Cationic antimicrobial peptides in clinical development, with special focus on thanatin and heliomicin.
Cationic host-defence antimicrobial peptides are recognised as an important component of the innate immune response in most multicellular organisms. In humans, several antimicrobial peptides have recently been recognised as key factors in the pathology of diseases such as cystic fibrosis, septic shock, atopic dermatitis and morbus Kostmann. To date, several hundred cationic antimicrobial peptides have been characterised. They are amphipathic peptides, comprising 20 to 50 amino acids, and exhibiting large structural diversity. These peptides display a broad spectrum of activity against bacterial, fungal and viral pathogens. Their mode of action is best known for cecropins and magainins, which act upon the cytoplasmic membrane of microorganisms, causing its disruption by a detergent-like activity and pore formation. In the last few years, several of these peptides or analogues (derived from magainin, protegrin, indolicidin and histatin) were in advanced clinical development, especially for localised infections (oral and cutaneous infections, pneumonias etc.). Several other molecules (rBPI, heliomicin and thanatin) are currently under development for various systemic infections (Staphylococcus sp., Aspergillus sp., Candida sp. etc.) and may represent important additions to the anti-infectious therapeutic arsenal.


Quercetin pretreatment increases the bioavailability of pioglitazone in rats: involvement of CYP3A inhibition.
Herbal antidiabetic preparations are often used as an add-on therapy in diabetes and such herbal preparations often contains quercetin that can inhibit cytochrome P450 (CYP) 3A4. This enzyme is responsible for metabolizing pioglitazone, a commonly used antidiabetic agent. Hence, it was speculated that quercetin may influence the bioavailability of pioglitazone, which could be particularly crucial, as any increment in its plasma levels may raise safety concerns. Thus, we first established the inhibitory influence of quercetin (2, 10 and 20 mg/kg, p.o.) on CYP3A activity by an in vivo method of estimating levels of midazolam in female rats pretreated with dexamethasone. It was further confirmed in vitro using erythromycin-N-demethylase (EMD) assay. These studies indicated potent inhibition of CYP3A activity by quercetin (10 and 20 mg/kg, in vivo; 1 and 10 microM, in vitro). In another experiment, pioglitazone was administered orally (10 mg/kg) and intravenously (5mg/kg) to quercetin (10 mg/kg) pretreated female rats and its plasma levels were determined at various time points (0.5, 1, 2, 4, 8 and 24 h after oral administration; 0.083, 0.5, 1, 2, 8, 12 and 18 h after i.v. administration) by HPLC. Quercetin pretreatment increased AUC(0-infinity) of pioglitazone after oral administration by 75% and AUC(0-infinity) after intravenous administration by 25% suggesting decreased metabolism, which could be due to inhibition of CYP3A by quercetin. In conclusion, add-on preparations containing quercetin may increase the bioavailability of pioglitazone, and hence should be cautiously used.


Microradiographic study of cerebral and ocular aneurysms in hypertensive rabbits.
Cerebral and ocular microaneurysms were produced in rabbits made hypertensive by surgically induced silk-turpentine perinephritis combined with contralateral nephrectomy 7 days later. The aneurysms distributed throughout the brain and iris were studied by microradiography; a few representative aneurysms selected from the microradiographs were studied histologically. The microradiographic findings and histological sections correlated well. Intracranial microaneurysms were multiple and frequently located in the basal ganglia and near the cortex. Arterial changes in the iris paralleled intracranial arterial changes in degree and type except for the addition of hemorrhage around some microaneurysms. The results of this study show the potential of microradiography and histological sections guided by microradiography for studying the natural history of hypertensive arterial lesions and support the contention that hypertension, microaneurysms and intracerebral hemorrhages are related.


Retrospective analysis on correlation factors of preserving the ligamentum flavum in microendoscopic discectomy.
The present study was conducted to investigate the correlation factors of successful preservation of the ligamentum flavum (LF) in microendoscopic discectomy. This retrospective review was carried out on 78 consecutive patients who underwent single segment microendoscopic discectomy projecting to preserve the LF between January 2012 to January 2013. The demographic and clinical data including age, gender, duration of disease, area of interlaminar space, disc level and position type of lumbar disc herniation were recorded. Intraoperative outcomes including duration of operation and peri-operative bleeding were recorded. Clinical outcomes were assessed by Oswestry disability index (ODI) and visual analog scale (VAS), and fibrosis formation was detected using computed tomographic scans with IV iopamidol injection. The follow up lasted six months. LF was successfully preserved in 54 patients (69.2%) and 24 patients (30.8%) underwent microendoscopic discectomy without preservation of LF. In multivariate logistic analysis, factors including age (OR, 1.17; 95% CI, 1.07-1.28; P<0.001) and area of the laminar space (OR, 0.09; 95% CI, 0.01-0.67; P=0.018) significantly associated with the preservation of LF. Other factors including gender, duration of ill course, disc level and position type seemed not associated with the preservation of the LF. In addition, there was no significant difference of the duration of the operation (P=0.689) as well as the peri-operative bleeding (P=0.147) between patients with preservation of the LF and patients without. However, patients with preservation of the LF showed significantly improved clinical outcomes (ODI: P=0.006, VAS: P=0.035) and less fibrosis formation than those without LF (P=0.018). Microendoscopic discectomy with preservation of the LF could achieve better clinical outcomes and less epidural fibrosis, while elder patients with smaller area of the laminar space should be deliberated on the preservation of the LF during the microendoscopic discectomy.


Anticancer properties of green-synthesised zinc oxide nanoparticles using Hyssopus officinalis extract on prostate carcinoma cells and its effects on testicular damage and spermatogenesis in Balb/C mice.
The unclear bio-safety issue and potential risk of nanoparticles (NPs) on various organelles can be considered as a major challenge. In the present study, we have assessed the green synthesis of ZnO nanoparticles using Hyssop (Hyssopus officinalis) extract and their effects on PC3 cell line and BALB/c mice model. The cytotoxicity of the ZnO-NPs was assessed on PC3 cell line by MTT test after characterisation. Apoptotic effect of ZnO-NPs was determined by in vitro AO/PI staining. The histopathological assessments and determination of LH and FSH levels carried out as in vivo analysis in BALB/c adult male mice. The expression of major genes involved in spermatogenesis and sperm maturation (Adam3, Prm1, Spata19, Tnp2, Gpx5) were also analysed. The obtained result demonstrated that the IC50 for PC3 cell line treated with green-synthesised ZnO-NPs during 24 and 48 hr was reported 8.07 and 5 µg/ml respectively. Meanwhile, the induced apoptosis was recorded 26.6% ± 0.05, 44% ± 0.12 and 80% ± 0.07 of PC3 cells. The results of gene expression analysis revealed that the increase in the concentration of ZnO-NPs significantly (p < .05) down-regulated the Adam3, Prm1, Spata-19, Tnp2 and Gpx5 genes. The overall results of this research elucidated that ZnO-NPs impaired spermatogenesis, sperm maturation process and sperm motility.


Compensation in Preclinical Huntington's Disease: Evidence From the Track-On HD Study.
Cognitive and motor task performance in premanifest Huntington's disease (HD) gene-carriers is often within normal ranges prior to clinical diagnosis, despite loss of brain volume in regions involved in these tasks. This indicates ongoing compensation, with the brain maintaining function in the presence of neuronal loss. However, thus far, compensatory processes in HD have not been modeled explicitly. Using a new model, which incorporates individual variability related to structural change and behavior, we sought to identify functional correlates of compensation in premanifest-HD gene-carriers. We investigated the modulatory effects of regional brain atrophy, indexed by structural measures of disease load, on the relationship between performance and brain activity (or connectivity) using task-based and resting-state functional MRI. Consistent with compensation, as atrophy increased performance-related activity increased in the right parietal cortex during a working memory task. Similarly, increased functional coupling between the right dorsolateral prefrontal cortex and a left hemisphere network in the resting-state predicted better cognitive performance as atrophy increased. Such patterns were not detectable for the left hemisphere or for motor tasks. Our findings provide evidence for active compensatory processes in premanifest-HD for cognitive demands and suggest a higher vulnerability of the left hemisphere to the effects of regional atrophy.


Effects of lactic acid bacteria fermentation on the phytochemicals content, taste and aroma of blended edible rose and shiitake beverage.
In this research, five commercial lactic acid bacteria (LAB) strains were selected for the fermentation of blended edible rose and shiitake beverage for their frequent application and strong adaptability in plant-based beverages. The viable counts of all strains reached above 10.0 log CFU/mL in the blended beverage after fermentation (48 h). Meanwhile, higher contents of total phenols, total flavonoids, free amino acids, and flavor nucleotide as well as stronger antioxidant capacity were observed in the fermented samples than the control. The results of electronic tongue (E-tongue) analysis showed that LAB fermentation significantly (p<0.05) lowered the signals of bitterness, astringency, aftertaste-A, and aftertaste-B, which ultimately improved the taste profiles of the fermented samples. Additionally, a total of 76 volatiles compounds were detected, among which alcohols were the dominant compounds. LAB fermentation also enriched the aroma complex of the fresh beverage, which resulted in new compounds, including 13 alcohols, 9 acids, 6 ketones, 4 esters, and 5 hydrocarbons in the fermented samples.


Cyclic nucleotides and ethanol tolerance and dependence.
Acute ethanol administration decreases cyclic AMP levels in brains of some animal strains. The ethanol-induced decrease of cyclic GMP levels cannot be blocked by intraventricular infusion of calcium but it can be prevented by pretreatment with pyrazole. Although cyclic AMP levels in the cerebral cortex change reciprocally after acute ethanol administration and during withdrawal, the increase observed during withdrawal is probably secondary to some withdrawal processes. Ethanol decreases cerebellar GABA levels only in rats which are stressed prior to sacrifice. Chronic intraventricular infusion of calcium increased the intensity of an acoustic startle response during ethanol withdrawal while EGTA infusion delayed the development of tolerance to the hypothermic effect of ethanol. The ratio of cyclic GMP to GABA levels is postulated to be important for expression of the withdrawal syndrome.


Clinical panurothelial disease in patients with superficial bladder tumors: therapeutic implications.
We established the prognostic and therapeutic implications of panurothelial involvement in patients with superficial bladder tumors for optimizing therapeutic approaches in those at risk for panurothelial involvement. We studied the records of 35 patients with clinical panurothelial disease. Since all of these patients presented with high risk superficial bladder cancer during followup, they were included in specific therapeutic and followup regimens. Radical procedures or conservative therapies were indicated mainly according to pathological examination and the recurrence pattern. Panurothelial involvement was a late stage of a recurrent and diffuse process that essentially developed in sequences, in which all patients presented with high risk superficial bladder tumors. This process involved continued relapse after panurothelial involvement developed. Notably 19 patients (79.1%) at risk for recurrence had repeat relapse in the urothelium. In the upper urinary tract 12 patients (34.3%) had bilateral involvement, including 7 (41.2%) of 17 patients after cystectomy. We identified 2 subgroups of patients. The subgroup with a better prognosis included 27 patients in whom late panurothelial disease developed step by step after a complete response to intravesical therapy, including 14 (51.8%) who were free of disease. The other subgroup with a poor prognosis included 8 patients with concurrent bladder carcinoma in situ and prostate involvement as well as early panurothelial disease, of whom only 2 (25%) were disease-free. All patients underwent many therapeutic approaches. A mean of 7.5 surgical procedures per patient were done, including a mean of 5.5 transurethral resections, a mean of 1 conservative approach to the upper urinary tract and a mean of 1.1 radical procedures. At a median followup of 111 months 10 patients (28.5%) were disease-free but only 7 (20%) retained the bladder, while 19 (54.3%) died of tumor. Patients with high risk superficial bladder multifocal tumors and associated bladder carcinoma in situ are at high risk for panurothelial involvement. Radical cystectomy may be recommended in these patients when initially or during followup, concurrent high risk superficial bladder tumors and prostate involvement develop or prostate involvement recurs. For the upper urinary tract conservative therapies may be advisable when noninfiltrating tumors are diagnosed even after cystectomy due to the high rate of bilateral new onset disease. When cystectomy is performed, extended excision of the upper urinary tract and pyelo-intestinal anastomosis may be considered.


Interstitial washdown during crystalloid fluid loading in graded hypovolemia -A retrospective analysis in volunteers.
"Interstitial washdown" is an edema-preventing mechanism that implies a greater redistribution of interstitial albumin occurs whenever the capillary filtration is increased. To study the effect of interstitial washdown on fluid distribution in normovolemic and hypovolemic volunteers. Capillary filtration was increased by infusing 25 mL/kg Ringer's acetate intravenously over 30 min 10 male just after withdrawal of 0, 450, and 900 mL of blood. Population volume kinetic analysis was used to assess the effects of washdown and hemorrhage on fluid distribution, using the difference in plasma dilution based on hemoglobin and albumin as biomarker of washdown. Blood withdrawal during 10-15 min recruited 100-150 mL of fluid of high albumin content to the plasma, which was probably lymph. The albumin recruitment was temporarily reduced during the fluid loading but increased from 40 min post-infusion and was then greater when preceded by hemorrhage. Simulations suggested that interstitial washdown decreased the extravascular fluid volume by 200 mL over 3 h. The plasma volume and urinary excretion both increased by approximately half this amount. Blood loss without hypotension probably recruited lymph to the plasma, but interstitial washdown played no major role in determining the distribution of crystalloid fluid after hemorrhage.


Can nutrition label recognition or usage affect nutrition intake according to age?
The aim of this study was to investigate the effect of the use of nutrition labeling on nutritional intake according to age groups, focusing on Korean elderly. Study participants (N = 5223) were adults at least 20 y of age and had participated in the Korean National Health Examination and Nutrition Survey in 2012. Data for recognition/use of nutrition labels were obtained by self-report. Nutrition intake also was estimated by 24-h dietary recall. Participants were categorized into three age groups: 20 to 39 y, 40 to 59 y, and ≥60 y. Generalized linear model was conducted to test mean differences between nutrition label recognition (NLR) and nutrition label use (NLU) groups for nutrient intake, according to the age groups. Results from this study indicated that younger individuals (age groups of 20-39 and 40-59 y) in the NLU group showed a significant association with nutrient intake compared to those in the NLR group. Additionally, nutrition intake status in the NLU group improved positively. Whereas older participants (≥60 y) in the NLR group showed a significant association with most nutrient intake compared with the NLU group. The study also found that protein intake was reduced in the NLU group compared with the non-NLU group across the age groups, except for older participants (age group 20-39 y: 79.16 versus 86.30 g, P = 0.050; age group 40-59 y: 69.97 versus 75.58 g, P = 0.040; age group of ≥60 y: 64.72 versus 64.89 g, P = 0.967). The present study revealed that nutrition labeling cannot be effective for the elderly, and there were several areas of misunderstanding. Therefore, more systematic education on the topic of nutrition labeling is required to help the elderly make healthier food decisions.


The combination of ınterleukin-10 -1082 and tumor necrosis factor α -308 or ınterleukin-6 -174 genes polymorphisms suggests an association with susceptibility to Hashimoto's thyroiditis.
The etiopathogenesis of Hashimoto's thyroiditis (HT) has not been clearly elucidated although the role of chronical inflammation and endothelial dysfunction has been established. The imbalance between pro- and anti-inflammatory cytokines may play a role in the etiology. The aim of the present study was to investigate whether cytokine gene polymorphisms are associated with HT, and to evaluate the relationship between genotypes and clinical/laboratory manifestation of HT. Tumor necrosis factor α (TNFα) G-308A (rs 1800629), interleukin-6 (IL-6) G-174C (rs 1800795) and IL-10 G-1082A (rs 1800896) single nucleotide polymorphisms (SNPs) in DNA from peripheral blood leukocytes of 190 patients with HT and 231 healthy controls were investigated by real-time PCR combined with melting curve analysis using fluorescence-labeled hybridization probes. There was no notable risk for HT afflicted by TNFα -308, IL-6 -174 and IL-10 -1082 polymorphisms alone. However, carriers of variant alleles of both IL-10 -1082 and TNFα -308 polymorphisms had four-fold times higher risk for HT in comparison with non-carriers. Additionally, concomitant presence of both mutant IL-10 -1082 A and IL-6 -174 C alleles raised three-fold the HT risk. Our results suggest that the combined effects of TNFα -308, IL-6 -174 and IL-10 -1082 variant alleles may be more decisive to induce functional differences and modify the risk for HT.


A functional polymorphism in the succinate-semialdehyde dehydrogenase (aldehyde dehydrogenase 5 family, member A1) gene is associated with cognitive ability.
Succinate-semialdehyde dehydrogenase (SSADH) deficiency is a rare cause of learning disability. We have investigated SSADH to assess its contribution to cognitive ability in the general population in both case-control- and family-based analyses. Sequence analysis of SSADH revealed four changes affecting the encoded protein, only one of which had a minor allele whose frequency is even moderately common. We genotyped this functional polymorphism in 197 high-IQ cases, 201 average-IQ controls and 196 parent high-IQ offspring trios. The minor allele was significantly less frequent in high-IQ cases and was significantly less frequently transmitted by parents to high-IQ subjects than chance expectation. A previous study has shown that the minor allele encodes a lower activity enzyme than the major allele. These data suggest that higher SSADH activity is associated with higher intelligence across the general population. The effect is small, with each allele having an effect size translating to about 1.5 IQ points.


Circular RNA Related to the Chondrocyte ECM Regulates MMP13 Expression by Functioning as a MiR-136 'Sponge' in Human Cartilage Degradation.
Circular RNAs (circRNAs) are involved in the development of various diseases, but there is little knowledge of circRNAs in osteoarthritis (OA). The aim of study was to identify circRNA expression in articular cartilage and to explore the function of chondrocyte extracellular matrix (ECM)-related circRNAs (circRNA-CER) in cartilage. To identify circRNAs that are specifically expressed in cartilage, we compared the expression of circRNAs in OA cartilage with that in normal cartilage. Bioinformatics was employed to predict the interaction of circRNAs and mRNAs in cartilage. Loss-of-function and rescue experiments for circRNA-CER were performed in vitro. A total of 71 circRNAs were differentially expressed in OA and normal cartilage. CircRNA-CER expression increased with interleukin-1 and tumor necrosis factor levels in chondrocytes. Silencing of circRNA-CER using small interfering RNA suppressed MMP13 expression and increased ECM formation. CircRNA-CER could compete for miR-136 with MMP13. Our results demonstrated that circRNA-CER regulated MMP13 expression by functioning as a competing endogenous RNA (ceRNA) and participated in the process of chondrocyte ECM degradation. We propose that circRNA-CER could be used as a potential target in OA therapy.


Desferrioxamine enhances the reactivity of vanadium (IV) and vanadium (V) toward ferri- and ferrocytochrome c.
Ligands, especially desferrioxamine, affect the rate at which vanadium reduces or oxidizes cytochrome c. Whether reduction or oxidation occurs, and how fast, depends on the nature of the ligand, the state of reduction of the vanadium, the pH (6.0, 7.0, or 7.4), and the availability of oxygen. In general, oxidation of ferrocytochrome c was favored by (1) low pH, (2) an oxidized state of the vanadium, (3) the presence of oxygen, and (4) more strongly binding ligands (desferrioxamine much greater than histidine = ATP greater than EDTA greater than albumin greater than aquo). Thus, at pH 6.0, desferrioxamine accelerated the V(V)-catalyzed ferrocytochrome c oxidation 160-fold aerobically, and 3500-fold anaerobically. In general, strongly binding ligands slowed oxidations, especially at higher pH. Desferrioxamine was unique among the five ligands in that it not only accelerated oxidation of ferrocytochrome c at pH 6.0, but at pH 7.4 the redox balance shifted to the point where it paradoxically reduced ferricytochrome c. V(V) is an improbable electron donor, but desferrioxamine will reduce cytochrome c, and V(V) accelerates this process. Oxidation of cytochrome c by V(V):desferrioxamine was faster anaerobically, and reduction by V(IV):desferrioxamine was faster aerobically. Although V(V) did not oxidize ferrocytochrome c at pH 7.4, V(IV) did, provided oxygen and desferrioxamine were both present. V(IV):desferrioxamine almost completely reduced ferricytochrome c, and this reduction was followed by a slow, progressive oxidation. This latter oxidation of cytochrome c is mediated by active species generated in the reaction between V(IV):desferrioxamine and oxygen, because none of these reagents alone can induce oxidation at a comparable rate. The mediating species were transient, and generated in reactions with oxygen.(ABSTRACT TRUNCATED AT 250 WORDS)


Nitric oxide release from coumarin 4-carboxamidoximes in the presence of thiols.
Coumarin 4-carboxamides with potent anti-inflammatory activity were studied for nitric oxide release in the presence of thiol cofactors in vitro. It was found that NO2- release was dependent on the nature of thiol.


Glucose uptake in Oesophagostomum dentatum and the effect of oxfendazole.
The uptake of 14C-glucose by adult Oesophagostomum dentatum was characterised. The uptake was a non-linear function of external glucose concentration. The maximum velocity of uptake (Vmax) was 0.964 nmol/100 mg dry weight (dw)/5 min, and the transport constant (Kt) was 10.02 microM. When phlorizin, phloretin and 3-O-methylglucose were tested for their effects on the uptake of 14C-glucose, phloretin and 3-O-methylglucose produced significant inhibitions, indicating that the uptake was mediated and occurred by facilitated diffusion. Exposure of the worms to oxfendazole prior to incubation with 14C-glucose did not affect the uptake of glucose. In another experiment worms were incubated with unlabelled glucose and the external glucose concentration was measured enzymatically. During a 7 h incubation period, the quantity of glucose remaining in the incubation medium of oxfendazole exposed worms was significantly greater than in the control group. It was concluded that oxfendazole did not influence the process of 14C-glucose uptake, but might induce changes in the parasite leading to a reduced ability to deplete the incubation medium of glucose.


An assessment of a collagen/vicryl composite membrane to repair defects of the urinary bladder in rabbits.
Collagen/vicryl (Polyglactin) composite membrane has been used to repair full-thickness defects in the urinary bladder of rabbits. The material has been shown to be biodegradable, prevent leakage of urine, and is readily replaced by collagenous scar tissue lined with a urothelium. Regeneration of smooth muscle has been observed in the repair area of some animals. The results suggest that such a material may well be of use to urologists wishing to augment contracted bladders or in the repair of bladder fistulae in human subjects, thereby avoiding the use of bowel or other material e.g. omentum.


Involvement of NMDA receptors in the hypotensive response to the injection of L-glutamate into the lateral hypothalamus of unanesthetized rats.
We report that microinjections of L-glutamate (L-glu) or N-methyl-D-aspartic acid (NMDA) in the lateral hypothalamus (LH) of unanesthetized rats caused a hypotensive response. Guide cannulas were stereotaxically placed in the LH 3 days before the experiments, under tribromoethanol anesthesia. One day before the experiments, the femoral artery was cannulated for pulsatile arterial pressure (PAP), mean arterial pressure (MAP) and heart rate (HR) measurements. In the first experiment, unanesthetized rats received microinjections of 2.5, 5.0 or 10.0 nmol/100 nL of L-glu in the LH. Dose-dependent hypotensive responses were observed, without significant concomitant changes in heart rate. In a second group of experiments, 5.0 nmol of L-glu was microinjected into the LH before and 10 min after pretreatment with glutamatergic antagonists. Pretreatments with the non-selective ionotropic glutamatergic-receptor antagonist kynurenic acid or the selective NMDA receptor antagonists AP-7 and LY235959 significantly reduced the hypotensive response to microinjection of L-glu in the LH. Pretreatment with the selective AMPA-receptor antagonist NBQX or with vehicle did not affect the hypotensive response. The present results suggest that the hypotensive response to the injection of L-glu into the LH is mediated by NMDA receptors.


First report of Sarcocystis halieti (Apicomplexa) in bearded vulture (Gypaetus barbatus).
At least three Sarcocystis species (S. falcatula, S. halieti and S. wobeseri-like) have been detected infecting raptorial birds. By histopathology and PCR-sequencing of the ITS1 marker, S. halieti was detected in a bearded vulture (Gypaetus barbatus) and a black kite (Milvus migrans) from the Catalonia region in North Spain. The 241 bp-long sequences obtained from the Sarcocystis organisms detected in both raptors showed 97.5-99.6% and 97.9-100% similarity with those of previously identified S. halieti; also, the phylogenetic trees generated placed the identified sequences together with other sequences of S. halieti available in GenBank. In sum, the description of the bearded vulture as a new intermediate host for S. halieti adds new insights on the complex epidemiology of the genus involving avian hosts.


Quantification of skeletal muscle mass: sarcopenia as a marker of overall health in children and adults.
Sarcopenia is defined as the loss of muscle mass or function and has been associated with increased morbidity and mortality in a variety of diseased populations. Sarcopenia results from a higher rate of muscle protein degradation compared to protein synthesis and is an important marker of metabolic status related to nutrition and physical activity. The diagnosis of sarcopenia is accomplished by clinical assessment demonstrating decreased muscle function and radiographic confirmation of decreased muscle mass, via dual X-ray absorptiometry, bioelectric impedance or cross-sectional imaging with CT or MRI. However, normative data for skeletal muscle mass are lacking, especially for children and young adults. Additionally, studies of skeletal muscle mass by cross-sectional imaging in children are scarce. Here, we review the concept of sarcopenia with an emphasis on its relevance in the pediatric population.


Domiciliary therapy during inpatient rehabilitation treatment for patients with an acquired brain injury: a preliminary study.
The objective was to assess the feasibility of additional domiciliary treatment for patients with an acquired brain injury while they are still inpatients at a rehabilitation centre. This cohort study included 22 patients with an acquired brain injury (mainly stroke) and with moderate to severe neuropsychological deficits. After two observational home visits for goal setting to determine the aim of the treatment, domiciliary treatment was given once a week in addition to the therapy at the rehabilitation centre. The number of home visits depended on the time interval between admission and time of inclusion in the study. The main outcome measures were (1) a semi-structured interview with patients and partners or children, (2) a questionnaire filled in by the attending therapist(s) before and after each therapeutic home visit and (3) the functional level of the patient. All but one of the patients and all partners and therapists were satisfied with the domiciliary treatment. Most of the main goals were attained during the treatment. The main goals during the domiciliary treatment were in the domains of domestic activities and leisure. A 90--120 min duration of the treatment sessions seems most feasible. It was concluded that domiciliary treatment for inpatients is feasible and seems useful for patients with moderate to severe cognitive impairments.


Flavone photoreactivity. UV-induced reactions in organic solvents and micellar systems.
Flavonoid photochemistry is a subject of interest in studies dealing with the role of phenolic compounds as screening pigments in plants. In order to contribute to the understanding of the processes involved in the interaction between flavonoids and UV radiation, we have studied UV-induced flavone photodegradation in both organic solvents and micellar systems. The results obtained show that flavone photosensitivity depends on the characteristics of the reaction environment and is influenced by the medium polarity and the charges on the micellar surface. Qualitative and quantitative differences in the photodegradation products were demonstrated by high performance liquid chromatography (HPLC) analysis.


miR-550a-3/NFIC plays a driving role in esophageal squamous cell cancer cells proliferation and metastasis partly through EMT process.
In this study, the functional role of miR-550a-3 and its direct target nuclear factor IC (NFIC) in esophageal squamous cell cancer (ESCC) cells were explored. Differential expression of miR-550a-3 in ESCC tissues was acquired from TCGA database, and Kaplan-Meier method was used to determine the relationship between miR-550a-3 expression and survival time of ESCC patients. Expression level of miR-550a-3 in several ESCC cell lines was measured by qRT-PCR. Two cell lines including Eca109 and JAR were used to perform proliferation, cloning, invasion and migration experiments. Targeted relationship between miR-550a-3 and NFIC was speculated by predication software and confirmed by dual luciferase assay. Additionally, potential relationship between miR-550a-3 and NFIC was analyzed by Spearman rank correlation analysis and western blot. Rescue assays were performed to explore the function of miR-550a-3/NFIC in ESCC cells biological behaviors. Expression levels of key proteins involved in epithelial-to-mesenchymal transition (EMT) process were determined by western blot. By consulting TCGA database, we found that high expression of miR-550a-3 was positively connected with the poor prognosis of patients with ESCC. In addition, overexpression of miR-550a-3 promoted the proliferation, colony formation and metastasis of ESCC cells. Moreover, rescue assays revealed that overexpression of NFIC attenuated the promoting effects of miR-550a-3 on ESCC cells malignant behaviors. While the promoting effects of miR-550a-3 on EMT process were inhibited by NFIC. Our results illustrate the importance of miR-550a-3/NFIC in regulation of ESCC cells growth and metastasis, which could contribute to developing novel target for early diagnosis or neoteric therapeutic target for ESCC.


Spatial-temporal characteristics and regional differences of the freight transport industry's carbon emission efficiency in China.
The freight transport industry is an important field in which to achieve the goal of carbon emission reduction within the transportation industry. Analyzing the spatial-temporal characteristics and regional differences in the freight transport industry's carbon emissions efficiency (CEE) is an essential prerequisite for developing a reasonable regional carbon abatement policy. However, few studies have conducted an in-depth analysis of the freight transport industry's CEE from the perspective of geographic space. This study combines the super-efficiency slack-based measure (SBM) model and the window analysis model to measure the freight transport industry's CEE in 31 Chinese provinces from 2008 to 2019. We then introduced a spatial autocorrelation analysis and the Theil index to analyze the spatial-temporal evolution characteristics and regional differences in the freight transport industry's CEE in China. The results show that (1) the overall level of the freight transport industry's CEE is low, with an average of 0.534, which showed a weak downward trend during the study period. This indicates that the freight industry's CEE has not improved, and there is a massive requirement for energy conservation and emission reduction. (2) From 2008 to 2019, CEE gradually shows a spatial distribution pattern of being "low in the west and high in the east," with a significant, positive spatial correlation (all passed the significance level test at P < 0.01). This indicates that the spatial diffusion and inhibition of the freight transport industry's CEE in adjacent areas cannot be ignored. (3) The overall differences in the freight transport industry's CEE show a fluctuating upward trend from 2008 to 2019. The inter-regional differences of the three regions (east, central, and west) are the main contributors of the total differences. Therefore, narrowing inter-regional gaps in CEE is one of the main ways to improve the freight transport industry's CEE.


Immunological identity between bovine preparations of thiol:protein-disulphide oxidoreductase, glutathione-insulin transhydrogenase and protein-disulphide isomerase.
Five preparations of bovine thiol:protein-disulphide oxidoreductase/glutathione-insulin transhydrogenase (EC 220.127.116.11) and one preparation of bovine liver protein-disulphide isomerase (EC 188.8.131.52) from four different laboratories showed immunological identity in double immunodiffusion and rocket-line immunoelectrophoresis. Consequently, thiol:protein-disulphide oxidoreductase/glutathione-insulin transhydrogenase and protein-disulphide isomerase, formerly classified as two separate enzymes, should be considered as alternative activities of the same enzyme.


Infantile spasms: ictal phenomena.
Ictal phenomena were studied during three separate six-hour video/polygraphic recording sessions in 10 patients with infantile spasms; 1,079 spasms occurred. Frequency during wakefulness (7.7 spasms/hour) was greater than that during sleep (2.5 spasms/hour); 46.6% of spasms occurred in clusters. Spasms were composed of one or more of three phases: a myoclonic contraction, a tonic contraction, and/or an arrest of activity. The most common types were myoclonic-tonic (40.3%) and myoclonic alone (36.3%). When classified by postural motor phenomena, 41.6% were "flexor", 16.3% "extensor", 39.0% "mixed", and 3.1% "arrest" alone. Electrographic monitoring revealed that myoclonic contractions were associated with an initial paroxysmal event. Tonic contractions and arrests were usually associated with suppression of electroencephalographic activity with or without rhythmic activity. Knowledge of these clinical and electrographic features is important for diagnosis and evaluation of proposed treatments.


Natural Variation in Stress Hormones, Comparisons Across Matrices, and Impacts Resulting from Induced Stress in the Bottlenose Dolphin.
Knowledge regarding stress hormones and how they vary in response to seasonality, gender, age, and reproductive status for any marine mammal is limited. Furthermore, stress hormones may be measured in more than one matrix (e.g., feces, blood, blubber), but the relationships between levels of a given hormone across these matrices are unknown, further complicating the interpretations of hormones measured in samples collected from wild animals. A study is underway to address these issues in a population of bottlenose dolphins trained for voluntary participation in sample collections from different matrices and across season and time of day.


Health-Related Quality of Life in Patients With Hodgkin Lymphoma: A Longitudinal Analysis of the German Hodgkin Study Group.
Many important details of health-related quality of life (HRQoL) after diagnosis and treatment of Hodgkin lymphoma (HL) are still unknown because large longitudinal studies of HRQoL are rare. Therefore, we analyzed a systematically assessed, comprehensive range of HRQoL domains in patients with HL of all stages from diagnosis up to 5 years of survivorship. We included patients with HL age 18-60 years at diagnosis from the German Hodgkin Study Group trials HD13, HD14, and HD15. We analyzed HRQoL using all functional and symptom scales of the European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire C30 including deviations from reference values. We estimated the effect of different disease, patient, and treatment characteristics using multiple regression and repeated measures analysis and computed correlations of HRQoL scores. We analyzed 4,215 patients with any HRQoL assessment within 5 years after treatment. Higher tumor burden at diagnosis was associated with impaired baseline scores in many HRQoL domains. During survivorship, cognitive, emotional, role, and social functioning and fatigue, dyspnea, sleep, and financial problems were severely and persistently affected. From year 2 on, mean deviations from reference values ranged between 12 and 29 points, with 10 points being a commonly used margin of clinical relevance. In all 3 trials, HRQoL domains 2 and 5 years after therapy were significantly influenced by baseline scores and age but not by randomized treatments. Fatigue was most closely correlated with other symptoms and scales. Our results show a high and persistent amount of different HRQoL deficits in survivors of HL that are largely independent of the applied chemotherapies. Our analysis underscores the high, unmet medical need of these rather young survivors of HL regarding the psychosocial adverse effects of the cancer experience.


Development and validation of a combined nomogram model based on deep learning contrast-enhanced ultrasound and clinical factors to predict preoperative aggressiveness in pancreatic neuroendocrine neoplasms.
This study aimed to develop and validate a combined nomogram model based on deep learning (DL) contrast-enhanced ultrasound (CEUS) and clinical factors to preoperatively predict the aggressiveness of pancreatic neuroendocrine neoplasms (PNENs). In this retrospective study, consecutive patients with histologically proven PNENs underwent CEUS examination at the initial work-up between January 2010 and October 2020. Patients were randomly allocated to the training and test sets. Typical sonographic and enhanced images of PNENs were selected to fine-tune the SE-ResNeXt-50 network. A combined nomogram model was developed by incorporating the DL predictive probability with clinical factors using multivariate logistic regression analysis. The utility of the proposed model was evaluated using receiver operator characteristic, calibration, and decision curve analysis. A total of 104 patients were evaluated, including 80 (mean age ± standard deviation, 47 years ± 12; 56 males) in the training set and 24 (50 years ± 12; 14 males) in the test set. The DL model displayed effective image recognition with an AUC of 0.81 (95%CI: 0.62-1.00) in the test set. The combined nomogram model that incorporated independent clinical risk factors, such as tumor size, arterial enhancement level, and DL predictive probability, showed strong discrimination, with an AUC of 0.85 (95%CI: 0.69-1.00) in the test set with good calibration. Decision curve analysis verified the clinical usefulness of the combined nomogram. The combined nomogram model could serve as a preoperative, noninvasive, and precise evaluation tool to differentiate aggressive and non-aggressive PNENs. • Tumor size (odds ratio [OR], 1.58; p = 0.02), arterial enhancement level (OR, 0.04; p = 0.008), and deep learning predictive probability (OR, 288.46; p < 0.001) independently predicted aggressiveness of pancreatic neuroendocrine neoplasms preoperatively. • The combined model predicted aggressiveness better than the clinical model (AUC: 0.97 vs. 0.87, p = 0.009), achieving AUC values of 0.97 and 0.85 in the training set and the test set, respectively.


Ultrasonic-assisted matrix solid-phase dispersion as an improved methodology for the determination of pesticides in fruits.
An ultrasonic-assisted matrix solid-phase dispersion (UA-MSPD) method has been developed for extracting and cleaning-up 15 organophosphorus pesticides (OPPs) and 9 triazines in fruits. Using reversed phase octasilyl-derivatised silica (C8) as dispersant and ethyl acetate as extraction solvent, two sonication devices, an ultrasonic bath and a sonoreactor, were tested for speeding and increasing the efficiency of the MSPD process. A standard MSPD and a heating-assisted MSPD procedure were also done. Gas chromatography-mass spectrometry was used for analyte determination. 1-min sonication with the sonoreactor at 50% amplitude provided the best results, with reproducibilities below 15% for the pesticides studied. The general low detection limits (1-42microgkg(-1)) ensure proper determination at maximum allowed residue levels set in current legislations, except for dimethoate and disulfuton. The analytical performance of the method was evaluated for apples, pear and apricot, showing little or no matrix effect.


Starch hydrolase inhibitors from edible plants.
Type 2 diabetes is a chronic disease with gradual deterioration in glucose metabolism, which causes multiple systemic complications. Postprandial hyperglycemia is a concern in the management of type 2 diabetes. Of all the available antidiabetic therapeutic methods, inhibition of α-glucosidase and α-amylase is postulated to be a preventive treatment. Many natural products and herbal medicines have been recommended as being beneficial for mitigation of postprandial hyperglycemia. In this review, recent discoveries of α-glucosidase and α-amylase inhibitors from edible plants are described along with their chemical structures. Their inhibition mechanisms, the type of each glucosidase and amylase, and measurement methods for the inhibitory activity are also given. Finally, recent progress on low glycemic index foods incorporated with plants containing starch hydrolase inhibitors is summarized.


Enzymatic Hydrolysis of Hydrotropic Pulps at Different Substrate Loadings.
Enzymatic hydrolysis of cellulosic raw materials to produce nutrient broths for microbiological synthesis of ethanol and other valuable products is an important field of modern biotechnology. Biotechnological processing implies the selection of an effective pretreatment technique for raw materials. In this study, the hydrotropic treatment increased the reactivity of the obtained substrates toward enzymatic hydrolysis by 7.1 times for Miscanthus and by 7.3 times for oat hulls. The hydrotropic pulp from oat hulls was more reactive toward enzymatic hydrolysis compared to that from Miscanthus, despite that the substrates had similar compositions. As the initial substrate loadings were raised during enzymatic hydrolysis of the hydrotropic Miscanthus and oat hull pulps, the concentration of reducing sugars increased by 34 g/dm(3) and the yield of reducing sugars decreased by 31 %. The findings allow us to predict the efficiency of enzymatic hydrolysis of hydrotropic pulps from Miscanthus and oat hulls when scaling up the process by volume.


Protoplast fusion of Schizosaccharomyces pombe Auxotrophic mutants of identical mating-type.
Protoplasts of methionine- and lysine-requiring h- mutants isolated from the L972 h- strain of Schizosaccharomyces pombe were fused. The protoplasts were obtained from the cells with enzymes produced by Trichoderma viride. When a mixture of the protoplasts was treated with 30% PEG 4000 solution containing 10 mM CaCl2, cell fusion and complementation was attained with a frequency of 0.17%. Both fusion partners were recovered among the spores after crossing of the fusion products with the strain M210 ade6 h+. Cytological and haploidization examinations showed that the fusion cells are not heterokaryons, and that the increased amount of genetic material is situated in one nucleus.


Population genetic differentiation of the ubiquitous brooding coral Pocillopora acuta along Phuket Island reefs in the Andaman Sea, Thailand.
The widespread Indo-Pacific coral species Pocillopora acuta Lamarck, 1816 displays varying levels of asexual versus sexual reproduction, with strong repercussions on genetic diversity, connectivity and genetic structuring within and among populations. For many geographic regions, baseline information on genetic diversity is still lacking, particularly in the Andaman Sea. The region suffered a massive heat-induced bleaching event in 2010 with high coral cover loss of branching coral species such as P. acuta. A subsequent bleaching in 2016, however, revealed a mild bleaching response in pocilloporids compared to other coral taxa in the region, suggesting that rare, heat tolerant genotypes had been selected by the 2010 bleaching event. In order to test whether this potential 'evolutionary rescue' event has led to a low genetic diversity, we conducted a population genetic survey covering a total of nine different P. acuta populations (336 individuals) along a 50 km coastal stretch around Phuket Island, Thailand. We used six microsatellite markers to assess genotypic diversity and to determine the prevalent mode of reproduction (i.e. sexual or asexual recruitment). In contrast to other Indian Ocean P. acuta populations, the majority of corals in this study adopted a sexual reproduction mode (75% across all populations). At the same time, substantial regional gene flow was observed around Phuket Island with strong genetic differentiation as indicated by three genetic clusters that were separated by only a few kilometers. Patterns of isolation by distance over 0.7 - 40 km suggest small-scale genetic barriers, such as changing currents throughout each monsoonal season, potentially contributing to locally restricted dispersal of P. acuta larvae. The occurrence of distinct genetic clusters within short coastal stretches suggests that the 2010 bleaching event has not led to extreme genetic impoverishment. While more in-depth genomic analyses are necessary to investigate changes in genetic diversity following extreme bleaching events, our results will help guide conservation efforts to maintain genetic diversity of a coral species that likely will be dominant in future, warmer Andaman Sea reefs.


Special Report: Frontline Nurses Say 'Never Again'.
Nurses share experiences from the pandemic and strategies to successfully manage future health care crises.


Biochemical and physiological effects of catecholamine administration in the horse.
Adrenaline was given intramuscularly to resting horses. It increased heart rate, sweating, blood levels of lactic dehydrogenase, aldolase, creatine kinase, glucose, lactate, free fatty acids and glycerol. Responses to isoprenaline, to noradenaline and to adrenaline after pretreatment with propranolol indicated that beta receptors were involved in stimulation of tachycardia, sweating, lipolysis and muscle glycogenolysis, and alpha receptors in stimulation of liver glycogenolysis and leakage of intracellular enzymes. The time course and relative magnitude of the effects on different enzymes was not identical to that produced by exercise.


Segmental pancreas transplantation in the canine model. A reappraisal.
Segmental vascularized grafts of pancreatic tissue have been recommended in the treatment of juvenile onset insulin-dependent diabetes mellitus. The method of intraperitoneal placement and unligated pancreatic duct provided a low incidence of exocrine complications, but clinical results were less than ideal. To further assess this technique, we used segmental pancreatic autografts in the canine model. Five of the 22 dogs survived with good function of the autograft as determined by glucose homeostasis and a slow return to normal of serum amylase levels. In 12 dogs, however, pancreatic dysfunction was evident within 24 hours and death followed within days. At reexploration or necropsy, necrotic pancreatic tissue and clotted vessels were found. We suggest from these findings that although few, if any, complications of exocrine drainage or endocrine function were encountered, a major technical problem with segmental pancreatic transplantation may involve inadequate arterial runoff to maintain patency of vessels and viability of the graft. Terminal arteriovenous fistulas do not seem to improve vascular patency in the canine model in our experience.


Higher pain sensitivity and lower muscle strength in postmenonpausal women with early rheumatoid arthritis compared with age-matched healthy women--a pilot study.
The purpose of the study was to examine muscle strength and pain sensitivity in postmenopausal women with and without RA. Ten women with and ten without early RA were recruited. All were postmenopausal, and did not use hormone replacement therapy. Measurements of isokinetic muscle strength in knee flexors/extensors, hand grip strength, timed standing, pressure pain thresholds (PPT), suprathreshold pressure pain, and segmental and plurisegmental endogenous pain inhibitory mechanisms during muscle contraction were assessed. Participants with early RA were weaker in knee flexors, in hand grip strength and they needed more time for the timed standing. Women with early RA had higher sensitivity to threshold pain and suprathreshold pressure pain compared to women without RA. PPTs increased in the contracting muscle as well as in a distant resting muscle during static contractions in both groups. Our results indicate differences in muscular strength between postmenopausal women with and without RA. Furthermore, women with RA had decreased PPT and hyperalgesia, but no dysfunction of segmental or plurisegmental pain inhibitory mechanisms during static exercise compared to healthy controls. The normal function of endogenous pain inhibitory mechanisms despite chronic pain in women with RA might contribute to the good effects of physical activity previously reported.


Disposition and Residue Depletion of Metronidazole in Pigs and Broilers.
Metronidazole (MNZ) is used in veterinary medicine for the treatment of anaerobic infections and a variety of protozoal and parasitic diseases. Current study has been conducted to examine the disposition and residue depletion studies of MNZ and its main metabolites in pigs and broilers. After a single oral administration of MNZ, the concentrations of MNZ and its main metabolites in the excreta of pigs and broilers were determined by LC-MS/MS. More than 75% of the drug was recovered within 14 days, of which MNZ accounted for about 40%, MNZ-OH 20-25% and MAA less than 10%. The residue depletion study showed that MNZ was the most predominant residue in all of the tissues and could be detected in liver, kidney and muscle up to the withdrawal time of 14 days. MNZ-OH concentrations were lower than MNZ in all of the tissues, but its elimination half-life was close to MNZ. It is proposed that kidney and muscle are appropriate residue target tissues and both MNZ and its hydroxylated metabolite, MNZ-OH, should be monitored in the routine surveillance of MNZ related residues in food of animal origin.


Towards biobanking technologies for natural and bioengineered multicellular placental constructs.
Clinical application of a large variety of biomaterials is limited by the imperfections in storage technology. Perspective approaches utilizing low-temperature storage are especially challenging for multicellular structures, such as tissues, organs, and bioengineered constructs. Placenta, as a temporary organ, is a widely available unique biological material, being among the most promising sources of various cells and tissues for clinical and experimental use in regenerative medicine and tissue engineering. The aim of this study was to analyse the mechanisms of cryoinjuries in different placental tissues and bioengineered constructs as well as to support the viability after low temperature storage, which would contribute to development of efficient biobanking technologies. This study shows that specificity of cryodamage depends on the structure of the studied object, intercellular bonds, as well as interaction of its components with cryoprotective agents. Remarkably, it was possible to efficiently isolate cells after thawing from all of the studied tissues. While the outcome was lower in comparison to the native non-frozen samples, the phenotype and expression levels of pluripotency genes remained unaffected. Further progress in eliminating of recrystallization processes during thawing would significantly improve biobanking technologies for multicellular constructs and tissues.


Clinical trials and treatment effects monitoring.
Treatment effects monitoring is the act of reviewing accumulated data by treatment group to determine whether a trial should continue unaltered. That monitoring is required for any trial where there is a risk of an aggregate form of harm for subjects because of continued use of an inferior treatment or because of failure to use a superior treatment. Institutional review boards (IRBs), in deciding whether to approve such research, require that (when appropriate) there is adequate provision for monitoring the data collected to ensure the safety of subjects. The focus here is on the conditions necessary to satisfy this requirement. The conditions discussed are timeliness of the monitoring, completeness of data for monitoring, competency of monitors, and freedom of monitors to act and recommend as they deem necessary, regardless of the wishes, desires, or dictates of sponsors.


Elevated CO2-Induced Responses in Stomata Require ABA and ABA Signaling.
An integral part of global environment change is an increase in the atmospheric concentration of CO2 ([CO2]) [1]. Increased [CO2] reduces leaf stomatal apertures and density of stomata that plays out as reductions in evapotranspiration [2-4]. Surprisingly, given the importance of transpiration to the control of terrestrial water fluxes [5] and plant nutrient acquisition [6], we know comparatively little about the molecular components involved in the intracellular signaling pathways by which [CO2] controls stomatal development and function [7]. Here, we report that elevated [CO2]-induced closure and reductions in stomatal density require the generation of reactive oxygen species (ROS), thereby adding a new common element to these signaling pathways. We also show that the PYR/RCAR family of ABA receptors [8, 9] and ABA itself are required in both responses. Using genetic approaches, we show that ABA in guard cells or their precursors is sufficient to mediate the [CO2]-induced stomatal density response. Taken together, our results suggest that stomatal responses to increased [CO2] operate through the intermediacy of ABA. In the case of [CO2]-induced reductions in stomatal aperture, this occurs by accessing the guard cell ABA signaling pathway. In both [CO2]-mediated responses, our data are consistent with a mechanism in which ABA increases the sensitivity of the system to [CO2] but could also be explained by requirement for a CO2-induced increase in ABA biosynthesis specifically in the guard cell lineage. Furthermore, the dependency of stomatal [CO2] signaling on ABA suggests that the ABA pathway is, in evolutionary terms, likely to be ancestral.


Immune system function. Implications for critical care nursing practice.
The purpose of the immune system is to protect the body from invasion from microorganisms. Natural defenses encompassing epithelial surfaces and secretions, nonspecific phagocytosis and inflammatory process, and the acquired defenses of humoral and cell-mediated immunity work together to defend the internal environment of the body. If microorganisms penetrate the external surfaces and enter the body, the inflammatory process is triggered. During phagocytosis by monocytes/macrophages, the invading antigen is identified as nonself. The antigen is presented to the T4-lymphocyte. This cell line orchestrates the activation of B-lymphocytes and a humoral response of antibody or Ig production and initiates the cytotoxic cell-mediated T cell response. Following an appropriate level of response, T8 cells suppress immune system response to the antigen, but not before memory B and T cells have formed. These memory cells protect the body against future exposure to the antigen, thus creating immunity. As concepts of immunology are identified and validated by future research, the knowledge base of immune system function and dysfunction will continue to grow. Critical care nurses must incorporate new knowledge related to immune system function into their practice, as patients in the ICU are at risk for immunocompromise from their underlying disease state and the therapeutic interventions used in their treatment. Nurses must expand their physiologic data base to include a nursing assessment of the immune system. A framework for organizing the data collection process would include: (1) a survey of factors affecting immune system function in each patient, (2) assessment of the cells and structures of the immune system, (3) monitoring of the status of first line defenses, (4) observing for activity of nonspecific defenses, and (5) evidence of specific acquired immune responses. After complete immune system assessment has identified areas of risk for compromise or the presence of immunocompromise, specific individualized nursing care plans can be developed to provide nursing support to maintain and enhance the patient's defenses.


Clinical and radiographic evaluation of the orbit.
A complete clinical examination of the orbit must include a detailed physical evaluation of the eye and its associated structures. Whether the examination is for a traumatic, infectious, inflammatory, or neoplastic condition of the orbit, the main concern is determination of the visual status and preservation of vision.


Immunologic and inflammatory mechanisms that drive asthma progression to remodeling.
Although histologic features of airway remodeling have been well characterized in asthma, the immunologic and inflammatory mechanisms that drive progression of asthma to remodeling are still incompletely understood. Conceptually, airway remodeling may be a result of persistent inflammation and/or aberrant tissue repair mechanisms. It is likely that several immune and inflammatory cell types and mediators are involved in mediating airway remodeling. In addition, different features of airway remodeling are likely mediated by different inflammatory pathways. Several important candidate mediators of remodeling have been identified, including TGF-beta and T(H)2 cytokines (including IL-5 and IL-13), as well as vascular endothelial growth factor, a disintegrin and metalloproteinase 33, and matrix metalloproteinase 9. Mouse models of airway remodeling have provided important insight into potential mechanisms by which TGF-beta activation of the Smad-2/3 signaling pathway may contribute to airway remodeling. Human studies have demonstrated that anti-IL-5 reduces levels of airway eosinophils expressing TGF-beta, as well as levels of airway remodeling as assessed by bronchial biopsies. Further such studies confirming these observations, as well as alternate studies targeting additional individual cell types, cytokines, and mediators, are needed in human subjects with asthma to determine the role of candidate mediators of inflammation on the development and progression of airway remodeling.


Histamine mediates the stimulatory action of ghrelin on acid secretion in rat stomach.
Ghrelin, a novel growth hormone-releasing peptide, is present in the rat and human stomach and is known to stimulate acid secretion and stomach motility. However, the mechanism of action of ghrelin is not fully understood. In the present study, we attempted to elucidate the role of histamine in ghrelin-induced acid secretion in rat stomach. Intravenous administration of ghrelin at 0.8 to 20 microg/kg dose dependently increased gastric acid secretion, as measured by the gastric lumen perfusion method. The maximum response was almost equal to that of gastrin (20 microg/kg). These actions were abolished by bilateral subdiaphragmatic vagotomy. Famotidine (0.33 mg/kg) also completely inhibited the effects of ghrelin. Furthermore, ghrelin increased histidine decarboxylase (HDC) messenger RNA (mRNA) levels, as measured by real-time reverse transcription-polymerase chain reaction using LightCycler. The action of ghrelin on HDC mRNA was abolished by vagotomy. Ghrelin did not affect histamine release from isolated vascularly perfused rat stomach. Taken together, these results suggest that ghrelin stimulates gastric acid secretion via a mechanism involving activation of vagal efferent nerve and histamine release from gastric enterochromaffin-like cells.


uPA/PAI-1 ratios distinguish benign prostatic hyperplasia and prostate cancer.
Urokinase plasminogen activator (uPA) and its inhibitor type 1 (PAI-1) are associated with tumour metabolism and are widely considered to be informative for the identification of cancer. We have analysed prostate tissue resections from patients with prostate cancer (PCa) and with benign prostatic hyperplasia (BPH) for protein levels of uPA and PAI-1, and searched for distinctions between these two clinical manifestations. Prostate tissue was deep frozen in liquid N2 and homogenized in a stainless steel punch homogenizer. The tissue powder was extracted with a pH 8.5 TRIS/Triton X-100 buffer, and the extract analysed by FEMTELLE assay to generate uPA and PAI-1 readings in ng/mg protein. The uPA/PAI-1 ratio was calculated for each sample, and the mean ratios for the two diagnostic groups were compared. The concentration of uPA (mean ± SD) was found to be 0.19 ± 0.04 ng/mg protein (range 0.05-0.72 ng/mg) and 0.15 ± 0.02 ng/mg protein (range 0.03-0.78 ng/mg) in PCa and BPH samples, respectively. The concentration of PAI-1 was found to be 4.93 ± 0.90 ng/mg (range 1.10-11.80 ng/mg) and 5.87 ± 0.70 ng/mg (range 0.2-25.0 ng/mg) in PCa and BPH samples, respectively. A consistent finding being that PAI-1 concentrations exceed uPA concentrations by far giving rise to characteristic uPA/PAI-1 ratios. In BPH samples, there was a trend of PAI-1 to increase with uPA content, while in PCa samples, PAI-1 remained fairly constant. The mean uPA/PAI-1 ratio in PCa samples was found to be 0.06 ± 0.01 and was significantly higher than in BPH samples where the mean uPA/PAI-1 ratio was 0.03 ± 0.003 (p = 0.0028). R(2) = 0.1389. Using a contingent of 62 patients of which 46 were BPH and 16 were PCa, we report definitive concentrations of uPA and PAI-1 in tumour tissue extracts and show that the uPA/PAI-1 ratio emerges as a candidate marker to distinguish between BPH and PCa.


Evaluation of apoptotic cell death on liver and kidney tissues following administration of levetiracetam during prenatal period.
Levetiracetam is a new generation antiepileptic drug used in treatment of patients with epilepsy and has adverse effects on different tissues. We aimed to evaluate the apoptotic effects of levetiracetam exposure during pregnancy on liver and kidney tissues of rat pups. We analyzed the newborn rat pups exposed to levetiracetam during prenatal period. Fifteen pregnant female rats were divided into three groups. The group 1 and 2 rats were treated with different doses of levetiracetam (25 mg/kg/d and 50 mg/kg/d, respectively) from gestational days 1-22 during pregnancy. Group 3 (control group) was treated with the same volume of saline. Apoptosis was evaluated by the terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL) method. Liver and kidney tissues from rat pups were used for investigation. The percent of TUNEL positive apoptotic cells in group 1 were 22 and 17.5 for kidney and liver, respectively. The percent of TUNEL positive apoptotic cells in group 2 were 20.9 and 20.9 for kidney and liver, respectively. The percent of TUNEL positive apoptotic cells in group 3 were 18.4 and 17.1, respectively, for kidney and liver. The apoptotic index was the same in kidney and liver tissues of all groups. Our results demonstrate that the prenatal exposure of levetiracetam has no apoptotic effects on liver and kidney of rat pups and, it has biosafety in pregnancy in terms of apoptosis. The first study evaluating the apoptotic effects on liver and kidney tissues following administration of levetiracetam during prenatal period.


The effect of preimplantation culture conditions on murine embryo implantation and fetal development.
Ham's F-10 medium (Gibco, Grand Island, NY) and medium T6 with and without 15% fetal calf serum (FCS) were compared for their ability to support development of murine blastocysts with the capacity to implant and produce normal fetuses when transferred to pseudopregnant females. All media supported equal rates of blastocyst development from 2-cell embryos. In addition, there were no differences in the rates of blastocyst implantation. However, once implanted, blastocysts grown in T6 produced a significantly higher proportion of normal fetuses (58.5% to 65.9%) than blastocysts grown in either Ham's F-10 (2.4%) or T6 with FCS (27.6%). These results demonstrate that the rate of murine blastocyst development from 2-cell embryos in vitro is not a good criterion of healthy embryos. Murine blastocysts transferred in medium with 0% versus 50% FCS implanted and developed into normal fetuses at equal rates.


Prostaglandin E2 and the renin-angiotensin system in primary aldosteronism and Cushing's syndrome.
8 patients with primary aldosteronism (PA) and 4 women with Cushing's syndrome (CS) were investigated with estimations of prostaglandin E2 (PGE2), plasma renin activity (PRA), and angiotensin-II (A-II) concentrations in renal venous blood, under resting and strictly standardized conditions and without pharmacological influence. In the PA group all hormone values were extremely low, but with a tendency to higher PRA and A-II concentrations in patients with idiopathic adrenal hyperplasia. Such low PGE2 concentrations have not previously been reported, and may be the consequence of decreased concentration of circulating A-II. However, a yet unidentified mechanism for the parallel suppression of the renin-angiotensin system (RAS) and of products of arachidonic acid metabolism cannot be excluded in hypertensive states where blood pressure is mainly dependent on volume factors. The number of patients in the CS group was too small to permit definitive conclusions, but a specific pattern of the PGE2 production is probably not seen in conjunction with hypercorticism. There seemed to exist only a covariation between PGE2 concentration and the degree of activation of the RAS.


Strategic consideration for effective chemotherapeutic transportation via transpapillary route in breast cancer.
Breast cancer is the most commonly occurring cancer in women and the second most common cancer overall. The current treatment option for breast cancer includes drug treatment (Chemotherapy and hormone therapy) or surgery (mastectomy and lumpectomy). The major drawbacks of chemotherapy include rapid metabolism, limited ductal exposure, lower bioavailability, higher elimination rate, undesirable side effects, and high dose of drug. Localized application of drug to breast achieves higher drug levels at the tumor site and reduces systemic drug distribution. Permeation of drug by local application is mainly by transepidermal and transductal routes. The multiple duct openings of mammary papilla provide a higher permeable surface for transport of drug. Various formulation approaches like conjugated nanocarriers, microemulsion, nanoemulsion, liposomes, microneedles and iontophoresis are used to achieve a tailored and targeted drug delivery to underlying breast tissue. Localized transport of drug via transpapillary route has developed a novel and effective non-invasive drug delivery approach to achieve a systemic drug level at the tumor site in the management of breast cancer.


Endosomal signaling and a novel pathway defined by the natural killer receptor KIR2DL4 (CD158d).
In addition to ligand-induced activation of receptors at the cell surface, certain internalized receptor-ligand complexes are activated in endosomes which are, now recognized as important intracellular platforms of signal transduction. The major receptor families that signal from endosomes and illustrate the diversity and complexity of endosomal signaling include receptor tyrosine kinases (RTKs), G-protein-coupled receptors (GPCRs) and toll-like receptors (TLRs). Natural killer (NK) cells, an important component of the innate immune system, not only provide a rapid defense against foreign invaders, such as bacteria and viruses, but also positively shape local responses by cytokine and chemokine secretion. The NK cell receptor KIR2DL4 (CD158d) utilizes a new mode of endosomal signaling after binding its ligand, soluble HLA-G, in the extracellular milieu. Internalization of the receptor and its ligand into endosomes and initiation of signaling at this site result in a proinflammatory and proangiogenic response with important functions at sites of ligand expression, such as at the maternal-fetal interface during early pregnancy. After a brief overview of the modes of endosomal signaling and its value in generating distinct physiological responses, this review will highlight the mechanism and physiological significance of a novel intracellular signaling pathway used by the endosome-resident immune receptor KIR2DL4.


Litopenaeus vannamei Toll-interacting protein (LvTollip) is a potential negative regulator of the shrimp Toll pathway involved in the regulation of the shrimp antimicrobial peptide gene penaeidin-4 (PEN4).
The Toll-like receptor (TLR)-nuclear factor (NF)-κB signaling pathway is evolutionarily conserved from insects to mammals as a regulator of the expression of immune-related genes. In mammals, TLR-NF-κB signaling is tightly controlled because excessive activation of this pathway can result in severe damage to the host. The mammalian Toll-interacting protein (Tollip) has an important function in the negative regulation of this pathway, but no reports about invertebrate Tollip have been published to date. In this study, we cloned Litopenaeus vannamei Tollip (LvTollip) and investigated its function in the regulation of the NF-κB pathway-controlled antimicrobial peptide genes (AMPs). The LvTollip full-length cDNA is 1231bp long and contains an open reading frame of 813bp that encodes a 270-amino acid protein. LvTollip shares significant similarities to mammalian Tollips, which contain a centrally localized protein kinase C conserved region 2 (C2) domain and a C-terminal CUE domain. After challenges with the white spot syndrome virus (WSSV) or Vibrio alginolyticus, the expression levels of LvTollip were altered in the gill, hemocyte, hepatopancreatic, intestinal, and muscle tissues. In Drosophila S2 cells, LvTollip localized in the membrane and the cytoplasm and significantly inhibited the promoter activities of the NF-κB pathway-controlled AMP penaeidin-4 (PEN4). In LvTollip-knockdown shrimp, the expression level of AMP PEN4 was increased. However, the mortality rates of LvTollip-knockdown shrimp in response to WSSV or V. alginolyticus infections were not significantly different from those of the control group. Our results suggested that LvTollip might be involved in the negative regulation of PEN4 and that LvTollip expression was responsive to microbial infections.


Hydroxychloroquine: mechanism of action inhibiting SARS-CoV2 entry.
Hydroxychloroquine (HCQ) has been proposed in the treatment of SARS-coronavirus 2 (SARS-CoV-2) infection, albeit with much controversy. In vitro, HCQ effectively inhibits viral entry, but its use in the clinic has been hampered by conflicting results. A better understanding of HCQ's mechanism of actions in vitro is needed to resolve these conflicts. Recently, anesthetics were shown to disrupt ordered monosialotetrahexosylganglioside1 (GM1) lipid rafts. These same lipid rafts recruit the SARS-CoV-2 surface receptor angiotensin converting enzyme 2 (ACE2) to an endocytic entry point, away from phosphatidylinositol 4,5 bisphosphate (PIP2) domains. Here we employed super resolution imaging of cultured mammalian cells to show HCQ directly perturbs GM1 lipid rafts and inhibits the ability of ACE2 receptor to associate with the endocytic pathway. HCQ also disrupts PIP2 domains and their ability to cluster and sequester ACE2. Similarly, the antibiotic erythromycin inhibits viral entry and both HCQ and erythromycin decrease the antimicrobial host defense peptide amyloid beta in cultured cells. We conclude HCQ is an anesthetic-like compound that disrupts GM1 lipid rafts similar to anesthetics. The disruption likely decreases viral clustering at both endocytic and putative PIP2 entry points.


[Effect of microwaves on the expression by thymocytes of various surface membrane markers].
A study was made of the effects of microwave irradiation of different intensity within decimeter and centimeter ranges in vitro on the guinea-pig thymocyte-induced receptor expression to their own and rabbit erythrocytes. Besides, effects of decimeter waves on mice thymocyte-induced expression of Thy-1 antigen were studied. Microwaves were found to modulate the thymocyte-induced expression of the membrane surface markers under study.


Deficient cholesterol side chain oxidation in patients without peroxisomes (Zellweger syndrome): evidence for the involvement of peroxisomes in bile acid synthesis in man.
The absence of peroxisomes in patients with the cerebro-hepato-renal (Zellweger) syndrome is accompanied by a number of biochemical abnormalities including the accumulation of the bile acid intermediates di- and trihydroxycoprostanoic acid. In this paper we show that there is a marked deficiency in the oxidative side chain cleavage of cholesterol in liver from Zellweger patients. These findings not only provide an explanation for the low levels of the major naturally occurring bile acids, cholic acid and chenodeoxycholic acid and the accumulation of di- and trihydroxycoprostanoic acid in Zellweger patients, but also suggest that peroxisomes are essential in bile acid synthesis in man.


Four-dimensional mapping of dynamic longitudinal brain subcortical development and early learning functions in infants.
Brain subcortical structures are paramount in many cognitive functions and their aberrations during infancy are predisposed to various neurodevelopmental and neuropsychiatric disorders, making it highly essential to characterize the early subcortical normative growth patterns. This study investigates the volumetric development and surface area expansion of six subcortical structures and their associations with Mullen scales of early learning by leveraging 513 high-resolution longitudinal MRI scans within the first two postnatal years. Results show that (1) each subcortical structure (except for the amygdala with an approximately linear increase) undergoes rapid nonlinear volumetric growth after birth, which slows down at a structure-specific age with bilaterally similar developmental patterns; (2) Subcortical local area expansion reveals structure-specific and spatiotemporally heterogeneous patterns; (3) Positive associations between thalamus and both receptive and expressive languages and between caudate and putamen and fine motor are revealed. This study advances our understanding of the dynamic early subcortical developmental patterns.


Fiber systems in the olfactory bulb and cortex: a study in adult and developing rats, using the timm method with the light and electron microscope.
The structure of the olfactory bulb and cortex of rats is described using the Timm method, in which endogenous transition or "heavy" metals are precipitated with sulfide and then stained with silver. With the light microscope, two types of staining, a fine, "colored" reaction and a coarse, "granular" reaction, are found in the neuropil of the adult forebrain; neuronal somata, fiber tracts, and some specific areas of the neuropil are almost unstained. With the electron microscope the Timm method consists of small silver granules, which are associated with axon terminals. These granules are located over the external surface of the presynaptic membrane, facing the synaptic cleft, and over synaptic vesicles, especially the internal surface of the vesicle membrane. This localization suggests that the reactive metals are related to a membrane component which is recycled during secretion of synaptic transmitters. The colored and granular reactions seen with the light microscope appear to be correlated with different intensities of staining of different types of axon terminals. The full extent of the olfactory cortex can be delineated by a bilaminar staining pattern in layer I, which coincides with the pattern of termination of fiber systems to the cortex demonstrated by axonal tracing experiments. The superficial part of layer I is very palely stained; this corresponds precisely to layer Ia, the zone of termination of fibers from the olfactory bulb. In contrast, the deep part of the layer is stained with an intense colored reaction; this corresponds precisely to layer Ib, the zone of termination of the major association fiber system within the olfactory cortex. A prominent granular reaction, which does not correspond to any known long axon system, is found in all layers of the cortex, although it is concentrated at the superficial edge of layer Ib and in layer II. Variations in staining in the several subdivisions of the olfactory cortex are illustrated and described. In the olfactory bulb, the most prominent staining is a relatively fine, granular reaction in the glomerular formations, which apparently corresponds to the olfactory nerve terminals. In immature rats, the Timm method stains the neuropil throughout the olfactory system, although the patterns and quality of staining differ from that found in adults. Before birth, the best-differentiated reaction is seen in glomeruli of the olfactory bulb and correlates well with the pattern of olfactory nerve ingrowth.(ABSTRACT TRUNCATED AT 400 WORDS)


Vulnerabilities for Exposure to Emerging Infectious Disease at Urban Settlements in Nepal.
In Nepal, rapid urbanization and rural-to-urban migration especially due to internal civil conflict have catalyzed the development of temporary settlements, often along rivers on undeveloped land. This study conducted surveillance for viruses in small mammals and assessed potential risks for virus transmission to people in urban settlements along rivers in Kathmandu, Nepal. We collected samples from 411 small mammals (100 rodents and 311 shrews) at four riverside settlement sites and detected six viruses from four virus families including Thottapalayam virus; a strain of murine coronavirus; two new paramyxoviruses; and two new rhabdoviruses. Additionally, we conducted surveys of 264 residents to characterize animal-human contact. Forty-eight percent of individuals reported contact with wildlife, primarily with rodents and shrews (91%). Our findings confirm that rodents and shrews should be considered a health threat for residents of temporary settlements, and that assessment of disease transmission risk coupled with targeted surveillance for emerging pathogens could lead to improved disease control and health security for urban populations. Additionally, interventions focused on disease prevention should consider the unique urban ecology and social dynamics in temporary settlements, along with the importance of community engagement for identifying solutions that address specific multi-dimensional challenges that life on the urban river margins presents.


Biological efficacy of silk fibroin nanofiber membranes for guided bone regeneration.
The favorable biological properties of silk fibroin (SF) nanofiber membrane make it a good candidate for clinical applications as a device in bone and periodontal regenerative therapy. The purpose of this study is to evaluate the biocompatibility of the SF nanofiber membrane, and to examine its effect on bone regeneration in a rabbit calvarial model. To examine the biocompatibility of the electrospun SF membrane, we investigated cell proliferation, morphology, and differentiation. The bone regenerative efficacy of the membrane was evaluated in the calvarial defect of rabbits. The cell numbers and osteocalcin production labels were significantly increased in accordance with culture period. Cells had a stellate shape and broad cytoplasmic extensions on the membrane. The cells showed activity of ALPase that was comparable to culture dishes, and were calcified similarly to culture dishes. In in vivo tests, a complete bony union across the defects was observed after 8 weeks. At 12 weeks, the defect had completely healed with new bone. In conclusion, the SF nanofiber membrane was shown to possess good biocompatibility with enhanced bone regeneration and no evidence of any inflammatory reaction. These results strongly suggest that the SF membrane should be useful as a tool for guided bone regeneration.


Synthesis and characterization of mead: from the past to the future and development of a new fermentative route.
In ancient times, mead was produced by fermentation of honey in water and presented low quality, undesired by-products, off-flavors, and the production was time consuming. In this study, nine experiments were performed to improve the fermentation and mead characteristics. Distillation was not part of the production process and it was performed in this work to produce a new spirit. The samples were characterized by gas chromatography coupled to mass spectrometry, high performance liquid chromatography, digital densimetry, titration, gravimetric method, pH, and refractometry. The results were compared to commercial beverages and legal limits. The meads presented high ethanol concentration, low by-products, fast fermentation, and high quality. The spirits showed high quality and the concentrations of acetic acid, ethyl acetate, methanol, higher alcohols, and ethyl carbamate were below the limits for safe consumption. In conclusion, it was possible to develop new conditions to produce high quality mead and mead spirit.


Universal screening for methicillin-resistant Staphylococcus aureus at hospital admission and nosocomial infection in surgical patients.
Experts and policy makers have repeatedly called for universal screening at hospital admission to reduce nosocomial methicillin-resistant Staphylococcus aureus (MRSA) infection. To determine the effect of an early MRSA detection strategy on nosocomial MRSA infection rates in surgical patients. Prospective, interventional cohort study conducted between July 2004 and May 2006 among 21 754 surgical patients at a Swiss teaching hospital using a crossover design to compare 2 MRSA control strategies (rapid screening on admission plus standard infection control measures vs standard infection control alone). Twelve surgical wards including different surgical specialties were enrolled according to a prespecified agenda, assigned to either the control or intervention group for a 9-month period, then switched over to the other group for a further 9 months. During the rapid screening intervention periods, patients admitted to the intervention wards for more than 24 hours were screened before or on admission by rapid, multiplex polymerase chain reaction. For both intervention (n=10 844) and control (n=10 910) periods, standard infection control measures were used for patients with MRSA in all wards and consisted of contact isolation of MRSA carriers, use of dedicated material (eg, gown, gloves, mask if indicated), adjustment of perioperative antibiotic prophylaxis of MRSA carriers, computerized MRSA alert system, and topical decolonization (nasal mupirocin ointment and chlorhexidine body washing) for 5 days. Incidence of nosocomial MRSA infection, MRSA surgical site infection, and rates of nosocomial acquisition of MRSA. Overall, 10 193 of 10 844 patients (94%) were screened during the intervention periods. Screening identified 515 MRSA-positive patients (5.1%), including 337 previously unknown MRSA carriers. Median time from screening to notification of test results was 22.5 hours (interquartile range, 12.2-28.2 hours). In the intervention periods, 93 patients (1.11 per 1000 patient-days) developed nosocomial MRSA infection compared with 76 in the control periods (0.91 per 1000 patient-days; adjusted incidence rate ratio, 1.20; 95% confidence interval, 0.85-1.69; P = .29). The rate of MRSA surgical site infection and nosocomial MRSA acquisition did not change significantly. Fifty-three of 93 infected patients (57%) in the intervention wards were MRSA-free on admission and developed MRSA infection during hospitalization. A universal, rapid MRSA admission screening strategy did not reduce nosocomial MRSA infection in a surgical department with endemic MRSA prevalence but relatively low rates of MRSA infection. isrctn.org Identifier: ISRCTN06603006.


Appointment systems.
At the heart of any dental practice is the appointment book. There cannot be a practice which does not depend upon appointments, yet many dentists seem to be unaware that they can control their appointments system. Practitioners may adopt an existing system which may no longer be appropriate and receptionists can remain oblivious to the pressures and burdens systems impose upon the dentist. In the third of his articles on practice management, Michael Wilkinson considers the problem.


Calibration-free concentration analysis of protein biomarkers in human serum using surface plasmon resonance.
In complex biological samples such as serum, determination of specific and active concentration of target proteins, independent of a calibration curve, will be valuable in many applications. Calibration-free concentration analysis (CFCA) is a surface plasmon resonance (SPR)-based label-free approach, which calculates active concentration of proteins using their known diffusion coefficient and observed changes in binding rates at different flow rates under diffusion-limited conditions. Here, for the first time we demonstrate the application of CFCA for determining protein biomarker abundance, specifically serum amyloid A (SAA), directly in the serum samples of patients suffering from different infectious and non-infectious diseases. The assay involves preparation of appropriate reaction surfaces by immobilizing antibodies on CM5 chips via amine coupling followed by serum sample preparation and injection over activated and reference surfaces at flow-rates of 5 and 100 μL/min. The system was validated in healthy and diseased (infectious and non-infectious) serum samples by quantifying two different proteins: β2-microglobulin (β2M) and SAA. All concentration assays were performed for nearly 100 serum samples, which showed reliable quantification in unattended runs with high accuracy and sensitivity. The method could detect the serum β2M to as low as 13 ng/mL in 1000-fold serum dilution, indicating the possible utility of this approach to detect low abundance protein biomarkers in body fluids. Applying the CFCA approach, significant difference in serum abundance of SAA was identified in diseased subjects as compared to the healthy controls, which correlated well with our previous proteomic investigations. Estimation of SAA concentration for a subset of healthy and diseased sera was also performed using ELISA, and the trend was observed to be similar in both SPR assay and ELISA. The reproducibility of CFCA in various serum samples made the interpretation of assay simple and reliable. This study illustrates a significant step forward in rapid monitoring of several protein markers in serum samples, with utility in biomarker validation and other therapeutic applications.


Non-synonymous SNPs variants of PRKCG and its association with oncogenes predispose to hepatocellular carcinoma.
PRKCG encodes PKC γ, which is categorized under the classical protein kinase C family. No studies have specifically established the relationship between PRKCG nsSNPs with structural and functional variations in PKC γ in the context of hepatocellular carcinoma (HCC). The present study aims to uncover this link through in-silico and experimental studies. The 3D structure of PKC γ was predicted. Molecular Dynamic (MD) Simulations were run and estimates were made for interactions, stability, conservation and post-translational alterations between wild and mutant structures. The association of PRKCG levels with HCC survival rate was determined. Genotyping analyses were conducted to investigate the deleterious PRKCG nsSNP association with HCC. mRNA expression of PKC γ, HIF-1 alpha, AKT, SOCS3 and VEGF in the blood of controls and HCC patients was analyzed and a genetic cascade was constructed depicting these interactions. The expression level of studied oncogenes was compared to tumour suppressor genes. Through Alphafold, the 3D structure of PKC γ was explored. Fifteen SNPs were narrowed down for in-silico analyses that were identified in exons 5, 10 and 18 and the regulatory and kinase domain of PKC γ. Root mean square deviation and fluctuation along with the radius of gyration unveiled potential changes between the wild and mutated variant structures. Mutant genotype AA (homozygous) corresponding to nsSNP, rs386134171 had more frequency in patients with OR (2.446), RR (1.564) and P-values (< 0.0029) that highlights its significant association with HCC compared to controls in which the wild genotype GG was found more prevalent. nsSNP rs386134171 can be a genetic marker for HCC diagnosis and therapeutic studies. This study has laid down a road map for future studies to be conducted on HCC.


Further evidence for the reliability and validity of the Fatigue and Daytime Sleepiness Scale.
Myotonic dystrophy type 1 (DM1) is an inherited neuromuscular disease causing, among other symptoms, fatigue and excessive daytime sleepiness, which are frequently undifferentiated by patients and/or clinicians. The Fatigue and Daytime Sleepiness Scale (FDSS) has been devised to measure these two overlapping symptoms as a single clinical entity. To further examine the reliability and the construct validity of the FDSS in patients with DM1. The scale was administered to 48 DM1 patients on two occasions at a 2week-interval. Intra-rater reliability and internal consistency were established by calculating the intraclass correlation coefficient (ICC) and Cronbach's alpha, respectively. Construct validity was assessed by using the known-group method. More precisely, the mean FDSS score of patients with and without subjective complaints of fatigue and/or sleepiness was compared. The FDSS showed good intra-rater reliability (ICC=0.83) and acceptable internal consistency (Cronbach's alpha =0.6). Also, the FDSS was able to distinguish between patients who had fatigue and sleepiness complaints from those who did not (mean FDSS score of 10.6 vs 8.0, p=0.01), suggesting good construct validity. Overall, the present study supports the continued use of the FDSS as a reliable and valid instrument to measure fatigue and daytime sleepiness in patients with DM1 for either clinical or research purposes.


Aquaporins in salivary glands and pancreas.
Salivary glands and pancreas are involved in saliva secretion, pancreatic fluid secretion and insulin secretion. These functions are essential for proper oral, pancreatic and glucose homeostasis. Aquaporins are water-permeable transmembrane protein involved in the physiology of these secretory gland functions. This review gives an overview of the morphology of salivary glands and pancreas, the expression and localization of aquaporins, the secretion roles and mechanisms, the physiological roles of aquaporins, and the role of aquaporins in pathophysiological conditions. Several aquaporins are expressed in salivary glands and pancreas, and some play important physiological roles. Modulation of aquaporin expression and/or trafficking may contribute to the pathogenesis of diseases affecting salivary glands and pancreas glands such as xerostomic conditions, pancreatic insufficiencies and diabetes. Aquaporins are involved in physiological and pathophysiological processes in salivary glands and pancreas. They could represent therapeutic targets for the treatment of diseases affecting the salivary glands and pancreas. This article is part of a Special Issue entitled Aquaporins.


Emotional SNARC: emotional faces affect the impact of number magnitude on gaze patterns.
The Spatial-Numerical Association of Response Codes (SNARC) effect is characterised by a spatial cognitive representation of low numbers to the left side of space and high numbers to the right side of space. This effect has been found using a diversity of stimuli and experimental paradigms. However, the influence of emotional stimuli on this effect remains unclear. In this study, the SNARC effect is analysed in relation to pairs of emotional facial stimuli (happy-neutral, sad-neutral and happy-sad pairs). Gaze patterns of 151 participants were analysed when exposed to a free-viewing eye-tracking paradigm consisting of pairs of emotional faces preceded by small and large numbers. Replicating previous results, a standard SNARC effect was found independently of the emotional expressions of the faces (i.e., there was a significant linear trend of number magnitude in the frequency of first fixations of the gaze to the left side space). However, specific slope analyses revealed that the SNARC effect was influenced by the spatial position of each emotion presented in the emotional pairs. Specifically, the effect disappeared in happy-neutral trials, when the happy faces were allocated in the right position and also in happy-sad trials when two emotional stimuli were simultaneously displayed. The study revealed that the SNARC effect is sensitive to the spatial position of emotional stimuli which further adds to other known limits of the phenomenon. The limitations of the study and its implications in the area of cognition and emotion are also discussed.


Parabens in urine of children and adolescents in Germany - human biomonitoring results of the german environmental survey 2014-2017 (GerES V).
Parabens are antimicrobial preservatives used in a wide range of consumer products such as personal care products, cosmetics, pharmaceuticals, and food. Consequently, the general population is ubiquitously exposed to these substances via dermal absorption, ingestion, and inhalation. Parabens promote estrogenic activity and are hence under assessment as endocrine disrupting substances. Urine samples from 3- to 17-year-old children and adolescents (N = 516) living in Germany were analysed for concentrations of nine parabens in the population representative German Environmental Survey for Children and Adolescents 2014-2017 (GerES V). Detection rates and urinary concentrations of the parabens decreased with increasing length of the alkyl chain. Methyl paraben was quantified in 97% of the samples with a geometric mean (GM) concentration of 7.724 μg/L (6.714 μg/gcreatinine), ethyl paraben was quantified in 69% (GM: 0.943 μg/L and 0.825 μg/gcrea), and n-propyl paraben in 31% (GM: 0.563 μg/L and 0.493 μg/gcrea). Concentrations of iso-propyl paraben, butyl paraben, iso-butyl paraben, and benzyl paraben were below the limit of quantification in most samples. Pentyl paraben and heptyl paraben were not detected in any of the samples. Paraben concentrations in urine were found to be associated with frequent usage of leave-on personal care products and cosmetics. Cumulative exposure to parabens (back-calculated daily intakes, expressed as hazard index) was found to be on a level raising concern in up to 14% of the population, mainly driven by n-propyl paraben, and depending on the level of conservativeness and point-of departures used for calculation.


Premature ovarian insufficiency (POI) is defined as loss of ovarian function in women less-than 40 years. This review summarises the causes and the possible treatment options. POI can be idiopathic, caused by genetic, autoimmune, or metabolic disease, or be induced by cancer therapy or surgery. POI causes infertility, increased morbidity and mortality, and decreased quality of life. Hormonal replacement therapy (HRT) can alleviate symptoms of POI and should be initiated at diagnosis. The benefit of HRT outweighs the minor side effects in most cases and should be continued until age of natural menopause.