Rapid overgrowth of a fetus after interruption of insulin therapy in a diabetic woman.
We experienced a case of fetal overgrowth probably due to maternal hyperglycemia following the interruption of insulin therapy in a woman with gestational diabetes mellitus (GDM), probably undiagnosed type 2 diabetes mellitus. A 27-year-old Japanese woman was referred to our center because of GDM at 15 weeks of gestation. During the course of her educational admission, diet control and insulin therapy resulted in favorable blood sugar levels and she was discharged at 17 weeks of gestation. During these periods, fetal growth remained below average at between -1 and -1.5 S.D. After a check-up in the 29th week, she failed to appear for any further appointments. After the onset of labor, at 38 weeks of gestation, she suddenly consulted her former physician who delivered a baby weighing 4852g by cesarean section on the 38th week. A noteworthy point in this case is the growth curve of the fetus. Even if rapid overgrowth is assumed, starting after the time of interruption of insulin at 30 weeks of gestation, fetal weight gain appears to be about 400g per week, suggesting a weight increase at twice the normal rate. This case prompted us to keep in mind that rigid glycemic control will prevent the development of fetal macrosomia in diabetic pregnant women.


High-rate biohydrogen production from xylose using a dynamic membrane bioreactor.
This study aimed a high-rate dark fermentative H2 production from xylose using a dynamic membrane module bioreactor (DMBR) with a 444-μm pore polyester mesh. 20 g xylose/L was fed continuously to the DMBR at different hydraulic retention times (HRTs) from 12 to 3 h at 37 °C. The maximum average H2 yield (HY) and H2 production rate (HPR) at 3 h HRT were found to be 1.40 ± 0.07 mol H2/mol xyloseconsumed and 30.26 ± 1.19 L H2/L-d, respectively. The short HRT resulted in the maximum suspended biomass concentration (8.92 ± 0.40 g VSS/L) along with significant attached biomass retention (7.88 ± 0.22 g VSS/L). H2 was produced by both butyric and acetic acid pathways. Low HY was concurrent with lactic acid production. The bacterial population shifted from non-H2 producers, such as Lactobacillus and Sporolactobacillus spp., to Clostridium sp., when HY increased. Thus, xylose from lignocellulose is a feasible substrate for dark fermentative H2 production using DMBR.


Metastasis of breast tumor cells to brain is suppressed by targeting RLIP alone and in combination with 2'-Hydroxyflavanone.
Brain metastasis is an important cause of morbidity and mortality in cancer-patients. Breast tumor cells frequently metastasize to brain and initiate severe therapeutic complications. In the present study, we evaluated the anti-metastatic effects of 2'-hydroxyflavanone (2HF) alone and in combination with RLIP targeted therapy in a novel murine model of breast tumor metastasis. The MDA-MB231Br (brain-seeking) breast cancer (BC) cells stably-transfected with luciferase were injected into the left-ventricle of NSG mouse heart and the migration of cells to brain was monitored using a non-invasive bioluminescent imaging system. To evaluate the tumor growth suppressive effects, mice were given 2HF (50 mg/kg, b.w., alternate days orally), RLIP-antibody (Rab; 5 mg/kg, b.w., weekly i.p.) or combination of 2HF+Rab starting day1 after intra-cardiac injection. Our results reveal that 2HF and Rab significantly prevented the metastasis of BC cells to brain. Further, mice treated with combination of 2HF+Rab exhibited no metastasis as compared to either or the single agent-treated mice. This study for the first time demonstrates the anti-metastatic effects of 2HF and RLIP-inhibition in-vivo in a novel breast tumor metastasis model and provides the rationale for further clinical investigation.


p38α in the preoptic area inhibits brown adipose tissue thermogenesis.
Elevation of energy expenditure through an increase of brown adipose tissue (BAT) thermogenesis is regarded as one of the most promising ways to prevent obesity development. The preoptic area (POA) of the hypothalamus is a critical area for control of BAT thermogenesis. However, the intracellular signaling cascades in the POA for regulation of BAT thermogenesis are poorly understood. Phosphorylation proteomics (phosphoproteomics) and bioinformatics approaches were used to disclose numerous hypothalamic signaling pathways involved in the regulation of BAT thermogenesis. Conditional manipulation of the p38α gene in mouse POA was performed by stereotaxic injection of adeno-associated virus 9 vector to explore the role of p38α in BAT thermogenesis. Multiple hypothalamic signaling pathways were triggered by cold exposure, especially the mitogen-activated protein kinase (MAPK) signaling pathway. The p38α activation, but not extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun NH2-terminal kinase (JNK), in the hypothalamus was significantly decreased during cold exposure. p38α deficiency in the POA dramatically elevated energy expenditure owing to a marked increase in BAT thermogenesis, resulting in significantly decreased body weight gain and fat mass. Overexpression of p38α in the POA led to a dramatic increase in weight gain. These results demonstrate that p38α in the POA exacerbates obesity development, at least in part owing to a decrease in BAT thermogenesis.


Fetal origin of adverse pregnancy outcome: the water disinfectant by-product chloroacetonitrile induces oxidative stress and apoptosis in mouse fetal brain.
Epidemiological studies indicate a relationship between water disinfectant by-products (DBP) and adverse pregnancy outcomes (APO) including neural tube defects. These studies suggest that fetal brain may be vulnerable to DBP during early stages of development. Therefore, we examined several molecular markers commonly known to indicate chemical-induced neurotoxicity during fetal brain development following prenatal exposure to the DBP; chloroacetonitrile (CAN). Pregnant mice, at gestation day 6 (GD6), were treated with a daily oral dose of CAN (25 mg/kg). At GD12, two groups of animals were treated with an i.v. tracer dose of [2-14C]-CAN. These animals were sacrificed at 1 and 24 h after treatment and processed for quantitative in situ micro-whole-body autoradiography. The remaining groups of animals continued to receive CAN. At GD18, control and treated animals were weighed, anesthetized, and fetuses were obtained and their brains were removed for biochemical and immunohistochemical analyses. Whole-body autoradiography studies indicate a significant uptake and retention of [2-14C]-CAN/metabolites (M) in fetal brain (cerebral cortex, hippocampus, cerebellum) at 1 and 24 h. There was a 20% reduction in body weight and a 22% reduction in brain weight of fetuses exposed to CAN compared to controls. A significant increase in oxidative stress markers was observed in various fetal brain regions in animals exposed to CAN compared to controls. This was indicated by a 3- to 4-fold decrease in the ratio of the reduced to oxidized form of glutathione (GSH/GSSG), increased lipid peroxidation (1.3-fold), and increased 8-hydroxy-2-deoxyguanosine levels (1.4-fold). Cupric silver staining indicated a significant increase in the number of degenerating neurons in cortical regions in exposed animals. In animals exposed to CAN there was increase in nuclear DNA fragmentation (TUNEL staining) detected in the cerebral cortex and cerebellum (2-fold increase in apoptotic indices). Caspase-3 activity in cerebral cortex and cerebellum of treated animals were also increased (1.7- and 1.5-fold, respectively). In conclusion, this study indicates that CAN/M crossed the placenta and accumulated in fetal brain tissues where it caused oxidative stress and neuronal apoptosis. These events could predispose the fetus to altered brain development leading to APO as well as behavioral and learning and memory deficits.


Secondary hyperalgesia is not affected by wound infiltration with bupivacaine.
The purpose of this study was to determine the effects of wound infiltration with bupivacaine on incisional pain and the zone of secondary hyperalgesia. Twenty-eight healthy parturients were studied in a double-blind randomized trial. At the time of Caesarean section one wound edge was infiltrated with saline 0.9% and the other with bupivacaine 0.25%. After 24 hr, visual analogue scores were obtained for incisional pain and the zone of secondary pain around the incision was measured. Patients served as their own controls. Visual analogue scores for the bupivacaine side of the wound were less than for the saline side (P < 0.05). The zone of secondary pain was similar overall for both sides of the wound. It is concluded that the bupivacaine-infiltrated side of the wound was less painful than the saline-injected side 24 hr postoperatively. The zone of pain measured around the wound edges was unaffected by bupivacaine or saline.


An improved method for detection and titration of antibodies cytophilic for macrophages.
A new microtechnique for detection and titration of antibodies cytophilic for homologous macrophages is introduced and experiments designed to determine the optimum conditions for its performance reported. The technique employs a series of small chambers formed on a microscope slide by the application of a plastic film in which holes have been punched. The procedure uses 5 x 10(4) cells and 25 microliter of serum per chamber, and each test can be completed in 2 h. A permanent preparation results which can be examined as and when convenient.


Qualitative user evaluation of a revised pharmacogenetic educational toolkit.
Pharmacogenetic (PGx) testing is a leading application for personalized and precision medicine; however, there are barriers, including limited provider and patient understanding, which affect its uptake. There is a need for tools that can enhance the patient and provider experience with testing and promoting the shared and informed decision-making. In this study, we sought to gather additional feedback on a PGx toolkit comprised of four educational tools that had been previously evaluated through an online survey by pharmacists. Specifically, we conducted semi-structured interviews with pharmacists and members of the public regarding their understanding and utility of the toolkit and its individual components. Participants found three of the four toolkit components, a test information sheet, flipbook, and results sheet, to be useful and important. The fourth component, results card, was viewed less favorably. Participants differed in their preference for medical jargon and detailed results nomenclature (namely star * alleles). User input during the development of educational materials is essential for optimizing utilization, effectiveness, and comprehension.


Comparison of diffusion-weighted MRI and [18F]FDG PET/MRI for treatment monitoring in pediatric Hodgkin and non-Hodgkin lymphoma.
To compare tumor therapy response assessments with whole-body diffusion-weighted imaging (WB-DWI) and 18F-fluorodeoxyglucose ([18F]FDG) PET/MRI in pediatric patients with Hodgkin lymphoma and non-Hodgkin lymphoma. In a retrospective, non-randomized single-center study, we reviewed serial simultaneous WB-DWI and [18F]FDG PET/MRI scans of 45 children and young adults (27 males; mean age, 13 years ± 5 [standard deviation]; age range, 1-21 years) with Hodgkin lymphoma (n = 20) and non-Hodgkin lymphoma (n = 25) between February 2018 and October 2022. We measured minimum tumor apparent diffusion coefficient (ADCmin) and maximum standardized uptake value (SUVmax) of up to six target lesions and assessed therapy response according to Lugano criteria and modified criteria for WB-DWI. We evaluated the agreement between WB-DWI- and [18F]FDG PET/MRI-based response classifications with Gwet's agreement coefficient (AC). After induction chemotherapy, 95% (19 of 20) of patients with Hodgkin lymphoma and 72% (18 of 25) of patients with non-Hodgkin lymphoma showed concordant response in tumor metabolism and proton diffusion. We found a high agreement between treatment response assessments on WB-DWI and [18F]FDG PET/MRI (Gwet's AC = 0.94; 95% confidence interval [CI]: 0.82, 1.00) in patients with Hodgkin lymphoma, and a lower agreement for patients with non-Hodgkin lymphoma (Gwet's AC = 0.66; 95% CI: 0.43, 0.90). After completion of therapy, there was an excellent agreement between WB-DWI and [18F]FDG PET/MRI response assessments (Gwet's AC = 0.97; 95% CI: 0.91, 1). Therapy response of Hodgkin lymphoma can be evaluated with either [18F]FDG PET or WB-DWI, whereas patients with non-Hodgkin lymphoma may benefit from a combined approach. Hodgkin lymphoma and non-Hodgkin lymphoma exhibit different patterns of tumor response to induction chemotherapy on diffusion-weighted MRI and PET/MRI. • Diffusion-weighted imaging has been proposed as an alternative imaging to assess tumor response without ionizing radiation. • After induction therapy, whole-body diffusion-weighted imaging and PET/MRI revealed a higher agreement in patients with Hodgkin lymphoma than in those with non-Hodgkin lymphoma. • At the end of therapy, whole-body diffusion-weighted imaging and PET/MRI revealed an excellent agreement for overall tumor therapy responses for all lymphoma types.


Comparative efficacy and tolerability of monotherapy with leflunomide or tacrolimus for the treatment of rheumatoid arthritis: a Bayesian network meta-analysis of randomized controlled trials.
We aimed to assess the relative efficacy and tolerability of monotherapy with leflunomide or tacrolimus at recommended dosages in rheumatoid arthritis (RA) patients. Randomized controlled trials (RCTs) examining the efficacy and tolerability of leflunomide 20 mg, leflunomide 10 mg, tacrolimus 3 mg, tacrolimus 1.5-2 mg, and placebo, based on the number of withdrawals of RA patients, were included. We performed a Bayesian random-effects network meta-analysis to combine direct and indirect evidence from the RCTs. Six RCTs including 1510 patients met the inclusion criteria. The proportion of patient withdrawals owing to lack of efficacy was significantly lower in the leflunomide 20 mg (OR 0.17, 95% credible interval (CrI) 0.08-0.34); leflunomide 10 mg (OR 0.16, 95% CrI 0.02-0.75); and tacrolimus 3 mg (OR 0.41, 95% CrI 0.21-0.74) groups than in the placebo group. Rank probability based on the surface under the cumulative ranking curve (SUCRA) values indicated that leflunomide 20 mg had the highest probability of being the best treatment based on the number of withdrawals owing to lack of efficacy (SUCRA = 0.8530), followed by leflunomide 10 mg (SUCRA = 0.8321), tacrolimus 3 mg (SUCRA = 0.4965), tacrolimus 1.5-2 mg (SUCRA = 0.3035), and placebo (SUCRA = 0.0150). Patient withdrawals owing to adverse events did not differ significantly among the groups; however, withdrawals in the placebo group were fewer than those in the leflunomide 20 mg group (OR 0.22, 95% CrI 0.07-0.74). Placebo had the highest probability of being the most tolerable treatment (SUCRA = 0.8161) followed by tacrolimus 3 mg (SUCRA = 0.6490), tacrolimus 1.5-2 mg (SUCRA = 0.4857), leflunomide 10 mg (SUCRA = 0.4651), and leflunomide 20 mg (SUCRA = 0.0841). Leflunomide 20 mg, leflunomide 10 mg, and tacrolimus 3 mg were more efficacious than placebo, while leflunomide 20 mg was less tolerable than placebo. Leflunomide is likely to be more efficacious but less tolerable than tacrolimus for RA treatment.


Riluzole ameliorates soluble Aβ1-42-induced impairments in spatial memory by modulating the glutamatergic/GABAergic balance in the dentate gyrus.
Soluble amyloid beta (Aβ) is believed to contribute to cognitive deficits in the early stages of Alzheimer's disease (AD). Increased soluble Aβ1-42 in the hippocampus is closely correlated with spatial learning and memory deficits in AD. Riluzole (RLZ), an FDA-approved drug for amyotrophic lateral sclerosis (ALS), has beneficial effects for AD. However, the mechanism underlying the effects remains unclear. In this study, its neuroprotective effect against soluble Aβ1-42-induced spatial cognitive deficits in rats was assessed. We found that intrahippocampal injection of soluble Aβ1-42 impaired spatial cognitive function and suppressed long-term potentiation (LTP) of the DG region, which was relevant to soluble Aβ1-42-induced shift of the hippocampal excitation/inhibition balance toward excitation. Interestingly, RLZ ameliorated Aβ1-42-induced behavioral and LTP impairments through rescuing the soluble Aβ1-42-induced excitation/inhibition imbalance. RLZ attenuated Aβ1-42-mediated facilitation of excitatory synaptic transmission by facilitating glutamate reuptake and decreasing presynaptic glutamate release. Meanwhile, RLZ attenuated the suppression of inhibitory synaptic transmission caused by Aβ1-42 by potentiating postsynaptic GABA receptor function. These results suggest that RLZ exerts a neuroprotective effect against soluble Aβ1-42-related spatial cognitive deficits through rescuing the excitation/inhibition imbalance, and it could be a potential therapy for AD.


Visualizing Cancer Cell Chemotaxis and Invasion in 2D and 3D.
We describe three chemotaxis assays-Insall chambers, circular invasion assays, and 3D organotypic assays-that are particularly appropriate for measuring migration of cancer cells in response to gradients of soluble attractants. Each assay has defined advantages, and together they provide the best possible quantitative assessment of cancer chemotaxis.


Men with biopsy-confirmed hepatocellular adenoma have a high risk of progression to hepatocellular carcinoma: A nationwide population-based study.
Hepatocellular adenoma is a benign liver tumour that may transform to hepatocellular carcinoma (HCC). We used data from Danish nationwide healthcare registries to investigate the incidence and prognosis of hepatocellular adenoma. We included all patients with a hospital discharge diagnosis for benign liver tumour (ICD-10: D13.4) in 1997-2012 and a liver biopsy confirming the hepatocellular adenoma diagnosis. Follow-up began 1 year after adenoma diagnosis, to minimise the possibility that the tumour was a misdiagnosed HCC. All patients were age- and gender-matched with 50 random controls from the Danish population. We followed patients and controls with respect to HCC development, adenoma resection, and death without HCC (ie, death without having been diagnosed with HCC) through 2013. HCC diagnoses were identified in the Danish Cancer Registry. We included 67 patients with hepatocellular adenoma, and 58 (87%) were women. The overall incidence rate of histologically verified hepatocellular adenoma in the Danish general population was 0.07 (95% CI: 0.06-0.09) per 100 000 population per year. Fifteen patients had their adenoma resected before follow-up began, leaving 52 patients for follow-up. Men with biopsy-confirmed hepatocellular adenoma had a 10-year cumulative HCC risk as high as 60.0% (95% CI: 15.3%-87.0%). All men who developed HCC were older than 50 years at adenoma diagnosis. By contrast, none of the 44 women in the follow-up analysis developed HCC. Histologically verified hepatocellular adenoma is rare in Denmark. It is a minor concern for women, but men have a very high risk of progression to HCC.


Primary repair of acute ascending aortic dissection.
Primary repair of acute ascending aortic dissection was performed in 14 patients. Repair included resection of the intimal tear where applicable, a circumferential suture line in the ascending aorta at the site of the tear, and wrapping of the intrapericardial ascending aorta with Teflon felt to contain the distal residual false channel. The aortic valve was resuspended in 6 patients. The single operative death was unrelated to the method of repair. Two late deaths at 4 and 6 years were due to preexisting multisystem disease. In 1 patient, new aortic insufficiency with an isolated aneurysm of the noncoronary sinus of Valsalva developed at 26 months, and was repaired successfully at another institution. These results compare favorably with those reported by others employing more extensive surgical procedures for repair of acute ascending aortic dissection.


Molecular characterization of pathogenic Fusarium species in cucurbit plants from Kermanshah province, Iran.
Fusarium is one of the important phytopathogenic genera of microfungi causing serious losses on cucurbit plants in Kermanshah province, the largest area of cucurbits plantation in Iran. Therefore, the objectives in this study were to isolate and identify disease-causing Fusarium spp. from infected cucurbit plants, to ascertain their pathogenicity, and to determine their phylogenetic relationships. A total of 100 Fusarium isolates were obtained from diseased cucurbit plants collected from fields in different geographic regions in Kermanshah province, Iran. According to morphological characters, all isolates were identified as Fusarium oxysporum, Fusarium proliferatum, Fusarium equiseti, Fusarium semitectum and Fusarium solani. All isolates of the five Fusarium spp. were evaluated for their pathogenicity on healthy cucumber (Cucumis sativus) and honeydew melon (Cucumis melo) seedlings in the glasshouse. F. oxysporum caused damping-off in 20-35 days on both cucurbit seedlings tested. Typical stem rot symptoms were observed within 15 days after inoculation with F. solani on both seedlings. Based on the internal transcribed spacer (ITS) regions of ribosomal DNA (rDNA) restriction fragment length polymorphism (RFLP) analysis, the five Fusarium species were divided into two major groups. In particular, isolates belonging to the F. solani species complex (FSSC) were separated into two RFLP types. Grouping among Fusarium strains derived from restriction analysis was in agreement with criteria used in morphological classification. Therefore, the PCR-ITS-RFLP method provides a simple and rapid procedure for the differentiation of Fusarium strains at species level. This is the first report on identification and pathogenicity of major plant pathogenic Fusarium spp. causing root and stem rot on cucurbits in Iran.


A pioneer study on the anti-ulcer activities of copper nicotinate complex [CuCl (HNA)2] in experimental gastric ulcer induced by aspirin-pylorus [corrected] ligation model (Shay model).
This study was planned to look for a more rational mechanism(s) that could explain the anti-ulcer activity of copper nicotinate complex, since the mechanism whereby it prevents ulceration still to be investigated. Ulcer and the preventive indexes were scored, mucin, juice volume, total acidity, luminal haemoglobin, total peroxide and total antioxidant as an oxidative stress index (OSI), as well as total DNA fragmentation (as an apoptotic marker) and its percentage ratio in the juice were evaluated. Results confirmed the antisecretory ability of copper nicotinate, as reflected from the significant reduction of gastric juice volume and acid output. Data confirmed also copper nicotinate has a gastrocytoprotective action against ulcer pathogenesis through; enhancement of mucin secretion, reduction each of ulcer index and the mucosal bleeding rate into the gastric lumen; antioxidant activity through scavenging reactive oxygen species (ROS); anti-apoptotic activity through attenuation of the DNA fragmentation in the ulcerous treated group when compared with each of control and aspirin ulcerous untreated groups. This study hypothesized; at least in part, aspirin in the acidic environment of gastric juice become un-ionized and freely penetrate the mucosal barrier reaching to gastric wall. Due to the weak basic nature of cytoplasm of gastric mucosal cells, it accumulates in high concentrations into mucosal cells, and yields a negatively charged anion that is unable to exit the cell. Thus, superficial or deeper erosions are produced and bleeding takes place, within minutes.


Optimization of management by analyzing ecosystem service value variations in different watersheds in the Three-River Headwaters Basin.
Variation analysis of ecosystem services (ESs) is an important means of regional watershed management, especially for the ecologically fragile Three-River Headwaters Basin (TRHB), which is an important part of the national ecological security barrier. In this study, meteorological data, remote sensing images and land use data from 2000 to 2020 in the TRHB were collected. Based on the estimation of ecosystem service values (ESVs), the spatial-temporal variations of ESVs in the three watersheds were analyzed via spatial autocorrelation analysis, one-way analysis of variance (ANOVA), and correspondence analysis. The study indicates that for the 20-year changes in the TRHB, the provisioning service (PS) and regulating service (RS) increased in most areas except for a small decrease in the northwest, while the supporting service (SS) value in most areas in the southwest showed a decreasing trend. Through the difference results of different watersheds, it can be seen that the differences in the PS and RS values among the three watersheds became increasingly more significant with the passage of time. Moreover, a significant correspondence was identified between each watershed and each ESV level, whereby the highest ESV levels (SS-VI, RS-VI, and PS-VI) mainly corresponded to the Yellow River watershed, the high values (SS-V, RS-V, PS-V, SS-IV, RS-IV, and PS-IV) mainly corresponded to the Lancang River watershed, and the low values (SS-I, RS-I, and PS-I) corresponded to the Yangtze River watershed. Our analysis shows the variation characteristics of ESVs to provide a guiding basis for ensuring the precise implementation of the management and protection of watersheds in the TRHB or similar areas.


Effect of propeptide mutations on post-translational processing of factor IX. Evidence that beta-hydroxylation and gamma-carboxylation are independent events.
Post-translational processing of Factor IX includes glycosylation, cleavage of the signal peptide and propeptide, vitamin K-dependent carboxylation of specific glutamic acid residues to form gamma-carboxyglutamic acid, and beta-hydroxylation of aspartic acid at residue 64 to form beta-hydroxyaspartic acid. The human Factor IX cDNA coding sequence was modified in the propeptide region (residue -18 to -1) using oligonucleotide-directed site-specific mutagenesis, and the altered Factor IX cDNA was expressed in Chinese hamster ovary cells. The effects of the mutations on proteolytic processing, gamma-carboxylation, and beta-hydroxylation were assessed by direct structural analysis. After purification, the molecular weight of each of the recombinant Factor IX species and its NH2-terminal amino acid sequence were shown to be identical to those of plasma Factor IX. gamma-Carboxyglutamic acid and beta-hydroxyaspartic acid analyses revealed that recombinant wild-type Factor IX contained 9.2 gamma-carboxyglutamic acid and 0.3 beta-hydroxyaspartic acid residues/molecule compared with 11.4 gamma-carboxyglutamic acid and 0.39 beta-hydroxyaspartic acid residues in plasma Factor IX. When the 18-residue propeptide was deleted or when the cells were grown in the presence of sodium warfarin, secreted Factor IX contained no detectable gamma-carboxyglutamic acid but 0.36 and 0.40 residues of beta-hydroxyaspartic acid, respectively. Point mutations leading to substitution of alanine for phenylalanine at residue -16 or glutamic acid for alanine at residue -10 contained 0.2 and 1.7 gamma-carboxyglutamic acid residues, respectively, and 0.2 residues of beta-hydroxyaspartic acid. These data confirm that the propeptide mutations made do not interfere with proteolytic processing and that the Factor IX propeptide contains a recognition site that designates the adjacent glutamic acid-rich domain for gamma-carboxylation. In contrast, beta-hydroxylation of aspartic acid 64 is an independent process which does not require vitamin K and is mediated through a hydroxylation recognition site in the mature Factor IX, not in the propeptide.


Proximal junctional kyphosis in pediatric spinal deformity surgery: a systematic review and critical analysis.
Proximal junctional kyphosis (PJK) is a commonly encountered clinical and radiographic phenomenon after pediatric and adolescent spinal deformity surgery that may lead to post-operative deformity, pain, and dissatisfaction. Understanding the risk factors of PJK can be useful for pre-operative informed consent as well as to identify any potential preventative strategies. We performed a systematic review and critical analysis following the PRISMA statement in July 2019 by searching the PubMed, Scopus, and Embase databases, including all prior published studies. We included articles with data on PJK in patients with operative pediatric and adolescent scoliosis and those that detailed risk factors and/or preventative strategies for PJK. Levels of evidence were determined based on consensus. Findings were summarized and grades of recommendation were assigned by consensus. This study was registered in the PROSPERO database; 202,457. Six hundred and thirty five studies were identified. Thirty-seven studies met criteria for inclusion into the analysis. No studies including neuromuscular scoliosis met inclusion criteria. No findings had Grade A evidence. There were 4 findings found to contribute to PJK with Grade B evidence in EOS: higher number of distractions, disruption of posterior elements, greater sagittal plane correction. There was no difference in incidence noted between etiology of the curvature. Five findings with Grade B evidence were found to contribute to PJK in AIS populations: higher pre-operative thoracic kyphosis, higher pre-operative lumbar lordosis, longer fusion constructs, greater sagittal plane correction, and posterior versus anterior fusion constructs. Greater sagittal plane correction has Grade B evidence as a risk factor for PJK in both EOS and AIS populations. In EOS patients, an increased number of distractions and posterior element disruption are Grade B risk factors. In AIS patients, longer fusion constructs, higher pre-operative thoracic kyphosis and lumbar lordosis, and posterior (as opposed to anterior) constructs also contributed to PJK with Grade B evidence. These findings can guide informed consent and surgical management, and provide the foundation for future studies.


FACTORS AND ESTIMATION OF RISK FOR CARDIOVASCULAR DISEASES AMONG PATIENTS IN PRIMARY HEALTH CARE IN CENTRAL SERBIA.
Cardiovascular diseases present major cause of morbidity and mortality in the world as well as in Central Serbia. The aim of this study is to analyze the frequency of risk factors and estimate risk for the development of cardiovascular diseases. The database was collected from health records of the Health Centre in Kragujevac. The following parameters were observed: gender and age, smoking, anthropometrical measurements and Body Mass Index, the value of systolic and diastolic blood pressure, diabetes, levels of cholesterol, and triglycerids in blood serum. The estimation of risk for developing the fatal cardiovascular disease during the period of ten years is determined according to SCORE system model. The statistical package SPSS version 18.0 was used for the statistical analysis. The study sample consisted of 1,053 people, 505 men and 548 women. 68.8% of people were overweight and obese, arterial hypertension was noted in 72.6%, and diabetes type II in 13.3% of people. 67.6% of people had a high level of total cholesterol in blood serum and 39.6% of people had a high level of triglycerids in blood serum. The metabolic syndrome was noted in 44.3%, and 3 or more risk factors for cardiovascular disease were noted in 65% of the population. We determined a significant statistic difference for age, smoking, nutritional status, dyslipidemia, and diabetes. The high incidence of potentially modifiable risk factors for cardiovascular disease gives a possibility to apply prevention measures for promotion and preservation of health.


The purification of cytochrome f and plastocyanin using affinity chromatography.
Both plastocyanin and cytochrome f were purified using a combination of affinity chromatography together with established methods. Plastocyanin was partially purified using the method of Davis and San Pietro (Anal. Biochem. 95 (1979) 254-259), after which it was further purified using a column of cytochrome c covalently attached to Sepharose 4B. The affinity column was prepared using the method of Godinot and Gautheron (Methods Enzymol. 54 (1979) 112-114). The final purity index ratio (A278/A597) was less than 1.2, which is equal to that obtained using the more expensive FPLC procedure (Anderson, G.P., Sanderson, D.G., Lee, C.H., Durell, S., Anderson, L.B. and Gross, E.L. (1987) Biochim. Biophys. Acta 894, issue 3). Cytochrome f was partially purified using a modification of the method of Matazaki et al. (Plant Cell. Physiol. 16 (1975) 237-246) and bound to an affinity column of plastocyanin covalently attached to Sepharose 4B. Cytochrome f purified using this procedure had a purity index ratio (A554.5/A277) of 1.2. Both proteins are tyrosine proteins containing no tryptophan residues. After the affinity chromatography step, the fluorescence emission spectrum of either plastocyanin or cytochrome f was typical of a tyrosine protein free from tryptophan contamination.


Recurrent versus new-onset depressive symptoms: Relationships with biomarkers of cardiovascular health following acute coronary syndrome.
New-onset depressive symptoms commonly arise among persons without a history of major depressive disorder (MDD) in the setting of acute medical illness. Although depressive symptoms in general are associated with alterations in prognostic biomarkers following acute coronary syndrome (ACS), the nature of specifically new-onset depressive symptoms is less well-characterized. It is unclear whether such symptoms neurobiologically resemble recurrent symptoms of MDD or instead represent a distinct condition. In this exploratory analysis, we aimed to examine the effects of prior MDD history on the relationships between post-ACS depressive symptoms and cardiovascular biomarkers. One-hundred sixty-four participants attended study visits 2 weeks and 6 months after ACS to complete self-report measures and provide biomarker samples. MDD history was identified by a psychiatrist through systematic electronic medical record review. Generalized estimating equations were performed to examine the moderating effects of MDD history on concurrent relationships between depressive symptoms and several biomarkers (endothelin-1, soluble intercellular adhesion molecule-1, high-sensitivity C-reactive protein, interleukin-6, and tumor necrosis factor-alpha). Twenty percent (n = 33) of participants had a history of MDD. Depressive symptoms were more strongly associated with levels of endothelin-1 in patients with prior MDD compared to those without (B = 0.024, 95% CI [0.005, 0.043], p = .012), adjusting for age, sex, medical factors, and anxiety. MDD history did not moderate relationships between depressive symptoms and other biomarkers. Recurrent post-ACS depressive symptoms are more strongly associated with elevated endothelin-1 levels than new-onset symptoms. Further work is needed to clarify the mechanism and clinical implications of this relationship.


Thoracic radiation therapy could give survival benefit to elderly patients with extensive-stage small-cell lung cancer.
To evaluate the effect of thoracic radiation therapy (TRT) in elderly patients (aged ≥65 years) with extensive-stage small-cell lung cancer (ES-SCLC). This study reviewed the records of 118 elderly patients with ES-SCLC (all with distant metastasis) through January 2006-December 2013. The patients were divided into either a chemotherapy (ChT)/TRT-combination group or a ChT-alone group. The median survival time and 3-year overall survival rates in the ChT/TRT group were significantly higher than those in the ChT-alone group (17.0 vs 11.7 months; 18.1 vs 14.9%; p = 0.014). The 3-year overall survival rates in patients who received prophylactic cranial irradiation were also higher than the rates in patients who did not receive prophylactic cranial irradiation (18.1 vs 5.1%; p = 0.708). TRT combined with ChT could provide a survival benefit to elderly patients with ES-SCLC.


Teacher's perceptions, institutional challenges, and educational sustainability during Covid-19 in Ecuador.
The Covid-19 pandemic has affected global educational systems, especially in developing countries. Therefore, governments and educational administrators have adopted contingency measures to maintain the functioning and the sustainability of their national educational systems. These measures involved the use of technologies and have enabled a significant amount of teachers to continue working with their students around the globe. This article aims to analyze the perception of Ecuadorian teachers about the contingency measures taken at the governmental and institutional levels while facing the COVID-19 crisis. Also, several aspects regarding the teachers' perceptions about their technological skills, access to electronic devices, and implications of teleworking in their mental health and performance are also analyzed. A cross-sectional study was carried out with 3,183 Ecuadorian teachers working at all levels of education. An online questionnaire which obtained a reliability of 0.84 in Cronbach's Alpha was used. The results show that despite the high levels of uncertainty, most teachers agreed with the measures that had been taken by the Government, the Ministry of Education as well as their institutions and reported feeling comfortable with teleworking. However, the lack of technological resources in rural areas and the lack of trust in the effectiveness of online classes are aspects that need to be analyzed before considering the viability of long-term online education in Ecuador.


Bone marrow transplantation enhances trafficking of host-derived myelomonocytic cells that rescue intestinal mucosa after whole body radiation.
Bone marrow (BM)-derived cells were demonstrated within intestines after radiation damage and were reported to be responsible for intestine repair. However, there was a discrepancy between intestine epithelial clonogenic regeneration, and mouse survival after BM transplantation (BMT) and radiation. The contribution of BM to acute intestine repair after radiation needed further investigation. Mouse survival, intestine microcolony assay, immunohistochemical studies of both intestine and BM were evaluated in mice after whole body irradiation (WBI) and BMT. Immunoblotting, flowcytometry, and double immunostaining were used to evaluate the amount and the character of stroma cells within intestines of recipient mice after receiving gender-mismatched BMT or BMT from green fluorescence donors. Stromal cell proliferation within the lamina propria correlated with the beneficial effect of BMT to intestine recovery and day-8 survival of mice. Few donor-derived cells were found before the completion of intestine repair. The number of host but not donor-derived myelomonocytic and stromal cells increased dramatically within one week after radiation and BMT. Depletion of myelomonocytic cells of recipient mice abolished the mitigating effect of BMT. Besides rescuing injured BM from aplasia, BMT triggers trafficking of host CD11b(+) myelomonocytic cells from the host marrow to the radiation-injured intestinal mucosa, enhancing the proliferation of intestinal stroma cells, leading secondarily to epithelial regeneration.


Use of the polymerase chain reaction to assess the success of visceral leishmaniasis treatment.
We investigated whether the polymerase chain reaction (PCR) performed with aspirates of bone marrow or lymph node can be used as a test of cure of visceral leishmaniasis (VL). Sixty-one VL patients who had received supervised treatment with sodium stibogluconate in the health centre of Médecins sans Frontières (MSF) Holland in Um-Kuraa, eastern Sudan, were studied. Immediately after treatment, no parasite could be demonstrated by microscopy in aspirates of bone marrow or lymph node. In contrast, PCR detected Leishmania deoxyribonucleic acid in 50 of the 61 lymph node aspirates (82%). Forty-nine patients were examined 3 and 6 months later; the other 12 were reported to be alive but had left the area. With 10 of these 49 patients, the PCR was negative and the patients remained free from signs and symptoms of VL; they were apparently cured. Of the 39 patients with a positive PCR after treatment, 14 (36%) developed post kala-azar dermal leishmaniasis and 9 (23%) had a recurrence of VL symptoms with reappearance of parasites in the aspirates. Four relapsed patients subsequently died of VL. We concluded that the PCR on lymph node aspirates can be used to assess treatment and cure of VL. The fact that 23 of 49 patients who received standard supervised treatment were not completely cured indicated that there is a need to investigate extended or alternative treatments.


Novel therapeutic applications of cardiac glycosides.
Cardiac glycosides are a diverse family of naturally derived compounds that bind to and inhibit Na+/K+-ATPase. Members of this family have been in clinical use for many years for the treatment of heart failure and atrial arrhythmia, and the mechanism of their positive inotropic effect is well characterized. Exciting recent findings have suggested additional signalling modes of action of Na+/K+-ATPase, implicating cardiac glycosides in the regulation of several important cellular processes and highlighting potential new therapeutic roles for these compounds in various diseases. Perhaps most notably, the increased susceptibility of cancer cells to these compounds supports their potential use as cancer therapies, and the first generation of glycoside-based anticancer drugs are currently in clinical trials.


Diabetes-specific variables associated with quality of life changes in young diabetic people: the type 1 diabetes Registry of Turin (Italy).
Type 1 diabetes (T1DM) affects young people during the most active years of their life. Our aim was to assess quality of life (QoL) and associated variables in a large cohort of adults with childhood-onset and adult-onset T1DM. A cohort of adult patients (18 years and older) from the T1DM Registry of Turin, Italy, was recruited. Clinical characteristics and Diabetes QoL (DQOL) questionnaire were assessed by standardized procedures. 310 adults completed the questionnaire. Age and diabetes duration at assessment (mean ± SD) were 32.8 ± 7.3 years and 17.3 ± 6.3 years, respectively. DQOL and its subscores were in the lower quartiles of their distributions, indicating a good level of QoL. However, scores were significantly higher in females than in males, particularly for the subscale of diabetes-related worries. In multivariate analysis, lower QoL was independently associated with female sex (β = 1.07, 95% CI 1.03-1.11, p = 0.003), higher age at onset (β = 1.03, 1.00-1.05, p = 0.009), lower schooling (β = 1.05, 1.00-1.09, p = 0.02), higher fasting plasma glucose (β = 1.03, 1.01-1.05, p = 0.008), daily SMBG >4 (β = 1.06, 1.01-1.10, p = 0.01), severe hypoglycemia over the last year (β = 1.06, 1.01-1.11, p = 0.02), lower numbers of diabetologic visits (β = 1.07, 1.01-1.13, p = 0.02) and hypertension (β = 1.06, 1.02-1.10, p = 0.005). Autonomic neuropathy was associated with diabetes impact. Female sex (β = 4.36, 2.43-7.83) and daily SMBG >4 (β = 3.77, 1.72-8.30) were independently associated with worst level and CSII with better level (β = 0.22, 0.07-0.68) of diabetes-related worries. The impact of T1DM on QoL may depend on demographic, metabolic control-related variables, presence of complications and insulin delivery modality.


Acute congestive heart failure: pathophysiological alterations.
The goal of a healthy heart is to transfer blood coming to the ventricles from the low pressure venous system into the high pressure arterial system. In acute congestive heart failure (CHF), impaired cardiac function leads to failure to empty venous reservoirs and therefore to reduce delivery of blood into the arterial circulation (Michaelson 1983). Once heart failure occurs it has a high mortality rate. According to the Framingham Heart Study, 50% of patients with New York Heart Association class II or III heart failure died within 5 years. With class IV patients, 50% survived less than 1 year (Cupples 1987). The purpose of this article is first to define acute CHF. Secondly, the pathophysiological alterations that occur as a result of strained and misdirected compensatory mechanisms will be discussed to help critical care nurses to recognise why certain patients are at risk of developing acute CHF.


Impaired Expression of GABA Signaling Components in the Alzheimer's Disease Middle Temporal Gyrus.
γ-aminobutyric acid (GABA) is the primary inhibitory neurotransmitter, playing a central role in the regulation of cortical excitability and the maintenance of the excitatory/inhibitory (E/I) balance. Several lines of evidence point to a remodeling of the cerebral GABAergic system in Alzheimer's disease (AD), with past studies demonstrating alterations in GABA receptor and transporter expression, GABA synthesizing enzyme activity and focal GABA concentrations in post-mortem tissue. AD is a chronic neurodegenerative disorder with a poorly understood etiology and the temporal cortex is one of the earliest regions in the brain to be affected by AD neurodegeneration. Utilizing NanoString nCounter analysis, we demonstrate here the transcriptional downregulation of several GABA signaling components in the post-mortem human middle temporal gyrus (MTG) in AD, including the GABAA receptor α1, α2, α3, α5, β1, β2, β3, δ, γ2, γ3, and θ subunits and the GABAB receptor 2 (GABABR2) subunit. In addition to this, we note the transcriptional upregulation of the betaine-GABA transporter (BGT1) and GABA transporter 2 (GAT2), and the downregulation of the 67 kDa isoform of glutamate decarboxylase (GAD67), the primary GABA synthesizing enzyme. The functional consequences of these changes require further investigation, but such alterations may underlie disruptions to the E/I balance that are believed to contribute to cognitive decline in AD.


Confinement Assembly of Terpolymer-Based Janus Nanoparticles.
While the confinement assembly of block copolymers into functional microparticles has been extensively studied, little is known about the behavior of Janus nanoparticles (JNPs) in spherical confinement. Here, the confinement self-assembly of JNPs in drying emulsion droplets is investigated and their behavior compared to their ABC triblock terpolymer precursors. Emulsions of both materials are prepared using Shirasu Porous Glass membranes leading to narrow size distributions of the microparticles with average hydrodynamic radii in the range of Rh = 250-500 nm (depending on the membrane pore radius, Rpore ). The internal structure of the microparticles is verified with transmission electron microscopy (TEM) on ultrathin cross sections and compared to the corresponding bulk morphologies. While the confinement assembly of terpolymers results in microparticles with ordered inner morphologies, order for JNPs diminishes when the Janus balance deviates from parity.


Movement discrimination in the rabbit.
In a two-choice discrimination box, rabbits were trained to discriminate vertical vs horizontal striations. After criterion was reached, the same striations were presented again, but now the patterns moved sinusoidally. All animals immediately transferred to moving patterns. After that, it was attempted to train the animals to discriminate between moving vs non-moving vertical striations. Even after prolonged training, none of the animals learned this discrimination task.


Advanced technological tools to study multidrug resistance in cancer.
The complexity of cancer biology and its clinical manifestation are driven by genetic, epigenetic, transcriptomic, proteomic and metabolomic alterations, supported by genomic instability as well as by environmental conditions and lifestyle factors. Although novel therapeutic modalities are being introduced, efficacious cancer therapy is not achieved due to the frequent emergence of distinct mechanisms of multidrug resistance (MDR). Advanced technologies with the potential to identify and characterize cancer MDR could aid in selecting the most efficacious therapeutic regimens and prevent inappropriate treatments of cancer patients. Herein, we aim to present technological tools that will enhance our ability to surmount drug resistance in cancer in the upcoming decade. Some of these tools are already in practice such as next-generation sequencing. Identification of genes and different types of RNAs contributing to the MDR phenotype, as well as their molecular targets, are of paramount importance for the development of new therapeutic strategies aimed to enhance drug response in resistant tumors. Other techniques known for many decades are in the process of adaptation and improvement to study cancer cells' characteristics and biological behavior including atomic force microscopy (AFM) and live-cell imaging. AFM can monitor in real-time single molecules or molecular complexes as well as structural alterations occurring in cancer cells induced upon treatment with various antitumor agents. Cell tracking methodologies and software tools recently progressed towards quantitative analysis of the spatio-temporal dynamics of heterogeneous cancer cell populations and enabled direct monitoring of cells and their descendants in 3D cultures. Besides, novel 3D systems with the advanced mimicking of the in vivo tumor microenvironment are applicable to study different cancer biology phenotypes, particularly drug-resistant and aggressive ones. They are also suitable for investigating new anticancer treatment modalities. The ultimate goal of using phenotype-driven 3D cultures for the investigation of patient biopsies as the most appropriate in vivo mimicking model, can be achieved in the near future.


Synthetic arabinofuranosyl oligosaccharides as mycobacterial arabinosyltransferase substrates.
A series of arabinofuranosyl oligosaccharides found as constituent parts of the polysaccharide portion of the cell wall of Mycobacterium tuberculosis have been chemically synthesized. Screening of these oligosaccharides as substrates for arabinosyltransferases present in mycobacterial membrane preparations suggests that modified oligosaccharide analogs as small as disaccharides may be inhibitors of glycan biosynthesis. Such inhibitors would be of potential utility as lead compounds in the identification of new drugs for the treatment of mycobacterial infections.


A predictive model for creep deformation following vertebral compression fractures.
Many vertebral compression fractures continue to collapse over time, resulting in spinal deformity and chronic back pain. Currently, there is no adequate screening strategy to identify patients at risk of progressive vertebral collapse. This study developed a mathematical model to describe the quantitative relationship between initial bone damage and progressive ("creep") deformation in human vertebrae. The model uses creep rate before damage, and the degree of vertebral bone damage, to predict creep rate of a fractured vertebra following bone damage. Mechanical testing data were obtained from 27 vertebral trabeculae samples, and 38 motion segments, from 26 human spines. These were analysed to evaluate bone damage intensity, and creep rates before and after damage, in order to estimate the model parameter, p, which represents how bone damage affects the change of creep rate after damage. Results of the model showed that p was 1.38 (R2 = 0.72, p < 0.001) for vertebral trabeculae, and 1.48 for motion segments (R2 = 0.22, p = 0.003). These values were not significantly different from each other (P > 0.05). Further analyses revealed that p was not significantly influenced by cortical bone damage, endplate damage, disc degeneration, vertebral size, or vertebral areal bone mineral density (aBMD) (P > 0.05). The key determinant of creep deformation following vertebral compression fracture was the degree of trabecular bone damage. The proposed model could be used to identify the measures of bone damage on routine MR images that are associated with creep deformation so that a screening tool can be developed to predict progressive vertebral collapse following compression fracture.


The behavioral neurobiology of self-injurious behavior in rhesus monkeys.
1. Self-injurious behavior (SIB) is prevalent among institutionalized children, but the efficacy of current behavioral and pharmacological treatments is marginal. 2. There is evidence that SIB in humans has a neurobiological basis. A better understanding of the neurobiological factors that may promote or cause SIB is necessary for the development of effective pharmacologic treatments. 3. SIB that is similar in some respects to SIB in humans occurs in nonhuman primates that have been deprived of social experience early in life. An analysis of the "cause" of SIB suggests that it is a relatively straight-forward example of the development of neurobiological and behavioral aspects of aggressive behavior in the absence of social factors that would normally bring the behavior under environmental control. Once induced, however, it becomes environmentally autonomous and its proximal cause is neurobiological in nature. 4. There are three lines of evidence that nonhuman primate SIB is linked to malfunctions in the norepinephrine (NE) and serotonin (5HT) neurotransmitter systems. The activity of these systems appears to be altered by psychosocial deprivation. The functional relationship between the two systems appears to be altered or absent in socially deprived monkeys. Pharmacologic agents that act on these systems alter SIB in monkeys. 5. Preliminary data from socially deprived rhesus monkeys are consistent in major respects with studies linking altered serotonin systems to self-injurious behavior and suicidal motivation in humans who also probably suffer from social deprivation. 6. Taken together, these findings indicate that developmental study of biogenic amine systems, particularly finding ways to circumvent deficits in, or restore functional linkages between, the 5HT and NE systems, will lead to a greater understanding of the neurobiologic basis of SIB in humans and animals and will enable us to develop more effective treatments of SIB.


Association between water, sanitation, and hygiene access and the prevalence of soil-transmitted helminth and schistosome infections in Wolayita, Ethiopia.
The Geshiyaro project is a 5-year intervention to assess the impact of community- and school-based water, sanitation, and hygiene (WaSH) interventions on reducing infection with soil-transmitted helminths (STH) and schistosome parasites in combination with deworming in Wolayita zone, Ethiopia. A population-based, cross-sectional census and parasitological mapping activity was conducted between 2018 and 2019. Individuals in the census were identified using either a registered study ID card or biometric fingerprint to enable linkage of their household WaSH data with baseline STH and schistosome prevalence for risk analysis. Prevalence of STH was 15.5% for any STH species, 9.47% for Ascaris lumbricoides, 1.78% for Trichuris trichiura, and 7.24% for hookworm. Intestinal schistosomiasis (Schistosoma mansoni) infection prevalence was 0.85% by Kato Katz, 21.6% by POC-CCA trace positive (Tr +), and 13.3% trace negative (Tr-). Microhaematuria was 2.77%, with 0.13% of people examined with S. haematobium eggs detected by urine filtration. At the household level, increased (> 30 min) time taken to collect drinking water, sharing a latrine, and lack of handwashing facilities were all associated with a greater risk of A. lumbricoides, hookworm, and S. mansoni infection. Not disposing of infant stool at the household and clothes washing/recreational freshwater contact were significantly associated with higher risk of schistosomiasis infection. Aggregating WaSH data at the community level showed odds of A. lumbricoides, hookworm, and T. trichiura infection were significantly lower as both community sanitation coverage and access to improved drinking water improved. The principal finding of this study is that lack of access to WaSH, such as improved drinking water and shared toilet and hand-washing facilities, were linked to an increased risk of infection with STH and schistosome parasites. These associations are difficult to establish at an individual household level because of wide variability in access between houses but are detectable when coverage is aggregated at the community level. Maintenance of WaSH facilities as well as increased access within the whole community is important in influencing the community-wide prevalence of infection with STH and schistosome parasites.


Do drains have an impact on the outcome after primary elective unilateral inguinal hernia repair in men?
The use of drains continues to be a controversial topic in surgery. In a review of that topic for incisional hernia it was not possible to find sufficient evidence of the need for a drain. Likewise, for inguinal hernia surgery the data available are insufficient. In a multivariable analysis of data from the Herniamed Registry for 98,321 patients with primary elective unilateral inguinal hernia repair in men, the role of a drain was investigated. A drain was used in 24.7% (n = 24,287/98,321) of patients. These patients were on average older, had higher BMI, longer operating time and received a smaller mesh. Drains were also used more often for patients with higher ASA score, risk factors, larger defects and scrotal hernia localization as well as for Lichtenstein, TEP and suture repair. The use of drains was highly significantly associated with intra- and postoperative complications as well as with complication-related reoperations. Hence, drains are used selectively in inguinal hernia repair for patients at higher risk of perioperative complications. Despite the use of drains, the outcome in this risk group is less favorable. It remains unclear if drains prevent further complications in high-risk patients. Drains are used selectively in high-risk men with primary elective unilateral inguinal hernia repair. Drains are associated with intra- and postoperative complications rates and complication-related reoperation rate. Drains can serve as an indicator for early detection of complications.


The demand for dependent health insurance: how important is the cost of family coverage?
From the mid-1980s to the mid-1990s, the proportion of non-elderly Americans with employment-based health insurance declined. Roughly 80% of this decline was due to the loss of coverage by dependent family members. During this period, workers became increasingly responsible for the costs of family coverage, while expanded Medicaid coverage provided low-income working families with an alternative to employment-based insurance. We examine the role of out-of-pocket premiums and expanded Medicaid eligibility in households' demand for employment-based family coverage. Cross-sectional results reveal that demand is affected by both factors. We find that between 1987 and 1996, the increase in out-of-pocket premium costs accounted for nearly half of the decline in dependent coverage while expanded Medicaid eligibility represented 14% of the decline.


Instability of integrated hepatitis B virus DNA with inverted repeat structure in a transgenic mouse.
We established four cell lines, from the liver cells of a transgenic mouse, constructed with hepatitis B virus DNA that had an inverted repeat structure. The integrated DNA patterns of the four established cell lines were different from one another and from the original pattern. These data show that the instability of integrated hepatitis B virus DNA would also occur in somatic cells during replication, apart from meiosis, which was previously reported.


Corticotropin-releasing factor binding protein within the ventral tegmental area is expressed in a subset of dopaminergic neurons.
Corticotropin-releasing factor (CRF) and related peptides play a role in mediating neuronal effects of stress. These peptides mediate stress responses by their interactions with the CRF receptors and the CRF-binding protein (CRF-BP). Because the CRF-BP is implicated in neurotransmission within the ventral tegmental area (VTA), we investigated whether the CRF-BP is expressed in VTA neurons. By in situ hybridization, we detected cellular expression of CRF-BP mRNA in the VTA; no such expression was seen in neighboring substantia nigra pars compacta (SNC) or substantia nigra pars reticulata. By double in situ hybridization, we determined that VTA neurons with CRF-BP mRNA coexpressed transcripts encoding either tyrosine hydroxylase [TH; a marker for dopamine (DA) neurons] or glutamic acid decarboxylase [GAD; synthesizing enzyme of gamma-aminobutyric acid (GABA)]. Neurons with CRF-BP mRNA represented 25% of the total population of TH-expressing neurons and 28% of the total population of GAD-expressing neurons, indicating that discrete subpopulations of dopaminergic and GABAergic neurons are present in the VTA. Within the total population of neurons containing CRF-BP mRNA, 70% coexpressed TH mRNA and only 27% coexpressed GAD mRNA. As far as we are aware, we provide the first anatomical evidence that a molecule, CRF-BP, is encoded by DAergic neurons of the VTA but not by those of the SNC. We propose, based on the observation that the majority of VTA neurons expressing CRF-BP mRNA are DAergic, that in the VTA interactions of CRF-BP with CRF, or with CRF-related peptides, are likely to be mediated predominantly by DAergic neurons.


Effect of continuous positive-pressure ventilation on oxygenation after pulmonary microemboli in dogs.
We infused starch microemboli (63 to 74-mu diameter) into the external jugular vein of 28 dogs, to observe the effects of continuous positive-pressure ventilation (CPPV) on gas exchange and hemodynamics during hypoxemia (PaO2 53 +/- 3 torr). CPPV at both 10 and 15 cm H2O end-tidal pressure improved PaO2. CPPV 15 returned PaO2 and pulmonary shunt to control values but reduced cardiac output, O2 transport, and O2 consumption. In spite of these changes suggesting inadequate tissue oxygenation with CPPV 15, mixed venous oxygenation was not reduced. We conclude that: (a) hypoxemia after microemboli infusion is improved by CPPV and therefore likely caused by ventilation/perfusion abnormalities; (b) the improvement in PaO2 produced by CPPV after microemboli is not beneficial if CPPV reduces perfusion and O2 transport; and (c) mixed venous oxygenation does not appear to be an adequate measure of oxygen transport to tissues when CPPV is applied in this high pulmonary vascular resistance setting.


Weak promoters to drive selection marker expression: Improvement of cell line development process for therapeutic protein production in CHO-K1 cells.
Chinese Hamster Ovary cells have been widely used as host cells for production of recombinant therapeutic molecules. Cell line development is a decisive step, which must be carried out with an efficient process. In particular, degree of selection stringency is an important parameter for identification of rare, high-producing cell lines. In the CHOZN® CHO K1 platform, selection of top-producing clones is based on puromycin resistance, whose expression is driven by Simian Virus 40 Early (SV40E) promoter. In this study, novel promoters have been identified to drive expression of selection marker. Decrease of transcriptional activity compared to SV40E promoter was confirmed by RT-qPCR. Selection stringency was increased, as seen by decreased surviving rate of transfected mini-pools and longer recovery duration of transfected bulk pools. Several promoters led to a 1.5-fold increase of maximum titer and a 1.3-fold increase of mean specific productivity of the monoclonal antibody over the clone generation. Expression level was maintained stable over long term cultivation. Finally, productivity increase was confirmed on several monoclonal antibodies and fusion proteins. Lowering the strength of promoter for expression of selective pressure resistance is an efficient strategy to increase selection stringency, which can be applied on industrial CHO-based cell line development platforms.


Two new myrmecophilous species of the genus Petalomium (Acari: Pygmephoroidea: Neopygmephoridae).
Two new myrmecophilous species of the genus Petalomium Cross, 1965 (Acari: Pygmephoroidea: Neopygmephoridae), P. crossi sp. nov. and P. reductus sp. nov. are described from ants Myrmicaria distincta Santschi, 1925 (Hymenoptera: Formicidae: Myrmicinae) in Ethiopia. The presence and shape of the postpalpal setae in species of the genus Petalomium are discussed.


Elevated incidence of sleep apnoea in acromegaly-correlation to disease activity.
An elevated prevalence of sleep apnoea (SA) in patients with acromegaly has been suggested. We performed polysomnographies in 52 patients with acromegaly (25 m, 27 f, age 51 years, range 19-82 years). Patients were defined having SA if they had more than five apnoeas or hypopnoeas per hour (respiratory disturbance index = RDI). The type of SA was divided into obstructive (OSA), central (CSA) or mixed (OSA+CSA). Seventeen patients had newly diagnosed disease, and 18 patients were treated with somatostatin analogues. Twenty-three patients had controlled disease activity (mean GH levels <1 μg/l during a 3-h profile and normalised IGF-1 levels). Twelve had active acromegaly despite medical treatment. Thirty patients (58%) had SA. Twenty-five of those had OSA, three had CSA, and two had mixed. Of the patients with active disease, 66% had SA, compared to 48% in the cured group. Significantly more patients with hypertension (n = 18) than without hypertension (n = 12, p = 0.041) had SA. Basal glucose was not significantly different between patients with (100 mg/dl, range 75-207 mg/dl) and without SA (92 mg/dl, range 74-120 mg/dl), but HbA1c was significantly higher in patients with SA (5.9% (4.9-9.0%) vs. 5.4% (4.3-6.1%), p = 0.001). A positive correlation between RDI and BMI (p = 0.04), RDI and age (p = 0.013) and RDI and disease activity (p = 0.014) was seen. No major correlation could be found between RDI and the duration of disease activity nor between RDI and GH levels. RDI correlates positively with disease activity but not with the duration of the disease. The parameters of the metabolic syndrome are positively associated to the degree of SA in acromegalic patients.


Corrigendum: Identifying single electron charge sensor events using wavelet edge detection (2015Nanotechnology26215201).
The simulated noise used to benchmark wavelet edge detection in this work was described incorrectly. The correct description is given here, and new results based on noise that matches the original description are provided. The results support our original conclusion, which is that wavelet edge detection outperforms thresholding in the presence of white noise and 1/fnoise.


Simulated diagnostic performance of low-field MRI: Harnessing open-access datasets to evaluate novel devices.
The purpose of this study is to demonstrate a method for virtually evaluating novel imaging devices using machine learning and open-access datasets, here applied to a new, low-field strength portable 64mT MRI device. Paired 3 T and 64mT brain images were used to develop and validate a transformation converting standard clinical images to low-field quality images. Separately, 3 T images were aggregated from open-source databases spanning four neuropathologies: low-grade glioma (LGG, N = 76), high-grade glioma (HGG, N = 259), stroke (N = 28), and multiple sclerosis (MS, N = 20). The transformation method was then applied to the open-source data to generate simulated low-field images for each pathology. Convolutional neural networks (DenseNet-121) were trained to detect pathology in axial slices from either 3 T or simulated 64 mT images, and their relative performance was compared to characterize the potential diagnostic capabilities of low-field imaging. Algorithm performance was measured using area under the receiver operating characteristic curve. Across all cohorts, pathology detection was similar between 3 T and simulated 64mT images (LGG: 0.97 vs. 0.98; HGG: 0.96 vs. 0.95; stroke: 0.94 vs. 0.94; MS: 0.90 vs 0.87). Pathology detection was further characterized as a function of lesion size, intensity, and contrast. Simulated images showed decreasing sensitivity for lesions smaller than 4 cm2. While simulations cannot replace prospective trials during the evaluation of medical devices, they can provide guidance and justification for prospective studies. Simulated data derived from open-source imaging databases may facilitate testing and validation of new imaging devices.


Ability of a polygenic risk score to refine colorectal cancer risk in Lynch syndrome.
Polygenic risk scores (PRSs) have been used to stratify colorectal cancer (CRC) risk in the general population, whereas its role in Lynch syndrome (LS), the most common type of hereditary CRC, is still conflicting. We aimed to assess the ability of PRS to refine CRC risk prediction in European-descendant individuals with LS. 1465 individuals with LS (557 MLH1, 517 MSH2/EPCAM, 299 MSH6 and 92 PMS2) and 5656 CRC-free population-based controls from two independent cohorts were included. A 91-SNP PRS was applied. A Cox proportional hazard regression model with 'family' as a random effect and a logistic regression analysis, followed by a meta-analysis combining both cohorts were conducted. Overall, we did not observe a statistically significant association between PRS and CRC risk in the entire cohort. Nevertheless, PRS was significantly associated with a slightly increased risk of CRC or advanced adenoma (AA), in those with CRC diagnosed <50 years and in individuals with multiple CRCs or AAs diagnosed <60 years. The PRS may slightly influence CRC risk in individuals with LS in particular in more extreme phenotypes such as early-onset disease. However, the study design and recruitment strategy strongly influence the results of PRS studies. A separate analysis by genes and its combination with other genetic and non-genetic risk factors will help refine its role as a risk modifier in LS.


The IgM antibody responses to the core antigen of hepatitis B virus.
Little is known about the immunoglobulin class of antibodies to HBcAg. In the present study sera containing anti-HBc were fractionated by surcose density-gradient centrifugation, and all serum fractions were tested against HBcAg by immunoelectro-osmophoresis. In addition selected fractions were examined by complement fixation test, immune adherence hemagglutination and immune electron microscopy. Anti-HBc activity in IgG serum fractions was demonstrated by all four techniques used, but HBcAg-specific IgM was detected only by immunoelectro-osmophoresis and by immune electron microscopy. In acute hepatitis B, HBcAg-specific IgM was detected for up to eight weeks after the onset of jaundice. It was also found transiently in two patients who developed chronic hepatitis B without an icteric episode and in one out of thirteen patients with HBsAg-positive chronic liver disease, but in none of eight healthy HBsAg carriers. The results suggested that HBcAg-specific IgM is formed transiently in response to primary HBV infection but is generally undetectable in established HBsAg carriers.


Self-awareness of executive dysfunction in Huntington's disease: comparison with Parkinson's disease and cervical dystonia.
This study assessed self-awareness of executive deficits in patients with Huntington's disease (HD) in comparison to patients with Parkinson's disease (PD) and with cervical dystonia (CD). Eighty-nine patient-proxy pairs participated in the study (23 with HD, 25 with advanced PD, 21 with mild PD and 20 with CD). Executive function was assessed with the Stroop test and the Dysexecutive Questionnaire. Insight into executive impairment in HD is mildly affected, when compared to PD and CD.


Comparison of the Nuss and sternal turnover procedures for primary repair of pectus excavatum.
Pectus excavatum (PE) is a common chest wall deformity. There are several surgical alternatives for the repair of PE. In our practice, the sternal turnover (STO) procedure had been performed for decades. In 2008, we started treating PE patients with the Nuss procedure. Our objective of this study is to compare these two procedures. A retrospective chart review was conducted on 50 patients undergoing pectus excavatum repairs from March 2005 to January 2013, including 20 patients with the STO procedure and 30 patients with the Nuss procedure. Patients were evaluated for type of repair performed, operating time, drainage after operation, length of postoperative stay, complications, and cosmetic results. The mean age of the STO group was 11.0 years and that of the Nuss group was 15.0 years (p = 0.353). The Nuss procedure had a much shorter mean operating time, a less mean drainage after operation, and a shorter mean time to drainage tube removal than those of the STO procedure. The rate of complication was 40.0% (8/20) in the STO group and 33.3% (10/30) in the Nuss group. Follow-up data indicated that 90% (18/20) of patients in the STO group and 96.7% (29/30) of patients in the Nuss group regarded the results as good or excellent (p = 0.965). Our data suggests that both the STO and Nuss procedures are equally safe and effective correction methods. However, less trauma, faster recovery, and better cosmetic results are the benefits of the Nuss procedure.


Take my breath away: TAM-ing anti-cancer immunity in hypoxic niches.
Hypoxia is a major driver of tumor aggressiveness and therapy resistance in GBM. In this issue of Immunity, Sattiraju et al. functionally link hypoxia with diminished anti-cancer immunity caused by sequestration of immunosuppressive TAMs and CTLs in pseudopalisades in GBM.


[Expression of sBCMA in Newly Diagnosed Multiple Myeloma Patients and Its Relationship with Prognosis].
To study the expression level of serum B-cell maturation antigen (sBCMA) in the peripheral blood of newly diagnosed multiple myeloma (MM) patients, and explore its relationship with the prognosis of MM patients. The peripheral blood of 31 newly diagnosed MM patients and 30 healthy volunteers were collected. The level of sBCMA in the peripheral blood was detected by enzyme-linked immunosorbent assay (ELISA). The correlation between the level of sBCMA and the prognosis of MM patients was analyzed. The level of sBCMA in newly diagnosed MM patients was significantly higher than that in healthy controls (P <0.05). The level of sBCMA was closely related to the plasma cells ratio in bone marrow, the M protein level and the treatment (P <0.05). The level of sBCMA was negatively correlated with the overall survival (OS) of MM patients (r =-0.47). MM patients with low expression of sBCMA had significantly longer OS than patients with high expression of sBCMA (P <0.05). The level of sBCMA is significantly increased in MM patients, which is expected to be a new indicator for evaluating the curative efficacy and prognosis of MM patients. Targeting sBCMA may provide new ideas for the treatment of MM.


Hemostatic and Coagulation Profile in Psoriasis: A Hospital-Based case control Study.
Psoriasis is a chronic inflammatory dermatosis associated with an increased risk of cardiovascular disease and atherothrombosis. This study was conducted to assess the levels of various hemostatic and coagulation parameters in psoriasis and their correlation with disease severity. This was a hospital-based observational study. Seventy-five patients with psoriasis and seventy controls were included in the study. History taking, clinical examination, and calculation of Psoriasis Area and Severity Index (PASI) were done. Blood analysis for Platelet count (PC), mean platelet volume (MPV), Vitamin B12, Thrombin Time (TT), Prothrombin time (PT) and Activated partial thromboplastin time (APPT) were done in both groups. MPV which is a marker of inflammation and platelet activation was significantly increased in cases and positively correlated with the disease severity. Vitamin B12 is an important cofactor in homocysteine (Hcy) metabolism and correlates inversely with serum Hcy which is a known atherothrombotic marker. Vitamin B12 levels were significantly decreased in the cases with a significant negative correlation between Vitamin B12 level and PASI. There was also a significant decrease in serum level of PT, aPTT and TT in cases as compared to controls; however they showed no significant correlation with PASI. Inflammation in psoriasis may drive the process of abnormal platelet activation and coagulation abnormalities thus predisposing psoriatic patients to an atherothrombotic state and increasing the cardiovascular risk in psoriatic patients.


Preliminary study on the diagnostic value of acoustic radiation force impulse technology for differentiating between benign and malignant thyroid nodules.
This study aimed to evaluate the diagnostic value of acoustic radiation force impulse (ARFI) elasticity imaging for differentiating between benign and malignant thyroid lesions. Hospitalized patients needing thyroid surgery were evaluated. After routine thyroid sonography, the patients underwent ARFI elasticity imaging. Virtual Touch tissue imaging (VTI) and Virtual Touch tissue quantification (VTQ; Siemens Medical Solutions, Mountain View, CA) were used to qualitatively and quantitatively analyze the elasticity and hardness of nodules. For statistical analysis, the Student t test, analysis of variance, and the χ(2) test were used to compare the elastic parameters. Of the 98 thyroid nodules observed in 72 hospitalized patients, 56 were nodular goiters, 16 thyroid adenomas, 4 thyroiditis, and 22 thyroid malignancies, with mean VTQ values ± SD of 2.034 ± 0.484, 1.835 ± 0.364, 2.293 ± 0.787, and 3.941 ± 1.393 m/s, respectively. The elastic parameters of malignant nodules were significantly higher than those of benign nodules (P < .001) and the surrounding thyroid parenchyma (P < .001). There was no significant difference between the VTQ value of benign nodules and that of the surrounding normal thyroid parenchyma (P > .05). For differentiating between benign and malignant nodules, the sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accordance rate were 86.36%, 93.42%, 79.17%, 95.95%, and 91.84% based on the standard VTQ value (2.555m/s). In total, 77.6% (59 of 76) of the benign nodules showed softer and equal images in the VTI mode, and 77.3% (17 of 22) of the malignant nodules showed stiffer images (P < .001). Acoustic radiation force impulse imaging has high sensitivity and specificity in evaluating benign and malignant thyroid nodules and therefore had good diagnostic value in clinical applications.


Education-based interventions for anxiety during the treatment and management of chronic disease: a systematic review protocol.
The objective of this systematic review is to evaluate the effectiveness of education-based interventions to decrease patient anxiety during the treatment and management of a chronic disease. Anxiety is a major contributor to poor patient outcomes in self-managed chronic disease. Health care manage anxiety prior or during education can adversely affect patient outcomes. By identifying interventions that effectively decrease anxiety, clinicians may be able to consider and implement strategies as standard practice within the education-based programs. The proposed systematic review will consider studies reporting the effectiveness of any intervention aimed at decreasing participant anxiety prior to a medical procedure or prior to undertaking an education-based program to address a technical aspect of self-management in a health care setting. It will consider studies whose participants are 18 years and older and who are diagnosed with a chronic disease. The systematic review aims to find published and unpublished studies in English from 1972 onward. Databases to be searched included MEDLINE, CINAHL, Embase, ERIC, Mosby's index, Cochrane Library, and Scopus. Studies will be reviewed and data extracted by two independent reviewers. The data will include details about the interventions, populations, study methods, and outcomes of significance to the review objectives. Where possible, data will be pooled in a statistical meta-analysis. PROSPERO CRD42019142260.


The lupus band test: A review of the sensitivity and specificity in the diagnosis of lupus erythematosus.
The lupus band test (LBT) is a direct immunofluorescence (DIF) technique which shows a band of localised immunoglobulins at the dermo-epidermal junction in lesional, non-sun-exposed skin of patients with both systemic and cutaneous lupus erythematosus (LE), and in perilesional skin of patients with systemic LE. However, low sensitivity and poor concordance between histological and clinical diagnoses warrant a review of the application of the LBT in the diagnosis of LE. To assess the sensitivity and specificity of the LBT in diagnosing LE following clinico-pathological correlation (CPC). All cases sent to our pathology department between 2011 and 2018 for DIF with a clinical query of LE were reviewed. Data collection included demographic details, pathology requests, histology and DIF reports, clinical reports and diagnoses, and serology. Of 256 histology requests, 9% (n = 23) had a positive LBT. This was discordant with the prevalence of LE diagnosis, as 46.3% were diagnosed with LE following CPC. The sensitivity and specificity of the LBT for LE was 17.6% and 98.8% respectively, with a positive predictive value of 92.9% and negative predictive value of 58.2%. The LBT is not a sensitive diagnostic test for LE, but is highly specific, and should be considered as a supportive diagnostic tool for LE. This is the largest reported case series evaluating the efficacy of the LBT in the diagnosis of LE.


Cell proliferation effects of S-allyl-L-cysteine are associated with phosphorylation of janus kinase 2, insulin-like growth factor type-I receptor tyrosine kinase, and extracellular signal-regulated kinase 2 in primary cultures of adult rat hepatocytes.
The cell proliferation effect of S-allyl-L-cysteine (SAC) and its mechanisms were examined in primary cultures of adult rat hepatocytes. In serum-free cultivation, SAC (10-6 M)-stimulated hepatocytes showed significant proliferation compared to control at 5-h culture; the effect was dependent on the culture time and the dose of SAC (EC50 value 8.58 × 10-8 M). In addition, SAC-stimulated hepatocytes significantly increased mRNA expression levels of c-Myc and c-Fos at 1 h and cyclin B1 at 3.5 and 4 h, respectively. In contrast, alliin and allicin, structural analogs of SAC, did not show these effects observed with SAC. The SAC-induced hepatocyte proliferation effects were completely suppressed by monoclonal antibodies against growth hormone receptor and insulin-like growth factor type-I (IGF-I) receptor, respectively. Furthermore, the Janus kinase 2 (JAK2) inhibitor TG101209, phospholipase C (PLC) inhibitor U-73122, IGF-I receptor tyrosine kinase (RTK) inhibitor AG538, PI3 kinase inhibitor LY294002, MEK inhibitor PD98059, and mTOR inhibitor rapamycin completely suppressed the SAC-induced hepatocyte proliferation. JAK2 (p125 kDa) phosphorylation in cultured hepatocytes peaked 5 min after SAC stimulation. SAC-induced IGF-I RTK (p95 kDa) and ERK2 (p42 kDa) phosphorylation had slower rises than JAK2, peaking at 20 and 30 min, respectively. These results indicate that SAC promoted cell proliferation by growth hormone receptor/JAK2/PLC pathway activation followed by activation of the IGF-I RTK/PI3K/ERK2/mTOR pathway in primary cultures of adult rat hepatocytes.


Concentration of pregnancy-associated plasma protein A (PAPP-A) in patients with pre-eclamptic toxaemia.
The plasma concentration of PAPP-A has been measured in 84 patients with pre-eclamptic toxaemia within 24 hours of their first admission to hospital. There were 51 patients who suffered from mild pre-eclampsia (late pregnancy hypertension without albuminuria). The PAPP-A values in these patients did not differ significantly from the normal value for the corresponding stage of gestation. On the other hand, the PAPP-A levels in albuminuric (severe) pre-eclampsia were significantly higher than normal; a finding which was more pronounced in those patients who presented with severe pre-eclampsia relatively early in pregnancy. It is unusual for a placental product to be raised in the presence of putative placental damage and it is suggested that PAPP-A might be involved with causal mechanisms in pre-eclampsia.


Mutual co-regulation between GPI-N-acetylglucosaminyltransferase and ergosterol biosynthesis in Candida albicans.
A novel co-regulation exists between the first step of GPI (glycosylphosphatidylinositol) anchor biosynthesis and the rate-determining step of ergosterol biosynthesis in Candida albicans. Depleting CaGpi19p, an accessory subunit of the enzyme complex that initiates GPI biosynthesis, down-regulates ERG11, altering ergosterol levels and drug response. This effect is specific to CaGpi19p depletion and is not due to cell wall defects or GPI deficiency. Additionally, down-regulation of ERG11 down-regulates CaGPI19 and GPI biosynthesis.


IFITM3 Enhances PI3K Pathway Signaling to Promote B-cell Malignancies.
Pre-B cells in Ifitm3 -/- mice were less able to transform into B-cell cancers than in wild-type mice.


Thromboinflammation in COVID-19: Can α2 -macroglobulin help to control the fire?
The complex COVID-19-associated coagulopathy appears to impair prognosis. Recently, we presented the hypothesis that children are to some extent protected by higher α2 -macroglobulin (α2 -M) levels from severe COVID-19. In addition to endothelial cells, thrombin, and platelets, neutrophil granulocytes also appear to play an important role. Neutrophils extrude extracellular nets, which are histone- and protease-coated web-like DNA structures; activate coagulation and platelets; and release radicals and proteases such as elastase. The unique phylogenetically ancient and "versatile" inhibitor α2 -M contributes particularly during childhood to the antithrombin activity of plasma, binds a broad spectrum of proteases, and interacts with other mediators of inflammation such as cytokines. It is suggested that the scope of basic research and clinical studies would include the potential role of α2 -M in COVID-19.


Hypovolemic hemorrhage induces Fos expression in the rat hypothalamus: Evidence for involvement of the lateral hypothalamus in the decompensatory phase of hemorrhage.
This study tested the hypothesis that the hypothalamus participates in the decompensatory phase of hemorrhage by measuring Fos immunoreactivity and by inhibiting neuronal activity in selected hypothalamic nuclei with lidocaine or cobalt chloride. Previously, we reported that inactivation of the arcuate nucleus inhibited, but did not fully prevent, the fall in arterial pressure evoked by hypotensive hemorrhage. Here, we report that hemorrhage (2.2 ml/100g body weight over 20 min) induced Fos expression in a high percentage of cells in the paraventricular, supraoptic and arcuate nuclei of the hypothalamus as shown previously. Lower densities of Fos immunoreactive cells were also found in the medial preoptic area (mPOA), anterior hypothalamus, lateral hypothalamus (LH), dorsomedial hypothalamus, ventromedial hypothalamus (VMH) and posterior hypothalamus. Bilateral injection of lidocaine (2%; 0.1 μl or 0.3 μl) or cobalt chloride (5mM; 0.3 μl) into the tuberal portion of the LH immediately before hemorrhage was initiated reduced the magnitude of hemorrhagic hypotension and bradycardia significantly. Lidocaine injection into the VMH also attenuated the fall in arterial pressure and heart rate evoked by hemorrhage although inactivation of the mPOA or rostral LH was ineffective. These findings indicate that hemorrhage activates neurons throughout much of the hypothalamus and that a relatively broad area of the hypothalamus, extending from the arcuate nucleus laterally through the caudal VMH and tuberal LH, plays an important role in the decompensatory phase of hemorrhage.


Involvement of PAF-acether in anaphylactic bronchoconstriction induced in guinea pigs by aerosolized antigen.
Bronchoconstriction following the aerosolization of PAF-acether or antigen to guinea pigs induces autodesensitization, but the responses to direct spasmogenic substances are not modified. Bronchoconstriction by PAF-acether is not reduced when it is aerosolized to passively sensitized animals previously desensitized by repeated inhalations of the allergen (ovalbumin). In contrast, when passively sensitized animals are initially desensitized to PAF-acether by repeated inhalations of this agonist, ovalbumin aerosolization induces a bronchoconstriction which is significantly reduced when compared with the response obtained in nondesensitized animals though, in this case, the response to aerosolized histamine is not modified. Thus, PAF-acether is released during intrapulmonary anaphylactic shock induced by aerosolized ovalbumin and can be a prime candidate for its development.


Analysis of available measures for malaria control in Africa south of the Sahara.
Africa south of the Sahara is not homogeneous and presents several extreme conditions where malaria persistence is ensured by a complex and highly adaptable vector system. Plasmodium falciparum is the most widespread and life threatening of the malaria parasites of man, particularly for young children and pregnant women. Large-scale residual spraying was not totally effective and was very costly, and mass chemoprophylaxis was not feasible. The spread of chloroquine resistance added arguments against uncontrolled use of drugs. Chemoprophylaxis is now recommended only for pregnant women, especially in their first pregnancy, whilst chloroquine 25 mg base/kg over 3 days is recommended for curative treatment in villages. Second line treatment regimens should be available, together with the possibility of referring severe malaria cases quickly to appropriate clinical facilities. Other control measures include self-protection against mosquito bites by bednets (especially those impregnated with synthetic pyrethroids), mosquito coils, repellents, window and door screening; other measures to prevent man-mosquito contact, such as careful siting of settlements and zooprophylaxis; anti-larval measures, i.e. source reduction, protection of wells and water reservoirs, larviciding, introduction of larvivorous fish; and sprays against adult mosquitoes. The elaboration of strategies for control and their application requires a study of the existing situation. A core of specialists is required in each country, to help with decentralized planning and evaluation of malaria control and to ensure quality control of services, training and applied field research. Additional measures may become available in the future, especially anti-malaria vaccines, and countries should be ready to study their application.


Antibodies to hepatitis E in Portuguese mothers and their newborns.
The high mortality of pregnant women is one of the main features of hepatitis E virus (HEV) infection in endemic countries. With the emergence of genotype 3 HEV in industrialized countries, attention should be given to pregnant women from these regions since the effect of genotype 3 HEV in pregnancy is not yet known. In this study, archived sera dating from 1993 from mothers with no history of traveling to HEV endemic countries and their newborns were tested for anti-HEV IgG antibodies. Four of the 12 maternal sera were positive for IgG anti-HEV, indicating exposure to this virus. Their newborns were also positive, showing higher anti-HEV IgG levels. Overall, these findings demonstrate not only an efficient transplacental transport of anti-HEV IgG, but also circulation of autochthonous HEV in Portugal in the early 1990s.


Neural correlates of automatic perceptual sensitivity to facial affect in posttraumatic stress disorder subjects who survived L'Aquila eartquake of April 6, 2009.
The "Emotional Numbing" (EN) constitutes one of the core symptoms in PTSD although its exact nature remains elusive. This disorder shows an abnormal response of cortical and limbic regions which are normally involved in understanding emotions since the very earliest stages of the development of processing ability. The aim of our study, which included ten physically healthy subjects with PTSD, diagnosed according to DSM-IV-TR, who survived L'Aquila earthquake of April 6, 2009, and 10 healthy controls matching for age, sex and education, was to examine automatic perceptual sensitivity to facial affect in PTSD, through an affective priming task that was administered during functional magnetic resonance (fMRI). Behavioural data revealed in the PTSD group a higher sensitivity to negative facial affect on an automatic processing level. FMRI data analysis revealed that PTSD subjects showed a significantly higher activation in right insula and left amygdala that we did not observe in healthy subjects; on the contrary, healthy controls showed a greater activation of left lingual gyrus. Our data support the hypothesis that PTSD appears to be sensitive to negative affect on an automatic processing level and correlates with the activation of specific areas involved in processing emotions. An elevated activation of these areas may underlie the emotion dysregulation in PTSD and could explain the Emotional Numbing symptom associated with this disorder. The present study suffers of a number of limitations, for instance, the relatively small sample size did not allow the application of alternative statistical models.


Patients treatment with neuroglioma by teniposide and semustine and its influence on Twist and E-cadherin expression.
This study focuses on curative effects of teniposide combining with semustine on patients with neuroglioma and the influences on the expression of Twist and E-cadherin in tissue. Sixty-eight patients with neuroglioma taking operation in our hospital were divided into two groups randomly. Single radiotherapy was given to 34 patients in group A, and teniposide (VM-26) and semustine (Me-CCUN) were added to radiotherapy for 34 patients in group B. Then, curative effects, survival rate, living quality and adverse reaction rate after operation were compared between two groups. Moreover, the difference in positive expression rate of Twist and E-cadherin before and after treatment between two groups was analyzed by immunohistochemistry. In group B, the effective rate of treatment was 88.2%, and the disease control rate was 70.6%, higher than 52.9% and 32.4% in group A with statistical significance (P < 0.05). Moreover, the survival rate in three years of group B was 44.1%, and the score of living quality was 67.11 ± 4.32, and also higher than 23.5% and 63.79 ± 4.53 in group A with statistical significance (P < 0.05). However, the difference between two groups in adverse reaction rate has no statistical significance (P > 0.05). In addition, the difference in positive expression rate of Twist and E-cadherin between group A and group B has no statistical significance before treatment (P > 0.05). After treatment, however, the positive rate of Twist in group B is lower than that in group A, while the positive rate of E-cadherin is higher. Both differences have statistical significance (P < 0.05). Chemotherapy of VM-26 combining with Me-CCNU can inhibit Twist expression and improve the expression rate of E-cadherin to help improving the curative effects and living quality and increasing survival rate.


Anti-Mullerian hormone concentrations in individual follicular fluids within one stimulated IVF cycle resemble blood serum values.
Anti-Mullerian hormone (AMH) is commonly known as the most potent marker for ovarian reserve due to its decline as female age increases. While serum AMH (sAMH) levels have been intensively investigated, there is less data regarding AMH concentrations in follicular fluid (FF), since FF has usually been designated as waste product during oocyte collection in assisted reproductive technologies. This pilot study investigated follicle AMH concentrations (fAMH) of several follicles per ovary, individually collected with the Steiner-Tan needle, and compared them to sAMH concentrations in women undergoing IVF treatment. We hypothesized that there is no difference of fAMH concentrations in individual follicles and that these concentrations resemble the sAMH value of the patient. Patients were stimulated with a gonadotropin-releasing hormone antagonist ovarian hyperstimulation protocol. On the day of oocyte retrieval, serum samples and FF from all individual follicles from one stimulated IVF cycle were collected and individually analyzed for AMH concentrations. Intracyclic mean fAMH values (n follicle = 2-14) were significantly correlated to sAHM values (ρ = 0.85, p < 0.001) and showed a trend to be negatively associated with age (ρ = -0.43, p = 0.06). Mean intrapatient fAMH concentrations differed significantly (p < 0.001). Furthermore, significant correlations of sAMH with individual fAMH values of the first five follicles of each patient were observed. In conclusion, our results clearly showed that individual fAMH concentrations reflected sAMH values and that fAMH concentrations did not significantly differ within one patient. In future studies, it will be interesting to correlate individual fAMH values to the respective embryo development and overall pregnancy outcome in order to improve IVF treatments and to refrain from embryo overproduction.


Completing the folate biosynthesis pathway in Plasmodium falciparum: p-aminobenzoate is produced by a highly divergent promiscuous aminodeoxychorismate lyase.
Enzymes that produce or recycle folates are the targets of widely used antimalarial drugs. Despite the interest in the folate metabolism of Plasmodium falciparum, the molecular identification of ADCL (aminodeoxychorismate lyase), which synthesizes the p-aminobenzoate moiety of folate, remained unresolved. In the present study, we demonstrate that the plasmodial gene PF14_0557 encodes a functional ADCL and report a characterization of the recombinant enzyme.