The role of chemokines in acute and chronic hepatitis C infection.
Hepatitis C imposes a significant burden on global healthcare. Chronic infection is associated with progressive inflammation of the liver which typically manifests in cirrhosis, organ failure and cancer. By virtue of elaborate evasion strategies, hepatitis C virus (HCV) succeeds as a persistent human virus. It has an extraordinary capacity to subvert the immune response enabling it to establish chronic infections and associated liver disease. Chemokines are low molecular weight chemotactic peptides that mediate the recruitment of inflammatory cells into tissues and back into the lymphatics and peripheral blood. Thus, they are central to the temporal and spatial distribution of effector and regulatory immune cells. The interactions between chemokines and their cognate receptors help shape the immune response and therefore, have a major influence on the outcome of infection. However, chemokines represent a target for modulation by viruses including the HCV. HCV is known to modulate chemokine expression in vitro and may therefore enable its survival by subverting the immune response in vivo through altered leukocyte chemotaxis resulting in impaired viral clearance and the establishment of chronic low-grade inflammation. In this review, the roles of chemokines in acute and chronic HCV infection are described with a particular emphasis placed on chemokine modulation as a means of immune subversion. We provide an in depth discussion of the part played by chemokines in mediating hepatic fibrosis while addressing the potential applications for these chemoattractants in prognostic medicine.


Outcomes of free vascularized fibular flaps for the treatment of radiation-associated femoral nonunions.
Nonunion is a known complication following fracture in the setting of radiotherapy. Free vascularized fibular (FVF) flaps have been used successfully in the treatment of segmental bone defects; however, their efficacy in the treatment of radiated nonunions is limited. The purpose of the study was to evaluate the outcome following FVFG for radiation-associated femoral fracture nonunions. 23 (11 male and 12 female; mean age 60 ± 12 years) patients underwent FVF for radiation-associated femoral fracture nonunions. The most common indication for radiotherapy was soft tissue sarcomas (n = 16). The mean follow-up was 5 ± 4 years. Mean radiation dose was 51 ± 14 Gy at a mean of 11 ± 3 years prior to FVF. The mean FVF length was 17 ± 4 cm and placed commonly with an intramedullary nail (n = 18). First time union was 52% (n = 12) following additional bone grafting, the overall union was 78% (n = 18) at a mean of 13 ± 6 months. Musculoskeletal Tumor Society scores improved from 30% preoperatively to 73% at latest follow-up (p < 0.0001). Five fractures failed to unite; 3 were converted to proximal femoral replacements. FVF are a reasonable treatment option for radiation-associated femoral fracture nonunions, providing a union rate of 78% and an improvement in functional outcome. Therapeutic Level IV.


Long-term ecology of euglossine orchid-bees (Apidae: Euglossini) in Panama.
Abundance patterns during 6-7 years and orchid visitation were determined for 51 species of the 57 local euglossine bees. Male bees were counted at 3 chemical attractants presented in the same manner each month. Sites were separated by 75 km but included wet Atlantic forest at 500 m elevation, moist forest at 180 m near Barro Colorado Island, and cloud forest at 900 m near the Pacific ocean. 1. From 15 to 30 euglossine species of 4 genera were active in each month and site; monthly species number and general bee abundance were positively correlated. Many species had 3 annual abundance peaks (range 1-4) and were active throughout the year, but peak annual abundances rarely occurred during late wet or early dry seasons. In contrast, Eufriesea generally were present as adults only 1-2 months in a year. 2. Euglossine populations were exceptionally stable. Species at each site were more stable than any known insect population, and stability and abundance were positively associated. However, year-to-year population stability and the degree of seasonality were not correlated. Among the three sites, the more diverse (species rich) bee assemblages displayed lower stability; these were the wetter and upland sites. 3. The most abundant bees visited more orchid species. Eg. and El. each visited and average of 4 orchid species (range 0-13); Ex. and Ef. visited 0-3. Stable populations did not visit more or fewer orchid species than did unstable populations. 4. Less than 68% of species at each site visited orchid flowers; less than a few dozen of the 100-800 bees counted in a day carried orchid pollinaria. Over 20% of the euglossine species never were seen with pollinaria at any site and probably seldom visit orchids in central Panama. 5. Most bee species visited 1 or no fragrance orchids in a given habitat. Orchids tended to utilize common pollinators that seldom included more than 1 species, and they utilized stable or unstable, seasonal or aseasonal bees. However, the most stable and abundant bee, Eg. imperialis, rarely pollinated orchids; fewer than 10 of ca. 20000 bees carried pollinaria. 6. Orchids may interact primarily with discrete seasonal bee population peaks-probably the emerging adults. Although specialized orchid preferences are implicated for species that visit few or no local orchids but pollinate other species and carry pollinaria in other areas, euglossine bees do not need orchids to survive or reproduce.


Effect of retinoids on carcinogen-induced mutagenesis in Salmonella tester strains.
The ability of retinol (Rol) in altering mutation frequencies induced by 7 carcinogens was studied in Salmonella/microsome assay using 4 tester strains namely TA98, TA100, TA102 and TA1535. The 7 carcinogens used were aflatoxin B1 (AFB), cyclophosphamide (CPP), 3-methylcholanthrene (MCA), benzo[a]pyrene (BP), benz[a]anthracene (BA), 9,10-dimethyl-1,2-benz[a]anthracene (DMBA) and mitomycin C (MMC). As reported previously, Rol significantly reduced the number of His+ revertants induced by AFB. It also reduced mutations induced by CPP or MCA but not that by BP, BA, DMBA or MMC. The abilities of Rol, retinoic acid, retinyl acetate and a known inhibitor for certain P-450 isozymes, 7,8-benzoflavone (BF) in inhibiting mutations caused by AFB and BP were studied and compared. All the 3 retinoids caused significant reduction of AFB-induced His+ revertants in a dose-dependent manner, but there was no effect on BP-induced mutation. BF strongly inhibited both AFB- and BP-induced revertants. The possibility of retinoids in exerting their effects on mutagenesis of precarcinogens by inhibiting only certain forms of cytochrome P-450 enzymes is discussed.


Safety and impact on diagnostic accuracy of early analgesia in suspected acute appendicitis: a meta-analysis.
The safety of early analgesia in patients suspected to have acute appendicitis (AA) is still controversial. Double blind randomized clinical trials comparing patients receiving or not receiving opiates for early analgesia in suspected AA were selected for meta-analysis according to PRISMA guidelines. Primary outcomes were the number of patients with AA confirmed by histology and the number of patients undergoing surgical intervention. Secondary outcomes were missed diagnoses, false positive AA and complication rate. Effect sizes were calculated using a Mantel-Haenszel fixed effects model. Previously published papers mostly analyzed surrogate end-points such as physician's confidence about the diagnosis or the alteration of clinical signs, subjective parameters dependent on personal perception. Our article focused on clinical outcome and specifically investigated those potentially related to AA instead of unspecified abdominal pain. Opiate administration did not have an impact on the number of histologically proven AA (OR = 1.196 [0.875-1.635]; P = 0.261). Differences in appendectomy rates were only slightly above the threshold for statistical significance (OR = 1.350 [0.966-1.887]; P = 0.079), suggesting that analgesia might influence the treatment approach. On the other hand missed diagnoses (OR = 0.509 [0.087-2.990]; P = 0.455) and false positive AA (OR = 1.071 [0.596-1.923]; P = 0.818) ascertained by histologic examination were unaffected, so diagnostic accuracy was retained. Safety was not compromised by opiates, as the difference in complication rates did not reach statistical significance (OR = 0.615 [0.217-1.748]; P = 0.372). Early analgesia with opiates in suspected AA might influence the approach to treatment, but does not appear to alter diagnostic accuracy or surgical outcome. To support our findings, further trials on larger sample sizes from different age groups and both genders are needed.


The acute and chronic effects of monosodium L-glutamate on serum iron and total iron-binding capacity in the jugular artery and vein of pigs.
We analyzed the effects of acute and chronic oral administration of monosodium L-glutamate (MSG) on serum iron (Fe) levels and total iron-binding capacity (TIBC) in piglets. In the first experiment, 12 piglets were randomly assigned to two groups: one fed a standard diet (SD) and the other fed an SD containing MSG (10 g/kg). On day 30, serum, liver, kidney, and spleen samples were collected to determine the Fe levels. In the second experiment, six pigs were surgically fitted with a catheter in the jugular artery and vein to investigate the dynamic changes of serum Fe and TIBC. Blood samples were taken from each pig via the catheter every 30 min, for a period of 4 h. The results show that MSG increases Fe levels in the spleen (P < 0.05) and in serum obtained from the jugular artery (P < 0.01). In addition, TIBC in serum obtained from the jugular artery demonstrated an increasing trend in pigs fed the MSG diet; however, this trend was not observed in the jugular vein. In conclusion, MSG increases Fe retention by enhancing TIBC in serum.


Cobalt destroys neurons without destroying fibers of passage in the lateral geniculate nucleus of the cat.
When several injections of 4 mM cobalt chloride are made into the lateral geniculate nucleus of the cat, cell somata are destroyed while fibers-of-passage are spared. Nissl and Wiel stains were used to assess the extent of cell and fiber damage, respectively, for cobalt lesions of layer A. Retrograde transport of horseradish peroxidase from the visual cortex to geniculate cells surrounding the lesions was used to demonstrate the integrity of fibers passing through the area depleted of cells. Cobalt provides a versatile and precise method of disrupting neural pathways. Depending upon the number and timing of the injections, the disruption can be temporary, resulting from a reversible inactivation of synaptic transmission, or permanent, caused by soma death.


Loss of type II calcium/calmodulin-dependent kinase activity correlates with stages of development of electrographic seizures in status epilepticus in rat.
Understanding the molecular basis of altered neuronal excitability in epilepsy is a major challenge in neuroscience research. The present study suggests an inverse correlation between changes in neuronal excitability in status epilepticus and the activity of type II multifunctional calcium/calmodulin-dependent kinase II (CaM kinase II), a major Ca(2+)-signal transducing system in brain. 'Continuous' hippocampal stimulation (CHS), a new model of non-convulsive limbic status epilepticus (SE), mimics the progression of electrographic changes characteristic in human SE and allows for quantitation of post-stimulus seizure severity. In the present study, hippocampus and anterior neocortex from CHS-stimulated rats and paired surgical controls were assayed for CaM kinase II activity by incorporation of radiolabeled phosphate from [gamma-32P]ATP into the 50-kDa subunit of the kinase itself (autophosphorylation). In all instances, CHS induced sustained interictal bursting and/or electrographic seizures. Decreased CaM kinase II activity was seen in all preparations from electrically stimulated hippocampus. CaM kinase II activity in CHS animals was diminished by 37% relative to controls (P less than 0.01; Student's paired t-test). The progressive intensity of the EEG discharges correlated directly with the decrement of CaM kinase II activity (P less than 0.05; Spearman's rank correlation test, n = 5). This is the first report of a dynamic modulation of a biochemical system that has been implicated in neuronal excitability in coordination with the characterized developmental stages of SE.


Demethylation of the type I procollagen genes in transformed fibroblasts treated with 5-azacytidine.
Transformation of the human embryonic lung fibroblast line, WI-38, with simian virus 40 (SV40) results in inactivation of the type I procollagen genes. No type I collagen or procollagen mRNA is detected in these transformed cells, as determined by polyacrylamide gel electrophoresis. Analysis of the methylation patterns of these genes showed the type I procollagen genes to be hypermethylated at certain cytosine residues in the transformed cells. However, several of the cytosine residues were methylated in the normal cells where these genes are expressed. These methylation patterns can be altered by treatment of the cells with 5-azacytidine or 5-azadeoxycytidine, but without a resultant activation of the type I procollagen genes. These results show that demethylation alone is not sufficient for gene activation, but that other signals are also required.


[Risk of injury to children's fingers in power-operated motor vehicle windows].
This study examines the dimensions of children's fingers and the risk of jam injuries in a 4-mm gap between glass and gasket of power-operated motor vehicle windows. The diameter of the proximal, middle, and distal phalanx and of the proximal and distal interphalangeal joint of each finger of the right hand of 160 children was measured in a cross-sectional investigation. Six different drawings in cross section of gaskets and glass window panes of current motor vehicle side door windows at a vertical gap of 4 mm were drawn in correct proportion. The larger actual width of the oblique gap between window glass and gasket was measured and related to the diameters of children's fingers. Almost all fingers and joints fit in the largest actual gap of 18 mm between glass and gasket of one seal design. The European guideline 74/60/EWG specifications currently pertaining to closing force restriction do not eliminate the risk of potentially serious injury to children's fingers in motor vehicle power windows.


Increasing risk of great floods in a changing climate.
Radiative effects of anthropogenic changes in atmospheric composition are expected to cause climate changes, in particular an intensification of the global water cycle with a consequent increase in flood risk. But the detection of anthropogenically forced changes in flooding is difficult because of the substantial natural variability; the dependence of streamflow trends on flow regime further complicates the issue. Here we investigate the changes in risk of great floods--that is, floods with discharges exceeding 100-year levels from basins larger than 200,000 km(2)--using both streamflow measurements and numerical simulations of the anthropogenic climate change associated with greenhouse gases and direct radiative effects of sulphate aerosols. We find that the frequency of great floods increased substantially during the twentieth century. The recent emergence of a statistically significant positive trend in risk of great floods is consistent with results from the climate model, and the model suggests that the trend will continue.


Phenotyping of proliferating cells in cultures of human lymphocytes.
Lymphocytes stimulated with mitogens, alloantigens or soluble antigens were pulsed with tritiated thymidine to label proliferating cells. Aliquots of the cells were labeled with OKT 4 or OKT 8 mouse monoclonal antibodies and then incubated in microtiter wells coated with goat anti-mouse immunoglobulin G (IgG). Unattached cells were flushed out of the wells: individual wells were then separated for counting in scintillation fluid in a B counter. This method permits simple characterization of the relative T cell subset proliferative responses and is clinically applicable.


Stimulation of Alexandrium fundyense growth by bacterial assemblages from the Bay of Fundy.
To evaluate the influence of marine bacterial assemblages from the Bay of Fundy on Alexandrium fundyense str. CB301 growth. Bacterial assemblages were collected from the Bay of Fundy during an Alexandrium spp. bloom, serially diluted to extinction and inoculated into axenic CB301 cultures. Bacterial assemblages dramatically enhanced CB301 growth. Retrieval and analysis of 16S rDNA fragments revealed an Alteromonas sp. strain to be the only detectable bacterium in all assemblages that promoted A. fundyense and the sole bacterium found in the most dilute inoculum that promoted A. fundyense. While this bacterium has not yet been isolated, other isolates obtained from the assemblages did not stimulate A. fundyense, indicating that the observed stimulation was not a general effect of marine bacteria. Bay of Fundy marine bacterial assemblages dominated by a member of the family Alteromonadaceae were found to dramatically stimulate growth of A. fundyense. These results show that native bacteria have the potential to dramatically promote the growth of A. fundyense and may play an important role in influencing A. fundyense dynamics in the Bay of Fundy.


Mesothelial Cells Covering the Surface of Primo Vascular System Tissue.
The primo vascular system (PVS) is reported to have a periductium composed of cells with spherical or spindle-shaped nuclei and abundant cytoplasm. However, little is known about these periductium cells. In this study, we examined the morphological features of cells covering the PVS tissue isolated from the surface of abdominal organs of rats. By hematoxylin and eosin (H&E) staining, we observed a layer of dark nuclei on the basement membrane at the borders of the sections of primo node (PN), primo vessel (PV), and their subunits. The nuclei appeared thin and linear (10-14 μm), elliptical (8-10 × 3-4 μm), and round (5-7 μm). The borders of the PVS tissue sections were immunostained with a selective antibody for mesothelial cells (MCs). Areas of immunoreactivity overlapped with the flattened cells are shown by hematoxylin and eosin staining. By scanning electron microscopy, we further identified elliptical (11 × 21 μm) and rectangular squamous MCs (length, 10 μm). There were numerous stomata (∼200 nm) and microparticles (20-200 nm) on the surface of the PVS MCs. In conclusion, this study presents the novel finding that the PVS periductium is composed of squamous MCs. These cells tightly line the luminal surface of the PVS tissue, including PNs, PVs, and small branches of the PVs in the abdominal cavity. These results will help us to understand the physiological roles such as hyaluronan secretion and the fine structure of PVS tissue.


Herbarium specimens reveal century-long trait shifts in poison ivy due to anthropogenic CO2 emissions.
Previous experimental studies have shown that poison ivy (Toxicodendron radicans; Anacardicaceae) responds to elevated CO2 with increased leaf production, water-use efficiency, and toxicity (allergenic urushiol). However, long-term field data suggest no increase in poison ivy abundance over time. Using herbarium specimens, we examined whether poison ivy and other species shifted leaf traits under natural conditions with increasing atmospheric CO2 (pCO2 ) over the past century. We measured stomatal density, leaf area, leaf N, leaf C:N, leaf carbon isotope discrimination (Δleaf ), and intrinsic water-use efficiency (iWUE) from 327 specimens collected from 1838 to 2020 across Pennsylvania. We compared poison ivy's responses to two evolutionarily related tree species, Toxicodendron vernix and Rhus typhina (Anacardiacae) and one ecological analog, Parthenocissus quinquefolia (Vitaceae), a common co-occurring liana. Stomatal density significantly decreased (P < 0.05) in poison ivy and the ecologically similar liana P. quinquefolia over the past century, but did not change in the related trees T. vernix and R. typhina. None of these species showed significant trends in changes in leaf N or C:N. Surprisingly, in poison ivy, but not the other species, Δleaf increased with increased pCO2 , corresponding to significant declines in iWUE over time. In contrast to the results of short-term experimental studies, iWUE decreased in poison ivy over the last century. Trait responses to pCO2 varied by species. Herbarium specimens suggest that realized long-term plant physiological responses to increased CO2 may not be reflected in short-term experimental growth studies, highlighting the value of collections.


Expression of the antioxidant stress protein heme oxygenase-1 (HO-1) in human leukocytes.
Inducible heme oxygenase (HO-1) is an antioxidant stress protein, that is mainly induced by reactive oxygen species (ROS), cytokines and hyperthermia. By using flow cytometry the present investigation demonstrated a rise in the cytoplasmic expression of HO-1 in lympho- (L), mono- (M) and granulocytes (G) of 9 endurance-trained male subjects after a half marathon run. The expression was more pronounced in M (median: 98.3% HO-1 positive cells/4.31 mfc) and G (94.8%/1.93 mfc) than in L (80.1%/1.51 mfc) when measured 3 h post-exercise. Additionally the exercise protocol caused a rise in the plasma levels of myeloperoxidase, TNF alpha and interleukin-8 (IL-8), indicating an inflammatory response. We could detect a correlation between IL-8 and HO-1, directly after exercise, that was apparent in G (r = 0.67, p < .05) and L (r = 0.80, p < .05), but did not reach significance in M (r = 0.65, p = 0.06). An additional detection of HO-1 at rest in 12 untrained subjects showed a higher baseline expression of HO-1 compared to the athletes. The regulatory pathways leading to an increased expression of HO-1 after endurance exercise are not completely clear, but a causal involvement of a cytokine-mediated generation of ROS must be discussed. We supposed that the down-regulation of the baseline expression of HO-1 in athletes reflects an adaptional mechanism to regular exercise training.


Structural elucidation studies of 15-membered azalide macrocycles using H/D exchange and ESI-MS(n.).
A comprehensive study of fragmentation pathways of 15-membered azalide macrocycles using electrospray ionisation with multistage mass spectrometry (ESI-MS(n)) is presented in this work. Hydrogen/deuterium (H/D) exchange experiments and high-resolution mass spectrometry were used to investigate the proposed fragmentation pathways. In addition, the fragmentation patterns of sodium adduct ions [M+Na](+) of the macrocycles were also investigated as the presence of an alkali metal interacting with the aglycone ring influences the product-ion spectra.


Integrated analysis of fibroblasts molecular features in papillary thyroid cancer combining single-cell and bulk RNA sequencing technology.
Papillary thyroid cancer (PTC) is the most common pathological type of thyroid cancer with a high incidence globally. Increasing evidence reported that fibroblasts infiltration in cancer was correlated with prognostic outcomes. However, fibroblasts related study in thyroid cancer remains deficient. Single-cell sequencing data of PTC were analyzed by Seurat R package to explore the ecosystem in PTC and identify fibroblasts cluster. The expression profiles and prognostic values of fibroblast related genes were assessed in TCGA dataset. A fibrosis score model was established for prognosis prediction in thyroid cancer patients. Differentially expressed genes and functional enrichment between high and low fibrosis score groups in TCGA dataset were screened. The correlation of immune cells infiltration and fibrosis score in thyroid cancer patients was explored. Expression levels and prognostic values of key fibroblast related factor were validated in clinical tissues another PTC cohort. Fibroblasts were highly infiltrated in PTC and could interact with other type of cells by single-cell data analysis. 34 fibroblast related terms were differentially expressed in thyroid tumor tissues. COX regression analysis suggested that the constructed fibrosis score model was an independent prognostic predictor for thyroid cancer patients (HR = 5.17, 95%CI 2.31-11.56, P = 6.36E-05). Patients with low fibrosis scores were associated with a significantly better overall survival (OS) than those with high fibrosis scores in TCGA dataset (P = 7.659E-04). Specific immune cells infiltration levels were positively correlated with fibrosis score, including monocytes, M1 macrophages and eosinophils. Our research demonstrated a comprehensive horizon of fibroblasts features in thyroid cancer microenvironment, which may provide potential value for thyroid cancer treatment.


Development of humanized mice as a model of inflammatory arthritis.
Clinically, rheumatoid arthritis (RA) is a chronic deforming disease characterized mainly by joint swelling and destruction. Although synovial inflammation and bone erosion are the hallmarks of this disease, the presentation of various features between patients is clearly heterogenous, suggesting that there are different variants of RA. Hence, the development of an animal model that has all of the elements of human RA has remained elusive. This review explores several different views on the etiology of RA and the recent data from various murine arthritis models which provide support for these theories. In addition to discussing the potential roles of CD4(+) T cell activation, autoantibodies, and lymphocyte-independent cytokine production, the role of CD4(+) T regulatory cells will be presented in the context of a newly developed humanized transgenic mouse model. This novel T cell receptor transgenic model is being characterized to enhance our understanding of the mechanisms involved in the breach of self-tolerance that occurs in autoimmune disorders such as RA.


Efficacy and safety of atorvastatin in children and adolescents with familial hypercholesterolemia or severe hyperlipidemia: a multicenter, randomized, placebo-controlled trial.
To determine the safety and efficacy of atorvastatin (10 to 20 mg) in children and adolescents with familial hypercholesterolemia or severe hypercholesterolemia. Subjects (n=187) were randomly assigned to 26 weeks of treatment with atorvastatin (10 mg) or placebo. Dosage was increased to 20 mg if LDL cholesterol (LDL-C) levels remained >3.4 mmol/L (130 mg/dL) at week 4. At week 26, subjects received 10 mg of atorvastatin for an additional 26 weeks. Efficacy variables included percent changes in LDL-C, total cholesterol, triglycerides, HDL cholesterol, and apolipoprotein B from baseline to week 26. Atorvastatin caused a highly significant reduction in LDL-C compared with placebo (-40% vs -0.4%, respectively; P<.001). Percent changes at week 26 also significantly favored atorvastatin for total cholesterol (-32% vs -1.5%; P<.001), triglycerides (-12% vs +1.0%; P=0.03), and apolipoprotein B (-34% vs +0.7%; P<.001), with a significantly greater increase in HDL cholesterol with atorvastatin compared with placebo (+2.8% vs -1.8%; P=.02). Atorvastatin was as well-tolerated as placebo. Treatment with atorvastatin for 12 months was effective and safe for pediatric subjects with known familial hypercholesterolemia or severe hypercholesterolemia.


Strong fish gelatin hydrogels double crosslinked by transglutaminase and carrageenan.
Gelatin hydrogels are usually soft and thermally unstable. Here, strong fish gelatin hydrogels were successfully prepared by double crosslinking gelatin with transglutaminase (TGase) and κ-carrageenan, and the mechanical properties and thermal stability of the double crosslinked gelatin hydrogels were significantly improved. Results showed that the gel strength, compression fracture stress and storage modulus of the double crosslinked gelatin hydrogels all reached the largest value when the concentration of TGase was 20 U/g gelatin. The double crosslinked gelatin hydrogels were also thermally stable due to the existence of the covalent crosslinks. The effect of TGase concentration on the physical properties and microstructure of the double crosslinked hydrogels were analyzed, and the differences between double crosslinked gelatin hydrogels and gelatin hydrogels single crosslinked by TGase or carrageenan were also systematically compared. This article is of great significance for expanding the application of natural polymer-based hydrogels.


Early identification of the critical view of safety in laparoscopic cholecystectomy using indocyanine green fluorescence cholangiography: A randomised controlled study.
Achieving critical view of safety (CVS) is vital during laparoscopic cholecystectomy (LC). There is no known study determining use of indocyanine green fluorescence cholangiography (ICGFC) in early identification of CVS during LC. This study aims to compare use of ICGFC in LC against conventional LC in early identification of CVS. Patients undergoing LC in a single centre were randomized into ICGFC-LC and conventional LC. Surgery was performed by a single surgeon and the time taken to achieve CVS from the time of gallbladder fundus retraction was measured. Difficulty level for each surgery was rated and analysed using a modified scoring system (Level 1- Easy to Level 4-Very difficult). 63 patients were recruited where mean time (min) to achieve CVS was 22.3 ± 12.9 in ICGFC-LC (n = 30) and 22.8 ± 14.3 in conventional LC (p = 0.867). The time taken to achieve CVS was shorter in ICGFC-LC group across all difficulty levels, although not significant (p > 0.05). No major complication was observed in the study. This study had shown ICGFC-LC reduces time to CVS across all difficulty levels but not statistically significant. ICGFC-LC maybe useful in difficult LC and in surgical training. Clinical Trials NCT04228835. UMMI Surgical - Karl Storz Distributor (Malaysia).


Involvement of low-calcium diet in the reduced bone mineral content of idiopathic renal stone formers.
The possibility that low-calcium intake in renal stone formers could lead to reduced bone mineral content was investigated in 123 male patients with idiopathic urolithiasis. Radius bone mineral content (BMC) was measured by single photon absorptiometry. Two groups of patients were analyzed: group 1 (n = 63) maintained on a free diet; group 2 (n = 60) maintained on a low-calcium diet (350 mg/day +/- 20 SEM) for 3.9 years +/- 0.6 SEM. The two groups of patients were investigated after a standard reduction of calcium intake for at least 1 week. The urinary excretion of calcium and of hydroxyproline, and the serum alkaline phosphatase activity were higher in both groups than in normal subjects submitted to the same low-calcium diet. Both groups of stone formers showed lowered radius BMC values at 3 cm (distal) and 8 cm (proximal) above the styloid process, but distal BMC was significantly lower in group 2 than in group 1. The results suggest that low-calcium intake could worsen the already decreased BMC of idiopathic renal stone formers.


Diagnostic performance of duplex ultrasound in patients suspected of carotid artery disease: the ipsilateral versus contralateral artery.
To evaluate duplex ultrasonographic thresholds for the determination of 70% to 99% stenosis of the ipsilateral and contralateral internal carotid artery in patients with symptoms of amaurosis fugax, transient ischemic attack (TIA), or minor stroke based on 2 criteria: maximizing accuracy and optimizing cost-effectiveness and to compare these with current recommendations. From January 1997 to January 2000, a prospective multicenter study was conducted including 350 consecutive patients with symptoms of amaurosis fugax, TIA, or minor stroke who underwent bilateral duplex ultrasonography and digital subtraction angiography. A linear regression analysis was performed to estimate the degree of angiographic stenosis as a function of the peak systolic velocity (PSV). PSV thresholds were calculated for the ipsilateral and contralateral carotid arteries based on maximizing accuracy and optimizing cost-effectiveness. The PSV measurements significantly overestimated the angiographic stenosis in the contralateral artery (9.5%; 95% CI, 6.3% to 12.7%) compared with the ipsilateral carotid artery. The recommended PSV threshold for the diagnosis of 70% to 99% stenosis is 230 cm/s. Maximizing accuracy, the optimal PSV threshold for the ipsilateral artery was 280 cm/s, and for the contralateral artery, 370 cm/s for diagnosing a 70% to 99% stenosis. Optimizing cost-effectiveness, the optimal PSV threshold was 220 cm/s for ipsilateral and 290 cm/s for contralateral carotid arteries. PSV measurements overestimate the degree of angiographic stenosis in the contralateral carotid artery in patients with symptoms of amaurosis fugax, TIA, or minor stroke. Separate PSV thresholds should be used for the ipsilateral and contralateral carotid artery. PSV thresholds that optimize cost-effectiveness differ from the recommended thresholds and from thresholds that maximize accuracy.


Caveolin: a possible biomarker of degradable metallic materials toxicity in vascular cells.
Iron-based materials could constitute an interesting option for cardiovascular biodegradable stent applications due to their appropriate ductility compared with their counterparts, magnesium alloys. However, the predicted degradation rate of pure iron is considered to be too slow for such applications. We explored manganese (35 wt.%) as an alloying element in combination with iron to circumvent this problem through powder metallurgical processing (Fe-35Mn). Manganese, on the other hand, is highly cytotoxic. We recently explored a new method to better characterize the safety of degradable metallic materials (DMMs) by establishing the gene expression profile (GEP) of cells (mouse 3T3 fibroblasts) exposed to Fe-35Mn degradation products in order to better understand their global response to a potentially cytotoxic DMM. We identified a number of up- and down-regulated genes and confirmed the regulation of a subset of them by quantitative real time polymerase chain reaction. Caveolin-1 (cav1), the structural protein of caveolae, small, smooth plasma membrane invaginations present in various differentiated cell types, was one of the most down-regulated genes in our GEPs. In the present study we further studied the potential of this 22 kDa protein to become a biomarker for cytotoxicity after exposure to degradable metallic elements. In order to better characterize cav1 expression in this context 3T3 mouse fibroblasts were exposed to either ferrous and manganese ions at cytostatic concentrations for 24 or 48 h. cav1 gene expression was not influenced by exposure to ferrous ions. On the other hand, exposure to manganese for 24h reduced cav1 gene expression by about 30% and by >65% after 48 h compared with control 3T3 cells. The cav1 cellular protein content was reduced to the same extent. The same pattern of expression of cav3 (the muscle-specific caveolin subtype) was also observed in this study. This strong and reproducible pattern of regulation of caveolins thus indicates potential as a biomarker for the toxicity of DMM elements.


A Method for Retrieval of Adverse Event Terms in Clinical Trial Databases Using Standardised MedDRA Queries.
Standardized MedDRA Queries (SMQs) are sets of terms determined by experts that are used to identify adverse events (AEs) related to different disease processes. Their use can be challenging because most SMQs have 50 to 100 preferred terms and AE databases can have many thousands of events. The aim of this study is to develop a technique where AEs corresponding to preferred terms in SMQs may be easily detected. The method I developed uses the Table Join function of the JMP® software program to quickly and easily probe clinical trial AE databases. The SMQ Severe cutaneous adverse reactions was used as a probe in a mock AE dataset. Potentially confounding demographic or study-specific factors were evaluated by combining these datasets with the dataset containing the AEs identified with the SMQs. AEs were successfully detected in an AE database using the method described. Cases with potential confounding factors, such as concomitant medications, were identified. The method developed allows for AEs to be found in clinical trial databases and evaluated using software programs that are readily available to clinical researchers.


Heme oxygenase-2 (HO-2) binds and buffers labile ferric heme in human embryonic kidney cells.
Heme oxygenases (HOs) detoxify heme by oxidatively degrading it into carbon monoxide, iron, and biliverdin, which is reduced to bilirubin and excreted. Humans express two isoforms of HO: the inducible HO-1, which is upregulated in response to excess heme and other stressors, and the constitutive HO-2. Much is known about the regulation and physiological function of HO-1, whereas comparatively little is known about the role of HO-2 in regulating heme homeostasis. The biochemical necessity for expressing constitutive HO-2 is dependent on whether heme is sufficiently abundant and accessible as a substrate under conditions in which HO-1 is not induced. By measuring labile heme, total heme, and bilirubin in human embryonic kidney HEK293 cells with silenced or overexpressed HO-2, as well as various HO-2 mutant alleles, we found that endogenous heme is too limiting a substrate to observe HO-2-dependent heme degradation. Rather, we discovered a novel role for HO-2 in the binding and buffering of heme. Taken together, in the absence of excess heme, we propose that HO-2 regulates heme homeostasis by acting as a heme buffering factor that controls heme bioavailability. When heme is in excess, HO-1 is induced, and both HO-2 and HO-1 can provide protection from heme toxicity via enzymatic degradation. Our results explain why catalytically inactive mutants of HO-2 are cytoprotective against oxidative stress. Moreover, the change in bioavailable heme due to HO-2 overexpression, which selectively binds ferric over ferrous heme, is consistent with labile heme being oxidized, thereby providing new insights into heme trafficking and signaling.


Analgesic effect of picenadol, codeine, and placebo in patients with postoperative pain.
A double-blind, parallel study was conducted to evaluate the analgesic effect and safety of a single 25 mg oral dose of picenadol, a centrally acting analgesic, and to compare it with a 60 mg dose of codeine and a placebo in patients with postoperative pain. Two sites using similar protocols enrolled a total of 178 inpatients with postoperative pain. Pain intensity, relief, and adverse experiences were then measured for up to 6 hours after administration of the test medications. Both picenadol and codeine were significantly more effective than placebo in reducing pain intensity (mean sum of pain intensity difference scores: picenadol 5.21, codeine 5.19, and placebo 2.82) and increasing total relief (mean total pain relief: picenadol 10.21, codeine 11.07, and placebo 6.96). Adverse experience profiles were similar among the three treatment groups.


Buprenorphine in primary care: risk factors for treatment injection and implications for clinical management.
Though the introduction of office-based buprenorphine has greatly contributed to stem the HIV epidemic since 1995, concerns have been raised about the intravenous use of buprenorphine even in patients on substitution treatment. The aim of this study was to identify which factors are predictive of buprenorphine injection in patients receiving office-based buprenorphine. Subazur is a survey consisting of two longitudinal assessments of 111 stabilized patients receiving office-based buprenorphine in southeastern France. Patients were interviewed by phone at enrolment and 6 months later about social characteristics, addictive behaviors, treatment experiences, overdoses and suicide ideation or attempt. A logistic regression based on generalized estimating equations (GEE) was used to identify factors associated with buprenorphine injection at any interview. Among the 111 patients (32% women, mean age 38 years), 36 reported buprenorphine injection after having started treatment initiation in 40 interviews. After adjustment for time since first injection, individuals perceiving their prescribed dosage as inadequate (OR=2.6 95%CI[1.2-5.7]) and those reporting a history of suicide ideation or attempt (OR=2.7 95%CI[1.1-7.0]) had approximately a three-fold higher risk of injecting buprenorphine. Providing adequate care for both drug dependence and psychiatric comorbidities in primary care is a major issue. Like heroin use during methadone treatment, buprenorphine injection should be regarded more as a response to inadequate care than simply as a "misuse". A re-assessment of the treatment efficacy through a possible dosage increase or a switch to methadone could potentially reduce diversion and assure sustained adherence to OST.


Correction to: Interventions to improve vaccine acceptance among rheumatoid arthritis patients: a systematic review.
The original version of this article contained error. Table 1 was shown in the wrong version, thus corrected table is shown in this article.


Role of orbitofrontal cortex in incubation of oxycodone craving in male rats.
One of the main challenges in treating opioid-use disorders is relapse during abstinence, triggered by re-exposure to drug-associated cues. Previous studies have demonstrated that drug-seeking in rats progressively increases over time during withdrawal (incubation of drug craving). Here, we used male rats and examined neural mechanisms underlying incubation of craving to oxycodone, a commonly abused prescription opioid, and we focused on orbitofrontal cortex (OFC), a brain region previously implicated in incubation of heroin craving. We first used neuronal activity marker Fos and measured neuronal activation in OFC (ventral and lateral OFC) associated with day-1 and day-15 relapse tests. Next, we determined the effect of pharmacological reversible inactivation of OFC on incubated oxycodone seeking on withdrawal day 15. Finally, we determined the effect of reversible inactivation of OFC on nonincubated oxycodone seeking on withdrawal day 1. We found that lever presses during relapse tests were higher on withdrawal day 15 than on withdrawal day 1 (incubation of oxycodone craving). Incubation of oxycodone craving is accompanied with a time-dependent increase of Fos protein expression in both ventral and lateral OFC. Lastly, OFC inactivation decreased oxycodone seeking on withdrawal day 15 but had no effect on withdrawal day 1. Together with the previous heroin study, results here show that OFC plays a critical role in incubation of opioid craving.


Acute leukopenia during topical burn therapy with silver sulfadiazine.
Leukopenia, with a mean white blood cell count of 2,680/mm3, was observed in nine patients with thermal injury early in the course of topical treatment with silver sulfadiazine. All manifested absolute neutropenia with a concomitant increase in immature band forms in the peripheral blood smear. The leukocyte counts returned to within normal limits within 48 to 72 hours of discontinuation of silver sulfadiazine therapy in four patients, and also did so in five patients in whom silver sulfadiazine therapy was continued. Leukopenia secondary to silver sulfadiazine application is currently believed to be an innocuous, self-limited phenomenon.


Separation and identification of desferrioxamine and its iron chelating metabolites by high-performance liquid chromatography and fast atom bombardment mass spectrometry: choice of complexing agent and application to biological fluids.
An HPLC-based method capable of separating desferrioxamine (DFO) and its iron chelating metabolites from uv-absorbing species present in biological fluids has been developed. This method relies on the use of nitrilotriacetic acid (NTA) as the complexing agent in the mobile phase, instead of EDTA, previously used in HPLC methods. The use of NTA ensures that iron contamination present in buffers and bound to the column does not interfere with analysis. The disadvantages of using EDTA are discussed. The identity of the iron chelating metabolites of DFO present in the urine of patients with beta-thalassemia major has been established using FAB mass spectrometry. The metabolism of DFO, reported in this study, takes place almost exclusively at the N-terminal region of the molecule and is in many respects similar to the degradation of the amino acid lysine. In addition, a metabolite which corresponds to N-hydroxylation of the terminal amino group has been identified.


Cost of Revision Total Knee Replacement: A Protocol for Systematic Review and Meta-Analysis.
A major limitation of total knee replacement (TKR), as with other joint replacements, is the risk of revision. Revision TKR is associated with high risk and economic burden to patients, healthcare providers, and societies. It will be worthwhile to assess the economic burden of revision TKR across countries or different study settings. This study aims to review the literature on the cost of revision TKR to assess costs across countries and studies, estimate a pooled cost estimate for homogenous data, and identify major cost components that contribute to the cost burden. We will conduct a search of the MEDLINE (OvidSp), EMBASE, Web of Science, Cochrane Library, EconLit, and Google Scholar databases to identify relevant studies, and will use an optimally designed search approach to search for relevant studies. EndNote library will be used to manage the searched studies. Selection will be undertaken in two phases-screening and eligibility. Study selection, data extraction, and assessment of the risk of bias will be performed in duplicate, after which the data will be analysed narratively and a meta-analysis performed for homogenous studies, if possible. This protocol provides a proposed stepwise plan for conducting a systematic review and meta-analysis of the cost of revision TKR. Findings from this systematic review will provide information about the cost across settings and identify the major cost drivers of revision TKR, which will, in turn, stimulate efforts to minimize the cost. PROSPERO CRD42020171988.


BDNF augments rat internal anal sphincter smooth muscle tone via RhoA/ROCK signaling and nonadrenergic noncholinergic relaxation via increased NO release.
Presently, there are no studies examining the neuromodulatory effects of brain-derived neurotropic factor (BDNF) on the basal internal anal sphincter (IAS) tone and nonadrenergic noncholinergic (NANC) relaxation. To examine this, we determined the neuromuscular effects of BDNF on basal IAS smooth muscle tone and the smooth muscle cells (SMCs) and the effects of NANC nerve stimulation before and after high-affinity receptor tyrosine kinase receptor B (TrkB) antagonist K252a. We also investigated the mechanisms underlying BDNF-augmented increase in the IAS tone and NANC relaxation. We found that BDNF-increased IAS tone and SMC contractility were TTX resistant and attenuated by K252a. TrkB-specific agonist 7,8-dihydroxyflavone, similar to BDNF, also produced a concentration-dependent increase in the basal tone, whereas TrkB inhibitors K252a and ANA-12 produced a decrease in the tone. In addition, BDNF produced leftward shifts in the concentration-response curves with U46619 and ANG II (but not with bethanechol and K+ depolarization), and these shifts were reversed by K252a. Effects of Y27632 and Western blot data indicated that the BDNF-induced increase in IAS tone was mediated via RhoA/ROCK. BDNF-augmented NANC relaxation by electrical field stimulation was found to be mediated via the nitric oxide (NO)/soluble guanylate cyclase (sGC) pathway rather than via increased sensitivity to NO. In conclusion, the net effect of BDNF was that it caused an increase in the basal IAS tone via RhoA/ROCK signaling. BDNF also augmented NANC relaxation via NO/sGC. These findings may have relevance to the role of BDNF in the pathophysiology and therapeutic targeting of the IAS-associated rectoanal motility disorders.NEW & NOTEWORTHY These studies for the first time to our knowledge demonstrate that increased levels of brain-derived neurotrophic factor (BDNF; conceivably released from smooth muscle cells and/or the enteric neurons) has two major effects. First, BDNF augments the internal anal sphincter (IAS) tone via tyrosine kinase receptor B/thromboxane A2-receptor, angiotensin II receptor type 1/RhoA/ROCK signaling; and second, it increases nonadrenergic noncholinergic relaxation via nitric oxide/soluble guanylate cyclase. These studies may have relevance in therapeutic targeting in the anorectal motility disorders associated with the IAS.


Use of COVID-19 Test Positivity Rate, Epidemiological, and Clinical Tools for Guiding Targeted Public Health Interventions.
India experienced a second wave of COVID-19 infection with an unprecedented upsurge in the number of cases. We have analyzed the effect of different restrictive measures implemented in six Indian states. Further, based on available national and international data on disease transmission and clinical presentation, we have proposed a decision-making matrix for planning adequate resources to combat the future waves of COVID-19. We conclude that pragmatic and well calibrated localized restrictions, tailored as per specific needs may achieve a decline in disease transmission comparable to drastic steps like national lockdowns. Additionally, we have underscored the critical need for countries to generate local epidemiological, clinical and laboratory data alongwith community perception and uptake of various non-pharmaceutical interventions, for effective planning and policy making.


The clinicopathological and prognostic impact of 14-3-3 sigma expression on vulvar squamous cell carcinomas.
14-3-3 sigma promotes G2/M cell cycle arrest by sequestering cyclin B1-CDC2 complex in cytoplasm. Down-regulation of 14-3-3sigma, which has been demonstrated in various carcinomas, may contribute to malignant transformation. However, the exact role of 14-3-3sigma in the pathogenesis of vulvar carcinoma is not fully characterized, and the prognostic impact of 14-3-3sigma protein expression is still unknown. We investigated the 14-3-3sigma expression in a series of 302 vulvar squamous cell carcinomas using immunohistochemistry and its associations with clinicopathological factors and clinical outcome. In cytoplasm, nucleus and cytoplasm/nucleus of vulvar carcinomas high 14-3-3sigma protein expression was found in 72%, 59% and 75% of the carcinomas, respectively, and low levels in 28%, 41% and 25% of the cases, respectively. High level of 14-3-3sigma in cytoplasm, nucleus and cytoplasm/nucleus was significantly correlated to large tumor diameter (p = 0.001, p = 0.002 and p = 0.001, respectively) and deep invasion (p = 0.01, p = 0.001 and p = 0.007, respectively). Variations of 14-3-3sigma protein expression were not associated to disease-specific survival. Our results indicate that 14-3-3sigma may be involved in the development of a subset of vulvar squamous cell carcinomas by down-regulation of 14-3-3sigma protein. Neither cytoplasmic nor nuclear level of 14-3-3sigma expression was associated with prognosis.


Splenogonadal fusion-limb defect "syndrome" and associated malformations.
Two types of splenogonadal fusion (SGF) have been proposed. In the continuous type the spleen is connected to the gonad, and often it is associated with limb defects and other anomalies such as micrognathia, microglossia, anal atresia, and hypoplastic lungs. Associated abnormalities are lower in the discontinuous type of SGF, where there is fusion between the gonad and accessory splenic tissue, without connection with the normal spleen. The cause of SGF and SGF with limb defects (SGFLD) is unknown. On the basis of associated orofacial limb abnormalities, some authors have suggested that SGFLD may be related to the Hanhart complex. Others have noted the overlap between SGFLD and femoral-facial syndrome (FFS). We consider SGFLD a developmental field defect that originates during blastogenesis. Although the cause is unknown, the earlier its action, the more severe the involvement resulting in SGFLD; later action may result only in SGF.


[Dobutamine stress echocardiography in patients with stable chronic angina and a low- or medium-risk on exercise testing: usefulness for assessing long-term prognosis].
The ability of stress echocardiography to provide prognostic information that supplements that obtainable from clinical data and exercise electrocardiography is still controversial. Our aim was to determine whether dobutamine stress echocardiography provides additional information on long-term prognosis after conventional exercise testing has indicated that a patient with chronic stable angina has a low or intermediate risk of a cardiac event. The study included consecutive patients with stable angina who were not found to be at high risk on a previous exercise test. All patients underwent dobutamine stress echocardiography. The mean follow- up period was 4.5+/-1.76 years. The single combined end-point was defined as death due to cardiac disease, nonfatal myocardial infarction, or hospitalization for unstable angina. Multivariate analysis was used to identify independent predictors of cardiac events. There were 24 (19%) cardiac events in the 124 participants: four deaths due to cardiac disease, 10 nonfatal myocardial infarctions, and 10 hospitalizations for unstable angina. Associations were found between a higher event rate during follow-up and previous myocardial infarction, Duke treadmill score, and the detection of regional wall motion abnormalities indicative of multivessel disease by stress echocardiography. In patients with stable angina who have undergone an exercise test that indicates that they have a low or intermediate risk of cardiac events, dobutamine stress echocardiography provides additional prognostic information to that obtainable from clinical data and exercise testing.


Interferon pretreatment regulates interferon and interleukin-6 production in L929 cells in a coordinated manner.
Two sublines of L929 cells with different interferon (IFN)-producing capacities synthesized IFN and interleukin-6 (IL-6) simultaneously in response to Sendai virus infection. IFN pretreatment primed the production of both cytokines. However, the difference in IL-6 production between the "high and low producer" L-cell sublines was about one magnitude of order larger than in the case of their IFN production. The determination of the corresponding mRNA levels also reflected this difference.


The occurrence of early atrial fibrillation after cardiac valve operation and the establishment of a nomogram model.
Postoperative atrial fibrillation (POAF) is a common complication after cardiac surgery, which is associated with age and massive bleeding. However, whether thyroid hormone (TH) level can affect POAF remains controversial. To investigate the occurrence and risk factors of POAF, in particular, the preoperative TH level of patients was introduced into this study as a variable for analysis, and a column graph prediction model of POAF was constructed. Patients who underwent valve surgery in Fujian Cardiac Medical Center from January 2019 to May 2022 were retrospectively analyzed and divided into POAF group and NO-POAF group. Baseline characteristics and relevant clinical data were collected from the two groups of patients. Independent risk factors for POAF were screened using univariate analysis and binary logistic regression analysis, and a column line graph prediction model was established based on the regression analysis results, and the diagnostic efficacy and calibration of the model were evaluated using the Receiver Operating Characteristic Curve (ROC) and calibration curve. A total of 2,340 patients underwent valve surgery, excluding 1,751 patients, a total of 589 patients were included, including 89 patients in POAF group and 500 patients in NO-POAF group. The total incidence of POAF was 15.1%. The results of the Logistics regression analysis showed that gender, age, leukocytes and TSH were risk factors of POAF. The area under the ROC curve of the nomogram prediction model for POAF was 0.747 (95% CI: 0.688-0.806, P < 0.001), with a sensitivity of 74.2% and specificity of 68%. Hosmer-Lemeshow test showed χ 2 = 11.141, P = 0.194 > 0.05, the calibration curve was well fitted. The results of this study show that gender, age, leukocyte and TSH are risk factors of POAF, and the nomogram prediction model has a good prediction effect. Due to the limited sample size and included population, more studies are needed to validate this result.


Tritium (3H) radiolabeling of protein A and antibody to high specific activity: application to cell surface antigen radioimmunoassays.
Staphylococcal protein A and several different immunoglobulins have been radiolabeled to high specific activities (greater than 10(6) cpm/microgram) by reductive methylation with tritiated (3H) sodium borohydride. The proteins retain excellent functional and antigenic properties. The utility of these reagents in a variety of assays for cell surface antigens is illustrated. The results indicate that this radiolabeling procedure may become the method of choice for many cell surface and solution immunoassays.


Preparation of a nitro-substituted tris(indolyl)methane modified silica in deep eutectic solvents for solid-phase extraction of organic acids.
A new sorbent for solid-phase extraction was synthesized by chemical immobilization of nitro-substituted tris(indolyl)methane on silica in new and green deep eutectic solvents. Elemental analysis results indicated that deep eutectic solvents could be an alternative to the traditional solvents in preparing nitro-substituted tris(indolyl)methane modified silica. Coupled with high performance liquid chromatography, the extraction performance of the sorbent was evaluated by using four organic acids as model analytes. The rebinding experiments results showed that the nitro-substituted tris(indolyl)methane modified silica sorbent had a good adsorption capacity towards the selected organic acids. Under the appropriate experimental conditions, good precision and wide linear ranges with coefficient of determination (R(2)) of higher than 0.9957 were obtained, and the limits of detection were in the range of 0.50-2.0μgL(-1) for the organic acids tested. The developed solid-phase extraction-high performance liquid chromatography-diode array detection (SPE-HPLC-DAD) method was successfully applied for the determination of organic acids in two drinking samples with recoveries ranging from 76.7% to 110.0% and 67.7% to 104.0% for all the selected organic acids, respectively.


Indirubin-3'-monoxime, a derivative of a Chinese anti-leukemia medicine, inhibits Notch1 signaling.
Notch proteins perform a critical function in cell-fate decisions and in differentiation. In this study, we determined that indirubin-3'-monoxime reduced Notch1 signaling to a remarkable extent. Indirubin-3'-monoxime has been shown to inhibit both constitutive active mutants of Notch1 and Notch1-IC-mediated transactivation activity. However, in such cases, neither the Notch cleavage pattern nor the protein stability of Notch1-IC was determined to have been significantly altered. Indirubin-3'-monoxime suppresses Notch1 transcriptional activity via the dissociation of the Notch1-IC-RBP-Jk complex. Notably, the transcriptional activity of Notch1-IC was not suppressed significantly in the GSK-3beta null cells by indirubin-3'-monoxime as compared to what was observed with GSK-3beta wild-type cells. In the previous study, we synthesized a series of indirubin derivatives. Interestingly, some of these indirubin derivatives were characterized as potent inhibitors of Notch1 signaling. Taken together, the results of this study indicate that indirubin-3'-monoxime downregulated Notch1 signaling in a GSK-3beta-dependent and proteosomal degradation-independent manner.


IMRT treatment planning:- a comparative inter-system and inter-centre planning exercise of the ESTRO QUASIMODO group.
The purpose of this work was a comparison of realistic IMRT plans based on the same CT-image data set and a common predefined set of dose objectives for the planning target volume and the organs at risk. This work was part of the larger European QUASIMODO IMRT verification project. Eleven IMRT plans were produced by nine different European groups, each applying a representative set of clinically used IMRT treatment planning systems. The plans produced were to be deliverable in a clinically acceptable treatment time with the local technical equipment. All plans were characterized using a set of different quality measures such as dose-volume histograms, number of monitor units and treatment time. Only one plan was able to fulfil all dose objectives strictly; six plans failed some of the objectives but were still considered to be clinically acceptable; four plans were not able to reach the objectives. Additional quality scores such as the number of monitor units and treatment time showed large variations, which mainly depend on the delivery technique. The presented planning study showed that with nearly all presently available IMRT planning and delivery systems comparable dose distributions could be achieved if the planning goals are clearly defined in advance.


Quality of marital life in Korean spinal cord injured patients.
The purposes of this study were to assess the quality of marital life and to investigate the most serious problem in the marital relationships of Korean spinal cord injured patients. 30 spinal cord injured (SCI) couples (SCI husbands and their non-disabled wives) who were married prior to injury and 30 able-bodied (AB) couples participated in a questionnaire study. The quality of marital life was measured with three parameters: marital stability, marital adjustment, and marital satisfaction. The results were as follows: (1) The marriage of chronic SCI couples was not noticeably unstable when compared with that of AB couples; (2) There was no significant difference in dyadic adjustment and marital satisfaction between SCI couples and AB couples; (3) There was no significant difference in marital stability, marital adjustment, and marital satisfaction between SCI husbands and their wives; (4) SCI couples had more cohesive marital relationships and SCI husbands expressed less affection to their wives than AB husbands and (5) Sex was the most serious problem in marriages of SCI couples. (1) the quality of marital life in chronic SCI couples is not highly different from that of AB couples; (2) There is no significant difference in the quality of marital life between chronic SCI husbands and their non-disabled wives; and (3) It is considered necessary that rehabilitation program for SCI patients should include information on the altered physiology of sexual function of SCI patients and subsequent mutual adaptation to changed sexual function.


[The effects of moderate acute normovolemic hemodilution on intestinal oxygen supply and consumption in anesthetized cats].
To study the effects of moderate acute normovolemic hemodilution on the intestinal blood perfusion and oxygen supply in cats. 15 domestic male cats were hemodiluted with Haemaccel. HR, MAP, lactate concentration in superior mesenteric vein (LAC), superior mesenteric artery blood flow (SMABF), superior mesenteric artery and venous blood gas analysis were recorded and the intestinal oxygen delivery (DO(2)), oxygen consumption (VO(2)) and oxygen extraction (EO(2)) were calculated before and after hemodilution. SMABF increased obviously (P < 0.05), intestinal DO(2) and EO(2) did not change (P > 0.05) but VO(2) increased (P < 0.05), and LAC in superior mesenteric vein did not increase. Moderate acute normovolemic hemodilution with Haemaccel do not alter intestinal oxygen delivery by increasing the superior mesenteric blood flow. It means that intestinal mucosa ischemia does not happen during moderate acute normovolemic hemodilution.


Acetylshikonin from Zicao exerts antifertility effects at high dose in rats by suppressing the secretion of GTH.
Zicao is being highlighted as a promising Chinese medicine due to all the beneficial effects that have been associated with its use. Unfortunately, studies on the toxicity of Zicao in different species are still missing and should be carried out. In this study, we investigated whether Acetylshikonin (AS) from Zicao has an anti-fertility effect through mating experiments and explored its underling mechanism. Sprague-Dawley rats received no treatment or were treated with 120, 360 or 1080 mg/kg AS extract by intragastric administration for 2 weeks. The rat pregnancy rate of the 1080 mg/kg dose group was significantly decreased relative to control group, while it recovered after a month of drug withdrawal, which indicated that the effect of antifertility is reversible. Serum follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels in rat were significantly decreased by AS. The secretion of FSH in rat anterior pituitary cells was decreased but the synthesis was not affected. AS reduced the number of developing follicle and mature follicle in rat ovarian cortical. Maybe all of these resulted from AS decreased the expression of synaptotagmin-1 and SNAP-25 which were the critical proteins of exocytosis. Our data suggested that AS at high dose can suppress the ability of pregnancy of the rats through decreasing serum FSH and LH levels by affecting exocytosis process of gonadotropic hormone (GTH).


Reinventing family therapy: teaching family intervention as a new treatment modality.
This article discusses the pedagogy of teaching family therapy in the new millennium. It draws on the strengths of "family systems therapy" but goes beyond it-suggesting a new paradigm, new terminology, and a new teaching perspective. It discusses the historical background of family therapy training, a scientific foundation for what residents should be taught, and an integrative clinical model for how it could be taught. The article is the synthesis of the perspectives and experience of a child and adolescent psychiatrist educator who began his career at the end of the systemic era in psychiatry and continues it through the developments of the neurobiologic era. It draws on selected literature from the fields of family therapy, child and adolescent psychiatry, developmental psychopathology, and general psychiatry. This article submits that the term "family therapy" should be replaced by the term "family intervention"; the evidence base indicates that family interventions are effective; family risk and protective factors influence the onset and course of disorders; families help shape and maintain cognitive schema; intervention must start with thorough case formulation; and family interventions should be coordinated with other interventions, include parent management training, and build on family strengths. Family intervention is an important clinical process in child and adolescent psychiatry, and contemporary education must address the multiple ways clinicians can assist families. Future models will be successful to the degree they build on the past contributions of systems thinking and include the perspectives of developmental psychopathology. Contemporary education should teach that family interventions are not optional but ideally can be integrated with other interventions in a sequential manner, emphasizing the interrelationship between self and system.


Scintigraphic localization of occult gastrointestinal bleeding using a combination of 111In-labelled platelets and 99mTc sulphur colloid.
Repeated scintigraphy was performed for 6 days after the injection of autologous 111In-labeled platelets in a patient showing signs of recurrent occult gastrointestinal bleeding. Colonic activity was observed 142 h post injection, at which time a superimposed study using 99mTc sulphur colloid pinpointed the source of bleeding in the transverse colon.


[Pheochromocytoma of the prostate: an unusual site].
Pheochromocytomas of the prostate are rare, with only two cases reported in adults. We report the uncommon case of a pheochromocytoma of the prostate in a 8-year-old boy.


Intraobserver reproducibility and interobserver agreement of ultrasonic flowmetry using the velocity profile color doppler method in the common carotid artery.
Application of the velocity profile method is recommended for reliable measurement of flow volume in larger vessels, and ultrasonic flowmetry is a useful clinical tool for this purpose. We used the velocity profile in conjunction with a minor modification in the conventional velocity profile method and examined the reproducibility of flowmetry from color Doppler data. Data of three examiners were allowed to analyze intraobserver reproducibility and interobserver agreement in the common carotid artery, and we measured flow volume in the peripheral vessels of healthy individuals. Estimated flow volumes in five healthy examinees were 350 to 550 ml/min and did not vary significantly between examiners. Interobserver correlation was good (r 1=0.63), but intraobserver correlations in two sonographers were excellent (r 1=0.85) in by one who was experienced in this method and poor (r 1=0.32) in the other. Good interobserver agreement and intraobserver reproducibility of experienced examiners suggests that this method is reliable. Values obtained with normal controls averaged 438±78 (mean±SD) ml/min and 449±64 ml/min, respectively, in the right and left carotid arteries (n=12) and 120±31 ml/min and 83±23 ml/min, respectively, in the right and left vertebral arteries (n=10). Normal values were 542±71 ml/min (n=11) in the right renal artery, 1210±211 ml/min in the abdominal artery (n=10), and 799±182 ml/min in the main portal vein (n=17). Arterial blood flow volume can measure reliably by the velocity profile color Doppler method.


An Ex vivo culture model for placental cytomegalovirus infection using slices of Guinea pig placental tissue.
Congenital infection with human cytomegalovirus (CMV) through the placenta is one of the major causes of birth and developmental abnormalities. Guinea pig CMV (GPCMV) causes in utero infection, which makes its animal models useful for studies on congenital diseases. Here, we established an ex vivo culture method for tissue slices prepared from guinea pig placentas and demonstrated that viral spread in the model resembles those in the placenta of GPCMV-infected animals and that the infection is independent of the pentameric glycoprotein complex for endothelial/epithelial cell tropism. Thus, this model affords a useful tool for pathobiological studies on CMV placental infection.


Space- and Time-Resolved Metabolomics of a High-Grade Serous Ovarian Cancer Mouse Model.
The dismally low survival rate of ovarian cancer patients diagnosed with high-grade serous carcinoma (HGSC) emphasizes the lack of effective screening strategies. One major obstacle is the limited knowledge of the underlying mechanisms of HGSC pathogenesis at very early stages. Here, we present the first 10-month time-resolved serum metabolic profile of a triple mutant (TKO) HGSC mouse model, along with the spatial lipidome profile of its entire reproductive system. A high-coverage liquid chromatography mass spectrometry-based metabolomics approach was applied to longitudinally collected serum samples from both TKO (n = 15) and TKO control mice (n = 15), tracking metabolome and lipidome changes from premalignant stages to tumor initiation, early stages, and advanced stages until mouse death. Time-resolved analysis showed specific temporal trends for 17 lipid classes, amino acids, and TCA cycle metabolites, associated with HGSC progression. Spatial lipid distributions within the reproductive system were also mapped via ultrahigh-resolution matrix-assisted laser desorption/ionization (MALDI) mass spectrometry and compared with serum lipid profiles for various lipid classes. Altogether, our results show that the remodeling of lipid and fatty acid metabolism, amino acid biosynthesis, TCA cycle and ovarian steroidogenesis are critical components of HGSC onset and development. These metabolic alterations are accompanied by changes in energy metabolism, mitochondrial and peroxisomal function, redox homeostasis, and inflammatory response, collectively supporting tumorigenesis.


Task shifting HIV care in rural district hospitals in Cameroon: evidence of comparable antiretroviral treatment-related outcomes between nurses and physicians in the Stratall ANRS/ESTHER trial.
Task shifting to nurses for antiretroviral therapy (ART) is promoted by the World Health Organization to compensate for the severe shortage of physicians in Africa. We assessed the effectiveness of task shifting from physicians to nurses in rural district hospitals in Cameroon. We performed a cohort study using data from the Stratall trial, designed to assess monitoring strategies in 2006-2010. ART-naive patients were followed up for 24 months after treatment initiation. Clinical visits were performed by nurses or physicians. We assessed the associations between the consultant ratio (ie, the ratio of the number of nurse-led visits to the number of physician-led visits) and HIV virological success, CD4 recovery, mortality, and disease progression to death or to the World Health Organization clinical stage 4 in multivariate analyses. Of the 4141 clinical visits performed in 459 patients (70.6% female, median age 37 years), a quarter was task shifted to nurses. The consultant ratio was not significantly associated with virological success [odds ratio 1.00, 95% confidence interval (CI): 0.59 to 1.72, P = 0.990], CD4 recovery (coefficient -3.6, 95% CI: -35.6; 28.5, P = 0.827), mortality (time ratio 1.39, 95% CI: 0.27 to 7.06, P = 0.693), or disease progression (time ratio 1.60, 95% CI: 0.35 to 7.37, P = 0.543). This study brings important evidence about the comparability of ART-related outcomes between HIV models of care based on physicians or nurses in resource-limited settings. Investing in nursing resources for the management of noncomplex patients should help reduce costs and patient waiting lists while freeing up physician time for the management of complex cases, for mentoring and supervision activities, and for other health interventions.


A replicator-specific binding protein essential for site-specific initiation of DNA replication in mammalian cells.
Mammalian chromosome replication starts from distinct sites; however, the principles governing initiation site selection are unclear because proteins essential for DNA replication do not exhibit sequence-specific DNA binding. Here we identify a replication-initiation determinant (RepID) protein that binds a subset of replication-initiation sites. A large fraction of RepID-binding sites share a common G-rich motif and exhibit elevated replication initiation. RepID is required for initiation of DNA replication from RepID-bound replication origins, including the origin at the human beta-globin (HBB) locus. At HBB, RepID is involved in an interaction between the replication origin (Rep-P) and the locus control region. RepID-depleted murine embryonic fibroblasts exhibit abnormal replication fork progression and fewer replication-initiation events. These observations are consistent with a model, suggesting that RepID facilitates replication initiation at a distinct group of human replication origins.


Plant adaptation to low phosphorus availability: Core signaling, crosstalks, and applied implications.
Phosphorus (P) is an essential nutrient for plant growth and reproduction. Plants preferentially absorb P as orthophosphate (Pi), an ion that displays low solubility and that is readily fixed in the soil, making P limitation a condition common to many soils and Pi fertilization an inefficient practice. To cope with Pi limitation, plants have evolved a series of developmental and physiological responses, collectively known as the Pi starvation rescue system (PSR), aimed to improve Pi acquisition and use efficiency (PUE) and protect from Pi-starvation-induced stress. Intensive research has been carried out during the last 20 years to unravel the mechanisms underlying the control of the PSR in plants. Here we review the results of this research effort that have led to the identification and characterization of several core Pi starvation signaling components, including sensors, transcription factors, microRNAs (miRNAs) and miRNA inhibitors, kinases, phosphatases, and components of the proteostasis machinery. We also refer to recent results revealing the existence of intricate signaling interplays between Pi and other nutrients and antagonists, N, Fe, Zn, and As, that have changed the initial single-nutrient-centric view to a more integrated view of nutrient homeostasis. Finally, we discuss advances toward improving PUE and future research priorities.


Ethonursing Study of Pain Management in Neonates in Indonesia.
Even though Indonesia is comprised of at least 1,300 ethnic groups, no specific traditional method for reducing pain, especially in neonates, has been elaborated. This study aimed to investigate the traditional methods used by the Sundanese people in Indonesia to manage pain in neonates. This ethnonursing study was conducted in the Pangandaran District, West Java Province, Indonesia. Thirty informants including health cadres, midwives, and community leaders in the health sector, such as trained traditional birth attendants and female elders, participated in this study. Data were collected through interviews and focus group discussions, as well as from field notes. Data were analyzed using the four phases of qualitative analysis based on Leininger's theory. Six themes were identified in this study: (1) Preference for the use of traditional methods to treat pain in neonates and infants, (2) Desire to obtain information on the safe use of traditional medicine, (3) Desire to use practical pain management methods, (4) Easy to get modern medicine, (5) Use of medicinal plants, (6) Physical methods. This study demonstrates the need for better harmonization between the use of traditional and modern medicines, in both research and practice, to determine the most appropriate mix that combines the strengths of each method based on accurate scientific evidence.


Probing the precision of the mitotic clock with a live-cell fluorescent biosensor.
Precise timing of mitosis is essential for high-fidelity cell duplication. However, temporal measurements of the mitotic clock have been challenging. Here we present a fluorescent mitosis biosensor that monitors the time between nuclear envelope breakdown (NEB) and re-formation using parallel total internal reflection fluorescence (TIRF) microscopy. By tracking tens to hundreds of mitotic events per experiment, we found that the mitotic clock of unsynchronized rat basophilic leukemia cells has a marked precision with 80% of cells completing mitosis in 32 +/- 6 min. This assay further allowed us to observe delays in mitotic timing at Taxol concentrations 100 times lower than previous minimal effective doses, explaining why Taxol is clinically active at low concentrations. Inactivation of the spindle checkpoint by targeting the regulator Mad2 with RNAi consistently shortened mitosis, providing direct evidence that the internal mitotic timing mechanism is much faster in cells that lack the checkpoint.


Natural progression of osteo-chondral defect in the femoral condyle.
We have tried to establish the course of an isolated osteochondral defect in a femoral condyle would take if it were not treated surgically. We studied 15 knees in 12 patients where a diagnosis of the condition had been made in the past and no surgical treatment instituted for it. Each patient was examined clinically and had an MRI scan. The average period of follow-up was 109 months, the range being 54-282 months. Six patients (seven knees) were under the age of 18 at the onset of symptoms. Of these, MRI of six knees showed either no abnormality or a healed lesion. Where the onset was after skeletal maturity, six of the eight knees showed features osteoarthritis on the MRI scan. Symptomatically, there was also a difference between the two groups.


Single-cell profiles reveal distinctive immune response in atopic dermatitis in contrast to psoriasis.
Understanding the complex orchestrated inflammation in atopic dermatitis (AD), one of the most common chronic inflammatory diseases worldwide, is essential for therapeutic approaches. However, a comparative analysis on the single-cell level of the inflammation signatures correlated with the severity is missing so far. We applied single-cell RNA and T-cell receptor (TCR) sequencing on immune cells enriched from skin biopsies and matched blood samples of AD in comparison with psoriasis (PS) patients. Clonally propagated skin-derived T cells showed disease-specific TCR motifs shared between patients which was more pronounced in PS compared to AD. The disease-specific T-cell clusters were mostly of a Th2/Th22 sub-population in AD and Th17/Tc17 in PS, and their numbers were associated with severity scores in both diseases. Herein, we provide for the first time a list that associates cell type-specific gene expression with the severity of the two most common chronic inflammatory skin diseases. Investigating the cell signatures in the patients´ PBMCs and skin stromal cells, a systemic involvement of type-3 inflammation was clearly detectable in PS circulating cells, while in AD inflammatory signatures were most pronounced in fibroblasts, pericytes, and keratinocytes. Compositional and functional analyses of myeloid cells revealed the activation of antiviral responses in macrophages in association with disease severity in both diseases. Different disease-driving cell types and subtypes which contribute to the hallmarks of type-2 and type-3 inflammatory signatures and are associated with disease activities could be identified by single-cell RNA-seq and TCR-seq in AD and PS.


Nursing workload in the acute-care setting: A concept analysis of nursing workload.
A pressing need in the field of nursing is the identification of optimal staffing levels to ensure patient safety. Effective staffing requires comprehensive measurement of nursing workload to determine staffing needs. Issues surrounding nursing workload are complex, and the volume of workload is growing; however, many workload systems do not consider the numerous workload factors that impact nursing today. The purpose of this concept analysis was to better understand and define nursing workload as it relates to the acute-care setting. Rogers' evolutionary method was used for this literature-based concept analysis. Nursing workload is influenced by more than patient care. The proposed definition of nursing workload may help leaders identify workload that is unnoticed and unmeasured. These findings could help leaders consider and identify workload that is unnecessary, redundant, or more appropriate for assignment to other members of the health care team.


Modified LDL Particles Activate Inflammatory Pathways in Monocyte-derived Macrophages: Transcriptome Analysis.
A hallmark of atherosclerosis is its complex pathogenesis, which is dependent on altered cholesterol metabolism and inflammation. Both arms of pathogenesis involve myeloid cells. Monocytes migrating into the arterial walls interact with modified low-density lipoprotein (LDL) particles, accumulate cholesterol and convert into foam cells, which promote plaque formation and also contribute to inflammation by producing proinflammatory cytokines. A number of studies characterized transcriptomics of macrophages following interaction with modified LDL, and revealed alteration of the expression of genes responsible for inflammatory response and cholesterol metabolism. However, it is still unclear how these two processes are related to each other to contribute to atherosclerotic lesion formation. We attempted to identify the main mater regulator genes in macrophages treated with atherogenic modified LDL using a bioinformatics approach. We found that most of the identified genes were involved in inflammation, and none of them was implicated in cholesterol metabolism. Among the key identified genes were interleukin (IL)-7, IL-7 receptor, IL- 15 and CXCL8. Our results indicate that activation of the inflammatory pathway is the primary response of the immune cells to modified LDL, while the lipid metabolism genes may be a secondary response triggered by inflammatory signalling.


Assessment of Mitophagy in iPS Cell-Derived Neurons.
Aberrant mitochondrial function is associated with many neurological diseases. Mitophagy is a key mechanism for the elimination of damaged mitochondria and maintenance of mitochondrial homeostasis. Induced pluripotent stem (iPS) cell technologies developed over the last decade have allowed us to analyze functions of the human neuron. Here we describe an efficient induction method from human iPS cells to neurons, followed by an image-based mitophagy assay.


Transgenic mice overexpressing human Bcl-2 are resistant to hepatic ischemia and reperfusion.
Apoptosis is a key mechanism of reperfusion injury in the ischemic liver. The apoptotic pathway is highly regulated by anti-apoptotic factors, such as Bcl-2. We evaluated the effect of Bcl-2 overexpression on apoptosis and the activation of the apoptotic cascade after hepatic ischemia and reperfusion. Ninety minutes of ischemia and reperfusion was performed in Bcl-2 transgenic and non-transgenic mice. Bcl-2 overexpression was determined by immunohistochemistry and Western blot. Liver injury was determined by aspartate aminotransferase (AST), Tunel test and the activation of the apoptotic cascade and animal survival. Bcl-2 overexpression was present in all hepatocytes and non-parenchymal liver cells in transgenic mice. Bcl-2 overexpression resulted in significant decreased AST levels after ischemic injury, and complete inhibition of apoptosis. After 90 min of total hepatic ischemia all control mice died, while four transgenic mice survived permanently. Bcl-2 overexpression was associated with inhibition of caspase 3 activation after reperfusion and increased baseline levels of cytoplasmic cytochrome c, caspase 3, and a reduction of Bcl-x(L) production. Bcl-2 overexpression protects against ischemic injury by inhibiting apoptosis. Extensive overproduction of Bcl-2 is associated with a compensatory increase of baseline levels of cytoplasmic cytochrome c and caspase 3, and a deletion of Bcl-x(L).


Altered immunologic reconstitution after standard-dose chemotherapy or high-dose chemotherapy with autologous bone marrow support.
Altered immunologic reconstitution is observed in patients treated with high-dose chemotherapy consisting of cyclophosphamide, cisplatin, and carmustine, melphalan, or etoposide with autologous bone marrow support, and it is similar to that seen in patients treated with high-dose chemoradiotherapy and allogeneic bone marrow transplantation. A decrease in the absolute number and percentage of B cell and CD4 antigen-positive cells and an increase in the absolute number and percentage of CD8 and Ia antigen-positive cells occur along with a decrease in the CD4/CD8 ratio that persists for 6-12 months after high-dose chemotherapy and autologous bone marrow support. These changes have been associated with four serious infectious episodes usually seen only in immunocompromised patients. The above changes were not seen in patients treated with high-dose busulfan, a drug that has relatively specific effects on granulocytes. It is postulated that these alterations result from effects of chemotherapy on the residual lymphocytes or on the environment of repopulating lymphocytes. Functional studies of lymphocyte populations during immunologic reconstitution after standard-dose combination chemotherapy and high-dose chemotherapy with autologous bone marrow support are needed.


Preparation and application of triangular silver nanoplates/chitosan composite in surface plasmon resonance biosensing.
This paper reports the utilization of triangular silver nanoplates (TSNPs) to enhance the sensitivity of surface plasmon resonance (SPR) biosensor. TSNPs modified with 3-mercaptopropinic acid (MPA) were simply mixed with chitosan and glutaraldehyde to form TSNPs/chitosan composite. The composite was deposited on Au film as immobilization substrate for SPR biosensor. The novel structures of TSNPs are preserved against etching by MPA and chitosan polymer. Moreover, chitosan cross-linked by glutaraldehyde enables antibody to be immobilized on fabricated substrate directly via Schiff alkali reaction. In the optimized conditions, the resulting biosensor based on TSNPs/chitosan composite shows a satisfactory response to bovine IgG in the concentration range of 0.075-40.00 μg mL(-1). While the biosensor based on chitosan without TSNPs shows a response in the concentration range of 0.6-40 μg mL(-1) and the biosensor based on Au film shows a response in the concentration range of 2.5-40 μg mL(-1). The experiment results show that the sensitivity of SPR biosensor based on TSNPs/chitosan composite was significantly enhanced and the immobilization procedure of antibody was simplified.


The Janus-face kynurenic acid.
Kynurenic acid is an endogenous product of the tryptophan metabolism. Studies on the mechanism of its action have revealed that kynurenic acid at high concentrations is a competitive antagonist of the N-methyl-D-aspartate receptor and acts as a neuroprotectant in different neurological disorders. This in vitro investigation was designed to show that kynurenic acid acts differently at low concentrations. In vitro electrophysiological examinations on the young rat hippocampus confirmed the well-known finding that kynurenic acid in micromolar concentrations exerts an inhibitory effect. However, in nanomolar concentrations, kynurenic acid does not give rise to inhibition, but in fact facilitates the field excitatory postsynaptic potentials. The results available so far are compatible with the idea that kynurenic acid in the concentration range between a few hundred nanomolar and micromolar displays different effects. Its probable action on different receptors, inducing the different mechanisms, is discussed. The findings strongly suggest the neuromodulatory role of kynurenic acid under both physiological and pathological circumstances.


Mutations in exon 17B of cartilage oligomeric matrix protein (COMP) cause pseudoachondroplasia.
Pseudoachondroplasia (PSACH) is a well characterized dwarfing condition mapping to chromosome 19p12-13.1. Cartilage oligomeric matrix protein (COMP), a cartilage specific protein, maps to the same location within a contig that spans the PSACH locus. Using single strand conformation polymorphism (SSCP) analysis and nucleotide sequencing we have identified COMP mutations in eight familial and isolated PSACH cases. All mutations involve either a single base-pair change or a three base-pair deletion in exon 17B. Six mutations delete or change a well conserved aspartic acid residue within the calcium-binding type 3 repeats. These results demonstrate that mutations in the COMP gene cause pseudochondroplasia.


Creating an Innovative Global Health Clinical Experience During the COVID-19 Pandemic.
When the COVID-19 pandemic hit and international travel was halted, nursing faculty were challenged to transform the in-person global health clinical experience with travel to a new virtual format. The virtual experience would need to meet the learning objectives and provide a global health perspective. This article describes the process of transforming the in-person clinical experience to a virtual format to provide a rich global learning opportunity for the students without travel to the host country. Virtual global health experiences can successfully help students understand the health of populations with a global perspective.


Effect of prostaglandin endoperoxide analogue on canine renal function, hemodynamics and renin release.
We studied the effect of a stable, cyclic ether analogue of prostaglandin endoperoxide (EPA) on canine renal function, hemodynamics, and renin release. Infusion of EPA into one renal artery decreased renal blood flow in a dose dependent manner. At a dose of 10(-7) g/kg/min the renal blood flow decreased from a baseline of 384 to 267 ml/min/100 g. This flow decrease was unaltered by phentolamine and saralasin, but was potentiated by prior treatment with indomethacin. Urine flow, glomerular filtration rate, sodium, and potassium excretion all decreased in a dose dependent manner; however, neigher fractional excretion of sodium nor free water clearance showed any significant change, making direct tubular effects of EPA unlikely. EPA caused a significant increase in renin release that was completely blocked by prior treatment with indomethacin. We conclude that EPA is a potent renal vasoconstrictor and that this vasoconstriction is responsible for the renal functional changes observed. Renin release is not a direct effect of EPA but probably is secondary to an endogenously generated prostaglandin. Since EPA mimics the effects of natural prostaglandin endoperoxides on smooth muscle in vitro, it is possible that prostaglandin endoperoxide-induced vasoconstriction in vivo modulates the effects of their vasodilatory products, prostaglandin E2 and I2.


The effect of neuroleptic treatments on executive function and symptomatology in schizophrenia: a 1-year follow up study.
Cognitive dysfunctions (as in memory, attention and executive function) have been recognized as fundamental features of schizophrenia. Executive dysfunction is a major obstacle to functional outcome, community functioning and rehabilitation success and it is crucial to assess the effects of so-called neuroleptic (NLP) medications in this domain of cognitive functioning. Risperidone, an atypical NLP, has been reported to improve executive function in schizophrenia (SZ), but there is controversy regarding these findings. The aim of the current study was to assess the differential effects of risperidone (2-6 mg) and conventional (2-40 mg haloperidol) NLPs on executive skills in 31 individuals with SZ over a 12-month period. The performance of both NLP groups was compared to the performance of 17 age- and education-matched healthy controls. In this randomized, double blind study, the Wisconsin Card Sorting Test (WCST) was administered at baseline, 3, 6, and 12 months after initiating medication. The relationship between executive functioning and the course of clinical symptoms, as assessed by the Positive and Negative Syndrome Scale (PANSS) was also investigated. Results showed that, relative to healthy controls, individuals with SZ showed marked impairment in WCST from baseline through 12 months of treatment. Also, participants under haloperidol or risperidone NLP medication performed similarly on the WCST at all assessment periods showing that risperidone and haloperidol do not differ in their effect on executive functioning. Risperidone treatment, however, was more effective in the reduction of negative symptoms. The differential efficacy of risperidone over negative symptoms and WCST performance strongly suggests that the executive impairments are to some extent the result of brain abnormalities independent of those that produce the major psychopathology manifestations seen in SZ.


Patterns of neuroglial proliferation in spinal cord white matter following exposure to ionizing radiation.
X-irradiation temporarily decreases the proliferative activity of neuroglia in immature rat spinal cord. Later, the proliferative activity in these irradiated regions surpasses that noted in control rats. Areas adjacent to the irradiated region have a greater than normal percentage of labelled neuroglia and may also be a source of neuroglia which re-populate the irradiated zone.


The bactericidal activity of a deuteroporphyrin-hemin mixture on gram-positive bacteria. A microbiological and spectroscopic study.
The combined antibacterial activity of various porphyrins with hemin on Gram-positive bacteria was studied. Protoporphyrin, hematoporphyrin derivative and deuteroporphyrin show only a marginal inhibitory effect in the dark. However, hemin has a strong cytotoxic effect which is independent of illumination and is equally strong in the dark. The disadvantage of hemin treatment is that it is temporary. In this study, we have demonstrated that a combination of deuteroporphyrin and hemin has a unique cytotoxic activity on Staphylococcus aureus, Streptococcus faecalis and Bacillus cereus. The effect of the combined compound is stronger than that of the separate constituents, and is as strong in the dark as in the light. Only 0.005% of the initial S. aureus population survive after a 2 h treatment. Absorption and fluorescence spectra of hemin-deuteroporphyrin mixtures in water and liposomes suggest the formation of a species with spectroscopic properties which are different from those of the two constituents.


Psychiatric symptoms and mental changes as major features of multiple sclerosis.
A patient is presented with clinically and laboratory supported definite multiple sclerosis who developed severe depression followed by mental deterioration. Magnetic resonance imaging MRI demonstrated multiple hemispherical lesions. It is suggested that the psychiatric and cognitive signs and symptoms observed in this patient are due to severe cerebral demyelination.