Identification of MKI67, TPR , and TCHH Mutations as Prognostic Biomarkers for Patients With Defective Mismatch Repair Colon Cancer Stage II/III.
Stage II/III disease is the most predominant form of colorectal cancer, accounting for approximately 70% of cases. Furthermore, approximately 15% to 20% of patients with stage II/III disease have deficient mismatch repair or microsatellite instability-high colorectal cancer. However, there are no identified significant prognostic biomarkers for this disease. To identify prognostic markers for patients with deficient mismatch repair/microsatellite instability-high colon cancer stage II/III. Retrospective study design. The study was conducted at a high-volume colorectal center, the Cancer Hospital, Chinese Academy of Medical Sciences. Patients diagnosed with stage II/III deficient mismatch repair/microsatellite instability-high colon cancer who underwent curative surgery at the Cancer Hospital at the Chinese Academy of Medical Sciences between July 2015 and November 2018 were included. The primary outcome measure was the influence of differentially mutated genes on progression-free survival. The retrospective deficient mismatch repair/microsatellite instability-high cohort involved 32 patients and The Cancer Genome Atlas-microsatellite instability-high cohort involved 45 patients. Patients with deficient mismatch repair/microsatellite instability-high colon cancer had higher mutational frequencies of MKI67 , TPR , and TCHH than patients with microsatellite stable colon cancer. MKI67 , TPR , TCHH , and gene combination were significantly correlated with prognosis. The biomarker mutation-type colon cancer group had a higher risk of recurrence or death than did the wild-type group. Moreover, biomarker mutation-type tumors had more mutations in the DNA damage repair pathway and tumor mutational burden than did biomarker wild-type tumors. This study was limited by its retrospective nature. MKI67 , TPR , and TCHH may serve as potential diagnostic and prognostic biomarkers for deficient mismatch repair/microsatellite instability-high colon cancer stage II/III. ANTECEDENTES:La enfermedad en estadio II/III es la forma más predominante de cáncer colorrectal y representa aproximadamente el 70% de los casos. Además, aproximadamente entre el 15% y el 20% de los pacientes con enfermedad en estadio II/III tienen reparación deficiente de errores de emparejamiento o inestabilidad de microsatélital alta. Sin embargo, no se han identificado biomarcadores pronósticos significativos para esta enfermedad.OBJETIVO:Este estudio tuvo como objetivo identificar marcadores pronósticos para pacientes con cáncer de colon con reparación deficiente de errores de emparejamiento/inestabilidad microsatelital alta en estadio II/III.DISEÑO:Diseño de estudio retrospectivo.ESCENARIO:El estudio se realizó en un centro colorrectal de alto volumen, el Hospital del Cáncer de la Academia China de Ciencias Médicas.PACIENTES:Pacientes diagnosticados con cáncer de colon en estadio II/III con reparación deficiente de errores de emparejamiento o inestabilidad de microsatélital alta que se sometieron a cirugía curativa en el Hospital del Cáncer de la Academia China de Ciencias Médicas entre julio de 2015 y noviembre de 2018.MEDIDAS DE RESULTADO PRINCIPALES:La medida de resultado primaria fue la influencia de los genes con mutaciones diferenciales en la supervivencia libre de progresión.RESULTADOS:La cohorte retrospectiva de reparación deficiente de errores de emparejamiento o inestabilidad de microsatélital alta y la cohorte de inestabilidad microsatelital alta del Atlas del Genoma del Cáncer involucraron a 32 y 45 pacientes, respectivamente. Los pacientes con de reparación deficiente de errores de emparejamiento/inestabilidad microsatélital alta tuvieron frecuencias mutacionales más altas de MKI67 , TPR y TCHH que los pacientes estables de microsatélites. MKI67 , TPR , TCHH , y la combinación de genes se correlacionaron significativamente con el pronóstico. El grupo de cáncer de colon de tipo mutación de biomarcador tenía un mayor riesgo de recurrencia o muerte que el grupo de mutación salvaje. Además, los tumores de tipo mutación de biomarcadores tenían más mutaciones en la vía de reparación del daño del ADN y la carga mutacional del tumor que los tumores de tipo salvaje de biomarcadores.LIMITACIONES:Este estudio estuvo limitado por su naturaleza retrospectiva.CONCLUSIONES:MKI67 , TPR , y TCHH pueden servir como posibles biomarcadores de diagnóstico y pronóstico para cáncer de colon en estadio II/III con reparación deficiente de errores de emparejamiento/inestabilidad microsatélital alta. (Traducción-Dr. Jorge Silva Velazco ).


Evidence that the Na+-Ca2+ exchange system is related to the antiarrhythmic action of disopyramide.
1. Disopyramide induced a concentration-dependent decrease in action potential amplitude and Vmax, and prolonged action potential durations (APD50 and APD90) in ventricular muscle. 2. Na+-loaded membrane vesicles isolated from canine ventricular muscle rapidly accumulated Ca2+. 3. Monensin (10(-5) M) abolished Na+-dependent Ca2+ uptake, and Na+ enhanced Ca2+ efflux. 4. Na+-Ca2+ exchange by membrane vesicles was more active in preparations pretreated with disopyramide (10(-5) M) than in control membranes. 5. The results suggest that disopyramide changes Na+ influx from Na+ channel mediated to Na+-Ca2+ exchange mediated. This is verified in part by increased Ca2+ efflux.


Motility abnormality in intestinal atresia.
This study was designed to investigate the motility of the small bowel of the lamb under the conditions of experimental intestinal atresia. Of 26 fetal lambs operated upon (50 to 90 days gestation), 13 came to term. All term lambs exhibited a type 3a atresia. Six were successfully repaired and had intestinal myoelectric activity monitored for periods from 2 to 27 hours. A slow wave pattern (12 to 13/min) was present in the bowel of control lambs and in the proximal dilated bowel and the microbowel of atretic lambs, confirming the presence of functional smooth muscle. Spike potentials, which indicated circular muscular contractions, occurred 10% of the time in the control intestine, 5% in the proximal dilated gut (P less than .2), and 0% in the microbowel (P less than .001). A previously undescribed myoelectric transient of variable amplitude and frequency (6 to 24/min) was identified in all lambs studied. Histologic evaluation demonstrated villous hyperplasia in the microbowel and mucosal flattening in the distended bowel. The data suggest that the quiescent microbowel distal to the atresia contributes significantly to the functional obstruction.


Histamine augments collagen content via H1 receptor stimulation in cultures of myofibroblasts taken from wound granulation tissue.
The inflammatory reaction influences the deposition of collagen within wound granulation tissue. The aim of the present study is to determine whether histamine acting directly on myofibroblasts derived from wound granulation tissue may influence collagen deposition. It also identifies the histamine receptor involved in this process. The experiments were carried out on cells isolated from the granulation tissue of a wound model (a polypropylene net inserted subcutaneously to rats) or intact rat skin. Collagen content was measured following the addition of different concentrations of histamine and treatment with histamine receptor antagonists (ketotifen - H1 inhibitor, ranitidine - H2 inhibitor) and a histamine receptor H1 agonist (2-pyridylethylamine dihydrochloride).The cells were identified as myofibroblasts: alpha-smooth muscle actin, vimentin, and desmin positive in all experimental conditions. Histamine increased the collagen level within both cell cultures, i.e., those isolated from granulation tissue or intact skin. It did not, however, influence the expression of either the collagen type I or III genes within the cultured myofibroblasts. Histamine activity was reduced by ketotifen (the H1 receptor inhibitor) and increased by the H1 receptor agonist, as demonstrated by changes in the levels of collagen in the myofibroblast culture. Histamine increased collagen content within the cultures, acting directly on myofibroblasts via H1 receptor stimulation.


CircRNA CircABCB1 Diminishes the Sensitivity of Breast Cancer Cells to Docetaxel by Sponging MiR-153-3p.
Resistance to docetaxel is a major problem to the success of docetaxel-based therapies for breast cancer. The present study was to identify the role of circABCB1 in altering the docetaxel resistance properties. Reverse transcription-quantitative PCR (qRT-PCR) was performed to quantify circABCB1 and miR-153-3p. The effects of circABCB1 on the viability, apoptosis and migration/invasion of docetaxel-resistant and -sensitive cells were investigated by cell function experiments, including Cell Counting Kit-8 and Transwell assays. Correlation between circABCB1 and the docetaxel-treated outcome was analyzed by multivariate Cox regression analysis, in addition to Kaplan-Meier analysis of time to treatment failure (TTF). The targeting relationship between circABCB1 and miR-153-3p was predicted and verified by dual-luciferase reporter assay and RNA immunoprecipitation. CircABCB1 was highly expressed in cancerous tissues, as well as the docetaxel-sensitive group and cells. The overexpression of circABCB1 contributed to cell viability, docetaxel-resistance and migration/invasion, but inhibited apoptosis. CircABCB1 can sponge miR-153-3p. CircABCB1 contributed to the docetaxel resistance of breast cancer, maybe via the miR-153-3p.


Circulating regulatory T cells (CD4+CD25+FOXP3+) decrease in breast cancer patients after vaccination with a modified MHC class II HER2/neu (AE37) peptide.
Regulatory T cells (T(Reg)), CD4(+)CD25(+)FOXP3(+), are implicated in suppressing tumor immune responses. We analyzed peripheral blood lymphocytes (PBL) from breast cancer patients receiving a modified HLA class II HER2/neu peptide (AE37) vaccine for T(Reg) cells and correlated their levels with vaccine-specific immune responses. The mean CD4(+)CD25(+)FOXP3(+) T(Reg) cells decreased in patients with vaccination with no significant difference in serum TGF-β levels. IFN-γ ELISPOT and DTH increased after vaccination with a good correlation between T(Reg) cell reduction and size of DTH to AE37. The T(Reg) cell reduction and associated immune response suggest that AE37 may be clinically useful.


Clinical relevance of low levels of preformed alloantibodies detected by flow cytometry in the first year post-kidney transplantation.
To determine the prevalence of transplants performed with a false-negative cytotoxicity cross-match and to analyze the clinical relevance of alloantibodies (Ab) detected only by flow cytometry (flow). We studied 66 patients undergoing kidney transplantation from a cadaveric donor. All patients had a simultaneous negative T+AHG+DTT and B+DTT. Pretransplant sera were retrospectively analyzed by flow cytometry according to an Emory University protocol: (1) T+ and B-: Ab anti-class I; (2) T- and B+: anti-class II; (3) T+B+: anti-class I + II. Chi-square, Fisher exact, Student t test, and Kaplan Meier analyses were employed with significance assigned at P < or = .05. The overall incidence of false-negative cytotoxicity was 33.3% (22/66), namely, 6.1% (n = 4) anti-class I; 9.1% (n = 6) anti-class II; and 18.2% (n = 12) anti-class I + II. Primary nonfunctioning grafts occurred in 6.8% (3/44) and 13.6% (3/22) negative and positive flow patients (two anti-class I + II and one class II; P = .39). The incidence of graft loss in the first year was respectively, 13.6% (6/44) and 18.2% (4/22; two anti-class II and two anti-class I + II; P = .72). Compared to flow-negative grafts, creatinine levels were significantly higher among flow-positive patients at 8 and 12 weeks. One-year graft survivals were 86.4% among negative versus 81.8% for the positive group (P = .67). We observed that 33% of kidney transplant recipients had low levels of alloantibodies detected only by flow. This single factor was associated with the worst graft function in the first trimester with a suggestion of a higher risk for non-functioning graft.


3D segmentation of abdominal aorta from CT-scan and MR images.
We designed a generic method for segmenting the aneurismal sac of an abdominal aortic aneurysm (AAA) both from multi-slice MR and CT-scan examinations. It is a semi-automatic method requiring little human intervention and based on graph cut theory to segment the lumen interface and the aortic wall of AAAs. Our segmentation method works independently on MRI and CT-scan volumes and has been tested on a 44 patient dataset and 10 synthetic images. Segmentation and maximum diameter estimation were compared to manual tracing from 4 experts. An inter-observer study was performed in order to measure the variability range of a human observer. Based on three metrics (the maximum aortic diameter, the volume overlap and the Hausdorff distance) the variability of the results obtained by our method is shown to be similar to that of a human operator, both for the lumen interface and the aortic wall. As will be shown, the average distance obtained with our method is less than one standard deviation away from each expert, both for healthy subjects and for patients with AAA. Our semi-automatic method provides reliable contours of the abdominal aorta from CT-scan or MRI, allowing rapid and reproducible evaluations of AAA.


Macronutrient (im)balance drives energy intake in an obesogenic food environment: An ecological analysis.
The protein leverage hypothesis (PLH) postulates that strong regulation of protein intake drives energy overconsumption and obesity when human diets are diluted by fat and carbohydrates. The two predictions of the PLH are that humans (i) regulate intake to maintain protein within a narrow range and that (ii) energy intake is an inverse function of percentage energy from protein because absolute protein intake is maintained within narrow limits. Multidimensional nutritional geometry was used to test the predictions of the PLH using dietary data from the Australian National Nutrition and Physical Activity Survey. Both predictions of the PLH were confirmed in a population setting: the mean protein intake was 18.4%, and energy intake decreased with increasing energy from protein (L = -0.18, p < 0.0001). It was demonstrated that highly processed discretionary foods are a significant diluent of protein and associated with increased energy but not increased protein intake. These results support an integrated ecological and mechanistic explanation for obesity, in which low-protein highly processed foods lead to higher energy intake because of the biological response to macronutrient imbalance driven by a dominant appetite for protein. This study supports a central role for protein in the obesity epidemic, with significant implications for global health.


[Acitretin-induced pneumonia: an underestimated complication of treatment of psoriasis].
Retinoids are known to induce side effects which can be severe. Alveolar and interstitial pneumonia of uncertain pathogenesis can rarely occur when retinoid are used among patients with psoriasis. We report an observation of acute respiratory distress beginning 26 days after introduction of acitretin, a second-generation retinoid. Treatment withdrawal and corticotherapy allow a spectacular amelioration of respiratory conditions. Pneumonia induced by retinoid must be known as a side effect of treatment of psoriasis.


Hurricane Elena and Pinellas County, Florida: some lessons learned from the largest evacuation of nursing home patients in history.
Prior planning for and problems associated with the evacuation of 1,860 nursing home patients in Pinellas County, Florida, due to Hurricane Elena in 1985 were studied through questionnaires administered to nursing home administrators and public documents. The most serious problems were transporting patients to shelters in a timely fashion, delayed passage of recalled staff through police checkpoints, and staff burnout.


Casein kinase 1G2 suppresses necroptosis-promoted testis aging by inhibiting receptor-interacting kinase 3.
Casein kinases are a large family of intracellular serine/threonine kinases that control a variety of cellular signaling functions. Here we report that a member of casein kinase 1 family, casein kinase 1G2, CSNK1G2, binds and inhibits the activation of receptor-interacting kinase 3, RIPK3, thereby attenuating RIPK3-mediated necroptosis. The binding of CSNK1G2 to RIPK3 is triggered by auto-phosphorylation at serine 211/threonine 215 sites in its C-terminal domain. CSNK1G2-knockout mice showed significantly enhanced necroptosis response and premature aging of their testis, a phenotype that was rescued by either double knockout of the Ripk3 gene or feeding the animal with a RIPK1 kinase inhibitor-containing diet. Moreover, CSNK1G2 is also co-expressed with RIPK3 in human testis, and the necroptosis activation marker phospho-MLKL was observed in the testis of old (>80) but not young men, indicating that the testis-aging program carried out by the RIPK3-mediated and CSNK1G2-attenuated necroptosis is evolutionarily conserved between mice and men.


Localization and migration route of Cephalopina titillator (Diptera: Oestridae) larvae in the head of infested camels (Camelus dromedarius).
The localization and migration routes of Cephalopina titillator larvae were studied in the heads of 45 naturally infested camels. Larvae were located in the pharyngeal cavity (95.6%), the labyrinth of the ethmoid bone (71.1%), the turbinates (28.9%) and the lower nasal meatus (6.7%). Out of 261 first-instar larvae, 73.7% were found in the labyrinth of the ethmoid bone and 26.3% in the turbinates, while 88.9% of second-instar larvae were observed in the pharyngeal cavity and 11.1% in the labyrinth of the ethmoid bone. Most third-instar larvae (91.9%) were found in the pharyngeal cavity, with a few larvae in the labyrinth of the ethmoid bone (6.5%) and lower nasal meatus (1.6%). The first moult occurred in the labyrinth of the ethmoid bone. The second moult was observed in both the labyrinth of ethmoid bone and the pharyngeal cavity. The migration route of these larvae in the infested head was discussed.


Importance of dose-schedule of 5-aza-2'-deoxycytidine for epigenetic therapy of cancer.
The inactivation of tumor suppressor genes (TSGs) by aberrant DNA methylation plays an important role in the development of malignancy. Since this epigenetic change is reversible, it is a potential target for chemotherapeutic intervention using an inhibitor of DNA methylation, such as 5-aza-2'-deoxycytidine (DAC). Although clinical studies show that DAC has activity against hematological malignancies, the optimal dose-schedule of this epigenetic agent still needs to be established. Clonogenic assays were performed on leukemic and tumor cell lines to evaluate the in vitro antineoplastic activity of DAC. The reactivation of TSGs and inhibition of DNA methylation by DAC were investigated by reverse transcriptase-PCR and Line-1 assays. The in vivo antineoplastic activity of DAC administered as an i.v. infusion was evaluated in mice with murine L1210 leukemia by measurement of survival time, and in mice bearing murine EMT6 mammary tumor by excision of tumor after chemotherapy for an in vitro clonogenic assay. Increasing the DAC concentration and duration of exposure produced a greater loss of clonogenicity for both human leukemic and tumor cell lines. The reactivation of the TSGs (p57KIP2 in HL-60 leukemic cells and p16CDKN2A in Calu-6 lung carcinoma cells) and the inhibition of global DNA methylation in HL-60 leukemic cells increased with DAC concentration. In mice with L1210 leukemia and in mice bearing EMT6 tumors, the antineoplastic action of DAC also increased with the dose. The plasma level of DAC that produced a very potent antineoplastic effect in mice with leukemia or solid tumors was > 200 ng/ml (> 1 microM). We have shown that intensification of the DAC dose markedly increased its antineoplastic activity in mouse models of cancer. Our data also show that there is a good correlation between the concentrations of DAC that reduce in vitro clonogenicity, reactivate TSGs and inhibit DNA methylation. These results suggest that the antineoplastic action of DAC is related to its epigenetic action. Our observations provide a strong rationale to perform clinical trials using dose intensification of DAC to maximize the chemotherapeutic potential of this epigenetic agent in patients with cancer.


Role of the endothelium in the vascular effects of the thrombin receptor (protease-activated receptor type 1) in humans.
The purpose of this study was to determine the role of the endothelium in the vascular actions of protease-activated receptor type 1 (PAR-1) activation in vivo in man. Thrombin is central to the pathophysiology of atherothrombosis. Its cellular actions are mediated via PAR-1. Protease-activated receptor type 1 activation causes arterial vasodilation, venoconstriction, platelet activation, and tissue-type plasminogen activator release in man. Dorsal hand vein diameter was measured in 6 healthy volunteers before and after endothelial denudation. Forearm arterial blood flow, plasma fibrinolytic factors, and platelet activation were measured in 24 healthy volunteers during venous occlusion plethysmography. The effects of inhibition of prostacyclin, nitric oxide (NO), and endothelium-derived hyperpolarizing factor on PAR-1 responses were assessed during coadministration of aspirin, the "NO clamp" (L-N(G)-monomethyl arginine and sodium nitroprusside), and tetraethylammonium ion, respectively. Endothelial denudation did not affect PAR-1-evoked venoconstriction (SFLLRN; 0.05 to 15 nmol/min). Although aspirin had no effect, SFLLRN-induced vasodilation (5 to 50 nmol/min) was attenuated by the NO clamp (p < 0.0001) and tetraethylammonium ion (p < 0.05) and abolished by their combination (p < 0.01). The NO clamp augmented SFLLRN-induced tissue-type plasminogen activator and plasminogen activator inhibitor type 1 antigen (p < 0.0001) release, but tetraethylammonium ion and aspirin had no effect. SFLLRN-induced platelet activation was unaffected by NO or prostacyclin inhibition. Acting via PAR-1, thrombin causes contrasting effects in the human vasculature and has a major interaction with the endothelium. This highlights the critical importance of endothelial function during acute arterial injury and intravascular thrombosis, as occurs in cardiovascular events including myocardial infarction and stroke.


Prompt Outpatient Care For Older Adults Discharged From The Emergency Department Reduces Recidivism.
Older adults present unique challenges to both emergency clinicians and health systems. These challenges are especially evident with respect to discharge after an emergency department (ED) visit as older adults are at risk for short-term, negative outcomes including repeat ED visits. The aim of this study was to evaluate characteristics and risk factors associated with repeat ED utilization by older adults. ED visits among participants in the Reasons for Geographic and Racial Differences in Stroke (REGARDS) study between 2003-2016 were examined using linked Medicare claims data to identify such visits and resulting disposition. Multilevel proportional hazards models examined associations of age, comorbidity status, race, gender, Medicaid dual eligibility status, social support characteristics (living alone or caregiver support), and use of ambulatory primary and subspecialty care with repeat ED utilization. Older adults discharged from the ED seen by a primary care provider (hazard ratio [HR] = 0.93, confidence interval [CI], 0.87-0.98, p = 0.01) or subspecialist (HR = 0.91, CI 0.86-0.97, P <0.01) after the ED visit were less likely to return to the ED within 30 days compared to those who did not have such post-ED ambulatory visits. Additionally, comorbidity (HR =1.14, 95% CI, 1.13-1.16, P <0.01) and dual eligibility for Medicare and Medicaid (HR = 1.34, 95% CI, 1.20-1.50, p<0.01) were associated with return to the ED within 30 days. Those who were older (HR = 1.10, 95% CI, 1.05-1.15), had more comorbidities (HR = 1.17, 95% CI 1.15-1.18), Black (HR = 1.23, 95% CI, 1.14-1.33,P <0.01), and dually eligible (HR =1.23, 95% CI, 1.14-1.33, P <0.01) were more likely to return within 31-90 days after their initial presentation. The association of outpatient visits with repeat ED visits was no longer seen beyond 30 days. Patients without a caregiver or who lived alone were no more likely to return to the ED in the time periods evaluated in our study. Both primary care and subspecialty care visits among older adults who are seen in the ED and discharged are associated with less frequent repeat ED visits within 30 days.


Activation of Nrf2 might reduce oxidative stress in human granulosa cells.
Nuclear factor-E2-related factor 2 (Nrf2)/Kelch-like ECH-associated protein 1 (Keap1)-antioxidant response element (ARE) signaling pathway is one of the most important defense mechanisms against oxidative stress (OS). It is well documented that equilibration status of OS plays fundamental roles in human reproductive medicine, and the physiological role of Nrf2 in ovarian granulosa cells (GCs) has not been determined yet. Herein we aimed to study the function of Nrf2 in GCs. Human ovarian tissues were subjected to immunohistochemistry to localize Nrf2 and Keap1 and we detected the expression of Nrf2 and Keap1 in the human GCs. Human luteinized GCs were isolated and cultured, and hydrogen peroxide (H2O2) or Dimethylfumarates (DMF), an activator of Nrf2, were added to GCs to analyze the relationship between Nrf2 and antioxidants by quantitative RT-PCR. The mRNA levels of Nrf2, catalase, superoxide dismutase 1 (SOD1), and 8-Oxoguanine DNA glycosylase (OGG1) were elevated by H2O2, and DMF treatment showed similar but pronounced effects through activation of Nrf2. To determine the relationship of Nrf2 and the generation of antioxidants, siRNAs were used and quantitative RT-PCR were conducted. Decreased expression of Nrf2 resulted in a decreased level of these antioxidant mRNA. Intracellular levels of ROS were investigated by fluorescence of 8-hydroxy-2'-deoxyguanosine and fluorescent dye, 2',7'-dichlorodihydrofluorescein diacetate after H2O2 and/or DMF treatment, and DMF treatment quenched intracellular ROS generation by H2O2. These results show that activation of Nrf2 might lead to alleviate OS in human GCs, and this could provide novel insight to conquer the age-related fertility decline that is mainly attributed to the accumulation of aberrant OS.


Segregation of transferrin to a mildly acidic (pH 6.5) para-Golgi compartment in the recycling pathway.
To study the intracellular sorting of internalized ligands and receptors, we examined the pathways of two ligands: transferrin, which is recycled, and alpha 2-macroglobulin (alpha 2M), which is degraded. In CHO cells the two ligands rapidly segregate into different intracellular compartments. Within 5 min fluorescein-labeled transferrin (F-Tf) is found in a large round juxtanuclear structure. Rhodamine-labeled alpha 2M is found in a punctate pattern. Ultra-structural localization studies demonstrate that colloidal gold-alpha 2M is found predominantly in endocytic vesicles, while ferritin-transferrin is found in small vesicles and tubular structures in a region adjacent to the Golgi complex. Using image intensified fluorescence microscopy and digital image analysis, we determined that the F-Tf containing structure has a pH of 6.4 +/- 0.2, while endocytic vesicles containing F-alpha 2M have a pH of 5.4 +/- 0.1. Our study defines a mildly acidic compartment, distinct from endocytic vesicles, that is involved in the recycling of internalized components back to the cell surface.


Diabetes and exercise.
The abnormal metabolic responses to exercise in insulin-dependent diabetes are in great part related to abnormal circulating plasma insulin concentrations. Exercising during relative insulin deprivation results in an increase in glycemia and ketosis. Exercise during insulin excess results in inhibition of hepatic glucose production and accelerated muscle glucose utilization and results in hypoglycemia. These responses can be significantly improved when insulin is administered more appropriately, as is the case with insulin infusion pumps. Self blood glucose monitoring before, during, and after exercise can provide important information that can be used to optimize the metabolic response to exercise in individual patients. A better understanding of the metabolic response to exercise in patients with diabetes will serve as the basis for developing specific recommendations to enable these individuals to have the freedom to take part in all forms of exercise with minimal restriction. However, the demonstration that exercise will have a long-term beneficial effect on the metabolic control of diabetes or prevent the development of the complications of diabetes remains to be established.


Small intestinal injury in a neonatal rat model of giardiasis is strain dependent.
The factors that determine the severity of giardiasis are poorly understood. Host factors are important, but parasite virulence may also play a role. The aim of this study was to compare the apparent virulence of three genotypically different Giardia isolates (PO1, VNB3, and WB). Infection rates, parasite loads, structural damage, disaccharidase activity, and water and electrolyte absorption were observed at 10 days after inoculation in a neonatal rat model of infection. DNA fingerprinting showed differences between isolates studied. The infective rate varied between 67% and 100%. There were no differences in intestinal parasite load. Infection with strains PO1 and WB, but not with VNB3, was associated with a reduction in villus height. There was precocious expression of sucrase at 10 days after inoculation in all infected groups. Water absorption of a plasma electrolyte solution was decreased in VNB3-infected animals when compared with PO1- and WB-infected animals and controls. Water absorption and lactose hydrolysis were impaired during perfusion with a lactose-containing solution in all infected groups. Three genotypically different Giardia isolates that infect neonatal rats with the same trophozoite load differ in their ability to cause functional mucosal damage. Infection with Giardia lamblia induced precocious expression of sucrase activity and impaired mucosal absorption.


A Patient's Journey to Pay a Healthcare Bill: It's Way Too Complicated.
The patient's financial experience is often complex and confusing.Recent regulations support bringing more transparency to healthcare billing processes.A patient journey map illustrates the patient experience in a healthcare process helping to identify improvement opportunities to better support patient-centered care.Price transparency can be increased by communicating the patient's financial responsibility early in the care process as well as simplifying patient communications.Digital health applications can help improve the patient financial experience and ultimately engagement in the payment process.Greater clarity and improved processes may occur if the patient's journey is visualized.


Arachidonic acid metabolomic study of BPH in rats and the interventional effects of Zishen pill, a traditional Chinese medicine.
Zishen pill (ZSP) is a traditional Chinese medicine (TCM) used to treat benign prostatic hyperplasia (BPH). The study used a metabolomic approach based on UHPLC-MS/MS to profile arachidonic acid (AA) metabolic changes and to investigate the interventional mechanisms of ZSP in testosterone- induced BPH rats. In order to explore the potential therapeutic effect of ZSP, rat models were constructed and orally administrated with ZSP. Plasma and urine samples were collected after four weeks and then eleven potential biomarkers (15-HETE, 12-HETE, TXA2, 5-HETE, AA, PGI2, PGF2α, 8-HETE, PGD2, PGE2 and LTB4) were identified and quantified by UHPLC-MS/MS. The chromatographic separation was carried out with gradient elution using a mobile phase comprised of 0.05% formic acid aqueous solution (pH=3.3) (A) and acetonitrile: methanol (80:20, V/V) (B), and each AA metabolites was measured using electrospray ionization source with negative mode and multiple reaction monitoring. The eleven biomarkers in BPH group rat plasma and urine were significant higher than those in sham group rats. Using the potential biomarkers as a screening index, the results suggest that ZSP can potentially reverse the process of BPH by partially regulating AA metabolism through refrain the expression of cyclooxygenase (COX) and lipoxygenase (LOX). This study demonstrates that a metabolomic strategy is useful for identifying potential BPH biomarkers and investigating the underlying mechanisms of a TCM in BPH treatment.


Rituximab identified as an independent risk factor for severe PJP: A case-control study.
Pneumocystis jirovecii pneumonia (PJP) was reported among immunosuppressed patients with deficits in cell-mediated immunity and in patients treated with immunomodulatory drugs. The aim of this study was to identify risk-factors for PJP in noninfected HIV patients. This retrospective, test negative, case-control study was conducted in six hospitals in Israel, 2006-2016. Cases were hospitalized HIV-negative patients with pneumonia diagnosed as PJP by bronchoalveolar lavage. Controls were similar patients negative for PJP. Seventy-six cases and 159 controls were identified. Median age was 63.7 years, 65% males, 34% had hematological malignancies, 11% inflammatory diseases, 47% used steroids and 9% received antilymphocyte monoclonal antibodies. PJP was independently associated with antilymphocyte monoclonal antibodies (OR 11.47, CI 1.50-87.74), high-dose steroid treatment (OR 4.39, CI 1.52-12.63), lymphopenia (OR 8.13, CI 2.48-26.60), low albumin (OR 0.15, CI 0.40-0.54) and low BMI (OR 0.80, CI 0.68-0.93). In conclusion, rituximab, which is prescribed for a wide variety of malignant and inflammatory disorders, was found to be significant risk-factor for PJP. Increased awareness of possible PJP infection in this patient population is warranted.


Effects of losartan and amlodipine on urinary albumin excretion and ambulatory blood pressure in hypertensive type 2 diabetic patients with overt nephropathy.
Few studies have assessed whether 24-h blood pressure control induced by antihypertensive agents improves macroalbuminuria in hypertensive type 2 diabetic patients with overt nephropathy. We evaluated the effects of losartan and amlodipine on 24-h blood pressure, autonomic nervous activity, and albuminuria in these patients. In this open-label, parallel-prospective, randomized study, 44 patients were treated with losartan and 43 with amlodipine for a 12-week titration phase and a maintenance phase for a maximum of 12 weeks. Twenty-four-hour blood pressure and urinary albumin excretion were measured before and during treatment. Simultaneously, power spectral analysis of heart rate was performed to evaluate low frequency (LF) and high frequency (HF) components and LF-to-HF ratios as an index of sympathovagal balance. Losartan decreased (P < 0.001) mean blood pressure from 162/91 to 150/82 mmHg during daytime and from 146/82 to 137/74 mmHg during nighttime (systolic/diastolic). Amlodipine also decreased (P < 0.001) blood pressure from 159/90 to 147/82 mmHg during daytime and from 143/81 to 131/72 mmHg during nighttime. LF and HF components and nighttime-to-daytime ratios for the LF-to-HF ratios did not differ during treatment in two groups, showing no changes in the diurnal autonomic nervous rhythm. Losartan decreased (P < 0.001) 24-h urinary albumin excretion from 810 mg/day (95% CI 780-1,140) to 570 (510-910). Amlodipine, however, did not decrease (P = 0.893) albuminuria (790 mg/day [780-1,170] vs.790 [710-1,260]). These results suggest that in type 2 diabetes with overt nephropathy, 24-h blood pressure regulation alone is inadequate to reduce macroalbuminuria and additional effects of losartan are crucial for antiproteinuric action.


Comparison of neuronal inositol 1,4,5-trisphosphate 3-kinase and receptor mRNA distributions in the human brain using in situ hybridization histochemistry.
The distribution of the messenger RNA coding for the recently cloned inositol 1,4,5-trisphosphate (InsP3) 3-kinase, the enzyme phosphorylating InsP3 to InsP4, was compared to the localizations of InsP3 receptor mRNA in the human brain using in situ hybridization histochemistry and oligonucleotide probes. InsP3 3-kinase and receptor mRNA levels were high in the cerebellar Purkinje cells. They were also observed, to a much lesser degree than in the cerebellum, in the hippocampal CA1 pyramidal cells and dentate gyrus granule cells, in the majority of the cortical neurons and in the striatal medium-sized neurons. Both mRNAs were not detected in the brainstem and in the glial cells.


Primo: integration of multiple GWAS and omics QTL summary statistics for elucidation of molecular mechanisms of trait-associated SNPs and detection of pleiotropy in complex traits.
To provide a comprehensive mechanistic interpretation of how known trait-associated SNPs affect complex traits, we propose a method, Primo, for integrative analysis of GWAS summary statistics with multiple sets of omics QTL summary statistics from different cellular conditions or studies. Primo examines association patterns of SNPs to complex and omics traits. In gene regions harboring known susceptibility loci, Primo performs conditional association analysis to account for linkage disequilibrium. Primo allows for unknown study heterogeneity and sample correlations. We show two applications using Primo to examine the molecular mechanisms of known susceptibility loci and to detect and interpret pleiotropic effects.


Novel Immunotherapy Combinations.
The last decade witnessed an explosion in immunotherapeutic agent approvals for various malignancies. The success of immune checkpoint inhibitors (CTLA-4 and PD-1/PD-L1) in melanoma quickly sprung to other cancer types and are considered the emerging face of oncology. Antibodies to CTLA-4 were first to enter the field, quickly followed by PD-1/PD-L1 inhibitors. Combination anti-CTLA4 and anti-PD-1/PD-L1 therapies were investigated, and after demonstrating improved responses, rapidly gained approval. Certain tumor types previously considered non-immunogenic also demonstrated durable responses which has been a remarkable discovery. However, not all tumor types respond to immunotherapies and it is widely recognized that tumor-specific immune inflammatory status predicts the best responders. Ongoing translational work indicates specific upregulation in additional immune checkpoints that circumvent response to anti-CTLA4 and anti-PD-1/PD-L1 antibodies. Here, we provide a comprehensive review of promising therapies on the horizon with unique combinations designed to overcome resistance or expand the pool of treatment responders.


The role of the neutrophil and phagocytosis in infection caused by Helicobacter pylori.
Recent advances in our understanding of Helicobacter pylori-phagocyte interactions indicate that these organisms actively modulate phagocyte function in order to retard phagocytosis, while simultaneously inducing a strong respiratory burst. The central players in this dynamic include H. pylori neutrophil activating protein and factors that are associated with the cag pathogenicity island type IV secretion apparatus. Additionally, catalase, alkyl hydroperoxide reductase, and factors that are unique to type I strains allow bacteria to resist phagocytic killing.


Structural and spectroscopic characterization of exogenous ligand binding to isolated factor F430 and its configurational isomers.
Binding of axial ligands to the nickel(II) of isolated factor F430 from the methyl reductase enzyme of Methanobacterium thermoautotrophicum is demonstrated. Evidence of bis-ligand coordination is obtained from the x-ray absorption, optical, and resonance Raman spectral characterization of F430 and its 12,13-diepimeric isomer in the presence of a large excess of cyanide, pyridine, or 1-methylimidazole. Significant broadening and 5-10-nm red shifts of the main 430-nm optical absorption band and shifts of up to 30 cm-1 for the high-frequency Raman lines are observed upon coordination of these axial ligands. The Raman spectra of native F430 and the diepimer with a particular axial ligand are nearly identical. Nickel x-ray absorption edge spectra of the diepimer in the absence and presence of these exogenous ligands are indicative of conversion from a square-planar to a tetragonally distorted octahedral geometry. Analyses of the nickel extended x-ray absorption fine structure data for the ligated diepimer complexes yield detailed structural information for these complexes. Implications of these data with respect to the enzymatic mechanism and the structure of the enzyme-bound factor are discussed.


Bilateral single system ectopic ureters in a male infant.
Bilateral single-system ureteral ectopia is a rare congenital malformation. We report a case of a male infant with bilateral single-system ectopic ureters without diffuse hydronephrosis, vesicoureteral reflux, or a history of recurrent urinary tract infections. The patient presented for an evaluation of hydronephrosis associated with the upper pole of an apparently duplex left kidney detected by prenatal and postnatal ultrasonography. Cystoscopy revealed two ureteral orifices distal to the bladder neck with a dilated left upper-pole calix and no duplex kidney. No surgical intervention has been planned until his continence status has been clarified.


Association between first-trimester maternal serum pregnancy-associated plasma protein-A and obstetric complications.
This study aimed to investigate the relationship between maternal serum pregnancy-associated plasma protein-A (PAPP-A) in the first trimester of pregnancy and the development of preeclampsia (PE), early PE, small-for-gestational age (SGA) fetus and preterm delivery (PD). This is a retrospective study of 12,355 pregnant women that delivered between 2008 and 2011. We define the first, third and fifth percentiles of maternal serum PAPP-A multiples of the median (MoM). The primary outcome measures were the occurrence of PE, early PE (PE requiring delivery before 34 weeks), SGA fetus (birth weight < 5th centile) and PD. The Mann-Whitney U-test and chi-squared test were used to analyze continuous and dichotomous variables, respectively. Maternal serum PAPP-A was significantly lower in women with PE, early PE, SGA fetus and PD (0.91, 0.74, 0.80 and 0.84 MoM, respectively) than in the study population (0.99 MoM) (p < 0.05). The lower the MoM percentile of PAPP-A, the higher are the odds ratio (OR) to develop PE, early PE, SGA fetus and PD. Maternal serum PAPP-A levels are lower in women who develop preeclampsia, those with SGA fetus and those who deliver preterm. However, on its own, maternal serum PAPP-A performs poorly (OR for PE between 1.76 and 2.41 with the lower percentile of PAPP-A) as a screening test for these conditions.


Organogold complexes probe a large beta-barrel cavity for human serum alpha1-acid glycoprotein.
Human alpha(1)-acid glycoprotein (AAG) is an acute phase component of the plasma, binding numerous drugs and natural compounds with high-affinity. Using circular dichroism (CD) spectroscopy, strong AAG binding of organogold complexes was found, the molecular size and chemical structure of which differ from known AAG binding agents. The 16-membered Au(2)P(4)C(8)O(2) macrocycles interconvert rapidly between two helical forms and produce enantiomeric conformations which are in dynamic equilibrium in solution. AAG binds preferentially one of the chiral conformers as indicated by strong Cotton effects generated by intramolecular exciton coupling between the pairs of hetercyclic chromophores. Lipophilic nature of the guest molecules suggests the dominant contribution of hydrophobic interactions in the AAG binding. Comparison of the main genetic variants of AAG revealed that both the 'F1/S' and 'A' variants bind with high-affinity the gold(I) macrocycles (K(a) approximately 10(6) M(-1)). CD/fluorescence displacement, and fluorescence quenching experiments indicated inclusion of the compounds into the central beta-barrel cavity of AAG of which exact tertiary structure is yet unknown. Molecular dimensions of the gold(I) macrocycles (13 x 14 x 14 A) indicate that the principal ligand binding cavity of both the 'F1/S' and 'A' variants must be larger compared to the models published to date. Based on these findings, a novel homology model of AAG 'F1' variant was constructed using the human neutrophil gelatinase-associated lipocalin as a template. The organogold complexes were successfully docked into the central cavity of this model.


Two years after lockdown: Longitudinal trajectories of sleep disturbances and mental health over the COVID-19 pandemic, and the effects of age, gender and chronotype.
Since the first lockdown of Spring 2020, the COVID-19 contagion waves pervasively disrupted the sleep and mental health of the worldwide population. Notwithstanding the largest vaccination campaign in human history, the pandemic has continued to impact the everyday life of the general population for 2 years now. The present study provides the first evidence of the longitudinal trajectories of sleep disturbances and mental health throughout the pandemic in Italy, also describing the differential time course of age groups, genders and chronotypes. A total of 1062 Italians participated in a three-time-point longitudinal study covering two critical stages of the emergency (the first lockdown in April 2020 and the second partial lockdown in December 2020) and providing a long-term overview 2 years after the pandemic outbreak (April 2022). We administered validated questionnaires to evaluate sleep quality/habits, insomnia, depression, stress and anxiety symptoms. Analyses showed a gradual improvement in sleep disturbances, depression and anxiety. Conversely, sleep duration progressively decreased, particularly in evening-type and younger people. Participants reported substantial earlier bedtime and get-up time. Stress levels increased during December 2020 and then stabilised. This effect was stronger in the population groups apparently more resilient during the first lockdown (older people, men and morning-types). Our results describe a promising scenario 2 years after the pandemic onset. However, the improvements were relatively small, the perceived stress increased, and the re-establishment of pre-existing social/working dynamics led to general sleep curtailment. Further long-term monitoring is required to claim the end of the COVID-19 emergency on Italians' sleep and mental health.


Effect of alkaline pH on the activity of rat liver phenylalanine hydroxylase.
The pH optimum of rat liver phenylalanine hydroxylase is dependent on the structure of the cofactor employed and on the state of activation of the enzyme. The tetrahydrobiopterin-dependent activity of native phenylalanine hydroxylase has a pH optimum of about 8.5. In contrast, the 6,7-dimethyltetrahydropterin-dependent activity is highest at pH 7.0. Activation of phenylalanine hydroxylase either by preincubation with phenylalanine or by limited proteolysis results in a shift of the pH optimum of the tetrahydrobiopterin-dependent activity to pH 7.0. Activation of the enzyme has no effect on the optimal pH of the 6,7-dimethyltetrahydropterin-dependent activity. The different pH optimum of the tetrahydrobiopterin-dependent activity of native phenylalanine hydroxylase is due to a change in the properties of the enzyme when the pH is increased from pH 7 to 9.5. Phenylalanine hydroxylase at alkaline pH appears to be in an altered conformation that is very similar to that of the enzyme which has been activated by preincubation with phenylalanine as determined by changes in the intrinsic protein fluorescence spectrum of the enzyme. Furthermore, phenylalanine hydroxylase which has been preincubated at an alkaline pH in the absence of phenylalanine and subsequently assayed at pH 7.0 in the presence of phenylalanine shows an increase in tetrahydrobiopterin-dependent activity similar to that exhibited by the enzyme which has been activated by preincubation with phenylalanine at neutral pH. Activation of the enzyme also occurs when m-tyrosine or tryptophan replace phenylalanine in the assay mixture. The predominant cause of the increase in activity of the enzyme immediately following preincubation at alkaline pH appears to be the increase in the rate of activation by the amino acid substrate. However, in the absence of substrate activation, phenylalanine hydroxylase preincubated at alkaline pH displays an approximately 2-fold greater intrinsic activity than the native enzyme.


Replication of feline coronaviruses in peripheral blood monocytes.
Feline infectious peritonitis virus (FIPV) (Coronaviridae) causes the most lethal viral infection in cats: FIP. The related feline enteric coronavirus (FECV) causes mild enteritis. Why these feline coronaviruses manifest so differently in vivo is not known. In this study, infection kinetics (titres and antigen expression) of FIPV 79-1146, and FECV 79-1683, were determined in peripheral blood monocytes from 3 donor cats and compared to those in Crandell feline kidney (CrFK) cells. The infection kinetics in monocytes were host dependent. Monocytes from 1 cat were resistant to both FIPV- and FECV-infection. Monocytes from the other 2 cats could initially be infected by both FIPV and FECV but FIPV infection was sustained in monocytes of only one cat. FECV-infection was never sustained and viral production was up to 100 times lower than in FIPV-infected monocytes. In CrFK cells, FIPV and FECV infection kinetics did not differ. In monocytes of a larger cat population (n = 19) the 3 infection patterns were also found. Considering all 22 investigated cats, 3/22 were not susceptible for FIPV and FECV. The rest could be infected with FECV and FIPV but 10/22 cats had monocytes that only sustained FIPV infection and 9/22 sustained neither FIPV nor FECV infection.


Food groups intake in relation to stunting among exceptional children.
Although several studies have examined the link between different food groups intake and stunting among children, no study, to our knowledge, was done on exceptional children. The aim of this study was assessed the association of dietary intake and stunting in Iranian exceptional children. This cross-sectional study was conducted on 470 exceptional children (226 mentally retarded, 182 deaf and 62 blind children), aged 5-15 years. Height was measured using standard tool. Stunting was defined as height-for-age z-score of <-1. A validated dietary habit questionnaire was applied to assess dietary intakes. Mean age of children was 10.02 ± 2.04 years. Stunting was prevalent among 50.6% of children. Compared with children in the lowest category of dairy consumption, those in the highest category had lower odds of stunting. This association remained significant even after adjusting for covariates (OR: 0.50, 95% CI: 0.29-0.87). In addition, moderate consumption of egg (1-3 time/wk) was inversely associated with stunting either before or after controlling for potential confounders (OR: 0.36, 95% CI: 0.21-0.64). Such finding was also seen among mentally retarded children (OR: 0.38, 95% CI: 0.16-0.89). No other significant association was seen between intakes of other food groups (including meat, fruits and vegetables) and stunting. We found that higher intake of dairy products and egg was associated with lower risk of stunting. However, intakes of other food groups including meat, fruits and vegetables were not significantly related to stunting.


Static and dynamic studies of lanthanum(III) ion adsorption/desorption from acidic solutions using chelating ion exchangers with different functionalities.
Increasing the number of applications of rare earth elements (REEs) has led to increased release of these metals into the environment. Removal of REEs from e-wastes is very important considering the increasing demand for these elements, the limited resource availability of them as well as the significant environmental issues. In this present study, optimization of the La(III) ions sorption from acidic solutions on chelating ion exchangers containing different functional groups, i.e. Amberlite IRC748, Purolite S930, Lewatit® Monoplus TP208, Amberlite IRC747, Purolite S940, and Purolite S950, was carried out. The sorption data was analyzed using the Lagergren pseudo-first order, Ho and McKay pseudo-second order, Weber-Morris intraparticle diffusion, Boyd kinetic models, pore and film diffusion coefficients as well as the Langmuir, Freundlich, and Temkin isotherm models. Additionally, thermodynamic parameters and regeneration abilities of chelating ion exchangers were evaluated. The maximum recovery of La(III) ions was found for HNO3 concentration equal to 0.2 mol/dm3. The La(III) ions sorption was fast and sorption equilibrium was achieved after about 60 min. The best fitting for the lanthanum(III) ions sorption was obtained using the pseudo-second order kinetic and Langmuir isotherm models. Moreover, breakthrough curves were obtained from dynamic studies. The physicochemical characterization places special emphasis on determination of chemical composition of ion exchangers using ATR/FT-IR and XPS spectroscopy. Furthermore, characterization parameters of ion exchangers such as surface area and porosity (pore size), point of zero charge, and thermal stability were estimated. Chelating ion exchangers with aminophosphonic functional groups are characterized by the best adsorption properties towards La(III) ions so they could be used for the recovery of rare earth elements from spent battery solutions.


Migratory behavior of leukemic cells from acute myeloid leukemia patients.
The chemokine stromal cell-derived factor-1 (SDF-1) and its receptor CXCR-4 contribute to stem cell homing and may play a role in the trafficking of leukemic cells. Therefore, we analyzed migration across Transwell filters of cells derived from bone marrow (BM) or peripheral blood (PB) from 26 acute myeloid leukemia (AML) patients. The presence of the extracellular matrix protein fibronectin (FN) strongly enhanced the spontaneous and SDF-1-induced migration of leukemic PB and BM cells. No differences in spontaneous, SDF-1-induced migration or CXCR-4 expression were observed between the different AML subtypes. Subsequently, it was determined whether SDF-1 preferentially promoted migration of subsets of leukemic cells. Leukemic cells expressing CD34, CD38 and HLA-DR were preferentially migrating, whereas cells expressing CD14 and CD36 showed diminished migration. Analysis of paired PB and BM samples indicated that significantly higher SDF-1-induced migration was observed in AML for CD34(+) BM-derived cells compared to CD34(+) PB-derived cells, suggesting a role for SDF-1 in the anchoring of leukemic cells in the BM or other organs. The lower percentage of circulating leukemic blasts in patients with a relatively high level of SDF-1-induced migration also supports this hypothesis. In conclusion, we have shown that primary AML cells are migrating towards SDF-1 independent of the AML subtypes.


Pharmacological tests of the mechanism of the periodic rhythm caused by veratramine in the sinoatrial node of the guinea pig.
1. We investigated the effects of several drugs and extracellular ions on the periodic sinoatrial node rhythm caused by high concentrations of veratramine (>2 microM) in isolated guinea pig sinus atria. 2. During the active phase of this rhythm, pacemaker activity appeared to be due to transient afterdepolarizations resembling the delayed afterdepolarizations attributed to Ca++-induced Ca++ release in cardiac tissue. 3. Ryanodine (200-2200 nM) did not decrease the transient afterdepolarizations, and instead increased the heart rate during the active phase, prolonged the active phase, and sometimes caused conversion to regular rhythm. 4. Dichlorobenzamyl (10-110 microM), a blocker of electrogenic Na+-Ca++ exchange, did not slow or stop beating during periodic rhythm, but rather increased average heart rate and, at a higher concentration, caused conversion to regular rhythm. 5. Ouabain (0.1 microM), an inhibitor of the sodium pump and electrogenic Na+-K+ exchange, had little effect on veratramine periodic rhythm, but at higher concentrations it caused increased average heart rate and conversion to regular rhythm. 6. The chronotropic effect of Ca++ was normally weakly positive; however, in the presence of veratramine, and before the appearance of periodic rhythm, the chronotropic effect of Ca++ was weakly negative, and was associated with destabilization of the heart rate, leading to frequency oscillations or periodic rhythm. 7. Veratramine changed the chronotropic effect of K+ from weakly negative to moderately positive. 8. When half the Na+ or Cl- in the bathing medium was replaced by an impermeant ion, in the absence of veratramine the average heart rate was slightly decreased, whereas, in the presence of veratramine and periodic rhythm the average rate was increased, although the increase was not statistically significant in the case of low Na+. 9. These observations indicate that Ca++-induced Ca++ release, Na+-Ca++ exchange, and probably electrogenic Na+-K+ exchange play no important role in generation of periodic rhythm. The increased K+ dependence suggests an altered pacemaker mechanism.


Simple and rapid reversed-phase high-performance liquid chromatographic determination of baclofen in human plasma with ultraviolet detection. Application to a pharmacokinetic study.
A sensitive, selective, and rapid reversed-phase high-performance liquid chromatographic method for separation and quantitation of intact racemic baclofen in human plasma has been described. Baclofen is very soluble in water and cannot be extracted efficiently by an organic solvent. Therefore, baclofen was isolated from plasma endogenous materials by adding 0.50 ml of acetonitrile and 50 mg of zinc sulfate to 1.0 ml of plasma. A 15 cm X 4.6 mm, 10-microns octyl (C8) column was slurry packed and used throughout the study. An isocratic mobile phase containing 0.01 M monobasic potassium phosphate (pH congruent to 3.5)-acetonitrile (80:20, v/v) was delivered at a flow-rate of 1.0 ml/min through the chromatographic system. Baclofen was detected with an ultraviolet-visible variable-wavelength detector at 220 nm and 0.50-0.005 a.u.f.s. The time needed to complete the analysis of one sample was approximately 15 min. The limit of detection for the assay of racemic baclofen was 35 ng/ml. After an oral dose of 20 mg of baclofen, blood samples were collected at several time points and plasma was analyzed using the method developed in this study. Various pharmacokinetics parameters were determined from the plasma concentration versus time profile of baclofen.


Synthesis and characterization of polypyridine ruthenium(II) complexes and anticancer efficacy studies in vivo and in vitro.
In this article, ligand IPP (IPP = 4-(1H-imidazo[4,5-f][1,10]phenanthrolin-2-yl)-N,N-diphenylaniline) and its three Ru(II) complexes: [Ru(bpy)2(IPP)](ClO4)2 (1) (bpy = 2,2'-bipyridine), [Ru(dmbpy)2(IPP)](ClO4)2 (2) (dmbpy = 4,4'-dimethyl-2,2'-bipyridine), and [Ru(phen)2(IPP)](ClO4)2 (3) (phen = 1,10-phenanthroline) were synthesized and characterized. The anticancer activity in vitro of the complexes was investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. The scratching and colony-forming experiments confirmed the complexes 1, 2, 3 interfered with the proliferation and migration ability of cells. The accumulation of the complexes in cells was researched and we found that these complexes directly accumulated in mitochondria, then the complexes cause a decline of the mitochondrial membrane potential and induce an increase of intracellular reactive oxygen species (ROS) levels. The growth of B16 cells were inhibited by 1, 2 and 3 at G0/G1 phase. Apoptosis was induced through mitochondrial pathway and the expression of apoptosis-related factors was regulated. In addition, the complexes promoted the transition of poly(ADP-ribose)polymerase (PARP) into the cleaved form (Cleaved PARP), downregulated the anti-apoptotic proteins, and upregulated the pro-apoptotic proteins. Consequently, complexes 1, 2 and 3 exerted their anticancer activity by regulating B-cell lymphoma-2 (Bcl-2) family proteins. Complex 2 showed excellent antitumor effects with a high inhibitory rate of 65.95% in vivo. Taken together, the complexes cause apoptosis in B16 cells through a ROS-mediated mitochondrial dysfunction pathway.


What is the role of locoregional anesthesia in breast surgery? A systematic literature review focused on pain intensity, opioid consumption, adverse events, and patient satisfaction.
Breast surgery in the United States is common. Pain affects up to 50% of women undergoing breast surgery and can interfere with postoperative outcomes. General anesthesia is the conventional, most frequently used anaesthetic technique. Various locoregional anesthetic techniques are also used for breast surgeries. A systematic review of the use of locoregional anesthesia for postoperative pain in breast surgery is needed to clarify its role in pain management. To systematically review literature to establish the efficacy and the safety of locoregional anesthesia used in the treatment of pain after breast surgery. Embase, MEDLINE, Google Scholar and Cochrane Central Trials Register were systematically searched in Mars 2020 for studies examining locoregional anesthesia for management of pain in adults after breast surgery. The methodological quality of the studies and their results were appraised using the Consensus-based Standards for the Selection of Health Measurement Instruments (COSMIN) checklist and specific measurement properties criteria, respectively. Nineteen studies evaluating locoregional anesthesia were included: 1058 patients underwent lumpectomy/mastectomy, 142 breast augmentation and 79 breast reduction. Locoregional anesthesia provides effective anesthesia and analgesia in the perioperative setting, however no statistically significant difference emerged if compared to other techniques. For mastectomy only, the use of locoregional techniques reduces pain in the first hour after the end of the surgery if compared to other procedures (p = 0.02). Other potentially beneficial effects of locoregional anesthesia include decreased need for opioids, decreased postoperative nausea and vomiting, fewer complications and increased patient satisfaction. All this improves postoperative recovery and shortens hospitalization stay. In none of these cases, locoregional anesthesia was statistically superior to other techniques. The results of our review showed no differences between locoregional anesthesia and other techniques in the management of breast surgery. Locoregional techniques are superior in reducing pain in the first hour after mastectomy.


Pancreatic schwannoma: a rare tumor of the pancreas.
Schwannomas are rarely detected in the pancreas. Preoperative diagnosis of this entity is still challenging so far, causing unnecessary radical resection, endoscopic ultrasonography or percutaneous biopsies. Presented here is a case of schwannoma occurring in the pancreatic head, confirmed by histopathology after pancreaticoduodenectomy.


Caesarean section rates in Mozambique.
The Caesarean section (C-section) rate is used as an indicator for availability and utilization of life-saving obstetric services. The purpose of the present study was to explore changes in C-section rates between 1995 and 2011 by area, place of delivery and maternal socioeconomic factors in Mozambique. Cross-sectional data from the Demographic and Health Surveys conducted in Mozambique in 1997, 2003 and 2011 were used, including women having a live birth within 3 years prior to the survey. Descriptive statistics and logistic regressions were used to identify factors associated with having a C-section. The C-section rate decreased slightly from 2.5% in 1995-1997 to 2.1% in 2001-2003 and then increased to 4.7% in 2009-2011. In 2009-2011, C-section rates ranged in urban areas from 4.6% in the northern region to 12.2% in the southern region and in rural areas from 1.6% in the northern region to 3.9% in the southern region. 12.3% of the richest women had had a C-section, compared to 1.7% of the poorest women. C-sections were the most common at public hospitals (12.6% in 2009-2011), but C-sections at health centers increased from the second to the third period. The likelihood of having a C-section was associated with living in urban areas and in the southern region, having a formal education and living in a rich household, even adjusting for age and parity (and study periods). The strongest relationship was for the richest household wealth quintile [OR (95% CI): 9.8 (6.3-15.3)]. The highest rate (20.6%) was found among the richest women giving birth at public hospitals in the southern region in 2009-2011. In Mozambique, underuse of C-section was likely among the poor and in rural areas, but overuse in the most advantaged groups seemed to be emerging.


Approximate entropy based on attempted steady isometric contractions with the ankle dorsal- and plantarflexors: reliability and optimal sampling frequency.
The aim of this study was to (1) examine the test-retest reliability of approximate entropy (ApEn) calculated for torque time-series from attempted steady isometric contractions performed at two different days, and (2) examine the significance of the sampling frequency for the ApEn values. Eighteen healthy young subjects (13+/-3 years, mean+/-1 S.D.) performed attempted steady isometric submaximal contractions with the ankle dorsal- and plantarflexors at two different days. Relative (ICC(3.1)) and absolute (standard error of measurement [S.E.M.], and S.E.M.%) test-retest reliability was assessed for the ApEn values calculated for torque time-series down-sampled to 30 and 100Hz, respectively. The relative reliability was generally moderate (0.360< or =ICC(3.1)< or =0.897), with an absolute reliability (S.E.M.%) of 6-14%. The mean ApEn values varied considerably depending on the applied down-sampling frequency (5-200Hz). When ApEn was used to quantify structure in the torque time-series, the relative and absolute reliability of steady isometric contractions with the ankle proved to be good in healthy young subjects. We propose that an optimal time-series down-sampling frequency exists for ApEn calculations, which will increase the sensitivity for biological system-changes, reduce adverse effects of random noise, and ensure that biological information in the signal is preserved. We recommend estimating this frequency using a variable high-pass filter-method for frequency analysis. Based on this method, the optimized time-series down-sampling frequency was around 30Hz for the isometric contractions performed with the ankle in the present study.


The first central nervous system autopsy in Southern Brazil.
The history of Anatomical Pathology in the state of Paraná, in southern Brazil, is closely linked with the foundation of the Universidade Federal do Paraná (UFPR). This study identified the first central nervous system (CNS) clinical autopsy performed by the Department of Anatomical Pathology of the UFPR. This study reviewed the autopsy report archives of the Hospital de Clínicas-UFPR from 1951 onward. The clinical anatomy interpretations of the autopsy report and possible etiologic agents were discussed. The first adult clinical autopsy with CNS study was performed on April 23, 1952 on a 45-year-old man with lobar pneumonia with abscesses complicated by bacterial meningitis. This case was the first CNS clinical autopsy performed in the state of Paraná and, possibly, in southern Brazil. The death was due to an infectious disease, which was the main cause of death in Brazil in the 1950s.


Iron-enriched mycelia of edible and medicinal basidiomycetes.
Iron bioaccumulation in basidiomycetes is an alternative to recover ferrous sulphate from titanium dioxide pigment production and to produce an iron-enriched mycelial biomass. This study aimed to evaluate iron bioaccumulation capacity in vegetative mycelium of edible and medicinal fungi grown in malt extract liquid medium with different ferrous sulphate contents. Five basidiomycetes were grown in malt extract liquid medium with different iron contents from 0.116 to 100 mg L-1 iron. The iron content of dried mycelial biomass bioaccumulated with iron was determined by flame atomic absorption spectrophotometry. All fungi grew on the iron culture media and the mycelial biomass growth ranged from 3.24 ± 0.65a mg mL-1 to 12.46 ± 0.29 mg mL-1. Iron addition to culture media increased the iron content in the mycelial biomass from 4000-13,000-fold compared with control. Pleurotus ostreatus (2181 ± 218 mg kg-1) presented the greatest iron content in the mycelial biomass, followed by Schizophyllum commune (1769 ± 131 mg kg-1), Agaricus subrufescens (1272 ± 8.84 mg kg-1), and Ganoderma lucidum (840 ± 75 mg kg-1). P. ostreatus, followed by S. commune, and G. lucidum at 90 and 100 mg L-1 iron in the culture medium are the best choices to produce iron-enriched mycelial biomass. This extensive study of several edible and medicinal basidiomycetes grown in different iron contents was effective in recovering ferrous sulphate byproduct and transferring it to mycelium to produce a new nutraceutical food of iron-enriched mycelial biomass.


Functional MRI techniques demonstrate early vascular changes in renal cell cancer patients treated with sunitinib: a pilot study.
To assess the early vascular effects of sunitinib in patients with renal cell carcinoma (RCC) with diffusion-weighted magnetic resonance imaging (DWI), dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) and T2* perfusion MRI. In 10 patients with abdominal RCC lesions, DWI, DCE-MRI and T2* perfusion MRI measurements at 3 Tesla were performed at baseline, 3 and 10 days after start of sunitinib. VEGF-A plasma levels were measured on days 0, 3 and 10. DWI showed a significant increase in the apparent diffusion coefficient (×10(-6) s/mm(2)) from baseline (mean 1158, range 814-2003) to day 3 (mean 1306, range 1008-2097, P = 0.015) followed by a decrease to baseline levels at day 10 (mean 1132, range 719-2005, P = 0.001). No significant changes were found in mean DCE-MRI parameters. T2* perfusion MRI showed a significant decrease in relative tumor blood volume (rBV) and relative tumor blood flow (rBF) at day 3 (rBV P = 0.037, rBF P = 0.018) and day 10 (rBV P = 0.006, rBF P = 0.009). VEGF-A plasma levels significantly increased after 10 days, but did not correlate with MRI parameters. Sunitinib induces antiangiogenic effects as measured by DWI and T2*-perfusion MRI, 3 and 10 days after the start of the initial treatment. DCE-MRI did not show significant changes. In the near future, early functional MRI-based evaluation can play an important role in tailoring treatment to the individual patient with RCC. Further investigation is warranted.


Skin grafts for circumferential coverage of perianal wounds.
A technique for circumferential skin grafting of the perianal skin after excision of Bowen's disease is described. The technique has proved satisfactory. The protocol for perioperative management of the bowel and the regimen of strict bed rest must be followed carefully.


Estimation of leaf area index using WorldView-2 and Aster satellite image: a case study from Turkey.
The objective of this study is to estimate the leaf area index (LAI) of a forest ecosystem using two different satellite images, WorldView-2 and Aster. For this purpose, 108 sample plots were taken from pure Crimean pine forest stands of Yenice Forest Management Planning Unit in Ilgaz Forest Management Enterprise, Turkey. Each sample plot was imaged with hemispherical photographs with a fish-eye camera to determine the LAI. These photographs were analyzed with the help of Hemisfer Hemiview software program, and thus, the LAI of each sample plot was estimated. Furthermore, multiple regression analysis method was used to model the statistical relationships between the LAI values and band spectral reflection values and some vegetation indices (Vis) obtained from satellite images. The results show that the high-resolution WorldView-2 satellite image is better than the medium-resolution Aster satellite image in predicting the LAI. It was also seen that the results obtained by using the VIs are better than the bands when the LAI value is predicted with satellite images.


Exposure of wood in floodplains affects its chemical quality and its subsequent breakdown in streams.
In stream ecosystems, coarse organic matter from the riparian vegetation, a key food resource, is often retained in the floodplains before reaching the channel. During floodplain exposure, organic matter can be affected by abiotic and biotic processes ("preconditioning"), which alter its quality and affect its subsequent decomposition in streams. We analyzed the effect of floodplain preconditioning on wood quality (lignin, C, N, P, K, among others), and its subsequent aquatic breakdown, paying special attention to microbial activity. We simulated preconditioned standard wooden sticks on one arid stream floodplain for 3 and 4 months, and then monitored their breakdown in three different streams, together with control (non-preconditioned) sticks. Preconditioning reduced lignin mass and C:N and lignin:N ratios, caused the leaching of soluble nutrients such as P and K, as well as N immobilization by microbes. These changes enhanced the breakdown of wood in the first week of immersion, but had no effect on breakdown rates after 4 months of incubation in the streams, although N immobilization was diminished. Our results suggest that terrestrial preconditioning could alter the role of wood as a long-lasting nutrients and energy source for freshwater ecosystem.


An improved computer model for 60cobalt dose distributions.
A previously developed model for the calculation of 60Co isodose contours exhibits poor fit to experimental data at certain points. Methods for improving the fit at these points are described. The model contains few parameters, and may be useful for treatment planning in cases for which data are not available.


Protease activities in cultured beef lens epithelial cells peak and then decline upon progressive passage.
Beef lens cells in culture are readily obtained and provide many opportunities to study phenomena related to cell differentiation and maturation, environmental stress, disease, and perhaps mechanisms of transformation. Although altered rates of proteolysis are known to accompany these phenomena, the proteolytic activities available in cultured beef lens epithelial cells have not been documented. In this work are documented the specific activities, based on protein and DNA content, of neutral exo- and endopeptidase, cathepsins B- and D-like enzymes and acid phosphatase in lens epithelial cortical and core tissue and in cultured epithelial cells at passages 1-43. Maximal activity of each protease occurs almost routinely at passage 5 or 9, reaching values of approx. 1400-, 0.77-, 4520-nmol min-1 per mg protein for neutral exopeptidase (passage 5), neutral endopeptidase (passage 5) and cathepsin B (passage 5) respectively, and 7.1 micrograms trichloroacetic acid soluble peptide min-1 per mg protein for cathepsin D (passage 15). On a microgram-1 DNA basis, the maximal specific activities for the same enzymes were 48 (passage 5), 0.03 (passage 5), 283 (passage 9), and 0.5 (passage 9) respectively. In subsequent passages, the specific activities declined to values which were similar to or lower than the specific activities observed for these proteases in lens epithelial tissue.


Renal vitamin D receptor expression and vitamin D renoprotection.
Therapy to enhance the renoprotective actions of vitamin D receptor (VDR) activation should safely overcome the distinct VDR content along the nephron to effectively control renal calcium reabsorption, control renal klotho levels for the phosphaturic actions of FGF23, and inhibit proteinuria and the activation of the renin-angiotensin system.


Pharmacokinetics, safety, and tolerability of dispersible and immediate-release abacavir, dolutegravir, and lamivudine tablets in children with HIV (IMPAACT 2019): week 24 results of an open-label, multicentre, phase 1-2 dose-confirmation study.
Child-friendly fixed-dose combination (FDC) antiretroviral therapy (ART) options are limited. We evaluated the pharmacokinetics, safety, and tolerability of dispersible and immediate-release FDC abacavir, dolutegravir, and lamivudine taken once per day in children younger than 12 years with HIV. IMPAACT 2019 was an international, phase 1-2, multisite, open-label, non-comparative dose-confirmation study of abacavir, dolutegravir, and lamivudine in children younger than 12 years. Participants were enrolled across five weight bands: those weighing 6 kg to less than 25 kg received abacavir (60 mg), dolutegravir (5 mg), and lamivudine (30 mg) dispersible tablets (three to six tablets depending on body weight), and those weighing 25 kg to less than 40 kg received abacavir (600 mg), dolutegravir (50 mg), and lamivudine (300 mg) in an immediate-release tablet. At entry, participants were ART naive or ART experienced and virologically suppressed on stable ART for 6 months or more. Dose confirmation was based on pharmacokinetic and safety criteria in the first five to seven participants in each weight band to week 4; all participants were followed up to week 48. We present the results for the primary objectives to assess pharmacokinetics, confirm dosing, and evaluate safety through 24 weeks across all weight bands. The trial is registered with ClinicalTrials.gov (NCT03760458). 57 children were enrolled and initiated study drug (26 [46%] female and 31 [54%] male; 37 [65%] Black, 18 [32%] Asian, and 1 [2%] had race reported as unknown). Within each weight band, 6 kg to less than 10 kg, 10 kg to less than 14 kg, 14 kg to less than 20 kg, 20 kg to less than 25 kg, and 25 kg or higher: the geometric mean dolutegravir area under the concentration time curve over the 24 h dosing interval (AUC0-24 h) was 75·9 h·μg/mL (33·7%), 91·0 h·μg/mL (36·5%), 71·4 h·μg/mL (23·5%), 84·4 h·μg/mL (26·3%), and 71·8 h·μg/mL (13·9%); dolutegravir concentrations 24 h after dosage (C24 h) were 0·91 μg/mL (67·6%), 1·22 μg/mL (77·5%), 0·79 μg/mL (44·2%), 1·35 μg/mL (95·5%), and 0·98 μg/mL (27·9%); abacavir AUC0-24 h was 17·7 h·μg/mL (38·8%), 19·8 h·μg/mL (50·6%), 15·1 h·μg/mL (40·3%), 17·4 h·μg/mL (19·4%), and 25·7 h·μg/mL (14·6%); lamivudine AUC0-24 h was 10·7 h·μg/mL (46·0%), 14·2 h·μg/mL (23·9%), 13·0 h·μg/mL (15·6%), 14·5 h·μg/mL (16·6%), and 21·7 h·μg/mL (26·2%), respectively. Pharmacokinetic targets and safety criteria were met within each weight band, and thus dosing of abacavir, dolutegravir, and lamivudine was confirmed at the originally selected doses. 54 (95%) of participants were treatment experienced and all who continued taking the study drug remained virologically suppressed (<200 copies per mL) through week 24. Virological suppression was achieved in two of three participants who were ART naive by week 24. There were no grade 3 or higher adverse events related to abacavir, dolutegravir, and lamivudine and no discontinuations because of toxicity to week 24. Both formulations were well tolerated. Dosing of abacavir, dolutegravir, and lamivudine was confirmed in children weighing 6 kg to less than 40 kg, and both FDC formulations were safe, well tolerated, and efficacious through 24 weeks of treatment. These findings support global efforts to expand the availability of FDC abacavir, dolutegravir, and lamivudine to children with HIV. National Institute of Allergy and Infectious Diseases, the Eunice Kennedy Shriver National Institute of Child Health and Human Development, the National Institute of Mental Health, ViiV Healthcare, and GlaxoSmithKline.


Serum osteoprotegerin (OPG) and the A163G polymorphism in the OPG promoter region are related to peripheral measures of bone mass and fracture odds ratios.
The purpose of this study is to investigate the association of serum osteoprotegerin (OPG) and the A163G polymorphism in the OPG promoter with peripheral measures of bone mass and with odds ratios for wrist and hip fracture in a case-control study of postmenopausal Danish women. The study included 66 women with lower forearm fracture, 41 women with hip fracture, and 206 age-matched controls. All had broadband ultrasound attenuation (BUA) and speed of sound (SOS) measured at the heel as well as bone mineral density (BMD) measured by DXA at the distal forearm. S-OPG was measured by ELISA. The A163G genotypes were determined by PCR-RFLP analysis. S-OPG levels correlated positively with age ( r = 0.45; P << 0.0001) and negatively with distal forearm BMD ( r = -0.31; P << 0.0001), heel BUA ( r = -0.23; P << 0.0001), and heel SOS ( r = -0.22; P << 0.0001). Comparing the highest quartile of S-OPG to the lowest, the odds ratio for osteoporotic fracture was 2.5 (95% CI, 1.3-4.7; P = 0.006). The G allele of the A163G was associated with significantly lower t-scores of both lower forearm BMD, heel BUA, and heel SOS as well as being significantly more frequent in the fracture patients compared to the controls. Patients with a combination of the highest quartile of S-OPG and presence of the G allele ( n = 23) had a significantly elevated fracture odds ratio, 4.0 (95% CI, 1.7-9.9). A significant negative association between S-OPG with peripheral measures of bone mass and with increased fracture odds ratios was found. Furthermore, the A163G mutation in the OPG promoter had a significant influence on bone mass and fracture status independently of S-OPG level.


Stable C isotope data of southern mixed-grass prairie vegetation from Oklahoma, United States.
Stable carbon isotopic data (δ13C) of 41 individual plant species was collected from long-term grazed and ungrazed pastures in Oklahoma, USA. These data can serve as a library of stable carbon isotope values for Southern mixed-grass prairie species. Seventeen warm-season (C4) and twenty-four cool-season (C3) plants were identified and collected from the United States Department of Agriculture (USDA) Southern Plains Experimental Range (SPER). Plant samples were dried at 55°C, and ground finely. The δ13C isotopic compositions were determined using a Europa Scientific automated nitrogen carbon analyzer (ANCA/NT) with a Solid/Liquid Preparation Module (Dumas combustion sample preparation system) coupled to a Europa 20-20 Stable isotope analyzer continuous flow isotope ratio mass spectrometer (Sercon Ltd, formerly Europa Scientific Ltd., Crewe, England). These data can be used as end members in isotopic mixing models or in paleoecology to correlate soil ages with plant species composition. Data from plant species provides information about soil organic carbon sequestration and possible long-term climate change.


Immunocytochemical demonstration of radial glia in the developing rat olfactory bulb with antibodies to glial fibrillary acidic protein.
Olfactory bulbs of 8-, 12-, 16- and 30-day-old rats were studied by means of immunocytochemistry using antibodies to glial fibrillary acidic protein (GFAP) and with the Toluidine blue-staining of semithin sections. Until day 12 the GFAP-reaction revealed a radial glia system, the fibres of which extended from the axial ventricular cleft to the surface. From day 16 onwards radial fibres were gradually replaced by typical astrocytes. The lack of proliferative activity within the bulb during the early postnatal period suggests that its cells are generated at and migrating from an external site. An intensely proliferating area was detected in the frontal lobe subventricular layer from where a bundle of migratory cells extends into the bulb. Radial glia may thus be of importance in guiding the migration of cells from this axial bundle to more peripheral regions of the olfactory bulb.


Speech sound production, speech intelligibility, and oral-motor outcomes of preterm children: Are they different from full term children?
We studied the speech sound abilities of preterm (PT) children. Thirty-one PT and twenty-nine full term (FT) children were recruited. Speech abilities were assessed in single word, story retelling, oral-motor, and intelligibility. PT group had poorer outcomes (Mean = 25.77, SD = 17.19) than FT ones (Mean = 5.9, SD = 4.97) for single word (p < 0.001). They obtained poorer results (Mean = 9.65, SD = 7.85) than FT peers (Mean = 2.95, SD = 3.34) in story retelling (p = 0.002) and intelligibility (Man-Whitney U = 89.50, p = 0.02). They obtained lower values for diadochokesis/patuku/(p < 0.001), isolated (p = 0.001), and sequenced movements (p = 0.02) but not for diadochokesis/patukejk/(p = 0.12). Significant values of correlation were found among single word scores with birth weight (r = -.54, p < .001) and gestational age (r = -0.67, p < .001) and story retelling scores with birth weight (r = -0.40, p = .013) and gestational age (r = -0.64, p < .001). The associations of single word score and maternal (r = -0.02, p = .85) and paternal education (r = -0.10, p = .41) were not significant. No significant relationships were obtained between story retelling score and maternal (r = 0.16, p = .34) and paternal education (r = 0.09, p = .59). The significant values were obtained for associations of intelligibility with isolated (r = 0.54, p = .001), sequenced movements (r = 0.59, p < .001), and diadochokesis/patukejk/(r = 0.39, p = .016) but not significant for intelligibility and diadochokesis/patuku/(r = 0.25, p = .13). Findings implied that speech abilities are weaker in PT children.


Effects of aluminium and low pH on growth and development in Rana temporaria tadpoles.
Rana temporaria tadpoles were raised to metamorphosis at 6 levels of pH (pH 3.6-6.5) and 2 levels of aluminium (800 and 1,600 μg l-1) at pH 4.4. Treatments involved both chronic and acute exposure to depressed pH. Decreasing pH reduced maximum body size and delayed metamorphosis. Growth was depressed and metamorphosis delayed by 800 μg l-1 Al, and several tadpoles died at foreleg emergence. At 1,600 μg l-1 Al, small tadpoles suffered arrested growth and development and eventually died, while larger tadpoles metamorphosed without delay, though at a very small size. There was no mortality among controls. The levels of pH and inorganic monomeric aluminium measured in the experiments were similar to field levels at a site in Scotland, and it is concluded that individual tadpoles will, in certain types of water body, be adversely affected by both acidity and aluminium. The impact of these factors on populations remains to be determined.


Finding the undiscovered roles of genes: an approach using mutual ranking of coexpressed genes and promoter architecture-case study: dual roles of thaumatin like proteins in biotic and abiotic stresses.
Regarding the possible multiple functions of a specific gene, finding the alternative roles of genes is a major challenge. Huge amount of available expression data and the central role of the promoter and its regulatory elements provide unique opportunely to address this issue. The question is that how the expression data and promoter analysis can be applied to uncover the different functions of a gene. A computational approach has been presented here by analysis of promoter regulatory elements, coexpressed gene as well as protein domain and prosite analysis. We applied our approach on Thaumatin like protein (TLP) as example. TLP is of group 5 of pathogenesis related proteins which their antifungal role has been proved previously. In contrast, Osmotin like proteins (OLPs) are basic form of TLPs with proved role only in abiotic stresses. We demonstrated the possible outstanding homolouges involving in both biotic and abiotic stresses by analyzing 300 coexpressed genes for each Arabidopsis TLP and OLP in biotic, abiotic, hormone, and light microarray experiments based on mutual ranking. In addition, promoter analysis was employed to detect transcription factor binding sites (TFBs) and their differences between OLPs and TLPs. A specific combination of five TFBs was found in all TLPs presenting the key structure in functional response of TLP to fungal stress. Interestingly, we found the fungal response TFBs in some of salt responsive OLPs, indicating the possible role of OLPs in biotic stresses. Thirteen TFBS were unique for all OLPs and some found in TLPs, proposing the possible role of these TLPs in abiotic stresses. Multivariate analysis showed the possibility of estimating models for distinguishing biotic and abiotic functions of TIPs based on promoter regulatory elements. This is the first report in identifying multiple roles of TLPs and OLPs in biotic and abiotic stresses. This study provides valuable clues for screening and discovering new genes with possible roles in tolerance against both biotic and abiotic stresses. Interestingly, principle component analysis showed that promoter regulatory elements of TLPs and OLPs are more variable than protein properties reinforcing the prominent role of promoter architecture in determining gene function alteration.


The trajectories of anxiety and depression during the COVID-19 pandemic and the protective role of psychological flexibility: A four-wave longitudinal study.
Published reports on the adverse mental health impacts of the initial phase of the COVID-19 pandemic suggest an emerging global mental health crisis. However, the trajectories of these mental health impacts over multiple COVID-19 peaks and corresponding lockdowns are unknown. This study explored the trajectories of anxiety and depression over two consecutive lockdowns during the first nine months of the pandemic in Europe (April 2020-January 2021) and examined whether they varied as a function of different psychological flexibility and inflexibility profiles. A total of 569 Italians completed online surveys at four assessment points. Trajectories of anxiety and depression were examined with latent growth modeling and according to different psychological flexibility and inflexibility profiles. Anxiety increased linearly throughout the study period, whereas depression displayed a quadratic trajectory evidencing a decrease with the easing of the first lockdown followed by an increase during the second lockdown. Furthermore, two profiles were identified that displayed different anxiety and depression trajectories. Compared to the psychologically flexible profile, the psychologically inflexible profile reported significantly higher anxiety and depression which remained higher across the study period. A reliance on self-report measures and convenience sampling constitute key study limitations. Results suggest that high psychological inflexibility is a risk factor for prolonged elevated anxiety and depression during the COVID-19 pandemic, whereas high psychological flexibility is a protective factor. Psychological flexibility and inflexibility should be targeted by preventive public health interventions that harness evidence-based strategies shown to effectively target these factors.


Population-based analysis of Alzheimer's disease risk alleles implicates genetic interactions.
Reported odds ratios and population attributable fractions (PAF) for late-onset Alzheimer's disease (LOAD) risk loci (BIN1, ABCA7, CR1, MS4A4E, CD2AP, PICALM, MS4A6A, CD33, and CLU) come from clinically ascertained samples. Little is known about the combined PAF for these LOAD risk alleles and the utility of these combined markers for case-control prediction. Here we evaluate these loci in a large population-based sample to estimate PAF and explore the effects of additive and nonadditive interactions on LOAD status prediction performance. 2419 samples from the Cache County Memory Study were genotyped for APOE and nine LOAD risk loci from AlzGene.org. We used logistic regression and receiver operator characteristic analysis to assess the LOAD status prediction performance of these loci using additive and nonadditive models and compared odds ratios and PAFs between AlzGene.org and Cache County. Odds ratios were comparable between Cache County and AlzGene.org when identical single nucleotide polymorphisms were genotyped. PAFs from AlzGene.org ranged from 2.25% to 37%; those from Cache County ranged from .05% to 20%. Including non-APOE alleles significantly improved LOAD status prediction performance (area under the curve = .80) over APOE alone (area under the curve = .78) when not constrained to an additive relationship (p < .03). We identified potential allelic interactions (p values uncorrected): CD33-MS4A4E (synergy factor = 5.31; p < .003) and CLU-MS4A4E (synergy factor = 3.81; p < .016). Although nonadditive interactions between loci significantly improve diagnostic ability, the improvement does not reach the desired sensitivity or specificity for clinical use. Nevertheless, these results suggest that understanding gene-gene interactions may be important in resolving Alzheimer's disease etiology.


The link between ego-resiliency, social support, SARS-CoV-2 anxiety and trauma effects. Polish adaptation of the Coronavirus Anxiety Scale.
Reports to date have shown that the SARS-CoV-2 pandemic may have a negative impact on individuals' mental health. The purpose of this study was to assess the relation between ego-resiliency, social support, coronavirus anxiety and trauma effects. The study employed the Polish adaptation of the Coronavirus Anxiety Scale (CAS). It involved 515 individuals aged 18-78. The Polish version of CAS revealed satisfactory internal consistency (α = 0.86). Structural equation modeling indicated that ego-resiliency (the Ego-Resiliency Scale) and social support (the Multidimensional Scale of Perceived Social Support) were correlated and negatively predicted the severity of the novel coronavirus anxiety (CAS). Moreover, the level of anxiety showed positive correlation with negative trauma effects (the short form of the Changes in Outlook Questionnaire). The scores indicate the need for practitioners to focus on interventions which elevate ego-resiliency and perceived social support to improve mental health during the SARS-CoV-2 pandemic.


Nitrate reductase of Dunaliella parva: electron donor specificity and heat activation.
Nitrate reductase of the salt tolerant alga Dunaliella parva, in contrast to that of most green algae, can use NADPH as well as NADH as electron donor. Extracts of cells contained various amounts of latent nitrate reductase. The latent enzyme could be activated at 45 degrees C but only in the presence of flavine adenine dinucleotide. The heat activated enzyme did not require flavine adenine dinucleotide for activity and was fully active with NADH, NADPH or reduced flavine mononucleotide as electron donors.


CO(2) enrichment reduces reproductive dominance in competing stands of Ambrosia artemisiifolia (common ragweed).
Plants growing in dense stands may not equally acquire or utilize extra carbon gained in elevated CO(2). As a result, reproductive differences between dominant and subordinate plants may be altered under rising CO(2) conditions. We hypothesized that elevated CO(2) would enhance the reproductive allocation of shaded, subordinate Ambrosia artemisiifolia L. (Asteraceae) individuals more than that of light-saturated dominants. We grew stands of A. artemisiifolia at either 360 or 720 muL L(-1) CO(2) levels and measured the growth and reproductive responses of competing individuals. To test whether elevated CO(2) altered size and reproductive inequalities within stands, we compared stand-level coefficients of variation (CV) in height growth and final shoot, root, and reproductive organ biomasses. Elevated CO(2) enhanced biomass and reduced the CV for all aspects of plant growth, especially reproductive biomass. Allocation to reproduction was higher in the elevated CO(2) than in the ambient treatment, and this difference was more pronounced in small, rather than large plant positive relationships between the CV and total stand productivity declined under elevated CO(2), indicating that growth enhancements to smaller plants diminished the relative biomass advantages of larger plants in increasingly crowded conditions. We conclude that elevated CO(2) stimulates stand-level reproduction while CO(2)-induced growth gains of subordinate A. artemisiifolia plants minimize differences in the reproductive output of small and large plants. Thus, more individuals are likely to produce greater amounts of seeds and pollen in future populations of this allergenic weed.


Loss of hepatic Nmnat1 has no impact on diet-induced fatty liver disease.
Nicotinamide adenine dinucleotide (NAD+), a biological molecule integral to redox reactions involved in multiple cellular processes, has the potential to treat nonalcoholic fatty liver diseases (NAFLDs) and nonalcoholic steatohepatitis (NASH). Nicotinamide mononucleotide adenylyltransferase (Nmnat1), one of the NAD+ biosynthesizing enzymes, plays a central role in all NAD+ metabolic pathways and it is vital to embryonic development. However, the function of Nmnat1 in metabolic pathology and, specifically, in the development and progression of NAFLD and NASH remains unexplored. First, we generated hepatic Nmnat1 knockout (H-Nmnat1-/-) mice to investigate the physiological function of Nmnat1 and found that NAD+ levels were significantly lower in H-Nmnat1-/- mice than control mice. However, H-Nmnat1-/- mice appeared normal with comparable metabolic activity. Next, we used three different diet-induced NASH models to assess the pathophysiological role of Nmant1 in metabolic disorders and discovered that hepatic loos of Nmnat1 decreased 35%-40% of total NAD+ in an obese state. Nevertheless, our analysis of phenotypic variations found comparable body composition, gene expression, and liver histology in all NASH models in H-Nmnat1-/- mice. We also found that aged H-Nmnat1-/- mice exhibited comparable liver phenotypes with control mice. These findings suggest that Nmnat1 has a redundancy to the pathophysiology of obesity-induced hepatic disorders.