Protective activity of propofol, Diprivan and intralipid against active oxygen species.
We separately studied the antioxidant properties of propofol (PPF), Diprivan (the commercial form of PPF) and intralipid (IL) (the vehicle solution of PPF in Diprivan) on active oxygen species produced by phorbol myristate acetate (10(-6) M)-stimulated human polymorphonuclear leukocytes (PMN: 5 x 10(5) cells/assay), human endothelial cells (5 x 10(5) cells/assay) or cell-free systems (NaOCl or H2O2/peroxidase systems), using luminol (10(-4) M)-enhanced chemiluminescence (CL). We also studied the protective effects of Diprivan on endothelial cells submitted to an oxidant stress induced by H2O2/MPO system: cytotoxicity was assessed by the release of preincorporated 51Cr. Propofol inhibited the CL produced by stimulated PMN in a dose dependent manner (until 5 x 10(-5) M, a clinically relevant concentration), while Diprivan and IL were not dose-dependent inhibitors. The CL produced by endothelial cells was dose-dependently inhibited by Diprivan and PPF, and weakly by IL (not dose-dependent). In cell free systems, dose-dependent inhibitions were obtained for the three products with a lower effect for IL. Diprivan efficaciously protected endothelial cells submitted to an oxidant stress, while IL was ineffective. By HPLC, we demonstrated that PPF was not incorporated into the cells. The drug thus acted by scavenging the active oxygen species released in the extracellular medium. IL acted in the same manner, but was a less powerful antioxidant.


Oral Crohn's disease revisited--a 10-year-review.
The clinical presentation of seven patients with oral lesions of Crohn's disease, but with no evidence of other gastrointestinal involvement was described by Tyldesley in 1979. The clinical progress of five of these patients available for review is reported after a minimum period of 10 years. The gross oedema of the lips and painful buccal lesions present in all five patients have shown gradual resolution from the time of initial diagnosis. The nomenclature for oral Crohn's disease and related conditions is discussed with specific reference to the term 'orofacial granulomatosis'.


Qualitative analysis of cancer care experiences among rural cancer survivors and caregivers.
Rural (vs urban) patients experience poorer cancer outcomes and are less likely to be engaged in cancer prevention, such as screening. As part of a community needs assessment, we explored rural cancer survivors' and caregivers' experiences, perceptions, and attitudes toward cancer care services. We conducted 3 focus groups (N = 20) in Spring 2021 in rural Nebraska. Three patterns of cancer diagnosis were regular care/screening without noticeable symptoms, treatment for symptoms not initially identified as cancer related, and symptom self-identification. Most participants, regardless of how diagnosis was made, had positive experiences with timely referral for testing (imaging and biopsy) and specialist care. Physician interpersonal skills set the tone for patient-provider communication, which colored the perception of overall care. Participants with physicians and care teams that were perceived as "considerate," "compassionate," and "caring" had positive experiences. Participants identified specific obstacles to care, including financial barriers, transportation, and lack of support groups, as well as more general cultural barriers. Survivors and caregivers identified organization-based supports that helped them address such barriers. Rural populations have unique perspectives about cancer care. Our results are being used by the state cancer coalition, state cancer control program, and the National Cancer Institute-designated cancer center to prioritize outreach and interventions aimed to reduce rural cancer disparities, such as revitalizing lay cancer navigator programs, conducting webinars for primary care and cancer specialty providers to discuss these findings and identify potential interventions, and collaborating with national and regional cancer support organizations to expand reach in rural communities.


Esophageal retention cyst: Esophagogastric junction outflow obstruction (EGJOO) as a potential etiology and management with endoscopic mucosal resection (EMR).
Esophageal retention cysts are acquired cysts with no known etiology. They are characterized by dilation of the submucosal glands. Symptomatic cysts are traditionally managed by surgical resection. We present a case of progressive dysphagia and chest pain secondary to esophageal retention cysts in the mid and distal esophagus with associated esophagogastric junction outflow obstruction (EGJOO) and jackhammer esophagus on high resolution manometry (HRM). The patient underwent staged endoscopic mucosal resection (EMR) with subsequent improvement in her symptoms. However, EGJOO persisted after resection, suggesting it was the primary pathology and not a consequence of the obstruction from the cysts. Esophageal retention cysts are rarely reported in the literature with most descriptions coming from incidental post-mortems. The presented case suggests EGJOO as a potential etiology of retention cysts. The proposed mechanism is that a significant rise in esophageal intraluminal pressure creates a state of stasis in the esophagus, ideal for the development of these cysts. Symptomatic or malignant retention cysts should be resected. We demonstrate the feasibility of EMR as an alternative to surgical resection. Esophageal retention cyst is a rare entity, which may arise as a result of EGJOO. The natural history and malignant potential of these cysts are unknown, and no formal guidelines have been established for follow-up for patients with asymptomatic retention cysts. Endoscopic mucosal resection can be used to successfully manage these cysts.


Successful Surgical Salvage for a Patient with Ruptured Coronary Sinus during Percutaneous Ablation for Atrial Fibrillation: A Case Report.
Herein, we describe the rare anatomy of an abnormal shunt from the left atrium to the coronary sinus, which ruptured during a percutaneous ablation for atrial fibrillation. The iatrogenic lesion was successfully repaired after emergent extracorporeal membrane oxygenation set up followed by surgical exploration. The patient's postoperative course was uneventful, and she was regularly followed up without any complications.


Brazilian normal static bone histomorphometry: effects of age, sex, and race.
Bone histomorphometry values for normal individuals within different populations have been well established. We studied iliac crest bone samples from 125 healthy Brazilian subjects. The effect of sex, race, and age variables on histomorphometric parameters was evaluated. Bone volume showed a trend to decrease with age in both sexes, being significantly higher in black females and Caucasian males. Interactions among sex, race, and age had no effect on trabecular thickness (Tb.Th) and trabecular separation (Tb.Sp). However, age had a significant effect on Tb.Th and Tb.Sp, and sex had an impact on Tb.Sp. Trabecular number (Tb.N) was higher in black females than in males and was higher in Asian males than in females. Among females, Tb.N was lower in Asians than in other races and was higher in blacks than in Caucasians and or in those of mulattos. In addition, Tb.N was higher in males under 10 than in males over 50 years old, was higher in females under 10 than in females in any other age bracket, and was lower in females in the 41-50 age bracket than in younger females. Osteoid volume and osteoid surface were significantly higher in males than in females, and a significant age-related difference in osteoid thickness was observed. No significant sex-related or race-related differences were found in terms of resorption, although eroded surface decreased with age. In conclusion, sex, race, and age, as well as interactions among these three variables, were found to affect some static histomorphometric indexes in healthy Brazilian subjects.


Chronic exposure to random restraint stress retards the development of estrogen-induced pituitary prolactinoma in rats.
The effects of stress on carcinogenesis have been equivocal. The present work examined the influence of an unpredictable pattern of chronic restraint stress on estradiol-induced pituitary hyperplasia in male Fischer 344 rats. The animals were grouped as follows: (1) control, (2) stressed, (3) estradiol (31 mg), subcutaneous implant for 40 days and (4) estradiol with stress, 1 h daily, randomly, for 40 days. The pituitaries increased markedly in mass in groups 3 and 4 compared to the first two groups. However, the gland weight in rats exposed to estradiol plus stress was significantly (P less than 0.04) reduced as compared to estradiol per se. Prolactin levels increased due to estradiol treatment and decreased following exposure to stress. Weights of adrenal glands increased as a result of estradiol treatment when compared to those in stressed or untreated animals. This adrenal hypertrophy correlated well with plasma corticosterone levels. The data demonstrate that chronic immobilization stress retarded estradiol-induced pituitary prolactinoma. Modulation of the immune response by stress-related factors may play a role in the inhibition of tumorigenesis.


Modulation of lipocalin-type prostaglandin D2 synthase expression in catfish seminal vesicles by thyroid disrupting agents and hormones.
Thyroid hormones play crucial role in several biological processes including reproduction. Disruption of normal thyroid status by environmental contaminants can cause severe impairment in reproductive functions. In our previous study, we reported down-regulation of a protein in seminal vesicular fluid of air-breathing catfish, Clarias gariepinus during experimentally induced hyperthyroidism. N-terminal amino acid sequence analysis followed by search in sequence database denoted it to be lipocalin-type prostaglandin D2 synthase (ptgds-b). In the present study, we cloned full-length cDNA of ptgds-b based on the N-terminal amino acid sequence. Surprisingly, Northern blot as well as RT-PCR analysis demonstrated the presence of ptgds-b transcript predominantly in seminal vesicles and developing testis. Further, ptgds-b mRNA significantly decreased in seminal vesicles following L-thyroxine overdose while there was an increased expression of ptgds-b after depletion of thyroid hormone by thiourea and withdrawal of the treatments reverted this effect. Treatment of catfish with human chorionic gonadotropin and estradiol significantly reduced ptgds-b expression. Taken together, we report ptgds-b as a thyroid hormone regulated protein in the seminal vesicles in addition to gonadotropin and estradiol. Further studies might explain the exclusive presence of ptgds-b in seminal vesicles and developing testis yet present data evaluated it as a putative biomarker for thyroid hormone disruption.


Mg²⁺-dependent modulation of BKCa channels by genistein in rat arteriolar smooth muscle cells.
Genistein, a protein tyrosine kinase (PTK) inhibitor, regulates ion channel activities. However, the mechanism of action of genistein on large-conductance calcium-activated potassium (BK(Ca)) channels is unclear. This study aimed to investigate whether the mechanism of Mg(2+)-dependent modulation of BK(Ca) channel activity in vascular smooth muscle cells involved inhibition of phosphorylation by genistein or direct interaction between genistein and BK(Ca) channels. The whole-cell and inside-out patch-clamp techniques were used to measure BK(Ca) currents and the effects of genistein on BK(Ca) channel activities in rat mesenteric arteriolar smooth muscle cells. We found that the effects of genistein on BK(Ca) currents were Mg(2+)-dependent. Genistein (50 μM) inhibited BK(Ca) currents if the intracellular free magnesium concentration ([Mg(2+)]i) was 2 μM or 20 μM, but amplified BK(Ca) currents if [Mg(2+)]i was 200 μM or 2000 μM. The inhibitory effect of genistein on BK(Ca) currents was reversed by the protein tyrosine phosphatase inhibitor sodium orthovanadate (0.5 mM). Daidzein (50 μM), an inactive analogue of genistein, also amplified BK(Ca) currents, and its amplification was insensitive to orthovanadate. Another PTK inhibitor, tyrphostin 23 (50 μM), reduced the open probability of BK(Ca) channels. This inhibitory effect was weaker at 200 μM [Mg(2+)]i than at 2 μM [Mg(2+) ]i, and was countered by orthovanadate. Our results suggest that genistein amplifies BK(Ca) currents at a high [Mg(2+)]i, but inhibits BK(Ca) currents at a low [Mg(2+)]i. The mechanism of this biphasic effects involves PTK-independent amplification and [Mg(2+)]i -PTK-dependent inhibition.


Genome Sequences of 42 Bacteria Isolated from Sorghum bicolor Roots.
Forty-two bacterial strains were isolated from root samples of Sorghum bicolor The strains spanned 17 genera, including Dechloromonas, Duganella, Dyella, Flavobacterium, Herbaspirillum, Lutibacter, Mucilaginibacter, Novosphingobium, Paraburkholderia, Pedobacter, Pleomorphomonas, Rhizobacter, Rhizobium, Rhizomicrobium, Rugamonas, Variovorax, and Xanthobacter Their whole-genome sequences revealed diverse metabolic processes, including biological nitrogen fixation, in sorghum root microbiota.


A visual schematic for clarifying the temporal relationship between the anion and osmol gaps in toxic alcohol poisoning.
Delayed treatment of ethylene glycol or methanol poisoning can result in life-threatening complications, but obtaining quantitative levels is delayed by several hours in most hospitals. Calculating the anion gap and the osmol gap are recommended to hasten identification and treatment in cases of suspected toxic alcohol poisoning. However, relying on the anion and osmol gap calculations without understanding the temporal relationship between these 2 gaps can lead to delayed identification and increased morbidity in cases of toxic alcohol poisoning. Our "Mountain" schematic illustrates the presence of an elevated osmol gap found early in toxic alcohol poisoning and the presence of an elevated anion gap found later in the course of poisoning. Using the "Mountain" diagram as a clarification of the temporal relationship between the anion and the osmol gap can improve the diagnostic use of these screening assays.


Detection of Langat virus by TaqMan real-time one-step qRT-PCR method.
Langat virus (LGTV), one of the members of the tick-borne encephalitis virus (TBEV) complex, was firstly isolated from Ixodes granulatus ticks in Malaysia. However, the prevalence of LGTV in ticks in the region remains unknown. Surveillance for LGTV is therefore important and thus a tool for specific detection of LGTV is needed. In the present study, we developed a real-time quantitative reverse-transcription-polymerase chain reaction (qRT-PCR) for rapid detection of LGTV. Our findings showed that the developed qRT-PCR could detect LGTV at a titre as low as 0.1 FFU/ml. The detection limit of the qRT-PCR assay at 95% probability was 0.28 FFU/ml as determined by probit analysis (p ≤ 0.05). Besides, the designed primers and probe did not amplify ORF of the E genes for some closely related and more pathogenic viruses including TBEV, Louping ill virus, Omsk hemorrhagic fever virus (OHFV), Alkhurma virus (ALKV), Kyasanur Forest Disease virus (KFDV) and Powassan virus (POWV) which showed the acceptable specificity of the developed assay. The sensitivity of the developed method also has been confirmed by determining the LGTV in infected tick cell line as well as LGTV- spiked tick tissues.


Germline deletion of Krüppel-like factor 14 does not increase risk of diet induced metabolic syndrome in male C57BL/6 mice.
The transcription factor Krüppel-like factor 14 (KLF14) has been associated with type 2 diabetes and high-density lipoprotein-cholesterol (HDL-C) through genome-wide association studies. The mechanistic underpinnings of KLF14's control of metabolic processes remain largely unknown. We studied the physiological roles of KLF14 in a knockout (KO) mouse model. Male whole body Klf14 KO mice were fed a chow or high fat diet (HFD) and diet induced phenotypes were analyzed. Additionally, tissue-specific expression of Klf14 was determined using RT-PCR, RNA sequencing, immunoblotting and whole mount lacZ staining. Finally, the consequences of KLF14 loss-of-function were studied using RNA sequencing in tissues with relatively high Klf14 expression levels. KLF14 loss-of-function did not affect HFD-induced weight gain or insulin resistance. Fasting plasma concentrations of glucose, insulin, cholesterol, HDL-C and ApoA-I were also comparable between Klf14+/+ and Klf14-/- mice on chow and HFD. We found that in mice expression of Klf14 was the highest in the anterior pituitary (adenohypophysis), lower but detectable in white adipose tissue and undetectable in liver. Loss of KLF14 function impacted on the pituitary transcriptome with extracellular matrix organization as the primary affected pathway and a predicted link to glucocorticoid receptor signaling. Whole body loss of KLF14 function in male mice does not result in metabolic abnormalities as assessed under chow and HFD conditions. Mostly likely there is redundancy for the role of KLF14 in the mouse and a diverging function in humans.


Hydrocephalus in tuberculous meningitis in children: treatment with acetazolamide and repeated lumbar puncture.
Increased intracranial pressure was found in 36 of 49 children with tuberculous meningitis. Four patients who presented with acute onset died early. Eight patients with obstructive hydrocephalus were treated surgically. In the 24 patients who had communicating hydrocephalus and were treated with acetazolamide and repeated lumbar punctures, treatment was successful in all but two, in whom shunt surgery was needed.


Trends in Ischemic Evaluation in New-Onset Heart Failure Without Known Coronary Artery Disease.
Guidelines recommend consideration of an ischemic evaluation (Class IIa-IIb) in new-onset heart failure (HF), but it is not well-known how often this is performed and leads to revascularization. The authors investigated temporal trends in ischemic evaluation and revascularization within 90 days of HF onset in Denmark 2008-2018. From the Danish nationwide administrative registries, diagnostic tests and revascularizations within 90 days were identified among patients with new-onset HF between 2008 and 2018, alive 90 days after diagnosis. Of 61,475 patients (mean age: 72.6 ± 13.8 years, 46% women), 12,503 (20%) underwent an ischemic evaluation, of whom 10,547 (84%) underwent invasive coronary angiography, and 1,956 (16%) underwent an initial noninvasive test, most frequently coronary computed tomographic angiography (n = 1,813, 93%). Of those who were initially referred for coronary computed tomographic angiography, 374 (21%) had a subsequent invasive coronary angiogram undertaken. Among individuals undergoing ischemic testing, percutaneous coronary intervention and coronary artery bypass graft surgery were performed in 1,354 (11%) and 619 (5%), respectively, corresponding to 2.2% and 1.0% of the entire sample. Between 2008 and 2018, the number of patients referred for ischemic evaluations increased, adjusted OR for 1.07 (95% CI: 1.06-1.07) per year high, and was greater among older versus younger individuals (OR: 1.01 [95% CI: 0.99-1.03], OR: 1.04 [95% CI: 1.03-1.06], OR: 1.06 [95% CI: 1.05-1.07], OR: 1.11 [95% CI: 1.09-1.12], and OR: 1.14 [95% CI: 1.10-1.18] per year increase for age group <50, 51-60, 61-75, 76-85, and >85 years, respectively, P for interaction <0.0001). In clinical practice, few patients with new-onset HF are referred for an ischemic evaluation and a minority undergo revascularization. Studies are needed to establish the appropriateness of this practice.


When is an ileal conduit indicated in the spinal cord injured patient?
Ileal loop diversion is only considered for persistent or progressive hydronephrosis (failed conservative treatment directed at the bladder). This conservative approach is based on a comparison of our ongoing prospective mortality study on 470 paraplegics plus 175 spinal cord patients that have survived at least 30 years v. an updated literature review of ileal conduits performed for benign disease.


Alterations in Ca(2+)-channels during the development of diabetic cardiomyopathy.
In order to examine the status of Ca2+ channels in heart sarcolemma during the development of diabetes, rats were injected intravenously with 65 mg/kg streptozotocin and hearts were removed 1, 3 and 8 weeks later. Crude membranes from the ventricular muscle were prepared and the specific binding of 3H-nitrendipine was studied by employing different concentrations of this Ca(2+)-antagonist. A significant decrease in both dissociation constant and maximal number of 3H-nitrendipine binding was observed in 3 and 8 weeks diabetic preparations. No such alterations were evident in diabetic brain membranes. Treatment of diabetic animals with insulin prevented the occurrence of these changes in the myocardium. The altered 3H-nitrendipine binding characteristics in diabetic heart membranes may not be due to the high levels of circulating catecholamines in this experimental model because no such changes were seen upon injecting a high dose (40 mg/kg) of isoproterenol in rats for 24 hr. The reduced number of 3H-nitrendipine binding sites may decrease Ca(2+)-influx through voltage sensitive Ca2+ channels and partly explain the depressed cardiac contractile force development in chronic diabetes whereas the increased affinity of Ca2+ channels may partly explain the increased sensitivity of diabetic heart to Ca2+.


Studies on the Transcorrelated Method.
We investigate the possibility of using a transcorrelated (TC) Hamiltonian to describe electron correlation. A method to obtain TC wavefunctions was developed based on the mathematical framework of the bi-variational principle. This involves the construction of an effective TC Hamiltonian matrix, which can be solved in a self-consistent manner. This was optimized using a method we call second-order-moment minimization and demonstrate that it is possible to obtain highly accurate energies for some closed-shell atoms and helium-like ions. The effects of certain correlator terms on the description of electron-electron and electron-nuclear cusps were also examined graphically, and some TC wavefunctions were compared against near-exact Hylleraas wavefunctions.


Tuberculosis and latent tuberculosis infection in close contacts of people with pulmonary tuberculosis in low-income and middle-income countries: a systematic review and meta-analysis.
Investigation of people exposed to cases of infectious tuberculosis (contact investigation) is key to tuberculosis control in countries with low tuberculosis incidence. However, in countries in which the incidence of tuberculosis is high, contact investigation is not commonly done. Increasing concerns about the failure to meet case-detection targets and about the spread of drug-resistant Mycobacterium tuberculosis have prompted a reassessment of the potential benefits of contact investigation. We did a systematic review to determine the yield of household contact investigation. The yield for all tuberculosis (bacteriologically confirmed and clinically diagnosed) was 4.5% (95% CI 4.3-4.8, I(2)=95.5%) of contacts investigated; for cases with bacteriological confirmation the yield was 2.3% (95% CI 2.1-2.5, I(2)=96.6%). Latent tuberculosis infection was found in 51.4% (95% CI 50.6-52.2, I(2)=99.4%) of contacts investigated. The substantial heterogeneity in all analyses indicated high variability among studies that was not accounted for by subgroup analyses. These results suggest that contact investigation merits serious consideration as a means to improve early case detection and decrease transmission of M tuberculosis in high-incidence areas.


SPRC19: A Database of State Policy Responses to COVID-19 in the United States.
SPRC19 is a new database that seeks to capture a wide range of state policy actions in response to COVID-19 in the United States. Since March 2020 we have monitored state governments' and multi-state associations' websites for executive orders, agency rules, new legislation, and court decisions. We categorize each policy action into one of 206 distinct policies, then document the branch of government, source document, announcement date, implementation date, and expiration date (if applicable). We also record whether the action represents the introduction of a new policy or the expansion or contraction of an existing policy. The current release of SPRC19, v3.0, captures over 13,000 distinct policy actions through April 2020, which constitutes thousands more actions than similar resources over the same time period.


Altered expression of pro-inflammatory and developmental genes in the fetal brain in a mouse model of maternal infection.
Human studies of unexplained cerebral palsy (CP) suggest an association with maternal infection. We used an established model of maternal infection, lipopolysaccharide (LPS) administration, to investigate the molecular changes in the fetal brain that may link maternal infection and CP. We compared gene expression in brains from mouse pups exposed to LPS in utero to those from saline-treated controls. Dams were injected with 50 microg LPS or saline on E18 with surgical delivery from 0.5 to 6h later. Differential gene expression was analyzed in the whole mouse brain using RT-PCR. When compared to control mice, pups exposed to LPS showed increased expression of pro-inflammatory genes monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6), and interleukin-1beta (IL-1beta), as well as VEGF, a regulator of vascular development and permeability, the anti-apoptotic protein Y-box-binding protein-1 (YB-1), and the neuronal differentiation factor necdin. LPS-exposed mice also showed downregulation of semaphorin 5b and groucho, involved in axon guidance and neurogenesis, respectively, providing evidence that LPS may disrupt normal developmental pathways. These data suggest possible mechanisms for adverse neurological outcomes following maternal infection involving elevated cytokine levels and altered expression of developmental genes in the fetal brain.


The effect of low level laser on condylar growth during mandibular advancement in rabbits.
It has been shown that Low Level Laser (LLL) has a positive effect on bone formation. The aim of this study was to evaluate the effect of low level laser on condylar growth during mandibular advancement in rabbits. Continuous forward mandibular advancement was performed in fourteen male Albino rabbits with the mean age of 8 weeks and the mean weight of 1.5 ± 0.5 kg, with acrylic inclined planes. The rabbits were randomly assigned into two groups after 4 weeks. LLL (KLO3: wave length 630 nm) was irradiated at 3 points around the TMJ, through the skin in the first group. The exposure was performed for 3 minutes at each point (a total of 9 minutes) once a day for 3 weeks. The control group was not exposed to any irradiation. The rabbits in both groups were sacrificed after two months and the histological evaluation of TMJ was performed to compare fibrous tissue, cartilage, and new bone formation in condylar region in both groups. Disc displacement was also detected in both groups. Student's t-test, Exact Fisher and Chi square tests were used for the statistical analysis. The formation of fibrous tissue was significantly lower, while bone formation was significantly greater in lased group as compared with control group. The thickness of cartilage did not differ significantly between two groups. Irradiation of LLL (KLO3) during mandibular advancement in rabbits, increases bone formation in condylar region, while neither increase in the cartilage thickness nor fibrous tissues was observed.


Kinetic studies of lipase from Candida rugosa: a comparative study between free and immobilized enzyme onto porous chitosan beads.
The search for an inexpensive support has motivated our group to undertake this work dealing with the use of chitosan as matrix for immobilizing lipase. In addition to its low cost, chitosan has several advantages for use as a support, including its lack of toxicity and chemical reactivity, allowing easy fixation of enzymes. In this article, we describe the immobilization of Candida rugosa lipase onto porous chitosan beads for the enzymatic hydrolysis of olive oil. The binding of the lipase onto the support was performed by physical adsorption using hexane as the dispersion medium. A comparative study between free and immobilized lipase was conducted in terms of pH, temperature, and thermal stability. A slightly lower value for optimum pH (6.0) was found for the immobilized form in comparison with that attained for the soluble lipase (7.0). The optimum reaction temperature shifted from 37 degrees C for the free lipase to 50 degrees C for the chitosan lipase. The patterns of heat stability indicated that the immobilization process tends to stabilize the enzyme. The half-life of the soluble free lipase at 55 degrees C was equal to 0.71 h (Kd = 0.98 h(-1)), whereas for the immobilized lipase it was 1.10 h (Kd = 0.63 h(-1)). Kinetics was tested at 37 degrees C following the hydrolysis of olive oil and obeys the Michaelis-Menten type of rate equation. The Km was 0.15 mM and the Vmax was 51 micromol/(min x mg), which were lower than for free lipase, suggesting that the apparent affinity toward the substrate changes and that the activity of the immobilized lipase decreases during the course of immobilization.


Clinical application of extracorporeal shock wave lithotripsy using microexplosions.
Extracorporeal stone disintegration using a chemical explosive (10 mg. lead azide) as an energy source of underwater shock waves was performed in 105 patients 11 to 72 years old who had stones in the upper urinary tract. We used a prototype disintegrator in this series. The over-all rate free of stones 3 months after treatment was 82 per cent. Shock wave therapy was performed alone in 77 patients (73 per cent), while the remainder required combined treatment with percutaneous and/or transurethral lithotripsy. The most common complications were colic pain (30 per cent) and fever (23 per cent). In 4 patients other complications, that is bacteremia, gastrointestinal bleeding, ureteral injury and subcapsular renal hematoma, were observed but they were treated conservatively with no serious adverse effects. Our study demonstrates the safe use of this method for clinical treatment.


Elimination of low steady-state concentrations of [5,6-3H2]prostaglandin E1 in the pulmonary and the systemic circulations of anaesthetized rats.
The elimination of [3H]prostaglandin E1 in anaesthetized rats was studied by continuous intravenous or intraarterial infusions, producing steady-state concentrations at the level of endogenous prostaglandin E2 in mixed venous blood. Blood samples (0.5 ml) were collected from the carotid artery or the right atrium, respectively. The levels of [3H]prostaglandin E1 were measured at different infusion time intervals and the 3H-labeled hydrophobic metabolites characterized. Cardiac output was estimated by a modification of the dye injection method, using 125I-labelled albumin as the marker. From the cardiac output and the rate of infusion, the fractional clearance of the lung and the systemic beds in the steady-state situation were estimated to 88.3 +/- 3.2% and 54.1 +/- 15.2% (mean +/- S.D.), RESPECTIVELY. The hydrophobic metabolites were characterized chromatographically on Sephadez LH-20 columns, using synthetically prepared [14C]prostaglandin metabolites as internal standards and markers. The identities of some metabolites were further established by derivative formation to a constant [3H]/[14C] ratio. The major metabolite was 15-keto-13,14-dihydro-[3H]prostaglandin E1, while 15-keto-[3H]prostaglandin E1 and 13,14-dihydro-[3H]prostaglandin E1 could not be demonstrated.


Impaired Distal Tubular Acidification, Renal Cysts and Nephrocalcinosis in Monogenic Hypertension.
Monogenic defects in tubular sodium handling contribute a small proportion to hypertension in childhood. Presentation varies from severe hypertension manifesting at birth to asymptomatic hypertension and hypokalemic metabolic alkalosis detected incidentally in adulthood. A 12-y-old girl presenting with polyuria, polydipsia, severe hypertension and seizures, was found to have hypokalemia, renal medullary cysts and nephrocalcinosis. Clinical exome revealed a homozygous variation of unknown significance in exon 5 of the HSD11B2 gene, indicating the diagnosis of apparent mineralocorticoid excess. Therapy with spironolactone was associated with resolution of hypokalemia and normal blood pressure during two-year follow up.


Rare-earth fluorescence thermometry of laser-induced plasmon heating in silver nanoparticles arrays.
The laser-induced plasmon heating of an ordered array of silver nanoparticles, under continuous illumination with an Ar laser, was probed by rare-earth fluorescence thermometry. The rise in temperature in the samples was monitored by measuring the temperature-sensitive photoluminescent emission of a europium complex (EuTTA) embedded in PMMA thin-films, deposited onto the nanoparticles array. A maximum temperature increase of 19 °C was determined upon resonant illumination with the surface plasmon resonance of the nanoarray at the highest pump Ar laser power (173 mW). The experimental results were supported by finite elements method electrodynamic simulations, which provided also information on the temporal dynamics of the heating process. This method proved to be a facile and accurate approach to probe the actual temperature increase due to photo-induced plasmon heating in plasmonic nanosystems.


A laboratory evaluation of the tensile bond strength of some orthodontic bonding resins to enamel.
In this in vitro study, the tensile bond strengths of three two-paste and four no-mix orthodontic bonding resins applied to etched enamel were determined. One hundred ninety-six extracted human teeth were used to prepare 14 bonded test specimens for each of the seven resin systems evaluated. The tensile bond strengths were expressed in MN.m-2 and the data were analyzed by a one-way analysis of variance at the 5% level of significance and with a procedure of multiple comparisons with the best resin. The tensile bond strengths were not significantly different (P = 0.5065).


Outcomes and predictors of patients with moderate or severe functional mitral regurgitation and nonischemic dilated cardiomyopathy.
Patients with functional mitral regurgitation (FMR) and nonischemic dilated cardiomyopathy (DCM) are associated with high mortality. Our study aimed to compare the clinical outcomes between different treatment strategies and identify predictors associated with the adverse outcomes. A total of 112 patients with moderate or severe FMR and nonischaemic DCM were included in our study. The primary composite outcome was all-cause death or unplanned hospitalization for heart failure. The secondary outcomes were individual components of the primary outcome and the cardiovascular death. In this study, the primary composite outcome occurred in 26 patients (44.8%) in mitral valve repair (MVr) group and 37 patients (68.5%) in medical group (hazard ratio [HR], 0.28; 95% confidence interval [CI], 0.14-0.55; p < .001). The 1-, 3-, and 5-year survival rates for patients with MVr were 96.6%, 91.8%, and 77.4%, respectively, which were significantly higher than that of medical group: 81.2%, 71.9%, and 65.1%, respectively (HR, 0.32; 95% CI, 0.12-0.87; p = .03). Left ventricular ejection fraction (LVEF) < 41.5% (p < .001) and atrial fibrillation (p = .02) were independently associated with the primary outcome. LVEF < 41.5% (p = .007), renal insufficiency (p = .003), and left ventricular end-diastolic diameter > 66.5 mm (p < .001) were independently associated with heightened risk for all-cause death. Compared with medical therapy, MVr was associated with a better prognosis in patients with moderate or severe FMR and nonischemic DCM. We observed that LVEF < 41.5% was the only independent predictor of the primary outcome and all individual components of secondary outcomes.


Impact of Endothelial Lipase on Cholesterol Efflux Capacity of Serum and High-density Lipoprotein.
Endothelial lipase (EL) is a potent modulator of the structural and functional properties of HDL. Impact of EL on cholesterol efflux capacity (CEC) of serum and isolated HDL is not well understood and apparently contradictory data were published. Here, we systematically examined the impact of EL on composition and CEC of serum and isolated HDL, in vitro and in vivo, using EL-overexpressing cells and EL-overexpressing mice. CEC was examined in a validated assay using 3H-cholesterol labelled J774 macrophages. In vitro EL-modification of serum resulted in complex alterations, including enrichment of serum with lipid-free/-poor apoA-I, decreased size of human (but not mouse) HDL and altered HDL lipid composition. EL-modification of serum increased CEC, in line with increased lipid-free/-poor apoA-I formation. In contrast, CEC of isolated HDL was decreased likely through altered lipid composition. In contrast to in vitro results, EL-overexpression in mice markedly decreased HDL-cholesterol and apolipoprotein A-I serum levels associated with a decreased CEC of serum. HDL lipid composition was altered, but HDL particle size and CEC were not affected. Our study highlights the multiple and complex effects of EL on HDL composition and function and may help to clarify the seemingly contradictory data found in published articles.


Circulating radioimmunoassayable inhibin during periods of transient follicle-stimulating hormone rise: secondary surge and unilateral ovariectomy.
We have measured changes in circulating immunoreactive (ir-) inhibin in male and female rats using an RIA with an antiserum raised against porcine inhibin alpha (1-26)-Gly-Tyr. The same synthetic peptide was used for standards and for the preparation of tracer. Serum ir-inhibin levels were significantly higher in intact female than in intact male rats (p less than 0.001). Immunoreactive inhibin was significantly reduced in both sexes 24 h after bilateral gonadectomy (p less than 0.0001). Unilateral ovariectomy (ULO) of female rats on metestrus caused a transient decrease in serum inhibin 8 h after surgery, but levels were not significantly different from those of sham-operated controls at later times after surgery. Increases in serum FSH and LH were observed for 8-18 h after ULO. Serum ir-inhibin levels were also measured on the early morning of estrus during the secondary FSH surge. At this time, ir-inhibin levels were low, while FSH levels were high and LH levels were low. These results show that serum ir-inhibin levels in rats are decreased at times when serum FSH levels are high.


Roald Dahl nurses' rose is a peach.
A nurse working for Roald Dahl's Marvellous Children's Charity launched a new rose at Chelsea Flower Show to raise money and mark Dahl's centenary.


Community based distribution of ivermectin in eastern Sudan: acceptability and early post-treatment reactions.
A study to monitor ivermectin acceptability and post-treatment reactions during mass community distribution was carried out in eastern Sudan, where severe reactive onchodermatitis is prevalent. Of 1081 individuals eligible for treatment, 1076 (99.5%) accepted the ivermectin. Post-treatment reactions were monitored by self reporting, 5 d after a single dose of about 150 micrograms/kg (range 103-200 micrograms/kg); 230 persons reported adverse events (21.4%). No reaction was rated as severe. The most common problem was itching with cutaneous papular eruptions (16.2%). Local oedematous swelling was the second most common and the most slowly resolving complaint (5.4%), followed by musculoskeletal pain. There was a high acceptance rate of the treatment and remarkable tolerance of the post-treatment effects, probably due to efforts made to prepare the community to expect reactions to ivermectin, widespread awareness of the beneficial effects of treatment by villagers who had participated in clinical trials previously, and the encouragement we gave to the population to become involved in improvement of their health care services. Single doses of ivermectin resulted in good clinical responses and created much goodwill among villagers. Improvements in physical fitness, ability to work, and freedom from musculoskeletal pain were reported at the 3 months' follow-up. We recommend that, during mass distribution of ivermectin, community involvement in planning overall health improvement should be included, since the treatment initiates the process well. In areas where sowda syndrome is prevalent, medical surveillance for 3 d or more should be considered.


O-GlcNAc transferase plays a non-catalytic role in C. elegans male fertility.
Animal behavior is influenced by the competing drives to maintain energy and to reproduce. The balance between these evolutionary pressures and how nutrient signaling pathways intersect with mating remains unclear. The nutrient sensor O-GlcNAc transferase, which post-translationally modifies intracellular proteins with a single monosaccharide, is responsive to cellular nutrient status and regulates diverse biological processes. Though essential in most metazoans, O-GlcNAc transferase (ogt-1) is dispensable in Caenorhabditis elegans, allowing genetic analysis of its physiological roles. Compared to control, ogt-1 males had a four-fold reduction in mean offspring, with nearly two thirds producing zero progeny. Interestingly, we found that ogt-1 males transferred sperm less often, and virgin males had reduced sperm count. ogt-1 males were also less likely to engage in mate-searching and mate-response behaviors. Surprisingly, we found normal fertility for males with hypodermal expression of ogt-1 and for ogt-1 strains with catalytic-dead mutations. This suggests OGT-1 serves a non-catalytic function in the hypodermis impacting male fertility and mating behavior. This study builds upon research on the nutrient sensor O-GlcNAc transferase and demonstrates a role it plays in the interplay between the evolutionary drives for reproduction and survival.


Beneficial effect of moderate weight loss in older patients with non-insulin-dependent diabetes mellitus poorly controlled with insulin.
Fifteen patients with non-insulin-dependent diabetes mellitus, poorly controlled on insulin, were enrolled in a weight loss program. Criteria included a fasting plasma glucose concentration greater than 200 mg/dl, age in excess of 65 years, and the presence of obesity. Twelve of the 15 patients were able to complete the program, and the average weight loss of the group was 9 kg. Prior to initiation of the weight loss program the 12 patients were receiving (mean +/- SEM) 52 +/- 5 units of insulin per day and had a mean (+/- SEM) fasting plasma glucose concentration of 258 +/- 10 mg/dl. Insulin treatment was discontinued in all subjects during the weight loss period, and did not need to be resumed when weight stabilization had occurred. On the other hand, reasonable diabetic control could not be maintained without medication, and all patients were started on chlorpropamide during the period of weight stabilization. With this approach, a mean (+/- SEM) fasting glucose concentration of 137 +/- 4 mg/dl was achieved with a daily chlorpropamide dose of 354 +/- 30 mg. This dramatic clinical change took place despite the fact that no patient achieved ideal body weight. These results document the ability of a moderate amount of weight loss to transform patients with non-insulin-dependent diabetes mellitus, who are overweight and older than 65 years of age, from treatment failures to treatment successes.


External Noise and External Signal Induced Transition of Gene Switch and Coherence Resonance in the Genetic Regulatory System.
The transition of gene switch induced by external noises (multiplicative external noise and additive external noise) and external signals is investigated in the genetic regulatory system. Results show that the state-to-state transition of gene switch as well as resonant behaviors, such as the explicit coherence resonance (ECR), implicit coherence resonance (ICR) and control parameter coherence biresonance (CPCBR), can appear when noises are injected into the genetic regulatory system. The ECR is increased with the increase of the control parameter value when starting from the supercritical Hopf bifurcation parameter point, and there exists a critical control parameter value for the occurrence of ECR. However, the ICR is decreased as the control parameter value is increased when starting from the subcritical Hopf bifurcation point. In particular, the coherence of ECR is higher and more sensitive to noise than that of ICR. When an external signal is introduced into the system, the enhancement or suppression of the CPCBR and the number of peaks strongly depend on the frequency and amplitude of the external signal. Furthermore, the gene regulation system can selectively enhance or decrease the noise-induced oscillation signals at preferred frequency and amplitude of an external signal.


[Combination of the ureteral dilation catheter and balloon catheter under the ureteroscope in the treatment of male urethral stricture].
To investigate the clinical application of the ureteral dilation catheter combined with the balloon catheter under the ureteroscope in the treatment of urethral stricture in men. Under the ureteroscope, 45 male patients with urethral stricture received placement of a zebra guide wire through the strictured urethra into the bladder and then a ureteral dilation catheter along the guide wire, followed by dilation of the urethra from F8 initially to F14 and F16. Again, the ureteroscope was used to determine the length of the strictured urethra, its distance to the external urethral orifice, and whether it was normally located. An F24 balloon catheter and then a metal urethral calibrator was used for the dilation of the strictured urethra. After removal of the F18-F22 urethral catheter at 8 weeks, the urinary flow rate was measured immediately and again at 3 months. All the operations were successfully performed without serious complications. The maximum urinary flow rate was (13.3-29.9) ml/s (mean [17.7 ± 3.2] ml/s) at the removal of the catheter and (15.2-30.8) ml/s (mean [19.8 ± 3.9] ml/s) at 3 months after it. Smooth urination was found in all the patients during the 6-24 months follow-up. The application of the ureteral dilation catheter combined with, the balloon catheter under the ureteroscope is a good option for the treatment of male urethral stricture for its advantages of uncomplicatedness, safety, effectiveness, few complications, less pain, high success rate, and repeatable operation.


Self-assembled micellar aggregates based monomethoxyl poly(ethylene glycol)-b-poly(ε-caprolactone)-b-poly(aminoethyl methacrylate) triblock copolymers as efficient gene delivery vectors.
Amphiphilic triblock copolymers monomethoxyl poly(ethylene glycol) (mPEG)-b-poly(ε-caprolactone) (PCL)-b-poly(aminoethyl methacrylate)s (PAMAs) (mPECAs) were synthesized as gene delivery vectors. They exhibited lower cytotoxicity and higher transfection efficiency in COS-7 cells in presence of serum compared to 25 kDa bPEI. The influence of mPEG and PCL segments in mPECAs was evaluated by comparing with corresponding diblock copolymers. The studies showed the incorporation of the hydrophobic PCL segment in triblock copolymers affected the binding capability to pDNA and surface charges of complexes due to the formation of micelles increasing the local charges. The presence of mPEG segment in gene vector decreased the surface charges of the complexes and increased the stability of the complexes in serum because of the steric hindrance effect. It was also found that the combination of PEG and PCL segments into one macromolecule might lead to synergistic effect for better transfection efficiency in serum.


Calpain activity and translational expression increased in spinal cord injury.
Calpain, a calcium-activated neutral proteinase, has been implicated in myelin and cytoskeletal protein degradation following spinal cord injury. In the present study, we examined the activity and transcriptional expression of calpain in spinal cord injury lesions via Western blotting analysis and RT-PCR, respectively. No increases in transcriptional expression of calpain or calpastatin, the endogenous inhibitor, were observed in the lesion at 1, 4, 24, and 72 h following injury. However, calpain activity (as measured by calpain-specific degradation of the endogenous substrate fodrin) was marginally increased at 4 h and significantly increased by 129.8% at 48 h compared to sham controls after injury. Calpain translational expression was localized in injured spinal cords using double immunofluorescent labeling which revealed increased calpain expression in astrocytes compared to sham controls. These results suggest that calpain produced by astrocytes located in or near spinal cord injury lesions may participate in myelin/axon degeneration following injury.


Cumulative labeling of embryonic mouse neural retina with bromodeoxyuridine supplied by an osmotic minipump.
I describe a method for providing cumulative label of bromodeoxyuridine (BrdU) to mouse embryos. Commercially available osmotic pumps, which release their contents at a steady rate, were implanted subcutaneously in the interscapular space of pregnant mice on embryonic day (E) 9.5-12.5. Survival times varied from 4h to 37 days. Tissues (embryonic and neonatal eyes and maternal intestine) were immunochemically labeled for BrdU and examined histologically. The first detectably labeled cells appeared 4-7h post-implantation (hpi) and all cycling cells were labeled for at least 7 days post-implantation (dpi). Retinal development appeared normal. Terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) stained retinas that had been exposed to BrdU showed no more apoptotic cells than those unexposed. I conclude that the maternally implanted osmotic pump successfully provides cumulative BrdU labeling in the mouse embryo.


Detection and quantification of Campylobacter jejuni and Campylobacter coli using real-time multiplex PCR.
We describe a real-time quantitative multiplex polymerase chain reaction (qmPCR) assay to identify and discriminate between isolates of Campylobacter jejuni and Campylobacter coli. Two novel sets of primers and hydrolysis probes were designed to amplify the unique DNA sequences within the hipO, ccoN and cadF genes that are specific to Camp. jejuni and Camp. coli. Using the designed optimized qmPCR assay conditions, the amplification efficiency is in range from 108 to 116%. These qmPCR assays are highly specific for Camp. jejuni and Camp. coli, as seen through testing of 40 Campylobacter strains and 17 non-Campylobacter strains. In chicken juice and tap water models spiked with known quantities of Camp. jejuni, qmPCR detected 10(2) -10(3) CFU ml(-1) within 4 h. The qmPCR assays developed in this study provide reliable and simultaneous detection and quantification of Camp. jejuni and Camp. coli, with good amplification reaction parameters. Following further validation, the qmPCR assay reported here has the potential to be applied to various sample types as an alternative and rapid methodology.


Thermodynamics of proton transport coupled ATP synthesis.
The thermodynamic H(+)/ATP ratio of the H(+)-ATP synthase from chloroplasts was measured in proteoliposomes after energization of the membrane by an acid base transition (Turina et al. 2003 [13], 418-422). The method is discussed, and all published data obtained with this system are combined and analyzed as a single dataset. This meta-analysis led to the following results. 1) At equilibrium, the transmembrane ΔpH is energetically equivalent to the transmembrane electric potential difference. 2) The standard free energy for ATP synthesis (reference reaction) is ΔG°(ref)=33.8±1.3kJ/mol. 3) The thermodynamic H(+)/ATP ratio, as obtained from the shift of the ATP synthesis equilibrium induced by changing the transmembrane ΔpH (varying either pH(in) or pH(out)) is 4.0±0.1. The structural H(+)/ATP ratio, calculated from the ratio of proton binding sites on the c-subunit-ring in F(0) to the catalytic nucleotide binding sites on the β-subunits in F(1), is c/β=14/3=4.7. We infer that the energy of 0.7 protons per ATP that flow through the enzyme, but do not contribute to shifting the ATP/(ADP·Pi) ratio, is used for additional processes within the enzyme, such as activation, and/or energy dissipation, due e.g. to internal uncoupling. The ratio between the thermodynamic and the structural H(+)/ATP values is 0.85, and we conclude that this value represents the efficiency of the chemiosmotic energy conversion within the chloroplast H(+)-ATP synthase.


Effects of age on left ventricular dimensions and filling dynamics in 117 normal persons.
To determine standard left ventricular Doppler measurements in a large reference group of various ages to reflect the senescence process. We prospectively studied the influence of aging on left ventricular diastolic filling by performing Doppler echocardiography in 117 normal healthy volunteers. Transthoracic pulsed-wave Doppler echocardiographic studies of pulmonary venous flow and left ventricular inflow were done in the 53 male and 64 female study subjects. For analysis, the study subjects were arbitrarily classified into two groups: those younger than 50 years of age (group 1; N = 61) and those 50 years old or older (group 2; N = 56). A striking difference was found in pulmonary venous flow and left ventricular inflow between group 1 and group 2 subjects. Group 2 had increased pulmonary venous peak systolic flow velocity (71 +/- 9 versus 48 +/- 9 cm/s), decreased peak diastolic flow velocity (38 +/- 9 versus 50 +/- 10 cm/s), increased peak atrial reversal flow velocity (23 +/- 4 versus 19 +/- 4 cm/s), and increased percentage of forward flow in systole (65 +/- 7 versus 55 +/- 8%) in comparison with group 1. In group 2, peak early filling velocity (62 +/- 14 versus 72 +/- 14 cm/s) and ratio of early filling to atrial filling (1.1 +/- 0.3 versus 1.9 +/- 0.6) were lower and peak atrial filling velocity (59 +/- 14 versus 40 +/- 10 cm/s) was higher than in group 1. Deceleration time (210 +/- 36 versus 179 +/- 20 ms) and isovolumic relaxation time (90 +/- 17 versus 76 +/- 11 ms) were prolonged in group 2 in comparison with group 1. This study confirms the major influence of the aging process on left ventricular diastolic function in a large series of normal subjects. This physiologic factor should be considered in Doppler assessment of left ventricular diastolic filling in future studies.


Epicardial oxygen tensions during changes in arterial PO2 in pigs.
Arterial hypoxemia decreased epicardial tissue PO2, measured by means of a multiwire surface electrode, as well as coronary venous PO2 and myocardial lactate extraction. Left ventricular blood flow increased, O2 delivery, O2 demand and O2 consumption of the left ventricle remained unchanged. Thus, epicardial and coronary venous PO2 indicated decreased capillary and interstitial PO2 rather than cellular hypoxia. A linear relation between mean epicardial PO2 and coronary venous PO2 proves both parameters equally effective in reflecting changes in myocardial tissue oxygenation. However, PO2 distribution curves provide additional information and epicardial PO2 is superior in models with regional changes of myocardial oxygenation.


Differential expression of CCR3 ligand mRNA in guinea pig lungs during allergen-induced inflammation.
The gene expression profile of CCR3 ligands, eotaxin, RANTES, and monocyte chemoattractant protein-3 (MCP-3), was examined in normal and inflamed guinea pig lungs. Male Hartley guinea pigs (n = 49). Pulmonary mRNA was obtained from naive animals, animals treated with intravenous lipopolysaccharide administration, and animals repeatedly exposed to aerosolized allergen (ovalbumin). Northern analysis was performed to quantify pulmonary expression of eotaxin, RANTES, and MCP-3 mRNA. Pulmonary eosinophil peroxidase (EPO) activity was measured to quantify eosinophil accumulation. Eotaxin and RANTES mRNAs, but not MCP-3 mRNA, were constitutively expressed in guinea pig lungs. Lipopolysaccharide treatment increased MCP-3 mRNA expression, but not eotaxin or RANTES mRNA. In contrast, allergen exposure in sensitized animals caused an increase in eotaxin mRNA, which demonstrated good temporal and quantitative correlation with pulmonary EPO activity, but not in MCP-3 or RANTES mRNA. Guinea pig CCR3 ligands demonstrated different gene expression profiles in normal and inflamed airways, suggesting that they play different physiological and pathophysiological roles in the airway.


Roles of the hypoxia response system in hematopoietic and leukemic stem cells.
Stem cells exhibit a number of characteristic features, including the capacity for self-renewal and differentiation into multiple cell types, stress resistance, and drug efflux activity. These specific biological characteristics are supported by signals from the surrounding niche and the stemcell-specific transcription factor set, including hypoxia and the machinery that senses low oxygen levels. These properties are essential for normal stem cells, and when defective may induce cellular senescence and tumorigenesis. In contrast, cancer stem cells in tumor tissue utilize these biological characters driven by stemcell-specific molecular mechanisms and acquire indefinite self-renewal capacity, drug resistance, and metastatic ability. A fuller understanding of the differences between normal and malignant stem cells in the biological and molecular context is, therefore, necessary to the development of therapies against cancer stem cells. In this review, we discuss the effect of hypoxic microenvironment on normal and malignant stem cells and describe their molecular machinery with an emphasis on hematopoietic stem cells and their malignant counterparts, leukemic stem cells.


What are the underlying reasons behind socioeconomic differences in doctor-patient communication in head and neck oncology review clinics?
To explore socioeconomic status (SES) differences in patterns of doctor-patient communication within head and neck cancer clinics and why such differences exist. Thirty-six head and neck cancer review appointments with five Physicians were observed and audio-taped, along with follow-up interviews involving 32 patients. Data were analysed using Thematic Analysis, and compared by patient SES (education, occupation and Indices of Multiple Deprivation). Three main themes were identified: (a) Physicians used more humour and small talk in their consultations with high SES patients; (b) Low SES patients were more passive in their participation, engaged in less agenda setting and information-seeking, and framed their clinical experience differently; (c) Low SES patients had different preferences for involvement, defining involvement differently to high SES patients and were seen to take a more stoical approach. Low SES patients take a more passive role in medical consultations, engage in less relational talk and are less likely to raise concerns, but were satisfied with this. Physicians may adapt their communication behaviour in response to low SES patients' expectations and preferences. A question prompt list may help low SES patients to raise concerns during their consultations. This may reduce inequalities in communication and health.


Tumor necrosis factor-alpha inhibits cell proliferation and induces class II antigens and cell adhesion molecules in cultured normal human keratinocytes in vitro.
The effects of recombinant human tumor necrosis factor-alpha (TNF) on cell proliferation, cell morphology, and on the expression of class II alloantigens and intercellular adhesion molecule-1 (ICAM-1) were assessed in human keratinocytes cultured in serum-free medium. TNF inhibited cell proliferation in a time- and dose-dependent manner with a minimum effective dose of 10 U/ml and a 50% inhibitory dose of 100 U/ml. However, TNF did not induce cell death, and the growth inhibition induced by TNF was completely reversible after its withdrawal. In vitro combination of TNF with interferon-alpha (IFN-alpha) and IFN-beta resulted in additive growth inhibitory effects, while IFN-gamma enhanced the TNF mediated growth inhibition in a synergistic way. Furthermore, TNF altered the morphology of the growing keratinocytes inducing the appearance of a fusiform, fibroblast-like population. Also, treatment with TNF over 6 days markedly induced the expression of ICAM-1 on the cultured keratinocytes with a minimal effective dose of 10 U/ml, while HLA-DR was only moderately expressed after 1,000 U/ml. TNF did not induce HLA-DQ, but reduced the IFN-gamma induced expression of HLA-DR and HLA-DQ. By immunoelectron microscopy, an intense membrane-bound labeling for ICAM-1 was found after treatment with TNF, clearly pronounced in areas of microvillous membrane protrusions. These results indicate that epidermal keratinocytes are a major target for various biological effects of TNF. We also found that TNF differentially modulates IFN-gamma-induced effects, thus suggesting its potential role in the regulation of inflammatory skin disorders.


Nebulized Amphotericin B in Mechanically Ventilated COVID-19 Patients to Prevent Invasive Pulmonary Aspergillosis: A Retrospective Cohort Study.
Despite high mortality rates of COVID-19-associated pulmonary aspergillosis (CAPA) in the ICU, antifungal prophylaxis remains a subject of debate. We initiated nebulized conventional amphotericin B (c-AmB) as antifungal prophylaxis in COVID-19 patients on invasive mechanical ventilation (IMV). To assess the CAPA incidence in COVID-19 patients on IMV treated with and without nebulized c-AmB as antifungal prophylaxis. Retrospective cohort study of consecutive COVID-19 patients admitted to our adult 17-bed ICU in a university-affiliated general hospital in Ede, The Netherlands, between January 25, 2021, and July 9, 2021. Patients not requiring IMV or transferred from or to another ICU were excluded. From April 9, 2021, daily nebulized amphotericin B in all patients on IMV was initiated. Bronchoscopy with bronchoalveolar lavage (BAL) was performed in case of positive cultures for Aspergillus from the respiratory tract and/or unexplained respiratory deterioration. Incidence of probable and proven CAPA was compared between patients treated with and without nebulized antifungal prophylaxis using Pearson chi-square test. A total of 39 intubated COVID-19 patients could be analyzed, of which 16 were treated with antifungal prophylaxis and 23 were not. Twenty-six patients underwent bronchoscopy with BAL. In patients treated with antifungal prophylaxis, the incidence of probable/proven CAPA was significantly lower when compared with no antifungal prophylaxis (27% vs 67%; p = 0.047). Incidence of tracheobronchial lesions and positive Aspergillus cultures and BAL-galactomannan was significantly lower in patients treated with antifungal prophylaxis (9% vs 47%; p = 0.040, 9% vs 53%; p = 0.044, and 20% vs 60%; p = 0.047, respectively). No treatment-related adverse events and no case of proven CAPA were encountered in patients receiving antifungal prophylaxis. Nebulization of c-AmB in critically ill COVID-19 patients on IMV is safe and may be considered as antifungal prophylaxis to prevent CAPA. However, a randomized controlled trial to confirm this is warranted.


Determinants of drug effects on punished responding.
The effects of drugs on punished responding depend on interactions among a large number of experimental variables. Among these variables are the drug history of the animal, the dose of the drug administered, the type of stimulus used to punish responding, the intensity and duration of the punishing stimulus, the schedule of presentation of the punishing stimulus, the control rate and pattern of punished responding, the schedule of positive reinforcement maintaining the punished responding, the species of animal, the deprivation state of the animal, the behavioral history of the animal, and the nature of the required response. Although it is not known how all of these variables interact to determine the effect of drugs on punished responding, there is evidence that many of these variables are important as determinants of drug effects. The task facing behavioral pharmacologists studying drug effects on punished responding is to determine under what conditions drugs produce their characteristic effects on punished responding.


Neurocognitive Performance is Not Degraded After Stellate Ganglion Block Treatment for Post-Traumatic Stress Disorder: A Case Series.
To measure key neurocognitive performance effects following stellate ganglion block (SGB) administered to treat post-traumatic stress disorder (PTSD) symptoms. Eleven patients diagnosed, screened, and scheduled for SGB to treat their PTSD symptoms were administered a panel of eight cognitive measures before and 1 to 3 weeks after undergoing this procedure. PTSD symptoms were evaluated using the Posttraumatic Stress Disorder Checklist-Military. One to three weeks post-SGB, none of the patients showed any statistically significant decline in neurocognitive performance. Rather, there was a clear trend in improvement, with four out of eight measures reaching statistical significance following SGB. All patients improved in PTSD symptoms with a mean improvement on Posttraumatic Stress Disorder Checklist-Military of 29. In this case series of 11 patients, SGB effectively treated PTSD symptoms and did not impair reaction time, memory, or concentration. Therefore, SGB should be considered as a viable treatment option for personnel with PTSD symptoms who will be placed in demanding conditions such as combat.


Plasmin generation and fibrinolysis in pediatric patients undergoing cardiopulmonary bypass surgery.
This prospective, single-centre cohort study aimed to evaluate plasmin generation and fibrinolysis during and after cardiopulmonary bypass (CPB) surgery in a cohort of children up to 6 years of age. Blood samples were drawn at eight time points: after induction of anesthesia, before unfractionated heparin (UFH), after UFH, after initiation of bypass, before protamine, after protamine, after chest closure, and 6 h after chest closure. The study identified an increase in fibrinolysis during CPB and particularly up to 6 h afterward in children. This could be the mechanism for the significant bleeding events observed in this young population after CPB. This study establishes the foundation for future studies in this area, particularly those focusing on clinical outcomes after CPB surgery.


A new synthetic chalcone derivative, 2-hydroxy-3',5,5'-trimethoxychalcone (DK-139), suppresses the Toll-like receptor 4-mediated inflammatory response through inhibition of the Akt/NF-κB pathway in BV2 microglial cells.
Microglial cells are the resident innate immune cells that sense pathogens and tissue injury in the central nervous system (CNS). Microglial activation is critical for neuroinflammatory responses. The synthetic compound 2-hydroxy-3',5,5'-trimethoxychalcone (DK-139) is a novel chalcone-derived compound. In this study, we investigated the effects of DK-139 on Toll-like receptor 4 (TLR4)-mediated inflammatory responses in BV2 microglial cells. DK-139 inhibited lipopolysaccharide (LPS)-induced TLR4 activity, as determined using a cell-based assay. DK-139 blocked LPS-induced phosphorylation of IκB and p65/RelA NF-κB, resulting in inhibition of the nuclear translocation and trans-acting activity of NF-κB in BV2 microglial cells. We also found that DK-139 reduced the expression of NF-κB target genes, such as those for COX-2, iNOS, and IL-1β, in LPS-stimulated BV2 microglial cells. Interestingly, DK-139 blocked LPS-induced Akt phosphorylation. Inhibition of Akt abrogated LPS-induced phosphorylation of p65/RelA, while overexpression of dominant- active p110CAAX enhanced p65/RelA phosphorylation as well as iNOS and COX2 expression. These results suggest that DK-139 exerts an anti-inflammatory effect on microglial cells by inhibiting the Akt/IκB kinase (IKK)/NF-κB signaling pathway.


Simulating alveoli-inspired air pockets in a ZnO/NiMoO4/C3N4 catalyst filter for toluene entrapment and photodecomposition.
Here, we propose an alveoli-inspired catalyst to address the susceptibility of photocatalytic air oxidation systems to fluctuations in volatile organic contaminant (VOC) loads. An alveoli structure was fabricated by covering ZnO nanorods grown on a stainless-steel mesh (SSM) with a porous NiMoO4/C3N4 layer. The alveoli catalyst regulates VOC mass transfer from the air to the catalyst surface using air pockets that capture VOC molecules by diffusion driven by a concentration gradient. Air pockets act as localized reservoirs of molecules that prevent scarcity and congestion at the catalyst surface at low and high VOC loads, respectively. The presence of air pockets in the catalyst assembly and its potential to capture VOC was confirmed by a distinct bimodal adsorption configuration. A ZnO/NiMoO4/C3N4@SSM (ZNC@SSM) catalyst with air pockets achieved a high degree of toluene adsorption (6.1 μmol·m-2). Toluene selectivity of ZnO controlled the delivery of molecules to active catalyst sites, resulting in 95% toluene conversion in 90 min. Synergetic toluene adsorption in air pockets and degradation on catalytic sites helped achieve a quantum yield of 4.14 × 10-05 molecules/photon. A figure of merit reflecting fundamental system parameters was compared with previous photocatalytic systems to evaluate the practicality of ZNC@SSM.


Hospitalisation for injuries among Finnish conscripts in 1990-1999.
To describe the incidence, trends and nature of hospitalisation for injuries among Finnish conscripts between 1990 and 1999. A hospital discharge register study on the hospitalisation for injuries with a follow-up of 213509-person-years. During the 10-year study period, altogether 20016 hospitalisations for injuries were registered. Over this period, the annual incidence of hospitalisation for injuries declined with 26%, from 113.3 (95% CI: 108.7-118.0) per 1000 conscripts in 1990 to 84.1 (95% CI: 80.0-88.2) in 1999. Summer and autumn were the seasons with the highest incidence rates. Half of the injuries were sprains, strains, or fractures, and in half of the injuries, the anatomical location involved was the lower limb. The leading external causes of injuries were falls (31.8%) and injuries inflicted by foreign objects or machinery (18.5%). Although hospitalisations for injuries declined between 1990 and 1999, injuries are still an important cause of morbidity in the Finnish Defence Forces. Further studies to evaluate the risk factors and mechanisms of injuries are needed to develop effective prevention strategies.


Functional-group modifications of dextran for linkage to a diazonium group. A potential vehicle for tumour targeting of antineoplastic triazenes.
Several derivatives of dextran, containing a triazene side-chain, which are analogues of the antitumour agent DTIC (Dacarbazine), were prepared. The extent of triazene incorporation was measured by determination of the nitrogen content, and the presence of triazene groups was corroborated by u.v.- and n.m.r.-spectroscopic measurements. Some dextran-triazenes exhibited cytotoxic activity against M21 tumour cells in vitro.


Non-covalent interactions of cysteine onto C60, C59Si, and C59Ge: a DFT study.
The study of intermolecular interactions is of great importance. This study attempted to quantitatively examine the interactions between cysteine (C3H7NO2S) and fullerene nanocages, C60, in vacuum. As the frequent introduction of elements as impurities into the structure of nanomaterials can increase the intensity of intermolecular interactions, nanocages doped with silicon and germanium have also been studied as adsorbents, C59Si and C59Ge. Quantum mechanical studies of such systems are possible in the density functional theory (DFT) framework. For this purpose, various functionals, such as B3LYP-D3, ωB97XD, and M062X, have been used. One of the most suitable basis functionals for the systems studied in this research is 6-311G (d), which has been used in both optimization calculations and calculations related to wave function analyses. The main part of this work is the study of various analyses that reveal the nature of the intermolecular interactions between the two components introduced above. The results of conceptual DFT, natural bond orbital, non-covalent interactions, and quantum theory of atoms in molecules were consistent and in favor of physical adsorption in all systems. Germanium had more adsorption energy than other dopants. The HOMO-LUMO energy gaps were as follows: C60: 5.996, C59Si: 5.309, and C59Ge: 5.188 eV at B3LYP-D3/6-311 G (d) model chemistry. The sensitivity of the adsorption increased when an amino acid molecule interacted with doped C60, and this capability could be used to design nanocarrier to carry cysteine amino acid.


Tag team simulation: An innovative approach for promoting active engagement of participants and observers during group simulations.
Active participation in immersive simulation experiences can result in technical and non-technical skill enhancement. However, when simulations are conducted in large groups, maintaining the interest of observers so that they do not disengage from the learning experience can be challenging. We implemented Tag Team Simulation with the aim of ensuring that both participants and observers had active and integral roles in the simulation. In this paper we outline the features of this innovative approach and provide an example of its application to a pain simulation. Evaluation was conducted using the Satisfaction with Simulation Experience Scale. A total of 444 year nursing students participated from a population of 536 (response rate 83%). Cronbach's alpha for the Scale was .94 indicating high internal consistency. The mean satisfaction score for participants was 4.63 compared to 4.56 for observers. An independent sample t test revealed no significant difference between these scores (t (300) = -1.414, p = 0.16). Tag team simulation is an effective approach for ensuring observers' and participants' active involvement during group-based simulations and one that is highly regarded by students. It has the potential for broad applicability across a range of leaning domains both within and beyond nursing.


National UK guidelines for the management of paediatric craniopharyngioma.
Although rare, craniopharyngiomas constitute up to 80% of tumours in the hypothalamic-pituitary region in childhood. Despite being benign, the close proximity of these tumours to the visual pathways, hypothalamus, and pituitary gland means that both treatment of the tumour and the tumour itself can cause pronounced long-term neuroendocrine morbidity against a background of high overall survival. To date, the optimal management strategy for these tumours remains undefined, with practice varying between centres. In light of these discrepancies, as part of a national endeavour to create evidence-based and consensus-based guidance for the management of rare paediatric endocrine tumours in the UK, we aimed to develop guidelines, which are presented in this Review. These guidelines were developed under the auspices of the UK Children's Cancer and Leukaemia Group and the British Society for Paediatric Endocrinology and Diabetes, with the oversight and endorsement of the Royal College of Paediatrics and Child Health using Appraisal of Guidelines for Research & Evaluation II methodology to standardise care for children and young people with craniopharyngiomas.


Experimental immunohistochemical studies on the distribution and origins of substance P in the medial preoptic area of the rat.
Distribution of substance P (SP)-positive fibers in the medial preoptic area (POM) of the rat and their origins were examined using indirect immunofluorescence. A very high density of SP-positive fibers was seen in the POM throughout its entire rostro-caudal extent. However, the distribution of these fibers was not even; the highest density was detected in the medial part of the POM, with less dense but still numerous fibers in the lateral part. On the other hand, in this area a small number of SP-positive cells could be found; a few cells were scattered in the rostral part and, in the caudal part, several cells could be seen in the ventral part of the POM. The origins of SP-positive fibers in the POM were experimentally examined. Since the destruction of the ventro-lateral part of the anterior hypothalamus (vlAH), where numerous SP-positive cells were seen, resulted in a marked decrease of SP-positive fibers in the POM on the operated side, the majority of these fibers may originate from SP-positive cells in the vlAH. The fine structure of SP-positive terminals in the POM were investigated by electron-microscopic immunohistochemical techniques. Immunoreactive terminals contained a few large granular vesicles together with numerous small vesicles, and they made synaptic contacts mainly with dendrites which were devoid of immunoreactive materials. Two different synaptic contacts could be distinguished: one asymmetrical (Gray's type I) and the other symmetrical (Gray's type II), with the latter being predominant.


Importance of Surgical Margin Status in Ductal Carcinoma In Situ.
The purpose of the study was to identify the effect of final surgical margin (SM) status and re-excision on outcomes in patients with ductal carcinoma in situ (DCIS) who underwent breast conservation therapy (BCT). The study population consisted of women diagnosed with DCIS who underwent BCT between 1989 and 2014. All women received adjuvant whole breast radiation and a boost. The primary end point was local control (LC). Final SMs were defined according to margin width: negative SM was defined as > 2 mm, close SM was defined as > 0 to ≤ 2 mm, and a positive SM was defined as tumor on ink. The Cox proportional hazards model was used to determine predictors of outcomes on multivariable analysis. Actuarial incidence of LC was estimated using the Kaplan-Meier method. A total of 498 patients were included; 400 patients had a final negative SM, 87 had a close SM, and 11 had a positive SM. A total of 172 patients received adjuvant hormonal therapy, 265 patients required ≥ 1 re-excision. Patients with positive or close SMs were more likely to receive a radiation dose > 60 Gy (P < .001) and undergo re-excision (P < .01). The 10-year LC rates were not significantly different between patients with a negative (93.5%), close (91.8%), or positive (100%) SM (P = .57). There was no difference in LC in patients who underwent re-excision for initial close or positive SMs (P = .55). This single-institution experience showed that risks of local recurrence remain poorly characterized. Re-excision and whole breast radiation with boost resulted in excellent LC for women with DCIS. Trials aimed at personalized deintensified local therapy are warranted.


What do we need to know about anatomy in gynaecology? An international validation study.
International validation of the Dutch Delphi study about which anatomical structures should be taught to ensure safe and competent practice among general gynaecologists. Validation study with gynaecologists and trainees in gynaecology from academic, non-academic teaching and non-academic, non-teaching hospitals worldwide. The relevance of 123 items included in the Dutch Delphi study was scored on a Likert scale between 1 (not relevant) and 5 (highly relevant). Consensus was defined when ≥70 % of the panellist scored the item as relevant or very relevant and the average rating was ≥4. A total of 192 gynaecologists and trainees from seven countries (Belgium, Germany, Norway, Oceania, Sweden, United Kingdom and United States) completed the questionnaire. Of the 123 structures, 72 (58.5%) were internationally relevant. When the 72 relevant structures from the international Delphi study were compared with the 86 relevant structures from the Dutch Delphi study, 70 (81.4%) structures matched. This study identified 70 anatomical structures that should be taught for safe and competent practice of general gynaecologists based on national and international validation. The results of our study identify the learning needs (i.e., the content) for an international anatomy curriculum. The development of the curriculum (i.e., the form) can be determined by each country and used to standardize and guide postgraduate training in gynaecology. This is an important step in the era of international teaching and training.


Bis(morpholin-4-ium) tetra-chlorido-cobalt(II).
The title compound, (C(4)H(10)NO)(2)[CoCl(4)], is an ionic compound consisting of two protonated tetra-hydro-1,4-oxazine (morpholine) cations and a [CoCl(4)](2-) dianion. The Co(II) ion is in a tetra-hedral coordination geometry. The cations exhibit chair-shaped conformations. A three-dimensional supra-molecular architecture is formed through N-H⋯Cl and C-H⋯Cl hydrogen bonds between the dianions and the cations.


Beneficial effects of celery seed extract (Apium graveolens), as a supplement, on anxiety and depression in hypertensive patients: a randomized clinical trial.
Anxiety and depression are crucial public health issues, affecting the rising in hospitalizations and death. Anxiety and depression can worsen hypertension and vice versa. The current study has investigated the effects of celery seed extract, as a drug supplement, with the active ingredient of 3-n-butylphthalide, on mental problems primarily anxiety and secondary depression in hypertensive patients. The current study was a randomized, triple-blind, placebo-controlled, cross-over, 4-week clinical trial with a 4-week washout period. Fifty hypertensive patients received 4 placebo or celery seed extract capsules (1.34 g per day) for 4 weeks as a supplement to their usual medication regimen. The blood pressure parameters were assessed using 24-h ambulatory blood pressure monitoring device. Anxiety and depression and their wide range of symptoms were evaluated using Beck anxiety and depression inventories (BAI and BDI). In the celery treatment step, the mean reduction in BAI and BDI scores were 6.78 (P < 0.001) and 3.63 (P < 0.01), respectively. Some symptoms of anxiety including unable to relax, nervousness, numbness, dizziness, flushed face, sweating, and breathing difficulty were significantly improved by celery consumption (P < 0.001). Celery could decrease symptoms of depression such as sadness, crying, loss of energy, insomnia, irritability, fatigue, loss of interest in sex, and punishment feeling (P < 0.01). The mean reduction in blood pressure parameters was also significant during celery therapy (P < 0.001). The psychometric properties of anxiety and depression were investigated and the results were promising. The results indicated the anti-anxiety and anti-depressive properties of celery seed extract as a supplement in hypertensive patients. Registry name: Iranian Registry of Clinical Trials (IRCT), Registration number: IRCT20130418013058N8, Registration link: https://www.irct.ir/trial/30021 . The study was carried out between 2018-09-21 and 2020-07-20.


Hex stimulates the hepatocyte nuclear factor 1alpha-mediated activation of transcription.
The homeodomain protein Hex can function either as a transcriptional repressor or activator in animals. Recent reports have indicated that Hex is involved in liver development. However, its target genes and interacting proteins are largely unknown. We found that Hex functionally interacted with hepatocyte nuclear factor (HNF) 1alpha to further stimulate its activity using reporter gene containing multiple copies of HNF1alpha-binding site of the L-type pyruvate kinase (L-PK) gene promoter or natural L-PK promoter. This stimulation required the homeodomain and the acidic carboxyl-terminal region of Hex. Over-expression of Hex in primary cultured hepatocytes resulted in stimulation of the L-PK gene expression. Glutathione S-transferase pull-down assay and co-immunoprecipitation revealed that Hex physically interacted with HNF1alpha in mammalian cells through the homeodomain of Hex and POU-homeodomain of HNF1alpha. Since HNF1alpha is an important liver-enriched transcription factor involved in liver differentiation, Hex may contribute to liver differentiation through interaction with HNF1alpha.


Mapping sites of interaction between rhodopsin and transducin using rhodopsin antipeptide antibodies.
Site-directed antipeptide antibodies generated against the predicted cytoplasmic sequences of rhodopsin were used to map the binding domains for transducin, the retinal G-protein, on the photoreceptor. Antibodies against synthetic peptides corresponding to loop 3-4, loop 5-6, and the serine/threonine-rich region of the COOH terminus recognize rhodopsin by immunoblot analysis and also recognize the native protein within the membrane, allowing these probes to be used for functional studies. Rhodopsin reconstituted into phospholipid vesicles binds transducin in the light which significantly reduces the binding of antipeptide antibodies corresponding to loop 3-4 and the COOH terminus of rhodopsin. However, the binding of the antibody raised against a 14-amino-acid peptide corresponding to a sequence within loop 5-6 of rhodopsin was unaffected by the presence of transducin. These results suggest a preferential involvement of regions in or near loop 3-4 and the COOH terminus in the binding of transducin to rhodopsin. In contrast, a significant portion of loop 5-6 does not form a binding domain for the G-protein.


Hypothalamic-pituitary-adrenal reactivity in prepubertal children with social phobia.
The aim of this study was to investigate whether a different pattern of HPA axis activity is found between children with social phobia (SP) and healthy control children. A total of 50 prepubertal subjects (25 children with SP and 25 healthy control subjects) were studied. The effects of stress were studied by comparing cortisol responses to a psychosocial stressor, consisting of a public speaking task. Children with SP showed an elevated cortisol response to the psychosocial stressor as compared with healthy controls. Trait but not state anxiety levels are associated with higher HPA axis activity. Limited sample size. The results indicate that a higher cortisol responsivity to stress may be a neurobiological marker for prepubertal children with SP. Directions for future research and clinical implications are discussed.


Large neostriatal neurons in the rat: an electron microscopic study of gold-toned Golgi-stained cells.
The large neurons in the rat neostriatum consist of at least two morphologically distinct types in Golgi preparations: Type I large cells had smooth somata whereas type II large cells had somatic spines. In the present study, the ultrastructural morphology of the Golgi-stained large neostriatal neurons in the rat was analyzed by the gold-toning technique. In addition, the ultrastructural morphology of type I medium (spiny) neurons was also studied for comparison. The results revealed that the type I medium neurons had unindented nucleus with small amount of organelles whereas both types of large neostriatal neurons had deeply indented nucleus and large amount of cytoplasmic organelles. The type I large neurons had much fewer synaptic contacts on the somata than the type II large neurons, the latter also had synapses on their somatic spines. Our data also support the possibility that some large neostriatal neurons may have long axons by the fact that some of these cells had myelinated axons. Our result should be very useful in interpreting morphological data obtained with techniques such as immunocytochemistry or retrograde labeling which enables only partial labeling of somata and dendrites.


Psychiatric mental health nursing: thriving in a changing environment through outcomes-based measurements.
This article focuses on the necessity of outcomes-based measurements in the behavioral health care field. The authors describe the reasons why outcomes research is essential to the survival of behavioral services and, even more specifically, psychiatric mental health nursing. An actual outcome study process using psychiatric patients treated by a clinical nurse specialist is described in detail. The authors reflect their orientation to outcomes-based measurements through their perspectives as a manager and an outpatient therapist.


Molecular responses to ocean acidification in an Antarctic bivalve and an ascidian.
Southern Ocean organisms are considered particularly vulnerable to Ocean acidification (OA), as they inhabit cold waters where calcite-aragonite saturation states are naturally low. It is also generally assumed that OA would affect calcifying animals more than non-calcifying animals. In this context, we aimed to study the impact of reduced pH on both types of species: the ascidian Cnemidocarpa verrucosa sp. A, and the bivalve Aequiyoldia eightsii, from an Antarctic fjord. We used gene expression profiling and enzyme activity to study the responses of these two Antarctic benthic species to OA. We report the results of an experiment lasting 66 days, comparing the molecular mechanisms underlying responses under two pCO2 treatments (ambient and elevated pCO2). We observed 224 up-regulated and 111 down-regulated genes (FC ≥ 2; p-value ≤ 0.05) in the ascidian. In particular, the decrease in pH caused an upregulation of genes involved in the immune system and antioxidant response. While fewer differentially expressed (DE) genes were observed in the infaunal bivalve, 34 genes were up-regulated, and 69 genes were downregulated (FC ≥ 2; p-value ≤ 0.05) in response to OA. We found downregulated genes involved in the oxidoreductase pathway (such as glucose dehydrogenase and trimethyl lysine dioxygenase), while the heat shock protein 70 was up-regulated. This work addresses the effect of OA in two common, widely distributed Antarctic species, showing striking results. Our major finding highlights the impact of OA on the non-calcifying species, a result that differ from the general trend, which describes a higher impact on calcifying species. This calls for discussion of potential effects on non-calcifying species, such as ascidians, a diverse and abundant group that form extended three-dimensional clusters in shallow waters and shelf areas in the Southern Ocean.


Systemic inflammation during the first year of life is associated with brain functional connectivity and future cognitive outcomes.
The first years of life are a sensitive period of rapid neural and immune system development vulnerable to the impact of adverse experiences. Several studies support inflammation as a consequence of various adversities and an exposure negatively associated with developmental outcomes. The mechanism by which systemic inflammation may affect brain development and later cognitive outcomes remains unclear. In this longitudinal cohort study, we examine the associations between recurrent systemic inflammation, defined as C-reactive protein elevation on ≥ 2 of 4 measurements across the first year of life, electroencephalography (EEG) functional connectivity (FC) at 36 months, and composite cognitive outcomes at 3, 4, and 5 years among 122 children living in a limited-resource setting in Dhaka, Bangladesh. Recurrent systemic inflammation during the first year of life is significantly negatively associated with cognitive outcomes at 3, 4, and 5 years, after accounting for stunting and family care indicators (a measure of stimulation in the home environment). Recurrent systemic inflammation is significantly positively associated with parietal-occipital FC in the Beta band at 36 months, which in turn is significantly negatively associated with composite cognitive scores at 3 and 4 years. However, FC does not mediate the relationship between recurrent systemic inflammation and cognitive outcomes.


The light-dependent control of chloroplast development in barley (Hordeum vulgare L).
The light-induced greening of etiolated barley plants is used as a model to study the light-dependent control of plastid development. Upon illumination a rapid transformation of etioplasts to chloroplasts is induced. The effect of illumination does not only include the light-dependent chlorophyll synthesis but also the appearance or decline of specific proteins within the plastid membrane fractions. So far two of these proteins have been studied in detail. The light-harvesting chlorophyll a/b protein (LHCP) is one of the major protein constituents of the thylakoid membrane of chloroplasts. However, this protein is not detectable among the membrane polypeptides of etioplasts. Illumination of dark-grown barley plants induces a massive insertion of the LHCP. The appearance of the protein is controlled by the cooperation of at least two distinct photoreceptors: protochlorophyllide and phytochrome. In dark-grown barley plants not only the LHCP but also its mRNA is not detectable. The light-dependent appearance of mRNA activity for the LHCP is under the control of phytochrome (Pfr). Even though the appearance of mRNA activity is induced via Pfr by a single red light pulse, the assembly of the complete LHCP takes place only under continuous illumination, which allows chlorophyll synthesis. The second protein analyzed so far is the NADPH-protochlorophyllide-oxidoreductase. This enzyme catalyzes the light-dependent reduction of protochlorophyllide to chlorophyllide and thus controls one of the first detectable light-dependent reactions during the greening period. It is generally assumed that this enzyme is responsible for the overall chlorophyll synthesis and accumulation during the greening period.(ABSTRACT TRUNCATED AT 250 WORDS)


Coexistence of cholecystokinin- and substance P-like peptides in neurons of the dorsal root ganglia of the rat.
Using the indirect immunohistochemical technique with antisera to cholecystokinin and to substance P, the spinal dorsal horn and dorsal root ganglia of normal and colchicine-treated rats were studied. In the spinal cord a similar distribution of substance P- and cholecystokinin-positive networks in the superficial layers of the dorsal horn was observed. In the dorsal root ganglia several cholecystokinin and substance P immunoreactive cell bodies were seen in colchicine-treated rats. After elution and restaining for substance P, of sections previously stained for cholecystokinin, it was found that all cholecystokinin-positive cells also contained substance P-like immunoreactivity and vice versa.


Saturating summation of the afterhyperpolarization conductance in spinal motoneurones: a mechanism for 'secondary range' repetitive firing.
Summation of the potassium conductance (GK) changes underlying the spike afterhyperpolarization (AHP) has been studied in cat spinal motoneurones. Cells were directly activated by one to five short current pulses at constant rate, each evoking an action potential. The analysis was restricted to cells displaying an approximately exponential decay of the AHP conductance. In these neurones the AHP conductances given by successive spikes were found to summate in a non-linear manner. This nonlinear summation seemed well described by a neurone model based on modified Hodgkin-Huxley equations. From the model equations the total AHP conductance in motoneurones could be calculated from values of GK measured experimentally at different times during the summation process. Adaptation and steady-state firing in motoneurones are assumed to be governed by summation of AHP conductance. The same model was then utilized for simulating neuronal repetitive firing in response to current steps. Such simulations were performed after substitution of the model parameters with values measured in individual motoneurones which had also been fired repetitively by intracellular injection of long-lasting current steps. The amount of adaptation and the shape and slopes of the steady-state frequency-to-current relation were found to coincide in the model and in the corresponding motoneurones.


Interaction of murine progesterone receptors with specific monoclonal antibodies to the avian progesterone receptor.
Monoclonal antibodies raised against purified chicken progesterone receptor (PgR) have been described and characterized recently. In this study we have screened these antibodies for cross-reactivity with murine PgR. Of the six anti-PgR antibodies tested, one (alpha PR6) precipitates murine PgR in an assay using protein A-sepharose as an absorbent for the antibody. The antibody is specific for PgR and does not react with the estrogen receptor or the glucocorticoid receptor in the same cytosol. In immunoblot experiments, both alpha PR6 and alpha PR11 recognize a 115,000 Da protein, however, alpha PR11 gives a weaker signal than alpha PR6. In photoaffinity labeling experiments, a 115,000 Da and an 83,000 Da protein covalently bind tritiated R5020 in a receptor-specific way. We conclude that the alpha PR6 antibody can be used as a tool to study the structure and function of the murine PgR.


Biodegradable polycaprolactone (PCL) nanosphere encapsulating superoxide dismutase and catalase enzymes.
Biodegradable polycaprolactone (PCL) nanosphere encapsulating superoxide dismutase (SOD) and catalase (CAT) were successfully synthesized using double emulsion (w/o/w) solvent evaporation technique. Characterization of the nanosphere using dynamic light scattering, field emission scanning electron microscope, and Fourier transform infrared spectroscopy revealed a spherical-shaped nanosphere in a size range of 812 ± 64 nm with moderate protein encapsulation efficiency of 55.42 ± 3.7 % and high in vitro protein release. Human skin HaCat cells were used for analyzing antioxidative properties of SOD- and CAT-encapsulated PCL nanospheres. Oxidative stress condition in HaCat cells was optimized with exposure to hydrogen peroxide (H2O2; 1 mM) as external stress factor and verified through reactive oxygen species (ROS) analysis using H2DCFDA dye. PCL nanosphere encapsulating SOD and CAT together indicated better antioxidative defense against H2O2-induced oxidative stress in human skin HaCat cells in comparison to PCL encapsulating either SOD or CAT alone as well as against direct supplement of SOD and CAT protein solution. Increase in HaCat cells SOD and CAT activities after treatment hints toward uptake of PCL nanosphere into the human skin HaCat cells. The result signifies the role of PCL-encapsulating SOD and CAT nanosphere in alleviating oxidative stress.