The mean numerical error (MNE) (the difference between the refractive outcome 3 months postoperatively and the predicted spherical equivalent) was 0.13 diopter (D) and 0.03 D for the ALM and IUS, respectively. The mean absolute error (MAE) (the absolute value of MNE) was 0.48 D (range 0.00 to 1.58 D) and 0.46 D (range 0.01 to 1.92 D) with the ALM and IUS, respectively. By recalculating the surgeon factor retrospectively to correct the Holladay formula to obtain a postoperative MNE of zero, a theoretical MAE of 0.46 D was obtained with both biometry techniques. Refractive outcome in cataract patients using PCI biometry was as good as that achieved with optimized IUS. However, the difference in axial length measured by the ALM compared to that measured by IUS must be considered when using the IOL A-constants supplied by the manufacturers.


Role of OH radicals in the formation of oxygen molecules following vacuum ultraviolet photodissociation of amorphous solid water.
Photodesorption of O(2)(X (3)Σ(g) (-)) and O(2)(a (1)Δ(g)) from amorphous solid water at 90 K has been studied following photoexcitation within the first absorption band at 157 nm. Time-of-flight and rotational spectra of O(2) reveal the translational and internal energy distributions, from which production mechanisms are deduced. Exothermic and endothermic reactions of OH+O((3)P) are proposed as plausible formation mechanisms for O(2)(X (3)Σ(g) (-) and a (1)Δ(g)). To examine the contribution of the O((3)P)+O((3)P) recombination reaction to the O(2) formation following 157 nm photolysis of amorphous solid water, O(2) products following 193 nm photodissociation of SO(2) adsorbed on amorphous solid water were also investigated.


Plural media ethics? Reformist Islam in India and the limits of global media ethics.
The transatlantic field of global media ethics is premised on a search for the conceptual foundations of plurality. This article is a critique of this very endeavor. I offer this critique through works authored by moral anthropologists of Islam and through a close reading of the Urdu text Cyberistan: Muslim Naujavan Aur Social Media (Cyberistan: Muslim Youth and Social Media) authored by Sadatullah Husaini, the current president of the Indian reformist Islamic organization Jamaat-e-Islami Hind. My article is a post-foundational critique of the implicit foundationalism through which "Islam" and "plurality" are related to each other within inquiries into the ethics of digital communication. I take on digital communication because of its increasingly global and synchronic nature that rendered questions concerning plurality in media ethics particularly urgent. I argue that even though it is important to ask what difference means conceptually for a global media ethics today, it can only make space for radical plurality via the negative, by way of its contradictions and structural constraints. If a global media ethics is supposed to be based on openness and plurality, it can be so only by limiting and weakening its own ontological claims - beyond positive metaphysical groundings, cultures, civilizations, Islam, etc. In other words, it requires a reflexivity to its own position as an academic discipline that produces knowledge under certain historical conditions and an understanding of its own political practice.


The neonatal blood count in health and disease. I. Reference values for neutrophilic cells.
Reference ranges for absolute total neutrophils/mm3, absolute immature neutrophils/mm3, and the fraction of immature to total neutrophils (I:T proportion) during the first 28 days of life are developed from 585 peripheral blood counts obtained from 304 normal neonates and 320 counts obtained from 130 neonates with perinatal complications demonstrated to have no statistically significant effect on neutrophil dynamics. Perinatal factors other than bacterial disease which significantly alter neutrophil dynamics include maternal hypertension, maternal fever prior to delivery, hemolytic disease, and periventricular hemorrhage. The predictive value of these reference ranges in identifying bacterial disease in the first week of age varies with the neutrophil factor evaluated and the clinical setting. Neutropenia in the presence of respiratory distress in the first 72 hours had an 84% likelihood of signifying bacterial disease, whereas neutropenia in the presence of asphyxia had a 68% likelihood of signifying bacterial disease. An abnormal I:T proportion had an accuracy of 82% and 61%, respectively, in the same clinical settings. Elevations of either immature or total neutrophils were less specific. Interpretation of abnormal neutrophil factors must include consideration of both infectious and noninfectious perinatal events.


Discharge and referral data exchange using global standards--the SCIPHOX project in Germany.
The goal of the German project "Standardization of Communication between Information Systems in Physician Offices and Hospitals using XML" (aka SCIPHOX) in its first phase is to provide information exchange based on the Extended Markup Language XML between Hospital Information Systems (HIS) and Physician Office Systems (POS). The Clinical Document Architecture (CDA), a standard developed by the Health Level Seven organization (HL7), was chosen to serve as the "backbone" specification. The CDA is an ANSI approved document architecture for exchange of clinical information using XML. In phase I of the SCIPHOX project the proposal specifies the use of the CDA as a generalized international standard in the national context of discharge and referral letters in Germany. The specification defines how to use the CDA header and associated vocabularies by providing a translation and interpretation of the CDA header tags and provides a solution for taking local needs (insurance information etc.) into account.


The free heme concentration in healthy human erythrocytes.
Heme, the prosthetic group of hemoglobin, may be released from its host due to an intrinsic instability of hemoglobin and accumulate in the erythrocytes. Free heme is in the form of hematin (Fe(3+) protoporphyrin IX OH) and follows several pathways of biochemical toxicity to tissues, cells, and organelles since it catalyzes the production of reactive oxygen species. To determine concentration of soluble free heme in human erythrocytes, we develop a new method. We lyse the red blood cells and isolate free heme from hemoglobin by dialysis. We use the heme to reconstitute horseradish peroxidase (HRP) from an excess of the apoenzyme and determine the HRP reaction rate from the evolution of the emitted luminescence. We find that in a population of five healthy adults the average free heme concentration in the erythrocytes is 21±2μM, ca. 100× higher than previously determined. Tests suggest that the lower previous value was due to the use of elevated concentrations of NaCl, which drive hematin precipitation and re-association with apoglobin. We show that the found hematin concentration is significantly higher than estimates based on equilibrium release and the known hematin dimerization. The factors that lead to enhanced heme release remain an open question.


Expression profiles of cytokines released in intestinal epithelial cells of the rainbow trout, Oncorhynchus mykiss, in response to bacterial infection.
To determine whether fish intestinal epithelial cells (IECs) contribute to mucosal immunity, we established a method for isolating IECs from the rainbow trout Oncorhynchus mykiss and examined cytokine production in these cells. Components of the intestinal epithelium were released by incubation of intestinal pieces with 1mM dithiothreitol (DTT)/ethylenediamine tetraacetic acid (EDTA). The IEC-rich fraction (purity >90%; survival rate approximately 95%) was obtained by centrifugation on a 35%/40% Percoll gradient, followed by magnetic cell sorting using an anti-trout IgM antiserum. The gene expression profiles of 14 cytokines in trout IECs were investigated after culturing the cells for 6h with or without the pathogenic bacterium Aeromonas salmonicida. Trout IECs could produce several cytokines, of which IL-1beta and TNFalpha2 were upregulated when the cells were stimulated with live A. salmonicida. Immunohistochemical analyses with the anti-trout TNF antibody confirmed that the TNF protein was present in the IECs of trout that were intra-anally challenged with live A. salmonicida. These results show that trout IECs are an important trigger of the intestinal immune system. Further, formalin-killed A. salmonicida, conditioned medium of this bacterium, or live nonpathogenic Escherichia coli could not upregulate the expression of these cytokines. These results indicate that the production of inflammatory cytokines by IECs is caused by the adhesion of A. salmonicida, but is not due to only simple ligand-receptor interactions between the surface molecules of IECs and the bacterium or in response to bacterial secretions.


Relevance of low specific IgE levels to egg, milk and peanut in infancy.
IgE sensitization is usually associated with allergy-related diseases, but may also occur in asymptomatic individuals. The clinical importance of IgE antibody concentrations in the interval 0.1-0.34 kU/L in early life in relation to allergy development is poorly evaluated. To assess the relevance of low specific IgE (s-IgE) to hen's egg, cow's milk and peanut at 6 months of age for development of sensitization and allergy-related disease during early childhood. s-IgE concentrations to relevant allergens from blood samples taken at 6 months and 1, 2 and 5 years from children in the prospective ALADDIN cohort were divided into three categories: non-sensitized (<0.1 kU/L), low sensitized (0.1-0.34 kU/L) and sensitized (≥0.35 kU/L) and allergy-related disease assessed. A total of 372 children were included in this study. Compared with non-sensitized children at 6 months of age, children with low levels of allergen specific IgE (0.1-0.34 kU/L) to food allergens, especially to egg, at 6 months of age were associated with development of sensitization to aeroallergens at 5 years of age (10/14 [71%] vs 39/250 [15%]). In addition, children with low levels to egg or milk at 6 months were more often sensitized to the respective allergen at 1 year of age and, regarding low levels to egg, also to the development of eczema (6/18 [33%] vs 29/292 [10%]). IgE antibody concentrations in the interval 0.1-0.34 kU/L to food allergens in infancy seem to increase the probability of sensitization to aeroallergens and, regarding low levels to egg, also of eczema during early childhood. Thus, IgE levels during the first year of life, although below 0.35 kU/L, can provide additional allergy-related prognostic information.


Phage-Bacteria Associations: Analyze. Match. Develop Therapies.
Although bacteriophages are highly abundant in the gut microbiome, little is known about their potential effects on gut bacteria. In this issue of Cell Host & Microbe, Hryckowian et al. (2020) and Fujimoto et al. (2020) combined metagenomic analysis and experiments to study phage-bacteria associations in order to develop future research tools and therapies.


Bacterial community analysis identifies Klebsiella pneumoniae as a native symbiotic bacterium in the newborn Protobothrops mucrosquamatus.
The study of the native microbiome of organisms is crucial. The connection between the native microbiome and the host affects the formation of the innate immune system and the organism's growth. However, the native microbiome of newborn venomous snakes has not been reported. Therefore, we aimed to determine the oral and skin microbiomes of newborn Protobothrops mucrosquamatus. We performed 16 S full-length sequencing on 14 samples collected from 7 newborn P. mucrosquamatus individuals, specifically targeting their oral and skin microbiomes. In terms of the oral and skin microbiome, the main species were Klebsiella pneumoniae lineages. According to subspecies/species analysis, the proportion from highest to lowest was K. quasipneumoniae subsp. similipneumoniae, K. pneumoniae subsp. pneumoniae, and K. pneumoniae subsp. rhinoscleromatis. These three bacteria accounted for 62.5% and 85% of the skin and oral activity, respectively. The oral microbiome of newborn P. mucrosquamatus did not comprise common bacteria found in snakebite wounds or oral cultures in adult snakes. Therefore, the source of other microbiomes in the oral cavities of adult snakes may be the environment or prey. Functional Annotation of the Prokaryotic Taxa analysis showed that the skin/oral native microbiome metabolism was related to fermentation and human infection owing to the dominance of K. pneumoniae lineages. The characteristics of K. pneumoniae may impact the development of venom in venomous snakes. The results of the native microbiome in the oral cavity and skin of newborn P. mucrosquamatus demonstrated that the habitat environment and prey capture may affect the composition of bacteria in adult snakes. We hypothesized that the native microbiome influences newborn venomous snakes and that K. pneumoniae lineages related to citrate fermentation may play a role in venom growth. However, further verification of this is required.


Comparison of metal levels in invertebrate detritivores and their natural diets: Concentration factors reassessed.
Concentration Factors (ppm in animal: ppm in diet) are presented for lead, zinc and cadmium in the snail Cepaea hortensis, and for lead and zinc in the woodlice Oniscus asellus and Philoscia muscorum, sampled at roadside sites. For each species such factors were found to be extremely variable, affected not only by season, and size and/or age of animals, but also by the choice of data used in estimating metal levels in the diet. It is concluded that factors other than seasonal changes in metal levels of senescent vegetation are primarily responsible for withinsite variation in the lead, zinc and cadmium concentrations of invertebrate detritivores.


Penicillin and Beta-Lactam Hypersensitivity.
Ten percent of patients report penicillin allergy, but more than 90% of these individuals can tolerate penicillins. Skin testing remains the optimal method for evaluation of possible IgE-mediated penicillin allergy and is recommended by professional societies, as the harms for alternative antibiotics include antimicrobial resistance, prolonged hospitalizations, readmissions, and increased costs. Removal of penicillin allergy leads to decreased utilization of broad-spectrum antibiotics, such as fluoroquinolones and vancomycin. There is minimal allergic cross-reactivity between penicillins and cephalosporins. IgE-mediated allergy to cephalosporins is usually side-chain specific and may warrant graded challenge with cephalosporins containing dissimilar R1 or R2 group side chains.


Mortality in patients with hepatic gas on point-of-care ultrasound in cardiac arrest: Does location matter?
Prior research has suggested an association of hepatic venous gas with mortality in cardiac arrest. As point of care ultrasound (POCUS) is frequently used in the context of resuscitation, we sought to evaluate if the presence of hepatic gas on POCUS had a similar mortality association. A retrospective review was conducted of patients who experienced nontraumatic cardiac arrest. Archived ultrasound images were independently reviewed to determine the presence of gas in the hepatic parenchyma and vasculature. Electronic medical records were then reviewed to collect remaining clinical data. From 1 January 2017 through 16 June 2019, 87 patients met inclusion criteria. Among them, 68 (78.2%) patients died. Among those who died, 40 (58.8%) had hepatic gas, while 28 (41.2%) had none. Only a single survivor demonstrated hepatic venous gas (11%). While the difference in mortality with respect to presence of undifferentiated hepatic gas was not significant (P = .37), there was a significant difference with respect to the presence of venous gas (P = .004). Our study demonstrated that the incidence of postarrest hepatic gas on POCUS was common, and that the presence of hepatic venous gas during cardiac resuscitation was associated with increased mortality, while hepatic parenchymal gas alone was not.


Prospective superficial EPR in-vivo dosimetry study during hypofractionated radiotherapy of breast cancer patients treated with helical tomotherapy.
In-vivo dosimetry (IVD) is a patient specific measure of quality control and safety during radiotherapy. With regard to current reporting thresholds for significant occurrences in radiotherapy defined by German regulatory authorities, the present study examines the clinical feasibility of superficial electron paramagnetic resonance (EPR) IVD of cumulative total doses applied to breast cancer patients treated with helical intensity-modulated radiotherapy (tomotherapy). In total, 10 female patients with left- or right-sided breast cancer were enrolled in this prospective IVD study. Each patient received a hypofractionated whole breast irradiation. A total median dose of 42.4 Gy in 16 fractions (5 fractions per week) was prescribed to the planning target volume. The treatments were completely delivered using helical tomotherapy and daily image guidance via megavoltage CT (MVCT). For each patient, three EPR dosimeters were prepared and placed at distinct locations on the patient's skin during the delivery of all fractions. Two dosimeters were placed next to the ipsilateral and contralateral mammilla and one dosimeter was placed ventrally to the thyroid (out-of-primary-beam). The total doses delivered to the dosimeters were readout after all fractions had been administered. The measured total dose values were compared to the planned dose values derived from the treatment planning system (TPS). Daily positional variations (displacement vectors) of the ipsilateral mammilla and of the respective dosimeter were analyzed with respect to the planned positions using the daily registered MVCT image. Averaged over all patients, the mean absolute dose differences between measured and planned total dose values (± standard deviation (SD)) were: 0.49 ± 0.85 Gy for the ipsilateral dosimeter, 0.17 ± 0.49 Gy for the contralateral dosimeter and -0.12 ± 0.30 Gy for the thyroid dosimeter. The mean lengths of the ipsilateral displacement vectors (± SD) averaged over all patients and fractions were: 10 ± 7 mm for the dosimeter and 8 ± 4 mm for the mammilla. Superficial EPR IVD is suitable as additional safeguard for dose delivery during helical tomotherapy of breast cancer. Despite positional uncertainties in clinical routine, the observed dose deviations at the ipsilateral breast were on average small compared to national reporting thresholds for total dose deviations (i.e. 10%/4 Gy). EPR IVD may allow for the detection of critical dose errors during whole breast irradiations.


Circ_0009910 sponges miR-491-5p to promote acute myeloid leukemia progression through modulating B4GALT5 expression and PI3K/AKT signaling pathway.
Acute myeloid leukemia (AML) is a heterogeneous group of leukemias with an overall poor prognosis. Circular RNAs (circRNAs) have been verified to play important regulatory roles in AML progression. However, the role and molecular mechanism of circ_0009910 in AML development have not be completely clarified. The expression levels of circ_0009910, microRNA-491-5p (miR-491-5p), and β-1, 4-galactosyltransferase 5 (B4GALT5) were measured by quantitative real-time polymerase chain reaction (qRT-PCR) or Western blot. Cell proliferation and self-renewal ability were assessed via Cell Counting Kit-8 (CCK-8) and sphere formation assay. Cell cycle distribution and cell apoptosis were evaluated by flow cytometry. Caspase-3 activity was tested by Caspase-3 Activity Assay Kit. Western blot was used to examine the protein levels of autophagy-related markers and PI3K/AKT pathway-related markers. The interaction between miR-491-5p and circ_0009910 or B4GALT5 was confirmed by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay, or RNA pull-down assay. Circ_0009910 was highly expressed in AML tissues and cells. Silenced circ_0009910 could significantly inhibit the proliferation, sphere formation, and autophagy and promoted the apoptosis of AML cells. Circ_0009910 bound to miR-491-5p in AML cells, and circ_0009910 promoted AML progression partly through sponging miR-491-5p in vitro. B4GALT5 was a target of miR-491-5p, and miR-491-5p overexpression-mediated influences in AML cells were effectually overturned by the addition of B4GALT5 overexpression plasmid. Furthermore, circ_0009910 could regulate the expression of B4GALT5 by downregulating miR-491-5p in AML cells. Additionally, circ_0009910 could activate the PI3K/AKT signaling pathway by sponging miR-491-5p. Circ_0009910 could suppress the proliferation, sphere formation, and autophagy and accelerated apoptosis by modulating B4GALT5 expression and activating the PI3K/AKT signaling pathway via sponging miR-491-5p in AML cells, suggesting that circ_0009910 might be a potential biomarker for the treatment of AML.


Photobiomodulation promotes angiogenesis in wound healing through stimulating the nuclear translocation of VEGFR2 and STAT3.
In recent years, Photobiomodulation (PBM) has gained prevalence as a kind of physical therapy for wound healing, however, concerning specific cellular mechanisms induced by PBM remains uncertain. The objective of this study is to evaluate the mechanisms of action of PBM (632.8 nm) on angiogenesis in wound healing in vitro and vivo. In the present work, we indicated that PBM with 1.0 J/cm2 irradiation dose exerts positive effects on cell viability, migration, proliferation and tube formation in human umbilical vein endothelial cells (HUVECs). Furthermore, we demonstrate that the VEGFA/VEGFR2/STAT3 pathway plays an important role in PBM effecting cellular function and promoting angiogenesis in wound healing. In addition, we also found that PBM activated the VEGFA/VEGFR2/STAT3 pathway by stimulating VEGFR2 and STAT3 nuclear translocation in the presence of importin-β. Our research offer a new insight into the potential molecular mechanisms in which PBM promotes angiogenesis in wound healing.


Multi-faceted quantitative proteomics analysis of histone H2B isoforms and their modifications.
Histone isoforms and their post-translational modifications (PTMs) play an important role in the control of many chromatin-related processes including transcription and DNA damage. Variants of histones H2A and H3 have been studied in depth and have been found to have distinct functions. Although 13 somatic histone H2B isoforms have been identified by various biochemical and mass spectrometric (MS) approaches, the distinct roles of these isoforms within human cells are as yet unknown. Here, we have developed quantitative MS techniques to characterize isoform-specific H2B expression across the cell cycle, in differentiated myogenic cells, and in different cancer cell lines to illuminate potential functional roles. Using the MS strategies that we developed, we identified differences in H2B isoform levels between different cancer cell types, suggesting cancer or tissue-specific H2B isoform regulation. In particular, we found large variations in the levels of isoforms H2B1B and H2B1M across the panel of cell lines. We also found that, while individual H2B isoforms do not differ in their acetylation levels, trends in the acetylation on all H2B isoforms correlated with acetylation on other histone family members in the cancer cell line panel. We also used the MS strategies to study H2B protein expression across the cell cycle and determined that H2B isoforms that are alternatively spliced to carry a polyadenylation signal rather than the standard histone downstream element are expressed independently of the cell cycle. However, the level of protein produced from the polyadenylated transcripts does not contribute significantly to the total pool of H2B isoforms translated across the cell cycle or in non-cycling myogenic cells. Our results show that H2B isoforms are expressed at varying levels in different cells, suggesting isoform-specific, and possibly cell-type-specific, H2B gene regulation. The bottom-up mass spectrometry technique we developed for H2B quantification is compatible with the current standard histone H3 and H4 bottom-up 'one-pot' analysis platform so that H2B isoforms and their modifications can be studied in future experiments at the same time as histone H3 and H4 modifications. Therefore, we have expanded the histone landscape that can be interrogated in future experiments.


Future microplastics in the Black Sea: River exports and reduction options for zero pollution.
The Black Sea receives increasing amounts of microplastics from rivers. In this study, we explore options to reduce future river export of microplastics to the Black Sea. We develop five scenarios with different reduction options and implement them to a Model to Assess River Inputs of pollutaNts to seA (MARINA-Global) for 107 sub-basins. Today, European rivers draining into the Black Sea export over half of the total microplastics. In 2050, Asian rivers draining into the sea will be responsible for 34-46% of microplastic pollution. Implemented advanced treatment will reduce point-source pollution. Reduced consumption or more collection of plastics will reduce 40% of microplastics in the sea by 2050. In the optimistic future, sea pollution is 84% lower than today when the abovementioned reduction options are combined. Reduction options affect the share of pollution sources. Our insights could support environmental policies for a zero pollution future of the Black Sea.


Sex Differences in Effects of Ketamine on Behavior, Spine Density, and Synaptic Proteins in Socially Isolated Rats.
The mechanistic underpinnings of sex differences in occurrence of depression and efficacy of antidepressant treatments are poorly understood. We examined the effects of isolation stress (IS) and the fast-acting antidepressant ketamine on anhedonia and depression-like behavior, spine density, and synaptic proteins in male and female rats. We used a chronic social IS paradigm to test the effects of ketamine (0, 2.5 mg/kg, and 5 mg/kg) on behavior and levels of synaptic proteins synapsin-1, postsynaptic density protein 95, and glutamate receptor 1 in male rats and female rats in diestrus. Medial prefrontal cortex spine density was also examined in male rats and female rats that received ketamine during either the diestrus or the proestrus phase of their estrous cycle. Male rats showed anhedonia and depression-like behavior after 8 weeks of IS, concomitant with decreases in spine density and levels of synapsin-1, postsynaptic density protein 95, and glutamate receptor 1 in the medial prefrontal cortex; these changes were reversed by a single injection of ketamine (5 mg/kg). After 11 weeks of IS, female rats showed depression-like behavior but no signs of anhedonia. Although both doses of ketamine rescued depression-like behavior in female rats, the decline observed in synaptic proteins and spine density in IS and in diestrus female rats could not be reversed by ketamine. Spine density was higher in female rats during proestrus than in diestrus. Our findings implicate a role for synaptic proteins synapsin-1, postsynaptic density protein 95, and glutamate receptor 1 and medial prefrontal cortex spine density in the antidepressant effects of ketamine in male rats subjected to IS but not in female rats subjected to IS, suggesting dissimilar underlying mechanisms for efficacy of ketamine in the two sexes.


Two DNA endonuclease activities from normal human and xeroderma pigmentosum chromatin active on psoralen plus ultraviolet light treated DNA.
DNA endonuclease activities from the chromatin of normal human and xeroderma pigmentosum, complementation group A (XPA), lymphoblastoid cells were examined on DNA treated with 8-methoxypsoralen (8-MOP) or 4,5',8-trimethylpsoralen (TMP) plus long wavelength ultraviolet (UVA) light, which produce monoadducts and DNA interstrand cross-links, and angelicin plus UVA light, which produces mainly monoadducts. 9 chromatin-associated DNA endonuclease activities were isolated from normal and XPA cells and assayed for activity on PM2 bacteriophage DNA that had been treated with 8-MOP or TMP in the dark and then exposed to UVA light. Unbound psoralen was removed by dialysis and a second dose of UVA light was given. Cross-linking of DNA molecules was confirmed by alkaline gel electrophoresis. In both normal and XPA cells, two DNA endonuclease activities were found which were active on 8-MOP and TMP plus UVA light treated DNA. One of these endonuclease activities, pI 4.6, is also active on intercalated DNA and a second one, pI 7.6, is also active on UVC (254 nm) light irradiated DNA. The major activity against angelicin plus UVA light treated DNA in both normal and XPA cells was found in the fraction, pI 7.6. The levels of activity of both of these fractions on all 3 psoralen-damaged DNAs were similar between normal and XPA cells. These results indicate that in both normal and XPA cells there are at least two different DNA endonucleases which act on both 8-MOP and TMP plus UVA light treated DNA.


The role of human interleukin-6 in B-cell isotype regulation and differentiation.
Human Interleukin-6 (IL-6) is a cytokine secreted by T cells, as well as a variety of other cell types, which exhibits B-cell differentiating activity. The recent cloning of the gene that codes for this molecule has allowed us the opportunity to study the function of this molecule alone and in conjunction with other lymphokines in human B-cell isotype-regulation and differentiation. Recombinant human IL-6 enhances immunoglobulin (Ig) M and G secretion by B-cells activated by Staphylococcal A Cowan strain (SAC) and enhances IgM, IgG, and IgA secretion by B-cells activated by pokeweed mitogen. IL-6 also augments immunoglobulin secretion of differing isotypes from various Epstein-Barr Virus transformed B-cell lines. However, IL-6 does not alter the secreted isotype of naive surface IgM-positive B-cells. As human T-cells secrete other lymphokines in association with IL-6 after activation we examined the interaction of Interleukin-2 (IL-2) gamma-interferon (IFN-gamma) and Interleukin-4 (IL-4) with IL-6 on B-cell immunoglobulin secretion. IL-2 and IL-4 synergized with IL-6 in augmenting immunoglobulin secretion by SAC-activated B-cells. IFN-gamma significantly inhibited the Ig secretion of SAC-activated B-cells cocultured with IL-6 alone or in combination with IL-2. These results demonstrate that human recombinant IL-6 augments immunoglobulin secretion of isotype-committed B-cells but it does not induce a change in the isotype secreted. In addition, this lymphokine synergizes with IL-2 and IL-4 in supporting Ig secretion. However, IFN-gamma significantly inhibits IL-6 induced Ig secretion.


A lipid viewpoint on the plant endoplasmic reticulum stress response.
Organisms, including humans, seem to be constantly exposed to various changes, which often have undesirable effects, referred to as stress. To keep up with these changes, eukaryotic cells may have evolved a number of relevant cellular processes, such as the endoplasmic reticulum (ER) stress response. Owing to presumably intimate links between human diseases and the ER function, the ER stress response has been extensively investigated in various organisms for a few decades. Based on these studies, we now have a picture of the molecular mechanisms of the ER stress response, one of which, the unfolded protein response (UPR), is highly conserved among yeasts, mammals, higher plants, and green algae. In this review, we attempt to highlight the plant UPR from the perspective of lipids, especially membrane phospholipids. Phosphatidylcholine (PtdCho) and phosphatidylethanolamine (PtdEtn) are the most abundant membrane phospholipids in eukaryotic cells. The ratio of PtdCho to PtdEtn and the unsaturation of fatty acyl tails in both phospholipids may be critical factors for the UPR, but the pathways responsible for PtdCho and PtdEtn biosynthesis are distinct in animals and plants. We discuss the plant UPR in comparison with the system in yeasts and animals in the context of membrane phospholipids.


Gene knockdown of gamma-glutamylcysteine synthetase by RNAi in the parasitic protozoa Trypanosoma brucei demonstrates that it is an essential enzyme.
The parasitic protozoa Trypanosoma brucei utilizes a novel cofactor (trypanothione, T(SH)2), which is a conjugate of GSH and spermidine, to maintain cellular redox balance. gamma-Glutamylcysteine synthetase (gamma-GCS) catalyzes the first step in the biosynthesis of GSH. To evaluate the importance of thiol metabolism to the parasite, RNAi methods were used to knock down gene expression of gamma-GCS in procyclic T. brucei cells. Induction of gamma-GCS RNAi with tetracycline led to cell death within 4-6 days post-induction. Cell death was preceded by the depletion of the gamma-GCS protein and RNA and by the loss of the cellular pools of GSH and T(SH)2. The addition of GSH (80 microM) to cell cultures rescued the RNAi cell death phenotype and restored the intracellular thiol pools to wild-type levels. Treatment of cells with buthionine sulfoximine (BSO), an enzyme-activated inhibitor of gamma-GCS, also resulted in cell death. However, the toxicity of the inhibitor was not reversed by GSH, suggesting that BSO has more than one cellular target. BSO depletes intracellular thiols to a similar extent as gamma-GCS RNAi; however, addition of GSH did not restore the pools of GSH and T(SH)2. These data suggest that BSO also acts to inhibit the transport of GSH or its peptide metabolites into the cell. The ability of BSO to inhibit both synthesis and transport of GSH likely makes it a more effective cytotoxic agent than an inhibitor with a single mode of action. Finally the potential for the T(SH)2 biosynthetic enzymes to be regulated in response to reduced thiol levels was studied. The expression levels of ornithine decarboxylase and of S-adenosylmethionine decarboxylase, two essential enzymes in spermidine biosynthesis, remained constant in induced gamma-GCS RNAi cell lines.


Use of autologous fat grafting for the correction of burn scar contracture in the hand: a case report.
Severe burn injuries to the hand represent a topic of great concern due to long-term complications such as hypertrofic burn scar and contracture, which may result in loss of function. We present a case of burn scar contracture in the hand of a child undergoing Z-plasty and lipofilling.


External electric field reduces the explosive sensitivity: a theoretical investigation into the hydrogen transference kinetics of the NH2NO2∙∙∙H2O complex.
Controlling the selectivity of detonation initiation reaction to reduce the explosive sensitivity has been a Holy Grail in the field of energetic materials. The effects of the external electric fields on the homolysis of the N-NO2 bond and initiation reaction dynamics of NH2NO2∙∙∙H2O (i.e., intermolecular and 1,3-intramolecular hydrogen transfers) were investigated at the MP2/6-311++G(2d,p) and CCSD/6-311++G(2d,p)//MP2/6-311++G(2d,p) levels. The results show that the N-NO2 bond is not the "trigger linkage." The notable transiliences of the activation energy of the intermolecular hydrogen transfer are found with the field strength of - 0.012 a.u. along the -x-direction, leading to the conversion of the main reaction between the intermolecular and 1,3-intramolecular hydrogen transference. The activation energies of two kinds of the hydrogen transferences are increased under the external electric fields along the -y-direction. In particular, due to the conversion of the main reaction, the activation energies of the overall reaction are increased significantly along the -x-direction, leading to the significant reduced explosive sensitivities. Therefore, by controlling the field strengths and orientations between the "reaction axis" and external electric field along the y- and x-directions, the selectivity of the initiation reaction could be controlled and the explosive sensitivity could be reduced. Employing AIM (atoms in molecules) and surface electrostatic potentials, the origin of the control of reaction selectivity and the reduction of sensitivity is revealed. This work is of great significance to the improvement of the technology that the external electric fields are added safely into the energetic material system to enhance the explosive performance. Graphical abstract.


Protected thiol-polyethylene glycol: a new activated polymer for reversible protein modification.
Monomethoxypolyethylene glycol (mPEG) of average molecular weight 5000 was transformed in a series of synthetic steps to a new activated form of PEG, a stable thiol-protected intermediary, for reaction with cysteine residues in proteins under mild conditions to produce PEG--protein conjugates as possible candidates for therapeutics. The modified protein has PEG polymer molecules attached to the backbone by the newly formed disulfide bonds, which are readily cleaved to regenerate the native protein under mild reducing conditions. The model protein papain, which has seven cysteine residues including a lone cysteine in its active site, was modified through the free available thiol. The resulting PEG--papain was characterized by matrix-assisted laser desorption mass spectrometry, SDS-PAGE, and high performance gel filtration chromatography. The major modified PEG--papain variant was demonstrated to be 5000 Da larger than the unreacted papain.


Ganglion cyst of the proximal tibiofibular joint causing foot drop-diagnostic problem of differentiating from neuritic leprosy.
A six-year old boy presented with pain around the knee joint and abnormal gait of one month duration. There was no history of hypopigmented anaesthetic patches, neuritis or family history of leprosy. Clinical examination revealed a localized cystic swelling of 1 x 1 cm in size in the region of left common peroneal nerve, with sensory loss on the lateral aspect of the left leg which was mistaken for a nerve abscess.


Hysteroscopy: choosing distending media.
The author reports 2,200 hysteroscopies in which three different distending media were used. He also reports the use of a new uterine fluid insufflator, as well as some modifications in the hysteroscope, to ease the whole endoscopic procedure.


Treatment of intractable epilepsy in a female with SLC6A8 deficiency.
A female heterozygous for a novel, disease causing, missense mutation in the X-linked cerebral creatine transporter (SLC6A8) gene (c.1067G>T, p.Gly356Val) presented with intractable epilepsy, mild intellectual disability and moderately reduced cerebral creatine levels. Treatment with creatine monohydrate, to enhance cerebral creatine transport, combined with L-arginine and L-glycine, to enhance cerebral creatine synthesis, resulted in complete resolution of seizures. Heterozygous SLC6A8 deficiency is a potentially treatable condition and should be considered in females with intractable epilepsy and developmental delay/intellectual disability.


Physiological changes in pregnancy and their influence on the endocrine investigation.
Physiological changes in pregnancy may result in significant alterations in endocrine hormone profiles, serum and urine electrolytes and endocrine gland morphology on imaging. Pregnancy-specific pathophysiological processes may also affect the results for endocrine tests. Investigation of endocrine disorders in pregnancy requires knowledge of these changes and awareness of the safety of dynamic hormone testing and imaging for the mother and foetus. This review seeks to outline the important physiological changes in pregnancy affecting reference intervals of basal and dynamic endocrine tests in pregnancy and the scenarios in which these changes are clinically significant, the pregnancy-specific disorders that may affect the investigation of endocrine disorders, and the safety of dynamic testing and imaging. Awareness of the effect of physiological changes, and the potential impact of pregnancy-specific disorders of endocrine tests, and the safety of imaging is crucial to the management of endocrine disorders in pregnancy.


Prediction of extraprostatic extension by MRI tumor contact length: difference between anterior and posterior prostate cancer.
Tumor contact length (TCL) is defined as the extent of contact between prostate cancer and the prostatic capsule, and its predictive value for microscopic extraprostatic extension (EPE) has been reported. However, the impact of the zonal origin (anterior or posterior tumor) of the tumor on the diagnosis of EPE is controversial. We retrospectively analyzed the records of 233 consecutive patients who underwent preoperative MRI and radical prostatectomy. We designated their tumors as anterior or posterior, and evaluated the correlation between the TCL measured by MRI and microscopic EPE in the radical prostatectomy specimen. Then, we created the predicted probability curves for EPE versus TCL for anterior and posterior prostate cancer. There were 109 patients (47%) with an anterior tumor and 124 patients (53%) with a posterior tumor. Postoperative pathological analysis confirmed pT3 in 18 patients (17%) with an anterior tumor and in 53 patients (43%) with a posterior tumor. Multivariate analysis demonstrated that the zonal origin of the tumor was an independent predictive factor for EPE. We developed separate probability curves of EPE versus TCL for anterior and posterior prostate cancer, which revealed that anterior tumors were less likely to invade the extraprostatic tissues. Among patients whose TCL was 10-20 mm, 9/32 patients (28%) with an anterior tumor had EPE compared with 24/45 patients (53%) with a posterior tumor (p = 0.036). The decision curve of this EPE predictive model had high clinical efficacy. Our results indicate that anterior tumors have more favorable pathological characteristics than posterior tumors with the same TCL measured by MRI. We constructed two separate predicted probability curves for EPE after discriminating anterior and posterior tumors, which will be useful for decision making in clinical practice.


Quality-of-life impact of lower urinary tract symptom severity: results from the Health Professionals Follow-up Study.
To estimate the magnitude of the quality-of-life impact of lower urinary tract symptom (LUTS) severity using a large (n = 8406) sample of U.S. men participating in the Health Professionals Follow-up Study. Multiple regression modeling was used to estimate the relative quality-of-life burden of different levels of LUTS severity (defined using the American Urological Association Symptom Index as none-mild symptoms, 0 to 7; low moderate, 8 to 14; high moderate, 15 to 19; and severe, 20 to 35) and to compare these with age-matched U.S. male norms. A standardized and validated measure of both LUTS severity (the American Urological Association Symptom Index) and a widely used, standardized, multidimensional measure of quality of life (Short Form 36) were used. In addition, the relative impact of severe LUTS on the quality of life was compared with that experienced for other chronic illness conditions (diabetes, angina, hypertension, gout). The results showed that high-moderate LUTS was associated with small to moderate deficits in anxious and depressed mood and poorer role functioning related to emotional problems arising from illness. Severe LUTS was associated with additional quality-of-life deficits related to vitality and the ability to work and carry out daily tasks as a result of illness. Comparisons of the severe LUTS patient group with four other chronic illness groups showed vitality/energy, in particular, but also role functioning and depressed and anxious feelings, to be poorer in the severe LUTS group. Men with high-moderate and severe LUTS identified in a large U.S. cohort have a poorer health status in several important quality-of-life dimensions. The detection and effective treatment of LUTS may substantially improve the health status for these men in these dimensions.


Functional properties of pulvinar-lateral posterior neurons which receive input from the superior colliculus.
Focal electrical stimulation of the superior colliculus induced short latency responses and/or a long duration suppression of spontaneous activity in single cells isolated in the pulvinar and lateral posterior (LP) nuclei of the cat. Cells which responded with one or more spikes at short latencies were relatively few in number, and these tended to be encountered in or near the principal tectorecipient zone of LP. In contrast, a suppression response was observed in many more cells which were widely distributed throughout the pulvinar-LP complex. Stimulation of the superficial layers of the colliculus was more effective in driving or suppressing pulvinar-LP cells than that of intermediate and deep layers. There was also an indication of a coarse topography in this projection. On the basis of their sensory response properties the cells responsive to tectal stimulation, at short latency or with a suppression, could not be differentiated from the overall sample of pulvinar-LP neurons.


A systematic review of clinical and preclinical evidences for Janus kinase inhibitors in large vessel vasculitis.
Corticosteroid-sparing disease-modifying anti-rheumatic drugs are an area of active exploration in large vessel vasculitis (LVV), i.e., Takayasu arteritis (TAK) and Giant Cell Arteritis (GCA). The role of Janus kinase (JAK) inhibitors has been recently identified in different inflammatory rheumatic diseases. We conducted a systematic review of the use of JAK inhibitors in LVV across MEDLINE, Scopus, Web of Science, EMBASE, PubMed Central, Cochrane database of controlled trials, clinicaltrials.gov, and major recent international conferences. We identified four cohort studies and ten case reports. The JAK inhibitors used in these studies were tofacitinib, baricitinib, and ruxolitinib. A cohort study in TAK compared 27 patients treated with tofacitinib with 26 others treated with methotrexate, with better clinical outcomes with tofacitinib but similar angiographic stabilization, relapses, corticosteroid-sparing effect, and adverse events in both groups. Most of the other studies favored clinical responses with JAK inhibitors in LVV but with a paucity of data on other outcomes. Most of the included studies were of moderate quality. Evidence from pre-clinical models of LVV as well as limited in vivo data in patients with TAK appears to suggest that JAK inhibition reduces adventitial fibrosis, intimal proliferation, and inflammatory T lymphocyte infiltration in the media as well as reduces resident memory T cells in the vascular wall (which are otherwise resistant to corticosteroids). Ongoing clinical trials of tofacitinib, baricitinib, and upadacitinib in LVV shall help to further clarify the potential promise of JAK inhibitors for LVV (PROSPERO registration number CRD42021273359). KEY POINTS : •Tofacitinib appeared to associate with better clinical outcomes than methotrexate in TAK. •JAKinibs reduce adventitial fibrosis, intimal proliferation, and inflammatory vascular infiltrate in pre-clinical models of LVV. •Tofacitinib downregulates resident memory vascular T lymphocytes in pre-clinical models of LVV.