25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 exert distinct effects on human skeletal muscle function and gene expression.
Age-associated decline in muscle function represents a significant public health burden. Vitamin D-deficiency is also prevalent in aging subjects, and has been linked to loss of muscle mass and strength (sarcopenia), but the precise role of specific vitamin D metabolites in determining muscle phenotype and function is still unclear. To address this we quantified serum concentrations of multiple vitamin D metabolites, and assessed the impact of these metabolites on body composition/muscle function parameters, and muscle biopsy gene expression in a retrospective study of a cohort of healthy volunteers. Active serum 1,25-dihydroxyvitamin D3 (1α,25(OH)2D3), but not inactive 25-hydroxyvitamin D3 (25OHD3), correlated positively with measures of lower limb strength including power (rho = 0.42, p = 0.02), velocity (Vmax, rho = 0.40, p = 0.02) and jump height (rho = 0.36, p = 0.04). Lean mass correlated positively with 1α,25(OH)2D3 (rho = 0.47, p = 0.02), in women. Serum 25OHD3 and inactive 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) had an inverse relationship with body fat (rho = -0.30, p = 0.02 and rho = -0.33, p = 0.01, respectively). Serum 25OHD3 and 24,25(OH)2D3 were also correlated with urinary steroid metabolites, suggesting a link with glucocorticoid metabolism. PCR array analysis of 92 muscle genes identified vitamin D receptor (VDR) mRNA in all muscle biopsies, with this expression being negatively correlated with serum 25OHD3, and Vmax, and positively correlated with fat mass. Of the other 91 muscle genes analysed by PCR array, 24 were positively correlated with 25OHD3, but only 4 were correlated with active 1α,25(OH)2D3. These data show that although 25OHD3 has potent actions on muscle gene expression, the circulating concentrations of this metabolite are more closely linked to body fat mass, suggesting that 25OHD3 can influence muscle function via indirect effects on adipose tissue. By contrast, serum 1α,25(OH)2D3 has limited effects on muscle gene expression, but is associated with increased muscle strength and lean mass in women. These pleiotropic effects of the vitamin D 'metabolome' on muscle function indicate that future supplementation studies should not be restricted to conventional analysis of the major circulating form of vitamin D, 25OHD3.


Homozygous protein C deficiency with late onset venous thrombosis: identification and in vitro expression study of a novel Pro275Ser mutation.
To identify the mutation and study the molecular mechanism of inherited protein C (PC) deficiency in a Chinese pedigree. The plasma levels of PC activity (PC:A) and antigen (PC:Ag) were measured by chromogenic assay and ELISA, respectively. The PROC gene was amplified and sequenced for mutational screening. Wild type and Pro275Ser mutant PC cDNA expression plasmids were constructed and transfected into HEK 293T cells and COS 7 cells, respectively. The expression and transcription of PC were investigated by ELISA, Western blot and real time RT-PCR. Immunofluorescence staining was utilised to analyse the intracellular distribution of PC, and pulse-chase experiments were used to detect the intracellular stability of the mutant PC. The proband's plasma PC:A and PC:Ag were 5% and 13.9%, respectively. A missense mutation (p.Pro275Ser) was identified in exon 9 of PROC gene. In vitro expression study showed that Pro275Ser variant was present at 22.6% and 78.9% of wild type levels in culture supernatants and cell lysates, respectively. No significant differences in the molecular weights, mRNA levels or intracellular stability were observed between the mutant and wild type PC. Immunofluorescence staining revealed that the mutant protein was mainly located in the endoplasmic reticulum. A homozygous Pro275Ser mutation was identified in a Chinese pedigree of PC deficiency. Impaired secretion of the mutant PC might be the molecular mechanism of PC deficiency caused by Pro275Ser mutation.


[Genetic polymorphisms of the renin-angiotensin system].
Essential hypertension is a multifactorial disease which is evoked by multiple environmental and genetic factors. Recent progress in molecular biology enables us a comprehensive understanding of the molecular pathogenesis of this disease. The genes of renin-angiotensin system such as renin, angiotensinogen, angiotensin-converting enzyme and angiotensin receptors genes are all reported as positive linkage to hypertension. Now ongoing progress of the human genome project will further accelerate the molecular studies on the rennin-angiotensin system and hypertension.


Investigation of adverse events following bacille Calmette-Guérin immunization using immunization safety surveillance system in Korea Centers for Disease Control and Prevention.
The report of adverse events following immunization (AEFI) in Korea has continued since 1994, and the most frequently reported cases of AEFI of Korea Centers for Disease Control and Prevention (KCDC) is bacille Calmette-Guérin (BCG). Meanwhile, various inoculation methods and strains have been used in the past 6 years in Korea. Therefore, we investigated AEFI of BCG by strain types and inoculation methods using immunization safety surveillance of KCDC. We reviewed BCG AEFIs registered in the KCDC from January 2013 to June 2018. There were 336 AEFI cases during the period, and average time interval from vaccination to symptom onset was within 2 months. AEFI proportion was 6.4 cases per 100,000 doses for BCG percutaneous Tokyo strain, 41.6 cases per 100,000 doses of BCG intradermal Danish strain, and 25.9 cases per 100,000 doses of BCG intradermal Tokyo strain. Intradermal type was more reported AEFI than percutaneous type in the same strain. The most common adverse events were local reaction like BCG lymphadenitis and severe adverse reactions such as osteomyelitis or disseminated BCG disease were 0.1 to 0.2 cases per 100,000 doses which are correlated with the range of World Health Organization published AEFI rates. The AEFI reporting rate does not equal the actual proportion of AEFI occurrence. Because AEFI monitoring is a passive surveillance system, various factors might influence the number of events reported. Nevertheless, it is important to analyze BCG AEFI by vaccine strains and inoculation method using surveillance data of KCDC.


Sequential screening for lung cancer in a high-risk group: randomised controlled trial: LungSEARCH: a randomised controlled trial of Surveillance using sputum and imaging for the EARly detection of lung Cancer in a High-risk group.
Low-dose computed tomography (LDCT) screening detects early-stage lung cancer and reduces mortality. We proposed a sequential approach targeted to a high-risk group as a potentially efficient screening strategy. LungSEARCH was a national multicentre randomised trial. Current/ex-smokers with mild/moderate chronic obstructive pulmonary disease (COPD) were allocated (1:1) to have 5 years surveillance or not. Screened participants provided annual sputum samples for cytology and cytometry, and if abnormal were offered annual LDCT and autofluorescence bronchoscopy (AFB). Those with normal sputum provided annual samples. The primary end-point was the percentage of lung cancers diagnosed at stage I/II (nonsmall cell) or limited disease (small cell). 1568 participants were randomised during 2007-2011 from 10 UK centres. 85.2% of those screened provided an adequate baseline sputum sample. There were 42 lung cancers among 785 screened individuals and 36 lung cancers among 783 controls. 54.8% (23 out of 42) of screened individuals versus 45.2% (14 out of 31) of controls with known staging were diagnosed with early-stage disease (one-sided p=0.24). Relative risk was 1.21 (95% CI 0.75-1.95) or 0.82 (95% CI 0.52-1.31) for early-stage or advanced cancers, respectively. Overall sensitivity for sputum (in those randomised to surveillance) was low (40.5%) with a cumulative false-positive rate (FPR) of 32.8%. 55% of cancers had normal sputum results throughout. Among sputum-positive individuals who had AFB, sensitivity was 45.5% and cumulative FPR was 39.5%; the corresponding measures for those who had LDCT were 100% and 16.1%, respectively. Our sequential strategy, using sputum cytology/cytometry to select high-risk individuals for AFB and LDCT, did not lead to a clear stage shift and did not improve the efficiency of lung cancer screening.


Rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.
Recent studies indicate that extracellular vesicles are an important source material for many clinical applications, including minimally-invasive disease diagnosis. However, challenges for rapid and simple extracellular vesicle collection have hindered their application. We have developed and validated a novel class of peptides (which we named venceremin, or Vn) that exhibit nucleotide-independent specific affinity for canonical heat shock proteins. The Vn peptides were validated to specifically and efficiently capture HSP-containing extracellular vesicles from cell culture growth media, plasma, and urine by electron microscopy, atomic force microscopy, sequencing of nucleic acid cargo, proteomic profiling, immunoblotting, and nanoparticle tracking analysis. All of these analyses confirmed the material captured by the Vn peptides was comparable to those purified by the standard ultracentrifugation method. We show that the Vn peptides are a useful tool for the rapid isolation of extracellular vesicles using standard laboratory equipment. Moreover, the Vn peptides are adaptable to diverse platforms and therefore represent an excellent solution to the challenge of extracellular vesicle isolation for research and clinical applications.


Clinical analysis of the related factors in acute appendicitis.
In order to determine reliable clues for early diagnosis of acute appendicitis, this study was conducted to examine the related factors in patients with clinically suspected acute appendicitis. We retrospectively analyzed 282 patients with the clinical diagnosis of acute appendicitis at China Medical College Hospital in Taiwan from January to December 2000. To study the significant related factors of acute appendicitis, the t-test, chi-square analysis, and multivariate logistic regression analysis were used. There were 153 males (54.3 percent) and 129 females (45.7 percent). The mean age was 30.3+/-17.4 years (range 1 to 81). The diagnostic rate of acute appendicitis was 86.2 percent. If the combination of elevated C-reactive protein, leukocytosis and elevated neutrophil ratio was used, satisfactory specificity and positive predictive value were achieved in diagnosing acute appendicitis. After controlling for the other covariates, the multivariate logistic regression analysis showed that the significant related factors of acute appendicitis were male sex (odds ratio = 3.4; 95 percent confidence interval = 1.6 to 7.3; p <0.01) and elevated neutrophil ratio (odds ratio = 4.6; 95 percent confidence interval = 2.0 to 10.6; p <0.001). If an elevated neutrophil ratio was observed, the probability of acute appendicitis was increased in patients with clinically suspected acute appendicitis. Thus, neutrophil ratio appears to be a good parameter for diagnosis of acute appendicitis in primary healthcare settings.


Reactions between resonance-stabilized radicals: propargyl + allyl.
This article describes a detailed theoretical analysis of the reaction between allyl and propargyl. In this analysis, we employ high-level electronic structure calculations to characterize the potential energy surface and various forms of transition-state theory (TST) to calculate microcanonical, J-resolved rate coefficients-conventional TST for isomerizations, and the variable reaction coordinate form of variational TST for the "barrierless" association/dissociation processes. These rate coefficients are used in a time-dependent, multiple-well master equation to determine phenomenological rate coefficients, k(T,p), for various product channels. The analysis indicates that the formation of (cyclic) c-C(6)H(7) and c-C(6)H(8) species is suppressed by elevated pressure. Overall, the results suggest that the formation of these five-membered rings from the reaction is not as important as previously thought. A simplified description of the kinetics of the reaction is discussed, and corresponding rate coefficients are provided.


The pathogenesis of brain abnormalities in the fetal alcohol syndrome: an integrating hypothesis.
The most obvious characteristics of the fetal alcohol syndrome are a cluster of minor, physical malformations that are rather nonspecific and that vary greatly in the frequency and severity of expression. The most common characteristics are general body and organ growth deficits, including microcephaly. The most debilitating characteristics, however, are central nervous system (CNS) dysfunctions, such as behavioral and intellectual impairments, which appear to be closely related to the growth deficits. The mechanisms whereby alcohol exerts its deleterious effects on intrauterine growth and development are being intensively investigated. Because of its pharmacological properties, maternal consumption of alcohol is associated with widespread effects throughout the maternal-placental-fetal organism, many of which can secondarily alter the in utero environment. Both direct and indirect mechanisms are discussed with respect to their contribution to alcohol-related effects on embryofetal growth and development. It is suggested that impairment in the protein synthetic machinery, resulting in cellular growth restriction at critical periods of development is the common mechanism underlying alcohol's teratogenicity. Because the development of the CNS is so prolonged and complex compared to other organ systems, it is exquisitely vulnerable to derangement throughout pregnancy.


State of the art: an integrated approach to pelvic floor ultrasonography.
Surgical management of pelvic floor disorders depends on a comprehensive understanding of the structural integrity and function of the pelvic floor. For visualizing this region, ultrasonography has emerged as a procedure that is relatively easy to perform, cost-effective and widely available. In this review, pelvic floor ultrasonography, including two-dimensional (2D), three-dimensional (3D) and 4D imaging as well as transvaginal, endoanal and transperineal techniques, is discussed from a global and multicompartmental perspective, rather than using a compartmentalized approach. The role of the different sonographic modalities in the major disorders of the pelvic floor-urinary and fecal incontinence, pelvic organ prolapse and obstructed defecation syndrome-is evaluated critically.


Not all Total Hip and Knee Arthroplasties Are the Same: What Are the Implications in Large Database Studies?
The use of claims databases for research after total hip and knee arthroplasty (THA, TKA) has increased exponentially. These studies rely on accurate coding, and inadvertent inclusion of patients with nonroutine indications may influence results. The purpose of this study was to evaluate the complexity of THA and TKA captured by CPT code and determine if complication rates vary based on the indication. The NSQIP database was queried using CPT codes 21730 and 27447 to identify patients undergoing THA and TKA from 2018 to 2019. The surgical indication was classified based on the ICD-10 diagnosis code as routine primary, complex primary, inflammatory, fracture, oncologic, revision, infection, or indeterminant. Patient factors and 30-day complications, readmission, reoperation, and wound complications were compared. A total of 86,009 THA patients had 703 ICD-10 diagnosis codes and 91.4% were routine primary indications. Complication rates were: routine primary 7.4%, complex primary 11.3%, inflammatory 12.5%, fracture 23.9%, oncologic 32.4%, revision 26.9%, infection 38.7%, and indeterminant 10.3% (P < .0001). 137,500 TKA patients had 552 ICD-10 diagnosis codes and 96.1% were routine primary cases. Complication rates were: routine primary 5.9%, complex primary 8.0%, inflammatory 7.2%, fracture 38.9%, oncologic 32.7%, revision 13.3%, infection 37.7%, and indeterminant 9.6% (P < .0001). Routine primary arthroplasty had significantly lower rates of reoperation, readmission, and wound complications. Using CPT code alone captures 10% of THA and 4% of TKA patients with procedures for nonroutine primary indications. It is essential to recognize identification of patients simply by CPT code has the potential to inadvertently introduce bias, and surgeons should critically assess methods used to define the study populations.


[Parkinson syndrome after CO poisoning. Unexpected late manifestation and favorable prognosis].
Although individual extrapyramidal symptoms as a consequence of carbon monoxide intoxication have frequently been reported, typical (complete) Parkinson's syndrome has very rarely been documented. This report presents for the first time a case of acute Parkinson's syndrome with delayed manifestation after the initially occurring organic psychosis. A 53-year-old man with a long history of schizoaffective psychosis first developed a severe acute organic psychosis with disturbances of orientation and marked amnestic disturbances after carbon monoxide intoxication as a result of a suicide attempt. On the 21st day after intoxication, acute Parkinson's syndrome with right accentuation was recorded, with CCT identification of lesions on both sides of the basal ganglia area. Full remission of the Parkinson's syndrome was achieved with rheological treatment and L-dopa therapy.


Esketamine Nasal Spray for Rapid Reduction of Depressive Symptoms in Patients With Major Depressive Disorder Who Have Active Suicide Ideation With Intent: Results of a Phase 3, Double-Blind, Randomized Study (ASPIRE II).
Patients with major depressive disorder (MDD) having active suicidal ideation with intent require immediate treatment. This double-blind study (ASPIRE II) randomized adults (aged 18-64 years) with MDD having active suicidal ideation with intent to esketamine 84 mg or placebo nasal spray twice weekly for 4 weeks, given with comprehensive standard of care (hospitalization ≥5 days and newly initiated or optimized oral antidepressant[s]). Change from baseline to 24 hours post-first dose in Montgomery-Asberg Depression Rating Scale total score (primary efficacy endpoint) was analyzed using ANCOVA. Clinical Global Impression-Severity of Suicidality-revised (key secondary endpoint) was analyzed using ANCOVA on ranks of change. Of 230 patients who were randomized (115 per arm), 227 received study drug and were included in efficacy/safety analyses; 184 (80.0%) completed double-blind treatment. Greater improvement in Montgomery-Asberg Depression Rating Scale total score was observed with esketamine (mean [SD]: -15.7 [11.56]) vs placebo (-12.4 [10.43]), each with standard of care, at 24 hours (least-squares mean difference [SE]: -3.9 [1.39], 95% CI: -6.60, -1.11; 2-sided P = .006). This was also noted at the earlier (4-hour) timepoint (least-squares mean difference -4.2, 95% CI: -6.38, -1.94). Patients in both treatment groups experienced rapid reduction in Clinical Global Impression-Severity of Suicidality-revised score; the between-group difference was not statistically significant. The most common adverse events among esketamine-treated patients were dizziness, dissociation, nausea, dysgeusia, somnolence, headache, and paresthesia. This study confirmed rapid and robust reduction of depressive symptoms with esketamine nasal spray in severely ill patients with MDD who have active suicidal ideation with intent. Trial Registration: Clinical Trials.gov identifier: NCT03097133.


Predicting osteoarthritis onset and progression with 3D texture analysis of cartilage MRI DESS: 6-Year data from osteoarthritis initiative.
In this study, we developed a gray level co-occurrence matrix-based 3D texture analysis method for dual-echo steady-state (DESS) magnetic resonance (MR) images to be used for knee cartilage analysis in osteoarthritis (OA) studies and use it to study changes in articular cartilage between different subpopulations based on their rate of progression into radiographically confirmed OA. In total, 642 series of right knee DESS MR images at 3T were obtained from baseline, 36- and 72-month follow-ups from the OA Initiative database. At baseline, all 214 subjects included in the study had Kellgren-Lawrence (KL) grade <2. Three groups were defined, based on time of progression into radiographic OA (ROA) (KL grades ≥2): control (no progression), fast progressor (ROA at 36 months), and slow progressor (ROA at 72 months) groups. 3D texture analysis was used to extract textural features for femoral and tibial cartilages. All textural features, in both femur and tibia, showed significant longitudinal changes across all groups and tissue layers. Most of the longitudinal changes were observed in progressors, but significant changes were observed also in controls. Differences between groups were mostly seen at baseline and 72 months. The method is sensitive to cartilage changes before and after ROA. It was able to detect longitudinal changes in controls and progressors and to distinguish cartilage alterations due to OA and aging. Moreover, it was able to distinguish controls and different progressor groups before any radiographic signs of OA and during OA. Thus, texture analysis could be used as a marker for the onset and progression of OA.


Acupuncture plus massage for cervicogenic headache: A protocol for systematic review and meta-analysis.
Cervicogenic headache (CGH), one of the most common headaches. It is characterized by pain starting from the neck and gradually involving the eyes, frontal and temporal regions. Acupuncture and massage therapy has been widely used in the treatment of CGH, the purpose of this study was to evaluate the effectiveness and safety of acupuncture combined with massage in the treatment of CGH. The databases of PubMed, Embase, Cochrane library, Medicine, Web of Science, China National Knowledge Infrastructure, Wan-Fang Database, China Biomedical Literature Service System, and Chongqing VIP Chinese Science will be searched. This study will include randomized controlled trials of acupuncture combined with massage in the treatment of CGH published before November 2021. The primary outcomes included the overall efficiency, visual simulation score, neck activity score, quality of life score, and adverse reactions as secondary outcomes were assessed. Cochrane bias risk assessment tool will be used for literature data screening and quality evaluation, and using RevMan5.4 to collect data for statistical analysis. We then will use the Grading of Recommendations Assessment, Development, and Evaluation approach to assess the overall quality of evidence supporting the primary outcomes. This systematic review will provide a high-quality synthesis to evaluate the efficacy and safety of acupuncture combined with massage therapy in the treatment of CGH, providing a reference for the safe and effective treatment plan. This study provides evidence that acupuncture combined with massage is effective and safe for patients with CGH. INPLASY2021120049U1.


Poor prognosis in multiple myeloma is associated only with partial or complete deletions of chromosome 13 or abnormalities involving 11q and not with other karyotype abnormalities.
Chromosomal abnormalities have major biologic and prognostic implications in leukemias. Cytogenetic information in typically hypoproliferative multiple myeloma (MM) is limited because of difficulties in obtaining analyzable metaphases. In this study, karyotypes and other known prognostic factors were analyzed in 155 newly diagnosed MM patients, entered on an intensive treatment program with two autotransplants. Complete remission (CR), event-free (EFS) and overall survival (OS) were analyzed using standard statistical methods. Abnormal cytogenetics were found in 39% of patients and were associated with a significantly lower CR rate (27% v 48%; P = .008). EFS and OS were inferior in patients with either partial or complete deletion of chromosome 13 or 11q abnormalities ("unfavorable" karyotype) when compared with the remaining patients (P < .001) who, as a group, had a similar prognosis irrespective of cytogenetic findings, ie, inevaluable, normal, or abnormal but without an "unfavorable" karyotype. The patients with abnormalities of both chromosomes 11 and 13 had a dismal prognosis with median EFS and OS of only 11 and 12 months, respectively. Significant associations were noted between an "unfavorable" karyotype and IgA isotype, elevated levels of beta-2 microglobulin (B2M, > or = 3 mg/L) and age > 60 years. On multivariate regression analysis, the absence of an "unfavorable" karyotype was the most significant variable associated with prolonged EFS and OS (P = .0001 and .0002, respectively). Other independent favorable variables were age less than 60 years, C-reactive protein (CRP) < or = 0.4 mg/dL and bone marrow plasmacytosis < or = 50% before treatment. On a multivariate analysis without cytogenetics, these same three standard parameters were identified as the only favorable variables. Patients not having all three standard favorable variables had a significantly lower CR rate (P = .03), EFS (P = .0001), and OS (P = .002) if an unfavorable karyotype was detected. We conclude that, in this program of uniformly treated MM patients, a poor prognosis was associated predominantly with abnormalities of chromosomes 11 and 13.


Galvanic-induced body sway in vestibular schwannoma patients: evidence for stimulation of the central vestibular system.
To investigate the various possible sites of excitation by galvanic stimulation by comparing the galvanic-induced body sway (GBS) in vestibular schwannoma (VS) patients with that in healthy subjects. This was a prospective experimental study conducted in a tertiary referral centre. In a previous study we developed a standardized procedure for reproducible quantification of GBS using a monaural continuous 1-cosinusoidal stimulus (0.5 Hz; 2 mA). In this study, 23 VS patients were tested before and 12 VS patients were also tested after surgical intervention (extirpation of tumour with concomitant vestibular neurectomy) and the results were compared with those obtained in 47 healthy subjects. There were no significant differences in mean total GBS gain between VS patients and healthy subjects (p>0.05) before surgical intervention. After vestibular neurectomy a significant difference in the mean total GBS gain was observed, but GBS was still present after surgery. These results prove that galvanic vestibular stimulation excites the central vestibular system even when conduction via the vestibular nerve is hampered.


Evidence for a light dependent increase of phosphoglucomutase activity in isolated, intact spinach chloroplasts.
Phosphoglucomutase (PGM) activity was measured in spinach (Spinacia oleracea L.) chloroplasts. Initial enzyme activity in a chloroplast lysate was 5 to 10% of total activity measured with 1 micromolar glucose 1,6-bisphosphate (Glc 1,6-P(2)) in the assay. Initial PGM activity increased 2- to 3-fold when chloroplasts were illuminated for 10 minutes prior to enzyme measurement and then decreased slowly in the dark. Measurements of total enzyme activity were unchanged by prior light treatment. Initial PGM activity from light treated chloroplasts was sufficient to account for in vivo rates of starch synthesis. Changes in PGM activity were affected by stromal pH and orthophosphate concentration. Photosynthetic inhibitors, dl-glyceraldehyde, glycolaldehyde, and glyoxylate, decreased and 3-phosphoglyceric acid increased light induced changes of PGM activity. Dark preincubation of chloroplasts with 10 millimolar dithiothreitol had no effect upon initial PGM activity, suggesting that light effects did not involve a sulfhydryl mechanism. Hexose monophosphate levels increased in illuminated chloroplasts. Activation of PGM in a chloroplast lysate by Glc 1,6-P(2) was maximal between pH 7.5 and 8.5. Stromal concentrations of Glc 1,6-P(2) were between 20 and 30 micromolar for both light and dark incubated chloroplasts and these levels should saturate PGM activity. Light dependent alterations of enzyme activity may be due to changes of phosphorylated PGM levels in the stroma or are the result of changes in residual activity by the dephosphorylated form of the enzyme. The above results indicate that PGM activity in spinach chloroplasts may be regulated by light, stromal pH, and Glc 1,6-P(2) concentration.


Hypomagnesaemia and hypocalcaemia in a patient with ovarian carcinoma.
A patient with disseminated ovarian carcinoma presented with symptoms of hypocalcaemia secondary to hypomagnesaemia. The low serum magnesium (0.4 mmol/l) appeared to be due to renal leakage with no evidence of ureteric obstruction or hydronephrosis on intravenous urogram. Parathyroid hormone activity (serum levels and cAMP response) was normal, despite hypomagnesaemia. The patient's complaints resolved after magnesium and calcium supplementation. Hypomagnesaemia of malignancy is a rare but important complication, and is both poorly recognized and understood.


Biological function of a DUF95 superfamily protein involved in the biosynthesis of a circular bacteriocin, leucocyclicin Q.
Biological functions of a DUF95 superfamily protein in the biosynthesis gene cluster of a novel circular bacteriocin, leucocyclicin Q (LcyQ), were characterized in this paper. Sequence analysis and database search of the regions flanking the LcyQ structural gene lcyQ revealed four open reading frames (lcyR, lcyB, lcyC, and lcyD) related to bacteriocin biosynthesis. LcyD shares some similarity to the DUF95 superfamily proteins, often found in the biosynthetic gene clusters of circular bacteriocins. Mass spectrometry analysis showed accumulation of active mature LcyQ inside lcyD knockout cells. Heterologous expression of lcyD demonstrated that it confers robust immunity against LcyQ. Peptide release/binding assay revealed that the immunity could be attributed to the secretion of LcyQ to the cell exterior. Thus, the DUF95 superfamily protein has a dual function in the biosynthesis of LcyQ, as an immunity-associated transporter and as a secretion-aiding agent. Accumulation of mature LcyQ inside the cell in lcyD knockout strains, further implied that cyclization occurs within the cell. To the best of our knowledge, this is the first report on LcyQ cyclization inside the cell and the dual role of a DUF95 superfamily protein in circular bacteriocin biosynthesis.


Arrhythmia Detection is Improved by 14-Day Continuous Electrocardiography Patch Monitoring and CHA2DS2-VASc Score.
Arrhythmias are not always easy to capture because they are often paroxysmal or asymptomatic. Using the CHA2DS2-VASc score for arrhythmia risk assessment, a 14-day electrocardiography monitor patch was used to evaluate patients with no documented history of arrhythmia. Ninety-three patients (mean age 59.8 ± 12.0 years, 46.2% female) received 14-day electrocardiography telemonitoring, and 14 patients (15%) were diagnosed with arrhythmias during a follow-up of 1004.4 person-days (mean recorded days 10.8 ± 4.1). The patients who were detected to have arrhythmias were older and had a higher prevalence of heart failure and chronic kidney disease. The result showed that arrhythmias were more likely to develop during a 14-day monitoring period in the patients with a CHA2DS2-VASc score of ≥ 3 or ≥ 4. Atrioventricular block was more likely to be detected in the patients with a CHA2DS2-VASc score of ≥ 3 or ≥ 4 during 7-day or 14-day monitoring periods. Ventricular tachycardia was also more likely to be detected in the patients with a CHA2DS2-VASc score of ≥ 4 or ≥ 5 during a 14-day monitoring period. When evaluating the risk of arrhythmia, a CHA2DS2-VASc score of ≥ 3 or ≥ 4 was associated with a higher risk of any arrhythmias during a 14-day monitoring period, while a CHA2DS2-VASc score of ≥ 4 was associated with a higher risk of any arrhythmias during a 7-day monitoring period. The results may suggest that a 14-day monitoring period is more favorable to detect arrhythmias. Atrioventricular block and ventricular tachycardia were more likely to develop in the patients with a higher CHA2DS2-VASc score.


Complications of endoscopic sinus surgery. The role of the radiologist in prevention.
Intranasal surgery via the endoscope has become an extremely popular technique for refractory rhinosinusitis, but like all procedures it carries a risk of complications. Mosher in 1912 stated that intranasal ethmoidectomy is "the blindest and most dangerous (operation) in all surgery." Complications of a surgical procedure are among the most devastating problems in the practice of medicine. A thorough knowledge on the part of the radiologist of what the otolaryngologist is attempting to do can only enhance patient care. All screening CT scans should be approached as if they are preoperative examinations. It is not enough for the radiologist to merely describe air-fluid levels or mucoperiosteal thickening. Instead, the radiologist is an important member of the health care team, and an active role should be assumed to describe anatomic variants that may place the patient at increased surgical risk and to assist the surgeon in the appropriate evaluation of the patient who has a surgical complication.


Transport mechanisms in capillary condensation of water at a single-asperity nanoscopic contact.
Transport mechanisms involved in capillary condensation of water menisci in nanoscopic gaps between hydrophilic surfaces are investigated theoretically and experimentally by atomic force microscopy (AFM) measurements of capillary force. The measurements showed an instantaneous formation of a water meniscus by coalescence of the water layers adsorbed on the AFM tip and sample surfaces, followed by a time evolution of meniscus toward a stationary state corresponding to thermodynamic equilibrium. This dynamics of the water meniscus is indicated by time evolution of the meniscus force, which increases with the contact time toward its equilibrium value. Two water transport mechanisms competing in this meniscus dynamics are considered: (1) Knudsen diffusion and condensation of water molecules in the nanoscopic gap and (2) adsorption of water molecules on the surface region around the contact and flow of the surface water toward the meniscus. For the case of very hydrophilic surfaces, the dominant role of surface water transportation on the meniscus dynamics is supported by the results of the AFM measurements of capillary force of water menisci formed at sliding tip-sample contacts. These measurements revealed that fast movement of the contact impedes on the formation of menisci at thermodynamic equilibrium because the flow of the surface water is too slow to reach the moving meniscus.


Subunit composition of synaptic AMPA receptors revealed by a single-cell genetic approach.
The precise subunit composition of synaptic ionotropic receptors in the brain is poorly understood. This information is of particular importance with regard to AMPA-type glutamate receptors, the multimeric complexes assembled from GluA1-A4 subunits, as the trafficking of these receptors into and out of synapses is proposed to depend upon the subunit composition of the receptor. We report a molecular quantification of synaptic AMPA receptors (AMPARs) by employing a single-cell genetic approach coupled with electrophysiology in hippocampal CA1 pyramidal neurons. In contrast to prevailing views, we find that GluA1A2 heteromers are the dominant AMPARs at CA1 cell synapses (approximately 80%). In cells lacking GluA1, -A2, and -A3, synapses are devoid of AMPARs, yet synaptic NMDA receptors (NMDARs) and dendritic morphology remain unchanged. These data demonstrate a functional dissociation of AMPARs from trafficking of NMDARs and neuronal morphogenesis. This study provides a functional quantification of the subunit composition of AMPARs in the CNS and suggests novel roles for AMPAR subunits in receptor trafficking.


Renal safety and efficacy of i.v. bisphosphonates in patients with skeletal metastases treated for up to 10 Years.
Bisphosphonates (BPs) delay the onset or reduce the incidence of skeletal complications in patients with bone metastases. However, there are few data on the renal safety and activity of i.v. BPs beyond 2 years of administration. We retrospectively analyzed serum creatinine (SCr) levels and skeletal-related events (SREs) in cancer patients receiving i.v. BPs for >or= 24 months. All patients received 90 mg pamidronate every 3-4 weeks. Pre- and post-treatment SCr levels and the peak levels attained were recorded. A notable SCr increase was defined as: an increase >0.5 mg/dl for patients with baseline SCr <1.4 mg/dl; an increase >1 mg/dl for patients with baseline SCr >1.4 mg/dl; or doubling over baseline. The following parameters were also analyzed: the proportion of patients with at least one SRE, the distribution of each type of SRE, the time to first SRE, and the skeletal morbidity rate (SMR). Fifty-seven patients with bone metastases resulting from breast cancer (BC) (n = 48), multiple myeloma (n = 7), renal cell carcinoma (n = 1), and prostate cancer (n = 1) were evaluated. The median age at the start of treatment was 57 years (range, 27-81); 25% of the patients were >70 years old. Forty-three patients received pamidronate then switched to zoledronic acid. The median overall duration of BP administration was 34 months (range, 24+ to 131+), with a median duration of zoledronic acid therapy of 25 months (range, 2-40). Twenty-seven of 48 BC patients received different chemotherapy regimens (median number of lines, 2; range, 1-6). The median SCr levels were: baseline, 0.82 mg/dl (range, 0.4-1.4); time of analysis, 0.89 mg/dl (0.4-2); highest level, 1.0 mg/dl (0.5-2). A notable SCr increase was observed in seven patients (12.2%; all grade 1). Twenty-six patients (45.6%) experienced SREs after starting BP treatment. The median time to first SRE was 911 days (95% confidence interval, 731; 1,023). The SMR was 0.20 events per year. Ten patients ceased treatment because of: an SCr level of 2 mg/dl (n = 1) physician decision (n = 6) and jaw osteonecrosis (n = 3). Ten patients died of progressive disease. i.v. BPs are safe and active during prolonged treatment administration, and renal function is maintained in patients receiving multiple cytotoxic therapies. Jaw osteonecrosis occurred in 5% of the study population, and its causal relationship with BP treatment requires further observation and study.


Mitochondrial channels permeable by calcium ions.
The aim of this work was to characterize inner mitochondrial membrane channels permeable to calcium cations (Ca(++)). Mitochondrial membranes isolated from rat heart were incorporated into the bilayer lipid membrane, and single Ca(++) channel currents were measured. The observed channels were selective for Ca(++) and barium cations (Ba(++)) (53 and 50 mM) over Tris(+) (113 mM), but single-channel currents in most cases were noisy and did not show the typical single-channel shape, as it is known, for example, in the ryanodine receptor or in inositol 1,4,5-trisphosphate (IP(3)R) Ca(++)-release channels. The most commonly observed single-channel currents, measured at the 0 mV and 53 mM Ca(++) gradient, were in the range of 1 pA or less. The channels responded to pharmacological modulators. Some of the channels were inhibited by ruthenium red and cyclosporin A, and others were modulated by ryanodine. This may indicate that the observed channels passing Ca(++) may originate from the mitochondrial Ca(++) uniporter, the permeability transition pore, and the ryanodine receptor calcium channel.


Commentary: methodological issues for quality management in long-term care.
The interest in quality management in long-term care has been rapidly escalating. This movement to assess and improve quality parallels the effort carried out by hospital management in the past 10 years. The methodological concerns of the 2 areas are similar. This essay identifies 10 issues to which quality management leaders should pay attention as they begin to expand the capability of addressing quality in long-term care: client-centered performance versus whole-organization performance; standardization of methods and instruments; reliability; and validity, multimethod thinking, the meaning of data, comparability of data across organizations, cost barriers, feedback mechanisms, management use of quality data, and public control of data.


Decidual lymphocytes of human spontaneous abortions induce apoptosis but not necrosis in JEG-3 extravillous trophoblast cells.
The human decidua contains an unusually high proportion of lymphocytes, mainly NK and T cells, which are potentially cytotoxic to the trophoblast when they are stimulated with certain cytokines. Given the high incidence of spontaneous abortion in humans and other species, our working hypothesis is that decidual lymphocytes are involved in immunological mechanisms that attack the trophoblast and induce abortion when any gestational problem arises. To test this hypothesis, flow cytometry was used to compare decidual lymphocyte populations in first-trimester spontaneous abortions and elective terminations of first-trimester pregnancy. We found significantly higher proportions of decidual lymphocytes that expressed activation markers, and of T cells (mainly T helper cells) in spontaneous abortions than in elective terminations of pregnancy. Decidual lymphocytes from spontaneous abortion, like decidual lymphocytes from elective termination of pregnancy and peripheral blood lymphocytes, were however, unable to lyse the JEG-3 extravillous cytotrophoblast cell line in a (51)Cr-release assay. Nevertheless, decidual lymphocytes from spontaneous abortion, unlike decidual lymphocytes from elective termination of pregnancy and peripheral blood lymphocytes, induced apoptosis in JEG-3 cells as determined by DNA fragment-release assay. Hematoxylin and eosin staining showed a significantly higher proportion of apoptotic JEG-3 cells when these cells were treated with decidual lymphocytes from spontaneous abortion than when JEG-3 cells were cultured with decidual lymphocytes from elective termination of pregnancy. The ultrastructural signs of apoptosis were confirmed by electron microscopy. These data support the hypothesis that activated decidual lymphocytes participate in human spontaneous abortion by inducing apoptosis but not necrosis of the trophoblast.


Research on experimentally induced cervical cancers and the reactivity of spleen cells to Con A.
We have investigated the spleen cell response to Con A in terms of the blastoformation response of lymphocytes, which is known to be an index of the nonspecific immune response of the host organism, in various stages of experimentally induced lesions of the murine uterine cervix. It was found that, under the controlled conditions of the present experiment, suppressor T cells are induced in histological samples exhibiting AT III or severer lesions, which correspond to carcinoma of the uterine cervix. This fact suggests the possibility that antibody synthesis is suppressed. In animals administered the immunopotentiator, ATSO, the development of malignant lesions of the uterine cervix was suppressed, the induction of suppressor T cells was suppressed and the response of spleen cells to Con A was found to decrease.


Ethical issues and accountability in pressure ulcer prevention.
Pressure ulcers represent a considerable cost, both in terms of healthcare spending and quality of life. They are increasingly viewed in terms of patient harm. For clinicians involved in pressure ulcer prevention, ethical issues surrounding accountability may arise from both policy and practice perspectives. It may be useful for clinicians to refer to ethical theories and principles to create frameworks when addressing ethical dilemmas. However, such theories and principles have been criticised for their simplicity and over-generalisation. Alternative theories, for example, virtue ethics and experiential learning, can provide more comprehensive guidance and promote a pluralistic approach to tackling ethical dilemmas.


Correlation of Stomatal Conductance with Photosynthetic Capacity of Cotton Only in a CO(2)-Enriched Atmosphere: Mediation by Abscisic Acid?
Some evidence indicates that photosynthetic rate (A) and stomatal conductance (g) of leaves are correlated across diverse environments. The correlation between A and g has led to the postulation of a "messenger" from the mesophyll that directs stomatal behavior. Because A is a function of intercellular CO(2) concentration (c(i)), which is in turn a function of g, such a correlation may be partially mediated by c(i) if g is to some degree an independent variable. Among individual sunlit leaves in a cotton (Gossypium hirsutum L.) canopy in the field, A was significantly correlated with g (r(2) = 0.41, n = 63). The relative photosynthetic capacity of each leaf was calculated as a measure of mesophyll properties independent of c(i). This approach revealed that, in the absence of c(i) effects, mesophyll photosynthetic capacity was unrelated to g (r(2) = 0.06). When plants were grown in an atmosphere enriched to about 650 microliters per liter of CO(2), however, photosynthetic capacity remained strongly correlated with g even though the procedure discounted any effect of variable c(i). This "residual" correlation implies the existence of a messenger in CO(2)-enriched plants. Enriched CO(2) also greatly increased stomatal response to abscisic acid (ABA) injected into intact leaves. The data provide no evidence for a messenger to coordinate g with A at ambient levels of CO(2). In a CO(2)-enriched atmosphere, though, ABA may function as such a messenger because the sensitivity of the system to ABA is enhanced.


Temporal Variations among Invasive Pneumococcal Disease Serotypes in Children and Adults in Germany (1992-2008).
Nationwide surveillance of invasive pneumococcal disease has been conducted in Germany since 1992. From 1992 to 2008, a total of 12,137 isolates from invasive pneumococcal disease were collected. Data on serotypes were available for 9,394 invasive isolates. The leading serotypes were serotypes 14 (16.5%), 3 (8.0%), 7F (7.6%), 1 (7.3%), and 23F (6.0%). Variations in serotype distribution over the years are particularly extensive, especially concerning serotype 14 (min 7.4%, max 33.5%) with the highest percentages among the isolates serotyped from around 1997 to 2006. Serotypes 1 and 7F increased over the last decade. No increase was observed concerning serotype 19A. Higher pneumococcal conjugate vaccine coverages were observed among children (7v, 57.3%; 10v, 72.8%; 13v, 83.5%) than among adults (7v, 39.9%; 10v, 55.5%; 13v, 73.5%). The temporal variations in serotype distribution have to be kept in mind when interpreting vaccine coverages reported in epidemiological studies.


Phospholipid-Dependence of Plant UDP-Glucose Sterol beta-d-Glucosyl Transferase : IV. Reconstitution into Small Unilamellar Vesicles.
The phospholipid dependence of the UDP-glucose sterol glucosyl transferase (UDPG-SGTase) from maize coleoptiles was previously demonstrated using the partially purified and highly delipidated enzyme, in the presence of the detergent Triton X-100 (P Ullmann, P Bouvier-Navé, P Benveniste [1987] Plant Physiol 85: 51-55). We now report the reconstitution of the enzyme activity into unilamellar lipid vesicles. This was achieved by adding phospholipids, sterols and beta-octylglucoside to the solubilized enzyme and passing the mixture through Sephadex G-50. The treatment led to almost complete removal of the detergents. The incorporation of UDPG-SGTase in the lipid vesicles was demonstrated by (a) coelution of the enzyme activity with the labeled lipid vesicles (average diameter: 260A) on a Sephacryl S-1000 column and (b) flotation experiments on metrizamide density gradients. Release of dithiobis-(2-nitro-benzoic acid) (DTNB) from DTNB-preloaded vesicles was very slow, indicating good membrane integrity of the vesicles. Treatment of the intact vesicles with the nonpermeant reagent p-chloro-mercuribenzene sulfonate led to more than 95% inactivation of the total enzyme activity, i.e. the activity measured in the presence of Triton X-100 at permeabilizing concentration. This suggests an outward orientation for the active site of the enzyme. Finally, the enzyme was incorporated into vesicles of various phospholipid compositions and the kinetic parameters of the reactions were determined. Our results clearly show that the reconstituted UDPG-SGTase activity is stimulated to a large extent by negatively charged phospholipids.


Thrombotic thrombocytopenic purpura in a human immunodeficiency virus (HIV)-seropositive homosexual man.
A case of thrombotic thrombocytopenic purpura (TTP) in a human immunodeficiency virus (HIV)-seropositive homosexual man is reported. The patient improved after corticosteroid and plasma exchange therapy was instituted. The case demonstrated most of the classic features of TTP, including histologic evidence of fibrin thrombi in small blood vessels. Atypical features included reduced number of megakaryocytes in the bone marrow and only partial resolution of thrombocytopenia after therapy was begun. An autoimmune thrombocytopenia has been well-characterized in HIV-antibody-seropositive homosexual men. Physicians caring for HIV-seropositive persons should also be aware of a possible association with TTP.


Production and evaluation of the recombinant antigen TES-30 of Toxocara canis for the immunodiagnosis of toxocariasis.
Toxocara canis is a pathogenic nematode of canines which can be accidentally transmitted to humans. Although serology is the most important diagnostic tool for this zoonosis, diagnostic kits use crude excretion/secretion antigens, most of them being glycoproteins which are not species-specific and may cross-react with antibodies generated against other parasites. To produce the rTES-30 recombinant antigen of Toxocara canis and evaluate it in the immunodiagnosis of toxocariasis. The gene that codes for TES-30 was cloned in the expression vector pET28a (+) using single-stranded oligonucleotides united by PCR. The protein rTES-30 was purified by Ni2+ affinity chromotography. Seroreactivity of rTES-30 was evaluated by immunoblot. Given that there is no gold standard test, the behaviour of the antigen was compared with the method that is routinely used to immunodiagnose toxocariasis, i.e., the conventional ELISA technique using excretion/secretion antigens. The rTES-30 was produced from an Escherichia coli LB culture which yielded 2.25 mg/L of the antigen with a purity of 95%. The results obtained showed 73% (46/63) concordance of reactivity between the rTES-30 immunoblot and the conventional ELISA, and 100% concordance with the nonreactive sera (21). Nineteen of the 21 sera positive for other parasitoses reacted with ELISA, while only seven of these were positive with the rTES-30 immunoblot. Concordance between the ELISA and the immunoblot was moderate (kappa coefficient: 0.575; 95% CI: 0.41- 0.74). The data presented show the potential of the rTES-30 inmunoblot for confirmation of possible ELISA positives, not only in epidemiological studies, but also as a candidate for the development of diagnostic tests for ocular toxocariasis in Colombia.


Bid, but not Bax, regulates VDAC channels.
During apoptosis, cytochrome c is released from mitochondria into the cytosol, where it participates in caspase activation. Various and often conflicting mechanisms have been proposed to account for the increased permeability of the mitochondrial outer membrane that is responsible for this process. The voltage-dependent anion channel (VDAC) is the major permeability pathway for metabolites in the mitochondrial outer membrane and therefore is a very attractive candidate for cytochrome c translocation. Here, we report that properties of VDAC channels reconstituted into planar phospholipid membranes are unaffected by addition of the pro-apoptotic protein Bax under a variety of conditions. Contrary to other reports (Shimizu, S., Narita, M., and Tsujimoto, Y. (1999) Nature 399, 483-487; Shimizu, S., Ide, T., Yanagida, T., and Tsujimoto, Y. (2000) J. Biol. Chem. 275, 12321-12325; Shimizu, S., Konishi, A., Kodama, T., and Tsujimoto, Y. (2000) Proc. Natl. Acad. Sci. U. S. A. 97, 3100-3105), we found no electrophysiologically detectable interaction between VDAC channels isolated from mammalian mitochondria and either monomeric or oligomeric forms of Bax. We conclude that Bax does not induce cytochrome c release by acting on VDAC. In contrast to Bax, another pro-apoptotic protein (Bid) proteolytically cleaved with caspase-8 affected the voltage gating of VDAC by inducing channel closure. We speculate that by decreasing the probability of VDAC opening, Bid reduces metabolite exchange between mitochondria and the cytosol, leading to mitochondrial dysfunction.


Inequalities of Suicide Mortality across Urban and Rural Areas: A Literature Review.
Suicide mortality is a major contributor to premature death, with geographic variation in suicide rates. Why suicide rates differ across urban and rural areas has not yet been fully established. We conducted a literature review describing the urban-rural disparities in suicide mortality. Articles were searched in five databases (EMBASE, PubMed, PsychINFO, Scopus, and Web of Science) from inception till 26 May 2021. Eligible studies were narratively analyzed in terms of the urban-rural disparities in suicides, different suicide methods, and suicide trends over time. In total, 24 articles were included in our review. Most studies were ecological and cross-sectional evidence tentatively suggests higher suicide rates in rural than in urban areas. Men were more at risk by rurality than women, but suicide is in general more prevalent among men. No obvious urban-rural pattern emerged regarding suicide means or urban-rural changes over time. Potential suicidogenic explanations include social isolation, easier access to lethal means, stigmatization toward people with mental health problems, and reduced supply of mental health services. For research progress, we urge, first, individual-level cohort and case-control studies in different sociocultural settings. Second, both rurality and urbanicity are multifaceted concepts that are inadequately captured by oversimplified typologies and require detailed assessments of the sociophysical residential environment.


AML1/Runx1 rescues Notch1-null mutation-induced deficiency of para-aortic splanchnopleural hematopoiesis.
The Notch1-RBP-Jkappa and the transcription factor Runx1 pathways have been independently shown to be indispensable for the establishment of definitive hematopoiesis. Importantly, expression of Runx1 is down-regulated in the para-aortic splanchnopleural (P-Sp) region of Notch1- and Rbpsuh-null mice. Here we demonstrate that Notch1 up-regulates Runx1 expression and that the defective hematopoietic potential of Notch1-null P-Sp cells is successfully rescued in the OP9 culture system by retroviral transfer of Runx1. We also show that Hes1, a known effector of Notch signaling, potentiates Runx1-mediated transactivation. Together with the recent findings in zebrafish, Runx1 is postulated to be a cardinal down-stream mediator of Notch signaling in hematopoietic development throughout vertebrates. Our findings also suggest that Notch signaling may modulate both expression and transcriptional activity of Runx1.


Epicatechin Gallate Protects HBMVECs from Ischemia/Reperfusion Injury through Ameliorating Apoptosis and Autophagy and Promoting Neovascularization.
Green tea is one of the most beverages with antioxidants and nutrients. As one of the major components of green tea, (-)-epicatechin gallate (ECG) was evaluated for its antioxidative properties in the present study. Cell proliferation assay, tube formation, cell migration, apoptosis, and autophagy were performed in human brain microvascular endothelial cells (HBMVECs) after oxygen-glucose deprivation/reoxygenation (OGD/R) to investigate potential anti-ischemia/reperfusion injury properties of ECG in vitro. Markers of oxidative stress as ROS, LDH, MDA, and SOD were further assayed in our study. Data indicated that ECG could affect neovascularization and promote cell proliferation, tube formation, and cell migration while inhibiting apoptosis and autophagy through affecting VEGF, Bcl-2, BAX, LC3B, caspase 3, mTOR, and Beclin-1 expression. All the data suggested that ECG may be protective for the brain against ischemia/reperfusion injury by promoting neovascularization, alleviating apoptosis and autophagy, and promoting cell proliferation in HBMVECs of OGD/R.


Interaction of calmodulin with the inducible murine macrophage nitric oxide synthase.
The inducible isoform of nitric oxide synthase (iNOS) isolated from murine macrophages copurifies with calmodulin (CaM) as a tightly bound subunit. The exact function of this tightly bound CaM, however, has not been elucidated. In order to probe the function of this unusually strong interaction between iNOS and CaM, a 30 amino acid peptide derived from the putative CaM binding site of iNOS was synthesized. Cross-linking and autoradiographic analyses demonstrated that the peptide and CaM form a 1:1 complex as well as several higher molecular weight complexes. When assayed in the presence of a 12-fold excess of peptide to iNOS, over 90% of the enzymatic activity was inhibited. This inhibition could be prevented with the addition of exogenous bovine CaM to the assay mixture, in a concentration-dependent manner. Native PAGE and Western blot analysis of iNOS treated with peptide revealed the formation of a peptide--CaM complex with CaM derived from iNOS. Moreover, EGTA (5 mM) caused a 30% maximal inhibition of activity that was reversed by the addition of exogenous Ca2+ in a concentration-dependent fashion, suggesting a role for Ca2+ in this interaction. EGTA also changed the native PAGE mobility of iNOS and increased the intensity of a band which comigrates with CaM. These results demonstrate that the binding interaction between CaM and iNOS is tight but reversible, requires Ca2+, and is atypical from other known CaM--enzyme interactions.


Cortical control of normal gait and precision stepping: an fNIRS study.
Recently, real time imaging of the cortical control of gait became possible with functional near-infrared spectroscopy (fNIRS). So far, little is known about the activations of various cortical areas in more complex forms of gait, such as precision stepping. From previous work on animals and humans one would expect precision stepping to elicit extra activity in the sensorimotor cortices (S1/M1), supplementary motor area (SMA), as well as in prefrontal cortices (PFC). In the current study, hemodynamic changes in the PFC, SMA, M1, and S1 were measured with fNIRS. In contrast to previous fNIRS gait studies, the technique was optimized by the use of reference channels (to correct for superficial hemodynamic interference). Eleven subjects randomly performed ten trials of treadmill walking at 3 km/h (normal walking) and ten trials of 3 km/h treadmill walking on predefined spots for the left and right foot presented on the treadmill (precision stepping). The walking trials of approximately 35 seconds were alternated with rest periods of 25-35 seconds consisting of quiet standing. The PFC revealed profound activation just prior to the onset of both walking tasks. There was also extra activation of the PFC during the first half of the task period for precision stepping. The SMA showed mainly increased activation prior to the start of both tasks. In contrast, the sensorimotor cortex did not show a change in activation during either task as compared to a condition of standing. The SMA, M1, and S1 revealed no significant differences between normal walking and precision stepping. It was concluded that fNIRS is suited to record the planning and initiation of gait. The lack of M1/S1 activation during gait suggests that even in the current precision stepping task the control of ongoing gait depended mostly on subcortical automatisms, while motor cortex contributions did not differ between standing and walking.


Relationship of abuse by religious authorities to depression, religiosity, and child physical abuse history in a college sample.
Discussion of sexual abuse by religious authorities has been plagued by allegations of false memories and misreports, often attributed to media attention. An analysis of a historical archive with information on abuse by religious and other authority figures and coexisting psychopathology is extremely useful to the current debate on outcomes of sexual abuse. The present study utilizes a database from the late 1970s that contains data on physical abuse and sexual abuse by various perpetrator types as well as on symptoms of depression, anxiety, and religiosity in a college population sample. Students alleging sexual abuse by religious authorities were as symptomatic (depressed and anxious) as students abused by parents and were more symptomatic than controls. Further, those abused by religious authorities showed greater variance in religiosity and a greater likelihood of breaking ties with their religious communities. Students self-labeling as physically abused by parents were more at risk for sexual abuse by religious authorities. Religious authorities often play complex roles in social and family life. These complexities then may produce parallel complexities in the patterns of symptoms. The intensity of the trauma from abuse by religious authorities may make it more difficult to study, treat, and recover from sexual abuse. Additional research is needed. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Break the fast? Update on patient preparation for cholesterol testing.
To provide an update on the clinical usefulness of nonfasting versus fasting lipid testing to improve patient compliance, patient safety, and clinical assessment in cholesterol testing. Recommendations are identified as supported by good, fair, and poor (conflicting or insufficient) evidence, according to the classifications adopted by the Canadian Task Force on Preventive Health Care. Screening for dyslipidemia as a risk factor for coronary artery disease and management of lipid-lowering medications are key parts of primary care. Recent evidence has questioned the fasting requirement for lipid testing. In population-based studies, total cholesterol, high-density lipoprotein cholesterol, and non-low-density lipoprotein cholesterol all varied by an average of 2% with fasting status. For routine screening, nonfasting cholesterol measurement is now a reasonable alternative to a fasting cholesterol measurement. For patients with diabetes, the fasting requirement might be an important safety issue because of problems with hypoglycemia. For the monitoring of triglyceride and low-density lipoprotein cholesterol levels in patients taking lipid-lowering medications, fasting becomes more important. Fasting for routine lipid level determinations is largely unnecessary and unlikely to affect patient clinical risk stratification, while nonfasting measurement might improve patient compliance and safety.


Surgical treatment of cystic neoplasms of the pancreas.
Diagnosis and treatment of cystic neoplasms of the pancreas increased significantly in the last decades. There are only a few Brazilian publications on these tumors. The majority of them are limited to reports of one or few cases. To present our experience with 27 patients with cystic neoplasms of the pancreas. Demographic data, clinical manifestations, diagnostic exams, surgical procedures, postoperative complications, and follow-up data of 27 patients with cystic neoplasms of the pancreas were analyzed, according to the histological type of the tumor. There were 10 (37%) serous cystic tumors, 10 (37%) mucinous cystic tumors, 4 (15%) intraductal papillary mucinous tumors, and 3 (11%) solid pseudopapillary tumors or Frantz tumor. All serous cystic tumors, 6 (60%) mucinous tumors, 2 (50%) intraductal papillary mucinous tumors, and 2 (67%) solid pseudopapillary tumors were benign. The age of the patients varied from 31 to 82 years and all tumors were more common in female. Two patients had been treated previously as a pseudocyst. Surgical procedures depended on the location and extension of the tumor. Two patients underwent only laparotomy with tumor biopsy, one cholecystectomy with Roux-en-Y hepaticojejunostomy for jaundice treatment, 6 pancreatoduodenectomy, and 18 partial pancreatectomy. The most common postoperative complication was pancreatic fistula (n = 5; 19%). One patient died of necrotic pancreatitis. Of the 10 patients with serous cystic tumor, only 1 had tumor recurrence at the section border. The three patients with mucinous cystoadenocarcinoma in which was not possible to resect the tumor, died 6 to 24 months after laparotomy. The six patients with benign mucinous tumors did not have tumor recurrence. The most common cystic neoplasms of the pancreas are serous and mucinous cysts. These tumors are more frequent in female. Although almost all serous cysts are benign, 40% of mucinous cysts are malign. Misdiagnosis may delay appropriate treatment and increase mortality.


[Epidemiological profile of lateral epicondylitis in rehabilitation department].
lateral epicondylitis, better known as tennis elbow, falls within the spectrum of upper extremity musculoskeletal disorders and it is a major public health problem. The purpose of our study was to determine the epidemiological and clinical profile and treatment modalities for patients with lateral epicondylitis followed at the Department of Physical Medicine and Rehabilitation. we conducted a retrospective study (2012-2017) of patients with lateral epicondylitis referred to the Department of Physical Medicine and Functional Rehabilitation at the Tahar Sfar Mahdia University Hospital over a period of 6 years. Socio-demographic and clinical features as well as treatment modalities were assessed for each patient. fifty patients were enrolled in the study. The average age of patients was 44.82 years, with predominance of the female sex (72%). Half of our patients were active and the majority of them (60%) worked in a traditional office environment. Right upper extremity was most affected (78% of cases). The median duration of symptoms was 14.66 months. Clinical examination showed positive tendon triad in the lateral epicondyle in 96% of cases. Radiological examination was performed in 8 patients, ultrasound in 6 patients. All patients received analgesics, 84% of patients received non-steroidal anti-inflammatory drugs, only 8 patients received corticosteroid injections. One patient underwent surgery after therapeutic failure. Ninety-two percent of patients were scheduled for rehabilitation sessions. Total improvement was noted in 42% of patients, 46% reported transient improvement and 12% progressed to chronicity. lateral epicondylitis is a common cause of elbow pain. Its management in physical medicine is based on medical treatment and appropriate functional rehabilitation. However no therapeutic option seems clearly superior to the other.


Effect of Photon Hormesis on Dose Responses to Alpha Particles in Zebrafish Embryos.
Photon hormesis refers to the phenomenon where the biological effect of ionizing radiation with a high linear energy transfer (LET) value is diminished by photons with a low LET value. The present paper studied the effect of photon hormesis from X-rays on dose responses to alpha particles using embryos of the zebrafish (Danio rerio) as the in vivo vertebrate model. The toxicity of these ionizing radiations in the zebrafish embryos was assessed using the apoptotic counts at 20, 24, or 30 h post fertilization (hpf) revealed through acridine orange (AO) staining. For alpha-particle doses ≥ 4.4 mGy, the additional X-ray dose of 10 mGy significantly reduced the number of apoptotic cells at 24 hpf, which proved the presence of photon hormesis. Smaller alpha-particle doses might not have inflicted sufficient aggregate damages to trigger photon hormesis. The time gap T between the X-ray (10 mGy) and alpha-particle (4.4 mGy) exposures was also studied. Photon hormesis was present when T ≤ 30 min, but was absent when T = 60 min, at which time repair of damage induced by alpha particles would have completed to prevent their interactions with those induced by X-rays. Finally, the drop in the apoptotic counts at 24 hpf due to photon hormesis was explained by bringing the apoptotic events earlier to 20 hpf, which strongly supported the removal of aberrant cells through apoptosis as an underlying mechanism for photon hormesis.


A minimum quantum chemistry CCSD(T)/CBS dataset of dimeric interaction energies for small organic functional groups.
We have performed a quantum chemistry study on the bonding patterns and interaction energies for 31 dimers of small organic functional groups (dubbed the SOFG-31 dataset), including the alkane-alkene-alkyne (6 + 4 + 4 = 14, AAA) groups, alcohol-aldehyde-ketone (4 + 4 + 3 = 11, AAK) groups, and carboxylic acid-amide (3 + 3 = 6, CAA) groups. The basis set superposition error corrected super-molecule approach using the second order Møller-Plesset perturbation theory (MP2) with the Dunning's aug-cc-pVXZ (X = D, T, Q) basis sets has been employed in the geometry optimization and energy calculations. To calibrate the MP2 calculated interaction energies for these dimeric complexes, we perform single-point calculations with the coupled cluster with single, double, and perturbative triple excitations method at the complete basis set limit [CCSD(T)/CBS] using the well-tested extrapolation methods. In order to gain more physical insights, we also perform a parallel series of energy decomposition calculations based on the symmetry adapted perturbation theory (SAPT). The collection of these CCSD(T)/CBS interaction energy values can serve as a minimum quantum chemistry dataset for testing or training less accurate but more efficient calculation methods. As an application, we further propose a segmental SAPT model based on chemically recognizable segments in a specific functional group. These model interactions can be used to construct coarse-grained force fields for larger molecular systems.


Protein Hydrolysis and Glycosylation as Strategies to Produce Bioactive Ingredients from Unmarketable Prawns.
The present work shows a procedure to valorize non-commercial boiled shrimp to produce functional ingredients, using a combined treatment based on enzymatic hydrolysis and subsequent glycation under mild conditions. Antioxidant and prolyl endopeptidase-inhibiting activities were determined as a function of hydrolysis and glycation times (0-120 min and 0-180 min, respectively). The reaction products were characterized by determining the degree of hydrolysis, browning, fluorescent compounds, free amino acids, phenol content, Fourier transform infrared spectroscopy (FTIR), and molecular weight of the different fractions obtained. Enzymatic hydrolysis generated hydrolysates with significant antioxidant and prolyl endopeptidase-inhibiting activities. Glycation under mild conditions was used as a strategy to improve the antioxidant and potential nootropic properties of the hydrolysates. During glycation, the free amino acid content decreased, total phenols and fluorescent compounds increased significantly, and low molecular weight melanoidins were formed. The presence of peptide-glucose conjugates was also confirmed by FTIR. Glycation increased the antioxidant activities of the hydrolysates; however, their prolyl-endopeptidase-inhibiting activity was lost. Results showed that compounds with promising antioxidant (hydrolysis and glycation) and potential nootropic (hydrolysis) activities and applications in food systems were obtained from the biotechnological strategy used.


Tumour metastasis to the liver, and the roles of proteinases and adhesion molecules: new concepts from in vivo videomicroscopy.
Most preclinical studies of tumour metastasis and effects of molecular interventions have been based on end point assays, and little is known about the fate of cells at sequential steps in the metastatic process. In vivo videomicroscopy permits direct observations of sequential steps in hematogenous metastasis as they occur in living animals over time. These steps include initial arrest of cells in the microcirculation, extravasation, postextravasation migration and growth in the target organ. In the mouse liver model, cells are arrested in periportal sinusoids based on size restriction, survive in the circulation and extravasate into the tissue by 48 to 72 h regardless of metastatic potential. Thereafter, cells may migrate to preferred sites for growth. Critical steps responsible for cell losses and metastatic inefficiency occur at the level of postextravasation cell growth. Many extravasated cells may remain dormant, and growth to form micrometastases is initiated in only a small subset of cells. Most early micrometastases may disappear after a few days, and only a small subset continue growth into macroscopic tumours. Angiogenesis is a prerequisite for continued growth of metastases, as shown previously by others. Integrin based interventions can modulate postextravasation cell migration and cell growth. Matrix metalloproteinase inhibitors can inhibit tumour angiogenesis and thus reduce growth. Key targets against which future therapeutic strategies should be directed include the initiation and maintenance of growth of micrometastases, and the activation of dormant solitary cells.


Development and clinical evaluation of a computerized limb volume measurement system (CLEMS).
Limb edema is a common problem in both rehabilitation and acute care settings. In the past, attempts to determine an optimal management strategy for limb edema have been limited by the lack of accurate, noninvasive, rapid, clinical tools for quantifying limb volumes. The water displacement method is slow and difficult to use in the clinical setting. Furthermore, water displacement requires that the limb be in a dependent position. The tape measure method is unreliable because it is difficult to position the tape measure on a swollen limb. The development and evaluation of a new tool called the computerized limb volume measurement system (CLEMS) is described. The shape and volume of a limb or limb segment can be rapidly measured by CLEMS, independent of limb position. The limb volumes generated by CLEMS were compared to volumes determined by water displacement and by a tape measure. Volumes of eighteen legs (plaster, nonedematous and edematous) were measured using CLEMS, water displacement, and the tape measure. In all cases, the CLEMS and water displacement methods showed close agreement. CLEMS was found to be a reliable and valid new method of determining limb volume; whereas, the tape measure method was found to be invalid. This new tool allows clinicians to measure the efficacy of different treatment strategies in the management of limb edema.


Long-term radioiodine retention and regression of liver cancer after sodium iodide symporter gene transfer in wistar rats.
Radioiodine therapy of nonthyroid cancers after sodium iodide symporter (NIS) gene delivery has been proposed as a potential application of gene therapy. However, it seems to be precluded by the rapid efflux of taken up iodine from most transduced xenografted tumors. We present an in vivo kinetic study of NIS-related hepatic iodine uptake in an aggressive model of hepatocarcinoma induced by diethylnitrosamine in immunocompetent Wistar rats. We followed the whole-body iodine distribution by repeated imaging of live animals. We constructed a rat NIS (rNIS) adenoviral vector, Ad-CMV-rNIS, using the cytomegalovirus (CMV) as a promoter. Injected in the portal vein in 5 healthy and 25 hepatocarcinoma-bearing rats and liver tumors in 9 hepatocarcinoma-bearing rats, Ad-CMV-rNIS drove expression of a functional NIS protein by hepatocytes and allowed marked (from 20 to 30% of the injected dose) and sustained (>11 days) iodine uptake. This contrasts with the massive iodine efflux found in vitro in human hepatic tumor cell lines. In vivo specific inhibition of NIS by sodium perchlorate led to a rapid iodine efflux from the liver, indicating that the sustained uptake was not attributable to an active retention mechanism but to permanent recycling of the effluent radioiodine via the high hepatic blood flow. Radioiodine therapy after Ad-CMV-rNIS administration achieved a strong inhibition of tumor growth, the complete regression of small nodules, and prolonged survival of hepatocarcinoma-bearing rats. This demonstrates for the first time the efficacy of NIS-based radiotherapy in a relevant preclinical model of nonthyroid human carcinogenesis.


Characterize the microstructure change after tendon enthesis injury using synchrotron radiation μCT.
The microstructure of the bone-tendon interface (BTI) deserves in-depth investigation. In this study, we first aimed to extend the application of synchrotron radiation μCT to characterize the gradient structure of supraspinatus tendon (SST) enthesis, from both tissue morphology to cell distribution. Second, to acquire detailed morphological information of SST enthesis when after injury. Our results showed that in normal enthesis, the phenotype of chondrocyte in BTI was dependent on its distance to subchondral bone. After injury, the fibrocartilage cells were disrupted, as evidenced by reduced lacunae size. Our observation may partly explain the loss of BTI mechanical properties after injury, and we believe the application of synchrotron radiation microcomputed tomography will have promising potential for characterizing the morphology changes in enthesis and for evaluating the therapeutic effects of interventions in preclinical studies.


Synthetic, structural, electrochemical, and theoretical studies of heterometallic aggregates with a [Pt(2)(mu-S)(2)M] core (M = Hg, Au).
Novel electroactive multimetallic compounds based on the [Pt(2)(mu(2)-S)(2)M] core, viz. [Pt(2)(PPh(3))(4)(mu(3)-S)(2)HgFc]PF(6) (1) [Fc = (eta(5)-C(5)H(4))Fe(eta(5)-C(5)H(5))] and [Pt(2)(PPh(3))(4)(mu(3)-S)(2)Hg(2)Fc'](PF(6))(2) (2) [Fc' = Fe(eta(5)-C(5)H(4))(2)], have been synthesized under the guide of electrospray mass spectrometry. The electrochemistry of these ferrocene funtionalized compounds together with the reported [Pt(2)(PPh(3))(4)(mu(3)-S)(2)HgPPh(3)](PF(6))(2) (3), [Pt(2)(PPh(3))(4)(mu(2)-S)(mu(3)-S)HgPh]PF(6) (4), and [Pt(2)(PPh(3))(4)(mu(2)-S)(mu(3)-S)AuPPh(3)]PF(6) (5) have been investigated using cyclic voltammetry and DFT calculations. These results point to a prominent ligand-based oxidation.


Adolescent development in interpersonal and societal contexts.
In this chapter we review theoretical and empirical advances in research on adolescent development in interpersonal and societal contexts. First, we identify several trends in current research, including the current emphasis on ecological models and the focus on diversity in and relational models of adolescent development. Next, we discuss recent research on interpersonal relationships, with an eye toward identifying major research themes and findings. Research on adolescents' relationships with parents, siblings, other relatives, peers, and romantic partners, and adolescents' involvement in community and society is reviewed. Future directions in research on adolescent development are discussed.


Intranasal delivery of morphine.
Morphine administered nasally to humans as a simple solution is only absorbed to a limited degree, with a bioavailability of the order of 10% compared with intravenous administration. This article describes the development of novel nasal morphine formulations based on chitosan, which, in the sheep model, provide a highly increased absorption with a 5- to 6-fold increase in bioavailability over simple morphine solutions. The chitosan-morphine nasal formulations have been tested in healthy volunteers in comparison with a slow i.v. infusion (over 30 min) of morphine. The results show that the nasal formulation was rapidly absorbed with a T(max) of 15 min or less and a bioavailability of nearly 60%. The shape of the plasma profile for nasal delivery of the chitosan-morphine formulation was similar to the one obtained for the slow i.v. administration of morphine. Furthermore, the metabolite profile obtained after the nasal administration of the chitosan-morphine nasal formulation was essentially identical to the one obtained for morphine administered by the intravenous route. The levels of both morphine-6-glucuronide and morphine-3-glucuronide were only about 25% of that found after oral administration of morphine. It is concluded that a properly designed nasal morphine formulation (such as one with chitosan) can result in a non-injectable opioid product capable of offering patients rapid and efficient pain relief.


Identification of a domain in Bordetella pertussis adenylyl cyclase important for subunit interactions and cell invasion activity.
Bordetella pertussis produces a calmodulin-stimulated adenylyl cyclase that invades animal cells and raises intracellular cAMP levels. The enzyme does not enter animal cells by receptor-mediated endocytosis, but the mechanism for invasion of animal cells has not been defined. We have proposed that the 177 kDa adenylyl cyclase is proteolyzed to a 45 kDa catalytic subunit and one or more polypeptides (invasive factor) that facilitate entry of the catalytic subunit into animal cells. In this study, we report the identification of a sequence of amino acids within the adenylyl cyclase catalytic subunit that is important for entry of the enzyme into eukaryotic cells. A synthetic peptide corresponding to amino acids 313-339 within the catalytic subunit was shown to inhibit invasion of neuroblastoma cells by the adenylyl cyclase. In addition, this peptide inhibited the association of the catalytic subunit with invasive factor. We propose that this domain is a site for interaction between the catalytic subunit and invasive factor.


[Prospects and challenges of exosomes as drug delivery systems].
Exosomes are nanoscale vectors with a diameter of 30~100 nm secreted by living cells, and they are important media for intercellular communication. Recent studies have demonstrated that exosomes can not only serve as biomarkers for diagnosis, but also have great potential as natural drug delivery vectors. Exosomes can be loaded with therapeutic cargos, including small molecules, proteins, and oligonucleotides. Meanwhile, the unique biological compatibility, high stability, and tumor targeting of exosomes make them attractive in future tumor therapy. Though exosomes can effectively deliver bioactive materials to receptor cells, there is a wide gap between our current understanding of exosomes and their application as ideal drug delivery systems. In this review, we will briefly introduce the function and composition of exosomes, and mainly summarize the potential advantages and challenges of exosomes as drug carriers. Finally, this review is expected to provide new ideas for the development of exosome-based drug delivery systems.


Tunable multiferroic order parameters in Sr1- x Ba x Mn1- y Ti y O3.
Responding to the rapidly increasing demand for efficient energy usage and increased speed and functionality of electronic and spintronic devices, multiferroic oxides have recently emerged as key materials capable of tackling this multifaceted challenge. In this paper, we describe the development of single-site manganese-based multiferroic perovskite materials with modest amounts of nonmagnetic Ti substituted at the magnetic Mn site in Sr1- x Ba x Mn1- y Ti y O3 (SBMTO). Significantly enhanced properties were achieved with ferroelectric-type structural transition temperatures boosted to ∼430K. Ferroelectric distortions with large spontaneous polarization values of ∼30μC/cm2, derived from a point charge model, are similar in magnitude to those of the prototypical nonmagnetic BaTiO3. Temperature dependence of the system's properties was investigated by synchrotron x-ray powder diffraction and neutron powder diffraction at ambient and high pressures. Various relationships were determined between the structural and magnetic properties, Ba and Ti contents, and T N and T C. Most importantly, our results demonstrate the large coupling between the magnetic and ferroelectric order parameters and the wide tunability of this coupling by slight variations of the material's stoichiometry.


A Physiological and Kinematic Comparison of two Different Lean Back Positions During Stationary Rowing on a Concept II Machine.
This study compared two different body positions at the finish of a stroke during stationary rowing exercise on physiological and kinematic measurements. Nine male and five female rowers volunteered for the study: mean age (± SD), body height and body mass were 27 ±9 yrs, 180.5 ±12.3 cm and 81.2 ±14.2 kg. The two body positions at the finish were controlled at an upright posture or a novel greater lean back position. All subjects completed 3 different experimental trials on a Concept IID rowing machine at 3 different exercise intensities and comparisons were made between the lean back position at the same stroke rate and the same power output as the upright trial. Power output, heart rate, oxygen uptake, energy expenditure and % efficiency were higher (p<0.05) with the greater lean back position at the same stroke rate compared to all other conditions. Range of motion at the hip, ankle, and elbow and the handle velocity and distance moved were greater (p<0.05) with the lean back position. In conclusion, a greater lean back posture at the finish during stationary rowing produces a higher power output and improved efficiency at the same stroke rate but at an elevated physiological cost compared to a more upright position. Despite the higher energy expenditure, the relative gain in power output and efficiency with no negative kinematic changes suggests that a greater lean back position at the finish will enhance performance during stationary rowing exercise.


Revision of the genus Dinotoperla Tillyard, 1921 (Plecoptera: Gripopterygidae) using morphological characters and molecular data: Establishes two new genera, three new species and updates the larval taxonomy.
The larval taxonomy of Australian stoneflies (Plecoptera) shows a large disparity in knowledge when compared to the adult taxonomy with many species having undescribed larval forms. The importance of stoneflies as an indicator group for monitoring aquatic ecosystems means knowledge of the larval taxonomy and the ability to identify species is essential. This study combined morphology and mitochondrial gene sequences to associate the adult and larval life-stages for species of Dinotoperla Tillyard. Morphological identification of adult males was recognised for 17 of the 35 Dinotoperla species and combining molecular data with morphology confirmed eight new adult-larval life stage associations. Further, molecular data supported the larval taxonomy for five morphospecies which remain unassociated. The combination of molecular and morphological methods enabled the larval morphology to be reassessed for the genus Dinotoperla and this has led to the establishment of two new genera, Odontoperla, gen. nov. and Oedemaperla, gen. nov., and the new species Dinotoperla aryballoi, sp. nov, D. tasmaniensis, sp. nov. and Oedemaperla shackletoni, sp. nov. as well as the new or updated descriptions of the larvae of 31 species and a comprehensive dichotomous key to these larvae.


Peripheral neuropathy in patients with CPEO associated with single and multiple mtDNA deletions.
To characterize peripheral nerve involvement in patients with chronic progressive external ophthalmoplegia (CPEO) with single and multiple mitochondrial DNA (mtDNA) deletions, based on clinical scores and detailed nerve conduction studies. Peripheral nerve involvement was prospectively investigated in 33 participants with CPEO (single deletions n = 18 and multiple deletions n = 15). Clinically, a modified Total Neuropathy Score (mTNS) and a modified International Cooperative Ataxia Rating Scale (mICARS) were used. Nerve conduction studies included Nn. suralis, superficialis radialis, tibialis, and peroneus mot. Early somatosensory evoked potentials were obtained by N. tibialis stimulation. Participants with multiple deletions had higher mTNS and mICARS scores than those with single deletions. Electrophysiologically in both sensory nerves (N. suralis and N. radialis superficialis), compound action potential (CAP) amplitudes and nerve conduction velocities were lower and mostly abnormal in multiple deletions than those in single deletions. Early somatosensory evoked potentials of N. tibialis revealed increased P40 latencies and decreased N35-P40 amplitudes in multiple deletions. Both sensory nerves had higher areas under the receiver operating characteristic curves for the decreased CAP amplitudes than the 2 motor nerves. The N. suralis had the best Youden index, indicating a sensitivity of 93.3% and a specificity of 72.2% to detect multiple deletions. Peripheral nerve involvement in participants with multiple mtDNA deletions is an axonal type of predominant sensory neuropathy. This is clinically consistent with higher mTNS and mICARS scores. Sensory nerve involvement in participants with multiple deletions was not correlated with age at onset and duration of disease.


Local confidential inquiry into avoidable factors in deaths from stroke and hypertensive disease.
To audit avoidable deaths from stroke and hypertensive disease. Details of care before death were obtained from general practitioners and other doctors, anonymised, and assessed by two experts against agreed minimum standards of good practice for detecting and managing hypertension. Health authority with population of 250,000. All patients under 75 years who died of stroke, hypertensive disease, or hypertension related causes during November 1990 to October 1991. Presence of important avoidable factors and departures from minimum standards of good practice. Adequate information was obtained for 88% (123/139) of eligible cases. Agreement between the assessors was mostly satisfactory. 29% (36/123, 95% confidence interval 21% to 37%) of all cases and 44% (36/81, 34% to 55%) of those with definite hypertension had avoidable factors that may have contributed to death. These were most commonly failures of follow up and continuing smoking. Assessment against standards of minimum good practice showed that care was inadequate but not necessarily deemed to have contributed to death, in a large proportion of patients with definite hypertension. Common shortcomings were inadequate follow up, clinical investigation, and recording of smoking and other relevant risk behaviours. This method of audit can identify shortcomings in care of patients dying of hypertension related disease.


Crystal structure and some properties of a major house dust mite allergen, Derf 2.
Pyroglyphid house dust mites are a major source of allergens in house dust. Mite allergens sensitize and induce asthma, rhinitis, and eczema in a large portion of patients with allergic diseases. Here, the crystal structure of a major mite allergen, Derf 2, derived from Dermatophagoides farinae was solved by single isomorphous replacement method with anomalous scattering (SIRAS) at 2.1A resolution. The present study also demonstrated that the conformation of the allergen was critical in the determination of Th1/Th2 shift based on physicochemical and immunological analyses. This indicates that rigidly folded and singly dispersed structure is essentially required for the generation of Th2 type cells by the allergen, while conformational variant protein leads to Th1 skewing, irrespective of the same amino acid sequence. This structure/function relationship may allow us to develop a novel strategy for hyposensitization therapy in patients with allergic diseases triggered by house dust mite allergens.


[Primary suture following the bile duct exploration in cholelithiasis].
Classical performance of bile duct exploration is always strictly connected with the problem of an incised bile duct closure. In majority of surgical centers an external T-tube drainage was carried out in last century. But is an external drainage the best method to be used? The aim of the paper was to establish criteria for including (or excluding) patients to groups undergoing primary suture of bile duct. The comparison of the patients in whom bile duct exploration was followed by primary suture with the group in which T-tube was left in bile duct lumen was performed. Recently in our Department a classical method has been used to operate on 65 patients with cholelithiasis. In 42 patients a preoperatively planned bile duct exploration has been performed. In 14 patients a T-tube was left after the exploration, in 24 patients bile duct was closed with a primary suture. In other 18 patients with residual and recurrent bile duct stones T-tube was left in bile duct only in two patients. Based on literature analysis, our observations, and surgical experience criteria for including and excluding patients for primary suture of bile duct have been established. Groups of patients with and without T-tube were compared, considering complications and economic aspects of hospitalization. Our opinion is that whenever in specialized surgical centers detailed criteria are established and followed precisely, primary bile duct closure is a simple and safe procedure, responsible for few complications. The cost of hospitalization in the case of primary suture is half as much as the price of the hospitalization in the case of T-tube.


Metastatic lung adenocarcinoma to the iris mimicking Cogan-Reese syndrome.
Case report of metastatic lung cancer to the iris mimicking Cogan-Reese syndrome. A 61-year-old female with a history of non-small cell lung cancer presented with unilateral elevated intraocular pressure (IOP) and multiple hyperpigmented iris nodules associated with an anterior iris membrane. The patient's IOP could not be controlled medically, so she underwent trabeculectomy with simultaneous iris biopsy. Histopathology revealed proliferation of glandular epithelium on the iris surface consistent with metastatic lung adenocarcinoma. Iris metastases have varied presentations, and one must consider metastatic disease in any patient with a history of cancer presenting with iris lesions. The formation of an iris membrane and nodules by metastatic adenocarcinoma to the iris may mimic Cogan-Reese syndrome.


Control of alpha-amylase mRNA accumulation by gibberellic Acid and calcium in barley aleurone layers.
Pulse-labeling of barley (Hordeum vulgare L. cv Himalaya) aleurone layers incubated for 13 hours in 2.5 micromolar gibberellic acid (GA(3)) with or without 5 millimolar CaCl(2) shows that alpha-amylase isozymes 3 and 4 are not synthesized in vivo in the absence of Ca(2+). A cDNA clone for alpha-amylase was isolated and used to measure alpha-amylase mRNA levels in aleurone layers incubated in the presence and absence of Ca(2+). No difference was observed in alpha-amylase mRNA levels between layers incubated for 12 hours in 2.5 micromolar GA(3) with 5 millimolar CaCl(2) and layers incubated in GA(3) alone. RNA isolated from layers incubated for 12 hours in GA(3) with and without Ca(2+) was translated in vitro and was found to produce the same complement of translation products regardless of the presence of Ca(2+) in the incubation medium. Immunoprecipitation of translation products showed that the RNA for alpha-amylase synthesized in Ca(2+)-deprived aleurone layers was translatable. Ca(2+) is required for the synthesis of alpha-amylase isozymes 3 and 4 at a step after mRNA accumulation and processing.


Progressive increase of FcεRI expression across several PBMC subsets is associated with atopy and atopic asthma within school-aged children.
Antigen-specific IgE binds the Fcε receptor I (FcεRI) expressed on several types of immune cells, including dendritic cells (DCs). Activation of FcεRI on DCs in atopics has been shown to modulate immune responses that potentially contribute to asthma development. However, the extent to which DC subsets differ in FcεRI expression between atopic children with or without asthma is currently not clear. This study aimed to analyse the expression of FcεRI on peripheral blood mononuclear cells (PBMCs) from atopic children with and without asthma, and non-atopic/non-asthmatic age-matched healthy controls. We performed multiparameter flow cytometry on PBMC from 391 children across three community cohorts and one clinical cohort based in Western Australia. We confirmed expression of FcεRI on basophils, monocytes, plasmacytoid and conventional DCs, with higher proportions of all cell populations expressing FcεRI in atopic compared to non-atopic children. Further, we observed that levels of FcεRI expression were elevated across plasmacytoid and conventional DC as well as basophils in atopic asthmatic compared to atopic non-asthmatic children also after adjusting for serum IgE levels. Our data suggest that the expression pattern of FcεRI on DC and basophils differentiates asthmatic from non-asthmatic atopic children. Given the significant immune modulatory effects observed as a consequence of FcεRI expression, this altered expression pattern is likely to contribute to asthma pathology in children.