Serological evidence of dengue fever and its associated factors in health facilities in the Borena Zone, South Ethiopia.
Dengue fever (DF) is a re-emerging public health threat in Ethiopia. Yet, little is known about the epidemiology and risk factors of dengue infection in the region. In this study, the seroprevalence and associated risk factors of dengue virus infection were assessed in the Borena Zone health facilities. A hospital-based cross-sectional study was conducted from July to August 2016. A total of 519 consecutive acute febrile patients attending the outpatient departments of Teltelle Health Center, Yabello and Moyale Hospital were enrolled. Data on socio-demographic and environmental risk factors were collected using a structured questionnaire. Three to five milliliter blood samples were collected from all participants and screened for dengue virus exposure using indirect immunofluorescent assay. The overall prevalence of anti-DENV IgG and IgM was 22.9% and 7.9%, respectively. DF serostatus was influenced by gender (adjusted odd ratio (AOR)=1.72; 95% CI 1.01-2.94), place of residence (AOR=2.69; 95%CL 1.55-4.64) that had a higher rate of exposure and recalling of a recent mosquito bite (AOR=2.98; 95% CI 1.51-5.89) probably imply recent and/or ongoing active transmission. This study showed that DF could potentially emerge as a public health threat in the study area. In addition to that, the observed low awareness of participants underlines the urgent need for further community-based studies to determine the environmental, and host factors that determine the extent of exposure to dengue virus infection in the area for appropriate control and prevention planning.


Mesophyll cell ultrastructure of wheat leaves etiolated by lead and selenium.
The ultrastructure of mesophyll cells was studied in leaves of the Triticum aestivum L. cv. "Trizo" seedlings after two weeks of growth on soil contaminated by Pb and/or Se. The soil treatments: control; (Pb1) 50mgkg-1; (Pb2) 100mgkg-1; (Se1) 0.4mgkg-1; (Se2) 0.8mgkg-1; (Pb1+Se1); (Pb1+Se2); (P2+Se1); and (Pb2+Se2) were used. Light and other conditions were optimal for plant growth. The (Se1)-plants showed enhanced growth and biomass production; (Pb1+Se1)-plants did not lag behind the controls, though O2 evolution decreased; chlorophyll content did not differ statistically in these treatments. Other treatments led to statistically significant growth suppression, chlorophyll content reduction, inhibition of photosynthesis, stress development tested by H2O2 and leaf etiolation at the end of 14-days experiment. The tops of etiolated leaves remained green, while the main leaf parts were visually white. Plastids in mesophyll cells of etiolated parts of leaves were mainly represented by etioplasts and an insignificant amount of degraded chloroplasts. Other cellular organelles remained intact in most mesophyll cells of the plants, except (Pb2+Se2)-plants. Ruptured tonoplast and etioplast envelope, swelled cytoplasm and mitochondria, and electron transparent matrix of gialoplasm were observed in the mesophyll cells at (Pb2+Se2)-treatment, that caused maximal inhibition of plant growth. The results indicate that Pb and Se effects on growth of wheat leaves are likely to target meristem in which the development of proplastids to chloroplasts under the light is determined by chlorophyll biosynthesis. Antagonistic effect of low concentration of Se and Pb in combination may retard etiolation process.


Case report of hypopituitarism with suspected syndrome of inappropriate VP secretion (SIADH) due to a large aneurysm of the internal carotid in the sellar region.
Hypopituitarism and hyponatremia, especially when severe, are infrequent findings particularly when the cause of hypopituitarism at presentation is unknown and untreated. Interestingly, hyponatremia is usually seen in elderly patients with hypopituitarism due to various causes. We present a case with unrecognized and untreated hypopituitarism due to a large aneurysm of the internal carotid artery in the sellar region causing the syndrome of inappropriate secretion of antidiuretic hormone (SIADH).


Determination of urinary malondialdehyde by isotope dilution LC-MS/MS with automated solid-phase extraction: a cautionary note on derivatization optimization.
A highly sensitive quantitative LC-MS/MS method was developed for measuring urinary malondialdehyde (MDA). With the use of an isotope internal standard and online solid-phase extraction, urine samples can be directly analyzed within 10 min after 2,4-dinitrophenylhydrazine (DNPH) derivatization. The detection limit was estimated as 0.08 pmol. This method was further applied to assess the optimal addition of DNPH for derivatization and to measure urinary MDA in 80 coke oven emission (COE)-exposed and 67 nonexposed workers. Derivatization optimization revealed that to achieve complete derivatization reaction, an excess of DNPH is required (DNPH/MDA molar ratio: 893-8929) for urine samples that is about 100 times higher than that of MDA standard solutions (molar ratio: 10-80). Meanwhile, the mean urinary concentrations of MDA in COE-exposed workers were significantly higher than those in nonexposed workers (0.23±0.17 vs 0.14±0.05 μmol/mmol creatinine, P<0.005). Urinary MDA concentrations were also significantly associated with the COE (P<0.005) and smoking exposure (P<0.05). Taken together, this method is capable of routine high-throughput analysis and accurate quantification of MDA and would be useful for assessing the whole-body burden of oxidative stress. Our findings, however, raise the issue that derivatization optimization should be performed before it is put into routine biological analysis.


Long-term outcome of operatively treated fracture-dislocations of the olecranon.
To report the long-term results of operative treatment of anterior and posterior olecranon fracture-dislocations and compare them with the results recorded fewer than 2 years after surgery. Retrospective case series with long-term evaluation. Level I trauma center. Ten patients with anterior olecranon fracture-dislocation and ten patients with posterior olecranon fracture-dislocation were evaluated after an average of 18 years (range, 11 to 28 years) after injury. Fifteen patients had an early follow-up available at an average 14 months (range, 6 to 24 months) after surgery. The average age at injury was 30 years (range, 14 to 53 years). Treatment included plate and screw fixation (11 patients), tension band wiring (8 patients), and radiocapitellar transfixation (1 patient). Six patients had additional elbow surgery before the final evaluation. Flexion arc, arthrosis, Mayo Elbow Performance Index (MEPI), Disability of Arm Shoulder and Hand questionnaire (DASH). The mean arc of elbow flexion was 105 degrees (range, 15 to 140 degrees) at 1 year and 122 degrees (range 10 to 145 degrees; P = 0.01) at final evaluation. Radiographic arthrosis was observed in 14 patients (70%): severe in 3, moderate in 2, and mild in 9 patients. Five patients (25%) had ulnar nerve dysfunction at the final evaluation. The MEPI was excellent in 13 patients, good in 4, fair in 2, and poor in 1. The mean DASH score was 9 points (range, 0 to 53 points). The initial results of operative treatment of fracture-dislocations of the olecranon are durable over time.


Prevalence of anemia and sociodemographic characteristics among pregnant and non-pregnant women in southwest China: a longitudinal observational study.
Globally, the prevalence of anemia among women of reproductive age is about 29.4%, and anemia impacts about 40% of pregnant women and more than 20% of non-pregnant women. We conducted a longitudinal observational study of anemia in pregnant and non-pregnant women, and analyzed the association between the prevalence of anemia and sociodemographic characteristics of women in southwest China. This study was a longitudinal observational study which involved 640,672 women aged 18-49 years from 129 counties in southwest China. Data were from databases of National Free Preconception Health Examination Project (NFPHEP) and electronic medical records of local hospitals. We adjusted the diagnostic thresholds of anemia for altitude. The prevalence of anemia was expressed in percentages and 95% confidence intervals (95% CI). The association between the prevalence of anemia and sociodemographic characteristics of pregnant and non-pregnant women were analyzed using univariate and multivariate logistic regression method, expressed in crude odds ratio (cOR), adjusted odds ratio (aOR) and 95%CI. Of the 640,672 participants, 121,254 women suffered from anemia, with the prevalence of 18.9% (95%CI: 18.8-19.0%). From 2014 to 2018, the prevalence of anemia declines from 23.0-16.4%.The prevalence was 21.6% in the first trimester, higher than women in non-pregnancy (17.4%) and women in the third trimester (10.5%). Results from the multivariable logistic regression showed that women aged 18-20 (aOR = 1.28) or over 35 years old (aOR = 1.07), being farmers (aOR = 1.42), being ethnic minorities (aOR: 1.19 ~ 1.73), during the first trimester (aOR = 1.32) were more likely to be anemic. Although the anemia prevalence of women of reproductive age has been decreasing in recent years, the prevalence of anemia is still high in pregnant and non-pregnant women in southwest China, especially during the first trimester. Women who were older or younger, being farmers, being ethnic minorities were at high risk of anemia. Anemia in women of reproductive age cannot be neglected.


Effects of different pelvic osteotomy surgeries on acetabular center and pelvic morphology.
To compare the effects of Salter pelvic osteotomy, Pemberton pelvic osteotomy, and triple pelvic osteotomy on the center of acetabulum and pelvic morphology in children with hip joint disease. The data of children treated with Salter pelvic osteotomy (2 males and 14 females with an average age of 2.49 years), Pemberton pelvic osteotomy (4 males and 11 females with an average age of 6.11 years), and triple pelvic osteotomy(4 males and 8 females with an average age of 9.59 years) between January 2011 and December 2020 were collected. After discharge, the outpatient review was followed up for at least 1 year. All patients underwent anterior-posterior pelvic X-ray scanning before surgery, three months after surgery in the first year and every six months after the first year. The following X-ray features were analyzed: bilateral pelvic height (PH), iliac crest inclination (ICI), a horizontal distance of the acetabulum center (HD), and vertical distance of the acetabulum center (VD). The mean follow-up time was 16.9 ± 4.9 months in the Salter group, 20.7 ± 5.1 months in the Pemberton group, and 18.0 ± 5.4 months in the triple group (all P > 0.05). No significant differences between PH, HD, and VD of both sides on the preoperative AP pelvic x-ray were found. However, at the last follow-up, PH, HD,VD, and ICI increased in the Salter group (all P < 0.05), PH and VD increased in the Pemberton group (all P < 0.05), and VD decreased in the Triple group (P < 0.05). Salter pelvic osteotomy may cause pelvic height to increase and the center of acetabulum to move outward and downward. In contrast, Pemberton pelvic osteotomy may cause pelvic height to increase and the center of acetabulum to move downward. Triple pelvic osteotomy only causes the center of acetabulum to move downward.


The impact of prospectively set hospital budgets on psychiatric admissions.
This article examines the impact of prospectively set hospital budgets on rates of admission of psychiatric patients in New York state, U.S.A. The analysis takes advantage of a natural experiment which took place in the early 1980s, whereby a geographic region adopted a prospective hospital budget reimbursement scheme that differed from the prospective per diem reimbursement scheme used in the rest of the state. The results indicate a strong decrease in psychiatric admissions attributable to the experimental payment method.


Pancreaticoduodenectomy and metastasectomy for metastatic pancreatic neuroendocrine tumors.
Various treatment options exist for patients with metastatic pancreatic neuroendocrine tumors (PNETs). Surgical resection with pancreaticoduodenectomy (PD) typically reserved for patients with limited disease. Definitive data are lacking to support either the resection of primary PNET in the metastatic setting or for surgical debulking of metastatic lesions. We conducted an analysis of the National Cancer Database (NCDB) using the pancreatic cancer Participant User File. Thirty- and 90-day mortality rates and survival rates were determined for patients undergoing PD for primary tumor resection and compared with patients who had no surgery or metastasectomy. The Kaplan-Meier method was used to compare survival time. Cox regression models were used to assess factors independently associated with overall survival time. Resection of the primary tumor or metastatic disease each significantly improved overall survival time compared with no resection. Adding metastasectomy to PD resulted in an incremental increase in overall survival time. Both PD and metastasectomy are independently associated with overall survival time. Our report highlights the potential for survival time benefit in appropriately selected patients who undergo PD in the setting of metastatic PNET.


Development of a recombinant TaSP-based Dot-ELISA for detection of Theileria annulata infection in cattle.
The study was conducted to develop and validate Dot-ELISA for the diagnosis of Theileria annulata infection in cattle using recombinant Theileria annulata surface protein (r-TaSP). The r-TaSP based indirect plate-ELISA was used as a reference test to compare the efficacy of the Dot-ELISA. The Dot-ELISA was optimized with 500 ng of antigen per dot, 1:150 dilution of serum and 1:1000 dilution of secondary antibody for positive and negative reaction. A total of 17 confirmed positive, 25 negative and 129 field sera samples were used to calculate the diagnostic accuracy of Dot-ELISA in comparison with indirect plate-ELISA. The diagnostic sensitivity and specificity of the Dot-ELISA was 95.8 per cent (95% CI, 93.1-97.2) and 80 per cent (95% CI, 48.1-96.2), respectively. The positive predictive value (PPV) of Dot-ELISA was 98.2 percent (95% CI, 95.5-99.7) and negative predictive value (NPV) was 61.6 percent (95% CI, 37-74). The positive and negative likelihood ratios were 4.79 (95% CI, 1.8-25.69) and 0.053 (95% CI 0.03-1.4), respectively. The Dot-ELISA showed moderate agreement (k value, 0.67, 95% CI, 0.36- 0.82) with indirect plate-ELISA. The developed Dot-ELISA is less expensive and convenient for the diagnosis of T. annulata infection in cattle under field conditions.


Behavior, genetics and biochemistry of an allele of the mutant mouse spastic, spaAlb.
A new autosomal recessive mutation, characterized by an early defect in righting reflex and stiffened gait, progression to severe spasticity, tremor and rigidity, and death before weaning, appeared spontaneously on the C57BL/6 background. It was shown to be an allele of the mutant spastic spa, and shall be known as spaAlb. Mutant levels of [3H]strychnine binding are less than 10% of control levels in the brainstem and spinal cord. Autoradiographic examination of the distribution of [3H]strychnine binding sites in the mutant confirm a greatly reduced level of binding compared to control in all areas of the spinal cord, brainstem, and midbrain.


Complete genome sequence of a novel virus belonging to the genus Badnavirus in jujube (Ziziphus jujuba Mill.) in China.
The genome sequence of a novel circular DNA virus related to members of the genus Badnavirus was identified in diseased jujube trees by high-throughput sequencing and verified by conventional Sanger sequencing of cloned PCR products. The name "jujube badnavirus WS" (JuBWS) is proposed for this virus. Diseased jujube leaves showed yellow mosaic and malformation symptoms, with round chlorotic spots found on diseased fruit. The genome of this virus has a length of 6450 nt and has a typical badnavirus genomic structure with three open reading frames (ORFs). JuBWS was identified as a novel badnavirus based on nucleotide differences in the RNase (RT + RNase H) coding region of ORF3. The JuBWS sequence showed 70.48-76.41% nucleotide sequence identity to other known badnaviruses, thus meeting the taxonomic criterion for establishing a new species within the genus Badnavirus. This study suggested that the novel badnavirus might be a pathogen associated with jujube mosaic disease, and this will be investigated in the future.


Top-bottom asymmetry in vertical orbwebs: a functional explanation and attendant complications.
The distribution of entangled insects (=actual prey) in symmetrical and asymmetrical orbwebs of four tropical araneid spiders was studied. Both web types show a inhomogeneous insect distribution between individual web sections. Possible explanations are discussed but do not completely explain the inhomogeneous insect distribution.


Performance and microbial community dynamics in anaerobic continuously stirred tank reactor and sequencing batch reactor (CSTR-SBR) coupled with magnesium-ammonium-phosphate (MAP)-precipitation for treating swine wastewater.
The impacts of magnesium-ammonium-phosphate (MAP) precipitation on the performance and microbial dynamics in an anaerobic continuously stirred tank reactor (CSTR) coupled with sequencing batch reactor (SBR) for swine wastewater treatment were investigated. In CSTR-SBR systems, an overall higher removal efficiency for COD, NH4+ and PO43-as 98.6%, 98.7% and 97.9% was achieved with MAP precipitation, compared to CSTR-SBR without MAP pretreatment (i.e., 97.5, 74.3% and 19.9% for COD, NH4+ and PO43-, respectively). With MAP precipitation, the high C/N ratio of 6.6 after anaerobic CSTR was observed. The increase in the richness and diversity of microbial communities in CSTR with MAP was conducive to nitrogen and phosphorus removal, as well as biogas production. The core community was affiliated with bacterial phyla Firmicutes, Bacteroidetes, Proteobacteria, Cloacimonetes, and Spirochaetae. The study provide a new insight into the potential application of MAP precipitation as pretreatment for dealing with nutrient recovery from high-strength swine wastewater.


Behavioural genetics of the serotonin transporter.
The serotonin transporter is a key regulator of the bioavailability of serotonin and therefore any modulation in the expression or action of the transporter would be expected to have consequences on behaviour. The transporter has therefore become a target for pharmaceutical intervention in behavioural and mood disorders. The search for polymorphic variants in the transporter that would associate with neurological disorders has been extensive but has become focused on two domains which are both termed variable number tandem repeat (VNTR)polymorphisms. Both of these VNTRs are in non-coding DNA and therefore proposed to be mechanistically involved in a disorder through their ability to modulate transcriptional or post-transcriptional regulation of the transporter. The most extensively studied is in the promoter and is a bi-allelic insertion/deletion found in the 50 promoter region of the gene 1.2 kb upstream of the transcriptional start site. This VNTR, termed, 5-HTTLPR was initially identified as two variants containing either, 14 (short/deletion) or 16 (long/insertion) copies of a 22 bp repeat. A second widely studied VNTR found in the non-coding region of the transporter is located within intron 2 and comprises 9, 10 or 12 copies of a16–17 bp repeat termed, STin2.9, STin2.10 and STin2.12, respectively. These VNTR polymorphisms have been associated with a range of behavioural and psychiatric disorders including depression, OCD, anxiety and schizophrenia, however often the lack of reproducibility in different cohorts has led to debate on the actual association of the polymorphisms with this extensive range of neurological conditions. Here we review these two polymorphic VNTRs in depth and relate that to pharmaceutical response, their ability to regulate differential transporter expression, their core involvement in gene-environment interaction and their genetic association with specific disorders.


Ecotoxicological effects of ciprofloxacin on freshwater species: data integration and derivation of toxicity thresholds for risk assessment.
Although antibiotics have been increasingly used and detected in natural samples, their ecotoxicological effects on aquatic wildlife are not yet extensively studied. Considering the environmental threat posed by the biological activity of antibiotics it is quite relevant to assess the resulting impact, especially on sub-lethal endpoints. As such, this study evaluated the effects of ciprofloxacin on Pseudokirchneriella subcapitata and Lemna minor growth, on the survival and reproduction of Daphnia magna and on Gambusia holbrooki survival. The risks associated with ciprofloxacin effects on non-target organisms were quantified through the calculation of the PEC/PNEC ratio. Overall, the toxicity values obtained (at the mg L(-1) level) were higher than the environmental concentrations. P. subcapitata and L. minor were more sensitive under short-term exposures than D. magna and G. holbrooki. No acute toxicity was observed for fish. The chronic assay with D. magna evidenced that long term exposures to lower concentrations of this antibiotic induced impairments on its life-history parameters. Such outcome may pre-empt potential damages on the long-term maintenance of natural populations continuously exposed to the input of antibiotics. Indeed, the PEC/PNEC ratios showed that ciprofloxacin represents a risk for the most sensitive aquatic organisms, since the defined threshold of an acceptable risk was considerably surpassed.


Frontalis muscle transfer technique for correction of severe congenital blepharoptosis in Chinese patients: An analysis of surgical outcomes related to frontalis muscle function.
The aim of this study is to evaluate the surgical outcomes related to frontalis muscle function using the frontalis muscle transfer technique in Chinese patients with severe congenital blepharoptosis and poor levator function. Between December 2003 and December 2011, using the frontalis muscle transfer technique, 37 patients of Chinese origin underwent surgery on 53 eyelids. The frontalis muscle function was assessed and categorized as poor (excursion ≤ 7 mm) and good (excursion > 7 mm) before the surgery. The results, including complications of the treatment procedure, were followed up and evaluated. Using the Cochran-Mantel-Haenszel statistic, the preoperative ptosis severity was compared with the degree of ptosis correction, and the preoperative frontalis muscle function was compared with the degree of ptosis correction using Fisher's exact test for paired data. A two-sided value of p < 0.05 was considered statistically significant. Of the total eyelids considered, sufficient postoperative correction of ptosis was achieved in 43 eyelids (81.1%), while the correction was insufficient in 10 eyelids (18.9%). Fifteen (28.3%) eyelids were either overcorrected (n = 5) or undercorrected (n = 10). The rate of lagophthalmos was 3.8%. Eyelids with preoperative frontalis muscle function >7 mm had a higher rate of sufficient correction in comparison to those with preoperative frontalis muscle function ≤7 mm (91.2% vs. 63.2%; p < 0.05). However, postoperative ptosis correction had no relationship with preoperative ptosis severity. The frontalis muscle is the main motor muscle used in the correction of severe blepharoptosis via the frontalis muscle transfer technique. Although the technique is considered to be an effective surgical method for the correction of severe blepharoptosis, the outcome of the correction procedure depends on the preoperative frontalis muscle function of the patient.


Association between postoperative delirium and mortality in elderly patients undergoing hip fractures surgery: a meta-analysis.
Postoperative delirium (POD) is a common complication in elderly surgical patients. Patients undergoing hip fractures surgery who are often characterized by advanced age could be particularly prone to suffering POD. We performed a meta-analysis to assess the association between POD and mortality in elderly patients undergoing hip fractures surgery. This meta-analysis included twenty-one cohort studies, and the pooled outcomes demonstrated that approximated one-fourth of patients undergoing hipfracture surgery would develop POD, and delirium increased the mortality in these patients. We searched PubMed, Embase, Web of Science, and Scopus for studies that investigated the effect of POD on mortality in elderly patients undergoing hip fracture surgery. Two reviewers independently selected studies, assessed quality, and extracted data. Statistical analyses were performed by STATA 14.0 and RevMan 5.3. Risk ratios (RRs) with 95% confidence intervals (CIs) were derived using random or fixed-effects model. Twenty-one cohort studies enrolling a total of 6288 patients were included, and the pooled prevalence (95% CI) of POD was 28% (23-34%). POD was associated with an increase in perioperative mortality (30-day or in-hospital mortality) [12 studies, 3123 patients, relative risk (RR) (95% CI) 2.79 (1.97-3.93)], 6-month mortality [6 studies, 1673 patients, 2.51 (1.99-3.16)], 1-year mortality [6 studies, 1896 patients, 1.98 (1.62-2.41)], and more than 1-year mortality [8 studies, 1926 patients, 2.06 (1.60-2.64)]. Our meta-analysis demonstrated that approximated one-fourth of patients undergoing hip fracture surgery would develop POD, and delirium increased the short-term and long-term mortality in these patients.


The systematic structure-activity relationship to predict how flavones bind to human androgen receptor for their antagonistic activity.
Although flavones act as potent androgen receptor (AR) antagonists, it remains unclear how flavones interact with AR. The aim of this in silico study was to investigate the molecular recognition processes of newly synthesized 5,4'-difluoroflavone with the highest activity (IC50 value=0.19 μM) in the AR-ligand binding domain (AR-LBD). The results demonstrated that at its 4'-position of 5,4'-difluoroflavone the substituents may face Arg752 and that in AR-LBD, the submolecular bulk of substituents is unfavorable for AR antagonists and the negative electrostatic interaction site prefers the stronger hydrogen bond capability of substituents of AR antagonists. The prediction model is a valuable tool for designing a novel AR antagonist.


Redox Regulation of Metabolic Syndrome: Recent Developments in Skeletal Muscle Insulin Resistance and Non-alcoholic Fatty Liver Disease (NAFLD).
Several new discoveries over the past decade have shown that metabolic syndrome, a cluster of metabolic disorders, including increased visceral obesity, hyperglycemia, hypertension, dyslipidemia and low HDL-cholesterol, is commonly associated with skeletal muscle insulin resistance. More recently, non-alcoholic fatty liver disease (NAFLD) was recognized as an additional condition that is strongly associated with features of metabolic syndrome. While the pathogenesis of skeletal muscle insulin resistance and fatty liver is multifactorial, the role of dysregulated redox signaling has been clearly demonstrated in the regulation of skeletal muscle insulin resistance and NAFLD. In this review, we aim to provide recent updates on redox regulation with respect to (a) pro-oxidant enzymes (e.g. NAPDH oxidase and xanthine oxidase); (b) mitochondrial dysfunction; (c) endoplasmic reticulum (ER) stress; (d) iron metabolism derangements; and (e) gut-skeletal muscle or gut-liver connection in the development of skeletal muscle insulin resistance and NAFLD. Furthermore, we discuss promising new therapeutic strategies targeting redox regulation currently under investigation for the treatment of skeletal muscle insulin resistance and NAFLD.


Measuring cellulase activity: application of the filter paper assay to low-activity enzyme preparations.
An approach is presented for obtaining relative filter paper activities for enzyme preparations having activities below that required for application of the traditional International Union of Pure and Applied Chemistry filter paper assay. The approach involves the utilization of protein stabilizers to retard the time-dependent enzyme inactivation that may occur under traditional filter paper assay conditions. Enzyme stabilization allows extended reaction times and the calculation of relative activities based on the time required for saccharification of 3.6% of the traditional substrate, making results proportional to those obtained in the traditional International Union of Pure and Applied Chemistry assay. The assay is demonstrated using a commercial cellulase preparation along with KCl and bovine serum albumin as protein stabilizers.


Stability of vaccinia virus DNA during persistent infections: accumulation of left-end deletions and of tandem repeats at both ends of the viral genome and prevention by interferon.
The stability of the large vaccinia virus genome (122 MDa) has been studied in long-term cultures of persistently infected Friend erythroleukemia (FEL) cells. Deletions of about 8 MDa at the left terminus of vaccinia DNA are generated at early passages, and are maintained even after 2 years of continuous cell passages. The generation of deletions is followed by a stable accumulation of tandem repeats up to 6 MDa at the left end and up to 2 MDa at the right end of vaccinia DNA. Neither translocations nor rearrangements of DNA are observed during persistent infection. A recombinational mechanism within the tandem repeats or a mechanism similar to that described for adenovirus might explain the reiteration of tandem repeats at both ends. Significantly, in persistently infected cultures continuously treated with interferon (IFN) both the deletion and the large accumulation of tandem repeats were completely blocked. We suggest that reiteration of tandem repeats at the termini of vaccinia DNA might provide signals for more efficient virus multiplication in FEL cells.


Observation of acoustic Dirac-like cone and double zero refractive index.
Zero index materials where sound propagates without phase variation, holds a great potential for wavefront and dispersion engineering. Recently explored electromagnetic double zero index metamaterials consist of periodic scatterers whose refractive index is significantly larger than that of the surrounding medium. This requirement is fundamentally challenging for airborne acoustics because the sound speed (inversely proportional to the refractive index) in air is among the slowest. Here, we report the first experimental realization of an impedance matched acoustic double zero refractive index metamaterial induced by a Dirac-like cone at the Brillouin zone centre. This is achieved in a two-dimensional waveguide with periodically varying air channel that modulates the effective phase velocity of a high-order waveguide mode. Using such a zero-index medium, we demonstrated acoustic wave collimation emitted from a point source. For the first time, we experimentally confirm the existence of the Dirac-like cone at the Brillouin zone centre.


Long-term continuous evolution of acetate resistant Acetobacter aceti.
Elevated concentrations of cytotoxic acetate are found in many environmental niches, and few species are relatively resistant to acetate. In particular the high-level acetate resistance of so-called acetic acid bacteria that occurs in industrial settings must be constantly selected for. To investigate the nature of such high-level resistance, we grew the moderately acetate-resistant Acetobacter aceti wild-type and acetate-sensitive Escherichia coli in long-term continuous cultures with increasing acetate concentrations at near neutral pH. While E. coli did not acquire any significant resistance after 125 generations of selection, A. aceti evolved the capability to grow at acetate concentrations exceeding 50 g/L within 240 generations. This phenotype was found to be stable for several generations in the absence of selective pressure, hence must be genetically determined. Intracellular acetate concentrations were significantly lower in evolved A. aceti, when compared to wild-type A. aceti and E. coli, indicating that cytoplasmatic anion accumulation is an important component of acetate toxicity.


Association between CASP7 and CASP14 genetic polymorphisms and the risk of childhood leukemia.
Current evidence suggests that apoptosis and the cell cycle system play an important role in cancer development. To identify susceptible genetic markers in these mechanisms, we did an association study in 63 patients and 148 controls. A total of 304 SNPs in 31 gene regions were selected. We evaluated an association at a gene region level by computing the minimum P-value (minP) and doing the false discovery rate (FDR) test. Both SNP and gene-based analyses presented associations with the risk of childhood leukemia for 5 genes: CASP7, CASP14, CASP8AP2, MYC, and RIPK1 (P(trend)<0.05). There were statistically significant associations for CASP7 (rs12416109 and rs3814231, P(trend) = 0.002 and 0.009, respectively, minP = 0.013, FDR = 0.042) and CASP14 (rs8110862, P(trend)<0.001, minP = 0.002, FDR = 0.027). This study suggests that genetic polymorphisms in apoptosis and cell cycle related genes might play a role in childhood leukemia development.


Chiral B-N AIEgens: Intense Blue Circularly Polarized Luminescence and Piezochromism.
We present a new class of blue circularly polarized luminescent emitters based on tetraarylaminoborane (TAAB) with considerable dissymmetry factor in the solid state. The chiral pendant 1-phenylethylamine in BN-RR and BN-SS imparts chirality to the core chromophore, resulting in circularly polarized luminescence signals (glum = 0.8 × 10-3) with a quantum yield of 33% in the crystalline state. This novel set of compounds also showcases intriguing thermally reversible piezochromism.


Physiologic features of hemolysis axxociated with altered cation and 2,3-diphosphoglycerate content.
A hemolytic disorder characterized by altered RBC cation composition (increases Na, decreases K), reduced monovalent cation content (decreased Na + K/liter RBC), and decreased levels of 2,3-diphosphoglycerate (2,3-DPG) is described. The etiology of these RBC abnormalities was not elucidated following extensive laboratory evaluation, although two important physiologic principles were manifested by this case: (1) Hemolysis was relatively well compensated (41% hematocrit) despite a significantly decreased RBC survival (51 Cr t 1/2 = 10.5 days). This effect presumably was due to reduced 2,3-DPG content (1.9 mumol/ml RBC) and the associated increase in whole blood oxygen affinity (P50 = 19.6 mm hg). (2) RBC size and water content were normal in spite of marked cation depletion. This anomaly was thought to reflect the osmotic effects of reduced polyvalent anion (2,3-DPG) content.


Environmental distribution and associated human health risk due to trace elements and organic compounds in soil in Jiangxi province, China.
The government of China launched its first national soil quality and pollution survey (NSQPS) during April 2006 to December 2013. Data gathered in several earlier soil surveys were rarely used to understand the status of pollution. In this study, the dataset collected at the provincial level was analyzed for the first time. Concentrations, distribution, diversity, and human health risks of trace elements (As, Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb, Se, V and Zn) and organic pollutants (benzene hexachloride (BHCs), dichlorodiphenyltrichloroethanes (DDTs), phthalic acid esters (PAEs), polycyclic aromatic hydrocarbon (PAHs), polychlorinated biphenyls (PCBs), and petroleum hydrocarbons (PHCs)) in surface soil samples collected across Jiangxi province,China were presented. The results showed that, the proportion of contaminants with concentrations higher than their corresponding regulatory reference value ranged from 0.12% to 17%. It is worth note that, the local residents are exposed to moderate non-carcinogenic and carcinogenic risks at some sites. The comprehensive analysis of soil pollutants provide baseline information for establishing a long-term soil environmental monitoring program in Jiangxi province, China.


Plasma amino acid response to single test meals in humans. III. Supplementary effect of dried skim milk and L-methionine on a vegetable protein mixture.
The plasma amino acid response to single test meals in young adults was used for human biological evaluation of the supplementary effect of dried skim milk powder (DSM) and synthetic L-methionine on a vegetable protein mixture. The protein sources in the vegetable mixture were wheat flour, defatted soya bean flour and pea flour. The plasma amino acid responses were evaluated both as PAA ratios (a modification of the Longenecker and Hause method), and as delta MR% (percentage change in the postprandial essential amino acid molar ratios according to Graham and Placko). Both evaluation methods indicated that there was an adequate supply of all the essential amino acids in the basic vegetable mixture, except for a small deficit of methionine. Supplementation (5% and 10%) with DSM did not significantly improve the low plasma amino acid response of methionine. The addition of synthetic L-methionine proved to be very effective in this respect. 1 g of L-methionine per 100 g of proteins from the vegetable mixture gave a plasma methionine response similar to that of the other essential amino acids. This implies that the deficit of methionine in the basic wheat/soya bean/pea mixture was about 30%.


Circadian rhythm in photoperiodic expressions of GnRH-I and GnIH regulating seasonal reproduction in the Eurasian tree sparrow, Passer montanus.
The present study investigates the involvement of circadian rhythm in photoperiodic expressions of GnRH-I and GnIH in the hypothalamus controlling seasonal reproduction in the Eurasian tree sparrow (Passer montanus). Groups of photosensitive birds were exposed for four weeks to resonance light dark cycles comprising of a light phase of 6 h (L) combined with dark phase of different durations (D) such that the period of LD cycles varied by 12 h increments viz. 12- (6 L/6D), 24- (6 L/18D), 36- (6 L/30D), 48- (6 L/42D), 60- (6 L/54D) and 72- (6 L/66D)h. In addition, a control group (C) was maintained under long day length (14 L/10D). Observations, recorded at the beginning and end of experiment, revealed significant testicular growth with corresponding increase in the hypothalamic expression of GnRH-I peptide but low levels of GnIH mRNA and peptide in the birds exposed to resonance cycles of 12, 36 and 60 h which were read as long days. On the other hand, birds experiencing resonance cycles of 24, 48 and 72 h read them as short days wherein they maintained their quiescent gonads and low levels of GnRH-I peptide but exhibited significant increase in GnIH mRNA and peptide expressions. Thus, sparrows responded to resonance light dark cycles differently despite the fact that each of them contained only 6 h of light. These findings suggest that an endogenous circadian rhythm is involved in photoperiodic expressions of above molecules and indicate a shift in their expressions depending upon whether the light falls in the photoinducible or non-photoinducible phase of an endogenous circadian rhythm.


BMI, body fat mass and plasma leptin level in relation to cardiovascular diseases risk factors among adolescents in Taitung.
To investigate the risk factors associated with cardiovascular diseases and its relation to BMI, body fat mass and plasma leptin level among adolescents in Taitung, Taiwan. A cross-sectional Taitung Children Heart Study for 500 young adolescents between ages 13 and 15 was conducted. Gender-specific regression models were used to determine the associations between BMI, percentage of body fat mass, plasma leptin level and seven CVDs risk factors (systolic and diastolic blood pressure, mean arterial pressure, triglycerides, total cholesterol, HDL-cholesterol and LDL-cholesterol) before and after adjusting for weight status and age. After adjusting for weight status and age, BMI was positively associated with systolic BP, triglycerides, LDL-cholesterol levels but negatively associated with HDL-cholesterol level in boys while positively associated with systolic and diastolic BP, mean arterial pressure, and LDL-cholesterol level in girls. The percentage of body fat mass was positively associated with triglycerides, total cholesterol, and LDL-cholesterol in boys while positively associated with systolic BP, total cholesterol, and LDL-cholesterol in girls. Plasma leptin was positively associated with triglycerides, total cholesterol and LDL-cholesterol in boys but no statistically significant associations with CVDs risk factors in girls. A strong relationship between the percentage of body fat mass and plasma leptin appeared among all participants (r=0.59, p<0.01). BMI, body fat mass and plasma leptin level may be used to identify certain CVDs risk factors among Taitung adolescents. Future researches could consider measuring body fat mass in the relationship of CVDs risk factors instead of plasma leptin among young adolescents.


Retention of cardiopulmonary resuscitation knowledge and skills by 11- and 12-year-old children.
This article reports the results of a cardiopulmonary-resuscitation training programme by way of the retention of the knowledge and skills of cardiopulmonary resuscitation by 11- and 12-year-old children. The results indicate that cardiopulmonary resuscitation can be taught to some individuals at this age but that performance declines quite markedly over a period of five months after training.


The decline of the Serengeti Thomson's gazelle population.
The population of Thomson's gazelles in the Serengeti National Park, Tanzania has declined by almost two thirds over a 13 year period. In the early 1970s, numbers stood at 0.66 million animals but had decreased to less than 0.25 million animals in 1985 as estimated by 5 different censuses using two different counting techniques. Predation, interspecific competition and disease are all factors that could have contributed to this decline, and at least one of these factors, predation, could now prevent the Thomson's gazelle population from increasing.


Density dependence and regulation in the systemNezara viridula (L.) (Hemiptera: Pentatomidae), host andTrichopoda giacomellii (Blanchard) (Diptera: Tachinidae), parasitoid.
The system formed byN. viridula and its almost specific parasitoidT. giacomellii has been studied in the Buenos Aires province, Argentina for 26 generations of the former and 20 generations of the latter. An intensive study over 9 host generations and key factor analysis made possible the identification and quantification of different mortality factors. The main factors determining the fluctuations in total generation mortality were: egg mortality due mainly to parasitism byT. basalis; nymphal predation on 1st, 2nd and 3rd instar nymphs and migration; and adult mortality and reduction on potential natality due to parasitism byT. giacomellii. Adult mortality and reduction of potential natality, due to parasitism byT. giacomellii, was density dependent. This factor is capable of maintaining the simulations generated by a simple model based on key factor analysis, within the range of observed densities. These results suggest that parasitism byT. giacomellii can regulate the population, even in a small area of 450 m2.


Ni-elimination from the active site of the standard [NiFe]‑hydrogenase upon oxidation by O2.
Hydrogenase is a key enzyme for a coming hydrogen energy society, because it has strong catalytic activities on both uptake and production of dihydrogen. We, however, have to overcome the sensitivity against O2 of the enzyme, because hydrogenase is, generally, easily inactivated in the presence of O2. In this study, we have revisited the crystal structures of [NiFe]‑hydrogenase from sulfate-reducing bacterium in the several oxidized and reduced conditions. Our results revealed that the Ni-Fe active site of the enzyme exposed into O2 showed two forms, Form-1 and Form-2. The Ni-Fe active site in Form-1 showed the typical Ni-B (inactive ready) structure, whereas those in Form-2 lost Ni with no relation to an exposure time to O2, and two cysteinyl sulfur ligands made a disulfide bond. On the other hand, the formation of sulfenylation of the cysteinyl ligand to Ni, which is often observed in the oxidized form, did not correlate with the Ni-elimination, but with exposure time to O2.


Rh-Catalyzed Highly Enantioselective Synthesis of Aliphatic Sulfonyl Fluorides.
Rh-catalyzed, highly enantioselective (up to 99.8% ee) synthesis of aliphatic sulfonyl fluorides was accomplished. This protocol provides a portal to a class of novel 2-aryl substituted chiral sulfonyl fluorides, which are otherwise extremely difficult to access. This asymmetric synthesis has the feature of mild conditions, excellent functional group compatibility, and wide substrate scope (51 examples) generating a wide array of structurally unique chiral β-arylated sulfonyl fluorides for sulfur(VI) fluoride exchange (SuFEx) click reaction and drug discovery.


Detection of various platelet-associated immunoglobulins by flow cytometry in idiopathic thrombocytopenic purpura.
In 1975, Dixson reported that anti-platelet IgG on platelets from patients with idiopathic thrombocytopenic purpura (ITP) is greater than in normal people, by determining anti-platelet antibodies directly on the platelet surface with a quantitative complement lysis-inhibition-assay. Since then, platelet-associated IgG (PAIgG) has been thought of as evidence of ITP. Although platelets from ITP patients show significantly higher PAIgG values than from normal control individuals, PAIgG is not specific for autoantibody because it increases in other than immune ITP patients. We analyzed positive platelet percentage with various platelet-associated immunoglobulins: IgG, IgM, IgA, and total immunoglobulins, in the blood from 17 normal donors and 23 ITP patients. The specificity for ITP disease was better in flow cytometry than in ELISA, because, other than ITP, only aplastic anemia was positive in flow cytometry; however, various disorders (aplastic anemia, chronic lymphocytic leukemia, acute myeloid leukemia, and myelodysplastic syndrome) showed positive in ELISA. Flow cytometry methods had the same sensitivity for ITP disease as ELISA. However, it is supposed that there was no nonimmune ITP in this study because the PAIgG negative patients (n = 1) showed positive results in flow cytometry. Flow cytometry method was effective for ITP screening, especially for specificity.


Gas deposition by counter-current multiplication in the eel swim-bladder: experimental verification of a mathematical model.
1. Blood pO2 along the rete of eel has been investigated with a microelectrode, the swim-bladder gas being replaced by air. 2. When the eel does not secrete gas, the pO2 profile is flat, whereas in actively secreting eel the pO2, along the rete has a maximum of approximately 1 atm near the bladder pole of the rete. 3. The findings confirm predictions of a mathematical model for counter current multiplication in the swim-bladder.


Toward Microbiome Engineering: Expanding the Repertoire of Genetically Tractable Members of the Human Gut Microbiome.
Genetic manipulation is necessary to interrogate the functions of microbes in their environments, such as the human gut microbiome. Yet, the vast majority of human gut microbiome species are not genetically tractable. Here, we review the hurdles to seizing genetic control of more species. We address the barriers preventing the application of genetic techniques to gut microbes and report on genetic systems currently under development. While methods aimed at genetically transforming many species simultaneously in situ show promise, they are unable to overcome many of the same challenges that exist for individual microbes. Unless a major conceptual breakthrough emerges, the genetic tractability of the microbiome will remain an arduous task. Increasing the list of genetically tractable organisms from the human gut remains one of the highest priorities for microbiome research and will provide the foundation for microbiome engineering.


The effects of telmisartan alone or in combination with hydrochlorothiazide on morning home blood pressure control: the SURGE 2 practice-based study.
SURGE 2, a large-scale, practice-based study in 10 countries, evaluated the effects of telmisartan alone or with hydrochlorothiazide (HCTZ) on morning (06:00-11:59) home blood pressure (HBP) control. Hypertensive patients (clinic blood pressure [BP] ≥ 140/90 mmHg) received telmisartan 40 or 80 mg either alone or in combination with HCTZ 12.5 mg for 8 weeks. Treatment could be adjusted if clinic BP remained ≥ 140/90 mmHg. Clinic BP was measured in the morning prior to medication, and seated HBP monitoring was performed, three times per day, 2 days per week. A total of 25,882 patients were included (71% were previously using antihypertensives). There was a statistically significant (all p < 0.001) reduction in mean morning, lunchtime and evening HBP following treatment with telmisartan/telmisartan plus HCTZ, and morning HBP control increased from 10.6-19.8% to 51.1-64.6%. Similar improvements were observed for lunchtime (from 20.6-26.0% to 57.7-70.5%) and evening (from 21.3-31.4% to 59.0-68.8%). The morning HBP response ranged from 62.6-67.5% (systolic BP) and from 81.4-87.0% (diastolic BP). Adverse events were reported by 1.2% of patients. Telmisartan alone or with HCTZ improved morning HBP control and maintained a smooth HBP profile throughout the day in a real-life setting.


Interaction of troponin components with F-actin and F-actin-tropomyosin complex.
1. Both TN-T and TN-I components of troponin interact with F-actin, causing its precipitation at 0.1 M KC1 and neutral pH in a form of highly ordered paracrystals, although the ability of TN-I component to precipitate of F-actin is much weaker. 2. F-actin paracrystals obtained in the presence of both TN-T and TN-I components consist of parallel arrays of F-actin filaments, although the fine structure is in each case different. 3. In the presence of tropomyosin in the proportion equal to that in muscle, less TN-T or TN-I component is needed to obtain full precipitation of F-actin. 4. Paracrystals of F-actin-tropomyosin-TN-T component and F-actin-tropomyosin-TN-I component show regular transverse striation spaced at about 380 A intervals. 5. The TN-C component of troponin solubilizes all precipitates of F-actin with TN-T or TN-I components, regardless of the presence of tropomyosin. 6. The results show that both TN-T or TN-I components can bind independently to F-actin-tropomyosin complex with the same periodicity, similar to that of the whole troponin in the living muscle.


Distinctive pretreatment features of bilateral nucleus accumbens networks predict early response to antidepressants in major depressive disorder.
The pretreatment neuroimaging markers from the resting-state brain network that could predict the early response to antidepressants are still unclear. The aim of the present study was to identify the performance of reward network features for discriminating patients with a dampened response to antidepressants. A total of 81 major depressive disorder (MDD) patients (44 patients with treatment-responsive depression (RD) and 37 patients with non-responding depression (NRD)) and 43 healthy controls (HC) underwent resting-state functional magnetic resonance imaging scans and clinical estimates. Bilateral nucleus accumbens (NAcc)-based networks were constructed for further functional connectivity (FC) analysis. The FC of the right superior frontal gyrus (SFG) (area under curve (AUC) = 0.837) and left parahippocampus (AUC = 0.770) within the left NAcc reward network, as well as the FC of the left SFG (AUC = 0.827) within the right NAcc reward network, could distinguish NRD subjects from RD subjects relatively well. Taken together, when considering the distinctive connectional pattern of the bilateral reward circuits, the synthetical differentiating effect was achieved to an optimal performance for discriminating NRD patients (AUC = 0.869), with balanced sensitivity (0.838) and specificity (0.818). The distinct pretreatment characteristics of the reward network make specific contributions to the early response to antidepressants and establish a promising imaging predictor for the classification of early efficacy.


Predictors and patterns of melanoma recurrence following a negative sentinel lymph node biopsy.
To analyse the patient demographics, tumour characteristics and follow-up data of patients with recurrence of melanoma following a negative sentinel lymph node biopsy. A retrospective review of a prospectively maintained melanoma database was conducted. Melanoma patients who had a negative sentinel lymph node were identified and we performed statistical analysis on their respective demographics, tumour histology characteristics and follow-up data. Of 164 patients studied, 40 (24%) had a recurrence of melanoma at a median of 39.5 months following diagnosis (range 1-92 months). Distant metastases were the most common form of disease recurrence (40% of all recurrences). Increasing tumour thickness was an independent predictor of recurrence on multivariate analysis while nodular histology approached significance. Median survival of 6 months was seen following disease recurrence (range 1-126 months). In the setting of a negative sentinel lymph node biopsy, there remains a risk of melanoma recurrence. Distant metastases were the most common form of recurrence. Disease recurrence occurred more frequently in those with thick primary tumours. Recurrences occurred at long intervals from diagnosis indicating the need to consider prolonged patient follow-up.


[Esophageal resection in treatment of achalasia. 4 case reports and review of the literature].
In the time period between 1985 and June 2001, four of 81 patients diagnosed with achalasia were surgically treated by means of transhiatal esophageal resection. Replacement with stomach and cervical anastomosis was performed in three patients aged 8, 19, and 27 years, respectively, after initial myotomy. In one case, the esophagus resection was the primary surgical measure and the colon was used as the replacement. The preoperative symptoms of the four patients showed improvement 51, 34, 15, and 8 months after the operation, respectively. Dysphagia and regurgitation no longer occurred. One patient died 4.5 years after esophageal resection due to ulcerous bleeding of the distal redundant colon interposition. In accordance with other studies investigating serious swallowing disorders, it could be shown that transhiatal resection with esophagus-replacement through stomach pull-up can lead to symptom-relief as well as an improvement in the quality of living.


Surgical outcomes of a breast cancer-screening program for low-income women.
Surgical outcomes from a breast cancer-screening program of low-income women are similar to those of other screening programs. Prospective cohort. Federally funded screening program. A total of 15730 women. A total of 23149 mammograms, 20396 with concomitant clinical breast examination, from January 1, 1997, through December 31, 2001. American College of Radiology scores; associated surgery consultations, biopsies, operations, and pathology results. Most (20868) of the 21296 mammograms assigned an American College of Radiology score were benign; only 428 (2%) were suspicious. Resulting from suspicious clinical breast examinations, the group with American College of Radiology scores of 1 to 3 accounted for 45%, 18%, and 10% of recommended surgical consultations, biopsies, and cancers detected, respectively. A rate of 12.3 cancers per 1000 women was found, greater than with other screening programs. Compliance with therapy was 97%. This screening program had a higher rate of advanced cancers. Clinical breast examination was an important component, and compliance with surgical recommendations was excellent.


Development and preclinical evaluation of multifunctional hydrogel for precise thermal protection during thermal ablation.
Image-guided thermal ablation (TA), which is less invasive, has been widely applied for treating various kinds of tumors. However, TA still poses the potential risk of thermal damage to sensitive tissue nearby. Therefore, an adjunctive thermoprotective hydrodissection technique with constant injection of 5% glucose (5% Glu) has currently been adopted for clinical application, but this may be hazardous to humans. In this study, a multifunctional hyaluronic acid-based hydrogel (HA-Dc) was developed for hydrodissection. Compared with 5% Glu (the most clinically used solution) and the previously reported F127 hydrogel, the HA-Dc hydrogel was studied in vitro in a porcine liver model and in vivo in a rabbit model and showed good injectability and better tissue retention, stability, and thermoprotective properties throughout the TA procedure. Furthermore, in the preclinical evaluation in a Macaca fascicularis (M. fascicularis) model, HA-Dc showed excellent performance in terms of stricter neuroprotection compared with 5% Glu. In addition, the HA-Dc hydrogel with good biocompatibility and controllable degradation behavior in vivo could be a promising platform for thermal protection during clinical TA procedures.


Crystallographic studies of the dynamic properties of lysozyme.
The patterns of atomic displacements in the crystals of hen and human lysozyme derived from independent crystallographic refinement are broadly similar. Analysis of the pattern indicates a close correlation with molecular structure, strongly suggestive of intramolecular motion. The active site of lysozyme is located in a region of high displacement. It is concluded that protein mobility may play a significant part in biological activity and that X-ray crystallography can contribute to its analysis.


Increased prostaglandin E2 levels in the airway of patients with eosinophilic bronchitis.
Eosinophilic bronchitis is a common cause of chronic cough, which like asthma is characterized by sputum eosinophilia, but unlike asthma there is no variable airflow obstruction or airway hyperresponsiveness. We tested the hypothesis that the different airway function in patients with eosinophilic bronchitis and asthma could be caused by an imbalance in the production of bronchoconstrictor (LTC(4)) and bronchoprotective (prostaglandin E(2); PGE(2)) lipid mediators. We measured cytokines levels, proinflammatory mediators and eicosanoids concentration in sputum from 13 subjects with nonasthmatic eosinophilic bronchitis, 13 subjects with asthma, and 11 healthy control subjects. Cytokines mRNA levels were measured by real time PCR, proinflammatory mediators, PGE(2), and LTC(4) were measured by enzyme immunoassays. The median sputum eosinophil count was not statistically different in patients with asthma (7.95%) and eosinophilic bronchitis (15.29%). The levels of mRNA specific to interleukin-5 (IL-5), IL-4, IL-10, IL-13, interferon gamma (IFN-gamma), IL-2, vascular endothelial growth factor and transforming growth factor beta were similar in both conditions. In addition, no differences were found between asthma and eosinophilic bronchitis in proinflammatory cytokines, such as IL-8, IFN-gamma and tumor necrosis factor alpha (TNF-alpha) levels. Sputum cysteinyl-leukotrienes concentration was raised both in eosinophilic bronchitis and asthma patients. We found that induced sputum PGE(2) concentrations were significantly increased in subjects with eosinophilic bronchitis (838.3 +/- 612 pg/ml) when compared with asthmatic (7.54 +/- 2.14 pg/ml) and healthy subjects (4 +/- 1.3 pg/ml). This data suggest that the difference in airway function observed in subjects with eosinophilic bronchitis and asthma could be due to differences in PGE(2) production in the airways.


Delirium and depression: inter-relationship and clinical overlap in elderly people.
Delirium and depression are complex neuropsychiatric syndromes common in the elderly and are associated with poor health-care outcomes. Accurate diagnosis is essential to the provision of optimum health care for individuals with these conditions but is complicated by substantial clinical overlap in symptoms and comorbidities. A careful assessment of the patient's symptoms, including their context and time course, is needed for accurate diagnosis. Previous depression is common in patients with delirium and depressive illness is a recognised sequelae of delirium. The two syndromes seem to be caused by similar pathophysiological mechanisms, involving disturbances in stress and inflammatory responses, monoaminergic and melatonergic signalling, which point to new avenues for therapeutic intervention. Improved methods to assess delirium and depression in populations at high risk by virtue of their age, diminished cognitive reserve and physical frailty is a key target to achieve improved health-care outcomes in elderly individuals.


Can the manipulation of urinary pH by beverages assist with the prevention of stone recurrence?
The formation of various types of stones in the urinary tract is strongly influenced by urinary pH. An acidic urinary pH promotes the crystallization of uric acid and cystine, respectively. Moreover, changes in systemic acid-base homeostasis alter urinary excretion of citrate, an important inhibitor of calcium oxalate stone formation. The effect of beverages on urinary pH and citrate excretion is mainly determined by the presence of bicarbonate and citrate. The bicarbonate content of mineral water can replace alkalization therapy with potassium citrate and contribute to urine inhibitory power by increasing urinary pH and citrate excretion. Citrus juices are rich sources of citrate. Oral citrate is absorbed in the intestine and nearly completely metabolized to bicarbonate, providing an alkali load, which in turn increases urinary pH and citrate excretion. However, data from observational and interventional studies on the effect of different types of citrus juices on the risk of urinary stone formation are conflicting. In conclusion, favourable changes in urinary pH and citrate excretion can be attained by various beverages. However, the long-term efficacy of certain beverages for the recurrence prevention of different types of stones has yet to be determined.


Model-predicted geometry variations to compensate material variability in the design of classical guitars.
Musical instrument making is often considered a mysterious form of art, its secrets still escaping scientific quantification. There is not yet a formula to make a good instrument, so historical examples are regarded as the pinnacle of the craft. This is the case of Stradivari's violins or Torres guitars that serve as both models and examples to follow. Geometric copies of these instruments are still the preferred way of building new ones, yet reliably making acoustic copies of them remains elusive. One reason for this is that the variability of the wood used for instruments makes for a significant source of uncertainty-no two pieces of wood are the same. In this article, using state-of-the-art methodologies, we show a method for matching the vibrational response of two guitar top plates made with slightly different materials. To validate our method, we build two guitar soundboards: one serving as a reference and the second acting as a copy to which we apply model-predicted geometry variations. The results are twofold. Firstly, we can experimentally validate the predictive capabilities of our numerical model regarding geometry changes. Secondly, we can significantly reduce the deviation between the two plates by these precisely predicted geometry variations. Although applied to guitars here, the methodology can be extended to other instruments, e.g. violins, in a similar fashion. The implications of such a methodology for the craft could be far-reaching by turning instrument-making more into a science than artistic craftsmanship and paving the way to accurately copy historical instruments of a high value.


Bioactive compounds and sensory quality of black and white mulberries grown in Spain.
The objective of this study was to evaluate and compare, for the first time, white and black mulberry species in terms of main phytochemical, volatile composition and sensory profile characteristics in eight Spanish clones. The results showed that black and white mulberry species displayed significant different characteristics. PLS analysis has allowed grouping of the clones into four groups (i) MA1, MA2 and MN2, (ii) MN3 and MN4, (iii) MA3 and MA4, and (iv) MN1. Experimental results proved that Spanish mulberries have potential for fresh consumption due to their high antioxidant capacity (10.7-86.1 mg Trolox 100 g(-1)), polyphenol (76.7-180 mg gallic acid 100 g(-1)) and ellagic acid content (8.7-15.5 mg 100 g(-1)) as well as considerable amount of volatile compounds (35) with desirable attributes, which were scored high by a trained panel. Cultivars from the Morus nigra species seem to provide fruits with higher content of bioactive compounds and better aptitude for fresh consumption than Morus alba. Differences among the species should be attributed to genetics because they were cultivated under same conditions.


Density dependence, population persistence, and largely futile arguments.
Wolda and Dennis (1993) suggest that no valid conclusions about population regulation can be drawn on the basis of statistical tests of density dependence in time series data of population abundance. They give some examples in which a 'population' persists even if it is not regulated by a density-dependent process: a sequence of independent, identically distributed random variables, the numbers of the migrant moth Autographa gamma in Britain, annual rainfall data. We suggest that such time series data may show persistence because of a static constraint, which compels the numbers to remain within finite, positive limits, or to fit some prescribed distribution. But this mechanism can explain persistence in a biological population only when the 'population' represents a sample from a regulated population (the case of A. gamma). We also comment on some suggestions made by Wolda and Dennis (1993) concerning the general value of statistical tests of density dependence, frequency of delayed versus non-delayed density dependence in natural populations, relative performance of different kinds of insect traps in sampling local populations, and the wider issue of how ecologists are likely to make progress in the study of population dynamics.


Prognostic Biomarkers for Survival in Nasopharyngeal Carcinoma: A Systematic Review of the Literature.
This systematic review aims to identify prognostic molecular biomarkers which demonstrate strong evidence and a low risk of bias in predicting the survival of nasopharyngeal carcinoma (NPC) patients. The literature was searched for on PubMed to identify original clinical studies and meta-analyses which reported associations between molecular biomarkers and survival, including ≥150 patients with a survival analysis, and the results were validated in at least one independent cohort, while meta-analyses must include ≥1000 patients with a survival analysis. Seventeen studies fulfilled these criteria-two studies on single nucleotide polymorphisms (SNPs), three studies on methylation biomarkers, two studies on microRNA biomarkers, one study on mutational signature, six studies on gene expression panels, and three meta-analyses on gene expressions. The comparison between the hazard ratios of high-risk and low-risk patients along with a multivariate analysis are used to indicate that these biomarkers have significant independent prognostic values for survival. The biomarkers also indicate a response to certain treatments and whether they could be used as therapeutic targets. This review highlights that patients' genetics, epigenetics, and signatures of cancer and immune cells in the tumor microenvironment (TME) play a vital role in determining their survival.


Redefining surgery for gastric cancer.
BACKGROUND: Despite encouraging retrospective and non-randomized trials, two large prospective, randomized trials of D1 vs D2 resections show double the mortality in the D2 group, with no increase in long-term survival. However, the D2 resection still offers the only hope of cure when N2 nodes are involved. We propose a reclassification of the International Union Against Cancer TNM "N" staging to a system with an anatomical basis that is useful in defining the surgery performed. Junctional nodes lying between the N1 and N2 tiers will act as a guide to surgery. Where these nodes are uninvolved, the probability of gastric bed (N2) involvement is low and the radical D2 dissection with its higher mortality and morbidity can be avoided.CONCLUSION: Such "stage-appropriate" surgery will reduce the number of D2 resections while ensuring that patients with N2 disease are not denied curative surgery. A prospective, randomized, controlled trial of targeted surgery is required.


Late chest wall toxicity after MammoSite breast brachytherapy.
Accelerated partial breast irradiation (APBI) with the MammoSite breast brachytherapy (MBB) system is being investigated as an alternative to whole breast radiation in breast conservation therapy (BCT) at multiple centers worldwide. The newness of MBB means a complete understanding of long-term toxicity, particularly involving the chest wall, has yet to be completely articulated. We report the first pathologic rib fractures associated with MBB and dosimetric analysis of the original treatment plans. As part of ongoing quality assurance, we reviewed the records of 129 sequential patients who underwent MBB for breast cancer and identified those who subsequently had clinically significant and radiographically documented rib fracture(s) involving the ipsilateral chest wall. Equivalent tolerance doses yielding a 5% and 50% risk of rib toxicity within 5 years from treatment with 10 fractions (as with MBB) were previously calculated using the linear quadratic equation based on 2Gy per fraction treatments delivered to one-third of the rib volume (TD5/5=37Gy; TD50/5=44Gy). The original radiation therapy plans were evaluated vis-à-vis the plane films or PET/CT images documenting the osseous abnormalities and presenting complaints to find the specific fractured ribs. The specific effected ribs were contoured on the planning CT in "bone windows" using the Nucletron MicroSelectron-classic V2 (Nucletron B.V., Veenendaal, The Netherlands) for this analysis and the original patient treatments. With these datasets, we determined the dose-volume characteristics of the effected ribs including maximal dose encompassing the entire rib on one CT slice, V(20Gy), V(30Gy), V(37Gy), V(44Gy), D(50), D(25), and D(5) (the mean dose to 50%, 25%, and 5% of the rib). Between May 2002 and August 2007, three of 105 patients with a minimum of 6-months follow-up who underwent adjuvant APBI by MBB were found to have a total of five treatment-related rib fractures. The average dose-volume characteristics from the original plans were as follows: D(50)=22.1Gy, D(25)=32.2Gy, D(5)=41.6Gy, max dose to 1cc=34.8, D(max) (to 0.1cc)=45.6Gy, V(20)Gy=57.4%, V(30)Gy=30.8%, V(37)Gy=15.9%, V(44)Gy=6.6%, and max dose through rib=35.8Gy. Two patients sustained two rib fractures and 1 patient had a single rib fracture. Four of five fractures occurred in postmenopausal patients and two of five fractures occurred in a patient with a history of osteoporosis and exposure to adjuvant chemotherapy. Fractures occurred in ribs with V(37)Gy and V(44)Gy each well below 33%. As long-term toxicity data accrue from APBI series, the traditional models for estimating the biologic equivalent dose may benefit from refinements that specifically address the unique radiobiologic and physical properties intrinsic to high-dose-rate brachytherapy for breast conservation therapy.


Aging of the extracellular matrix and its pathology.
Recent concepts on the mechanisms of aging of extracellular matrix (EM) are reviewed as well as its involvement in age-associated diseases. Cell differentiation, histogenesis and organogenesis can be analyzed in terms of the program of the biosynthesis of EM macromolecules during development, maturation and aging. The most important biological role of EM is the integration of cells in tissues, of tissues in organs and of organs in the whole organism. EM can directly influence cell behavior through the contact between EM and the genome mediated by structural glycoproteins (fibronectin, laminin, elastonectin, etc.) interacting with other EM macromolecules (collagen, proteoglycans, elastin) and the cytoskeleton by trans-membrane receptors (integrins). Most age-associated diseases exhibit a deviation (qualitative or quantitative) from the normal program of EM biosynthesis. Three examples are analyzed in some detail: atherosclerosis, diabetes and malignant tumors. The degradation of elastic fibers catalyzed by cellular elastase-type enzymes is observed in atherosclerosis and also in emphysema and skin aging. Several of these enzymes were isolated and characterized from platelets, fibroblasts, smooth muscle cells and lipoproteins. The biosynthesis of some of them increases with age and facilitates cell migration. Plasma fibronectin increases with age exponentially. This increase is absent or strongly attenuated in diabetes and some cancers. Tissue fibronectin increases in diabetes, Werner syndrome and in the peritumoral desmoplastic reaction while most tumor cells can no more retain fibronectin on their membrane facilitating their movement in the organism. These examples demonstrate the importance of the study of cell matrix interactions for gerontology.


Postdischarge opioid use and persistent use after general surgery: A retrospective study.
Opioid overprescribing after surgery is a significant public health issue in most developed countries, including New Zealand. However, there is a lack of literature on the patterns and risk factors for postoperative opioid use among general surgical patients in New Zealand. This study aimed to examine opioid use in patients undergoing general surgery at Auckland District Health Board between January and December 2019 and to identify factors associated with opioid use after surgery and persistent opioid use (defined as having filled ≥1 opioid prescription in the 91 to 180 days after surgery). This is a retrospective cohort study. Data from patients' electronic clinical records and community pharmacy dispensing records were extracted to obtain data on sociodemographics, surgical characteristics, comorbidities, co-prescribed medications, and opioid use. A total of 1,110 patients were included in the study, with 42.4% dispensed an opioid following discharge after surgery. Of opioid-naïve patients who filled opioids after surgery (n = 401), 9.5% became persistent opioid users. Preoperative use of nonopioid analgesics, longer hospital stays, higher operation severity, procedure type, and higher pain scores were positively associated with opioid use, whereas older age was a negative predictor. Longer hospital stays, an initial discharge prescription with high opioid load, and female sex increased the risk of persistent opioid use. Conversely, a higher severity of surgery was associated with lower risk of persistent opioid use. The findings suggest that a considerable proportion of patients become persistent opioid users after surgery. The risk factors identified can guide clinicians to prescribe in a manner that reduces opioid-related adverse outcomes and help guide future interventions.


Effect of Nocturnal Dialysis on Prognosis in Adult Patients: A Review of the Literature.
Compared to conventional hemodialysis dialysis, nocturnal hemodialysis represents a more effective modality for receiving hemodialysis and has been associated with a 25% reduction in risk of death and improved quality of life. This article identifies the evidence about the mortality and morbidity risk for patients undergoing conventional hemodialysis vs. nocturnal hemodialysis.


Validity of demirjian and nolla methods for dental age estimation for Northeastern Turkish children aged 5-16 years old.
To evaluate the applicability of Demirjian and Nolla methods for northeastern Turkish population. A retrospective study was performed on panoramic radiographs of 673 subjects aged 5-15.9 years. The mean dental age (DA) according to the Demirjian and Nolla methods were compared to the mean chronological age (CA). The mean CA of the study sample was 10.37±2.90 and 10.03±2.81 years for females and males, respectively. Using the Demirjian method, the mean estimated DA was 11.26±3.02 years for females and 10.87±2.96 years for males. For Nolla method, the mean estimated DA was 9.80±3.41 and 9.53±3.14 years for females and males, respectively. The mean differences between the CA and DA according to the Demirjian and Nolla methods were 0.86 and -0.54 years for total study sample. Nolla method was found to be a more accurate method for estimating DA in northeastern Turkish population.


Structure of RNA and RNA binding site in tobacco mosaic virus from 4-A map calculated from X-ray fibre diagrams.
The structure of tobacco mosaic virus has been solved to a resolution of 4 A using fibre diffraction methods. The general fold of the protein is clear, the conformation may be deduced in many places, and the conformation of the RNA and its interaction with the protein can be seen.


Comparative studies on the structure of biliary immunoglobulins of some avian species. I. Physico-chemical properties of biliary immunoglobulins of chicken, turkey, duck and goose.
In pooled bile of chicken, turkey, duck and goose immunoglobulins (Igs)+ were found in relatively high amounts between 4.5 and 15.0 mg/ml (corresponding to 28 to 36% of the total protein contents). All biliary Ig fractions studied possess (H2L2)n structure estimated by SDS-PAGE. In chicken and turkey L chains are assumed to be non-covalently linked to the polymeric H chains. In duck and goose L chains are completely linked to H chains. The Mr of H chains are rather similar: 64 kDa for the Igs of chicken and turkey bile and 67 kDa for the biliary Igs of duck and goose. There are slight differences in the electrophoretic mobility: Chicken and turkey Igs are beta 1/alpha 2-globulins and the corresponding Ig fractions from duck and goose are beta 2-globulins.


Effects of eccentric training with different training frequencies on blood circulation, collagen fiber orientation, and mechanical properties of human Achilles tendons in vivo.
The purpose of the present study was to compare the effects of eccentric training with different training frequencies on the blood circulation, collagen orientation, and mechanical properties of the human Achilles tendon in vivo. Ten healthy males completed 12 weeks of a unilateral eccentric training program {(15 repetitions with knee straight and 15 repetitions with knee slightly bent) × 6 sets in a single session} for the plantar flexor muscles. They performed training three times per week on one side (3TW) and six times per week on the other side (6TW). Before and after training, changes in blood volume, coefficient of variation (CV) of echogenicity (reflects collagen fiber orientation), and stiffness of the Achilles tendon were compared by two-way analysis of variance. The tendon blood volume tended to increase after 3TW and 6TW (p = 0.064). Tendon stiffness did not change after 3TW and 6TW, whereas the elongation of tendon structures at three force levels (50, 100, and 150 N) significantly decreased with 3TW, but not 6TW. The CV of echogenicity significantly decreased after 3TW and 6TW. However, no significant differences were observed in the relative changes in these measured variables between 3TW and 6TW. The present results demonstrated an increase in blood volume, the alignment of collagen fibers, and unchanged stiffness of the Achilles tendon after 12 weeks of eccentric training. Furthermore, the training frequency did not influence these training-induced changes in the tendon properties.


Quantification of natural killer cell polarization and visualization of synaptic granule externalization by imaging flow cytometry.
Defining immunological mechanisms underlying NK cell biology is crucial for the treatment and prevention of immune deficiency and malignancy. The limited availability of human biological specimens presents a challenge to the study of human immunobiology. The use of high throughput, multi-parametric assays will not only aid in the definition and diagnosis of complex human immune disorders affecting NK cell function but also advance NK cell biology through population-based assessment of molecular signaling. In an effort to garner the most information from limited numbers of human cells, we designed a quantitative method to study NK cell function using imaging flow cytometry (IFC), which combines multiparametric flow cytometry and fluorescence microscopy. Specifically, we developed IFC as a tool to measure polarization and secretion of lytic granules at the immunological synapse formed between an NK cell and a susceptible target. We have further validated our approach through quantitative comparison with high-resolution confocal microscopy. We show that IFC can be used as a quantitative, high throughput measure of NK cell biological function possessing greater dimensionality than standard flow cytometry.


Rapalog resistance is associated with mesenchymal-type changes in Tsc2-null cells.
Tuberous Sclerosis Complex (TSC) and Lymphangioleiomyomatosis (LAM) are caused by inactivating mutations in TSC1 or TSC2, leading to mTORC1 hyperactivation. The mTORC1 inhibitors rapamycin and analogs (rapalogs) are approved for treating of TSC and LAM. Due to their cytostatic and not cytocidal action, discontinuation of treatment leads to tumor regrowth and decline in pulmonary function. Therefore, life-long rapalog treatment is proposed for the control of TSC and LAM lesions, which increases the chances for the development of acquired drug resistance. Understanding the signaling perturbations leading to rapalog resistance is critical for the development of better therapeutic strategies. We developed the first Tsc2-null rapamycin-resistant cell line, ELT3-245, which is highly tumorigenic in mice, and refractory to rapamycin treatment. In vitro ELT3-245 cells exhibit enhanced anchorage-independent cell survival, resistance to anoikis, and loss of epithelial markers. A key alteration in ELT3-245 is increased β-catenin signaling. We propose that a subset of cells in TSC and LAM lesions have additional signaling aberrations, thus possess the potential to become resistant to rapalogs. Alternatively, when challenged with rapalogs TSC-null cells are reprogrammed to express mesenchymal-like markers. These signaling changes could be further exploited to induce clinically-relevant long-term remissions.


Characteristics of the THERAFLEX UV-Platelets pathogen inactivation system - an update.
Considerable progress has been made in the last decade in producing purer, safer, leucocyte and plasma reduced platelet concentrates (PC) with an extended shelf life. The development of different pathogen inactivation technologies (PIT) has made a substantial contribution to this trend. Preceding platelet PIT (INTERCEPT Blood System/Cerus Corporation, Concord, CA, USA; MIRASOL/Caridian BCT, Lakewood, CO, USA) are based on adding a photosensitive compound to PC. The mixture is then activated by UV light in the UVB and/or UVA spectral regions. A novel procedure, THERAFLEX UV-Platelets (MacoPharma, Mouvaux, France), was recently developed that uses short-wave ultraviolet light (UVC), without addition of any photoactive agent. This technology has proven to be highly effective in sterilising bacteria (the major cause of morbidity/mortality after platelet transfusion) as well as inactivating other transfusion transmitted DNA/RNA containing pathogens and residual leucocytes. Any PIT reflects a balance between the efficacy of pathogen inactivation and preservation of platelet quality and function. A broad spectrum of in vitro tests have become available for the assessment of platelet storage lesion (PSL), aiming to better predict clinical outcome and untoward effects of platelet therapy. Recent paired studies on the release of platelet-derived cytokines, as new platelet performance indicators, revealed a parallel increase in both THERAFLEX UV-treated and control PC throughout storage, supporting the notion that the bioavailability of platelet function is not grossly affected by UVC treatment. This is corroborated by some newer technologies for proteomic analysis, showing that the THERAFLEX UV-Platelets system results in limited disruption of integrin-regulating extracellular disulfide bonds and minimal protein alterations when compared to UVB and gamma irradiation. Moreover, standard in vitro parameters reflecting activation, metabolic activity and function of platelets are useful indicators of the overall performance of processing and storage and may be used as surrogate markers of platelet quality in vivo. However, there is some doubt as to what degree each marker alone or in combination reflects the true clinical outcome of transfused platelets. Therefore, an appropriate clinical programme has been initiated. The preclinical evaluation demonstrated tolerability and immunological safety of THERAFLEX UV-Platelets using an animal model. Additionally, the system has successfully completed two autologous Phase I trials on recovery and survival. Preliminary results suggest that the recovery and survival rates are consistent with other pathogen reduced platelet products that are licensed and in use. The method is currently under evaluation for safety and tolerability of UVC-treated platelets in healthy volunteers. Presently the THERAFLEX UV-Platelets system is the simplest and purest PIT easily adaptable to the existing blood bank setting. In the future, extension of the application range of the THERAFLEX UV-Platelets system is expected, in order to make this new technology compatible with a broad spectrum of collection and processing platforms, and with other blood products.


Epstein-Barr virus-encoded small RNAs (EBERs) do not modulate interferon effects in infected lymphocytes.
The recent derivation of otherwise isogenic Epstein-Barr virus (EBV) recombinants carrying or lacking the EBV small RNA (EBER) genes enabled us to test whether EBERs are similar to adenovirus VA RNAs in modulating interferon (IFN) effects on virus infection. EBER-positive and -negative EBV recombinants did not differ in their sensitivity to alpha interferon (IFN-alpha)- or IFN-gamma-mediated inhibition of lymphocyte growth transformation. In addition, EBERs did not decrease the inhibitory effects of IFN on vesicular stomatitis virus replication in EBV-transformed lymphocytes. EBER deletion also did not render EBV-transformed B lymphocytes susceptible to an IFN effect on cell proliferation or EBV replication.


Association of VMAT2 gene polymorphisms with alcohol dependence.
Alcohol-related diseases cause significant harm in the western world. Up to 65 % of the phenotypic variance is genetically determined. Few candidate genes have been identified, comprising ADH4, ALDH2, COMT, CRHR1, DAT (SLC6A3), GABRA2 and MAOA. While abnormalities in the dopaminergic mesolimbic reward system are considered important mediators of alcoholism, studies analyzing variants of dopamine receptors showed conflicting results. Other modulators of the reward system are synaptosomal genes. Among candidate genes, polygenic variants of the Vesicular Monamine Transporter 2 (VMAT2) gene locus associated with alterations of drinking behavior were published. These variants comprise single nucleotide polymorphisms (SNPs) within the promoter region and the open reading frame. In this study, we confirm the association of VMAT2 SNP rs363387 (allelic association: p = 0.015) with alcohol dependence. This SNP defines several haplotypes including up to four SNPs (minimal p = 0.0045). In addition, numeric effects in the subgroups of males and patients with positive family history were found. We suggest that several rs363387 T-allele containing haplotypes increase the risk of alcohol dependence (OR 1.53), whereas G-allele containing haplotypes confer protection against alcohol dependence. Taken together, there is supporting evidence for a contribution of VMAT2 gene variants to phenotypes of alcohol dependence.


Role of thyroid hormone and nerve growth factor in the development of choline acetyltransferase and other cell-specific marker enzymes in the basal forebrain of the rat.
The effects of treatment with L-thyroxine (subcutaneously 0.3 microgram/g body weight daily from birth, i.e., day 1) and 2.5S nerve growth factor (NGF; intraventricularly 2 micrograms on 1, 3, 5, 7, and 9 postnatal days), separately and together, were studied on the biochemical development of different cell types in the basal forebrain of 10-day-old rats. The development of cholinergic, gamma-aminobutyric acid-ergic (GABAergic), and glutamatergic neurons was monitored respectively in terms of choline acetyltransferase (ChAT), glutamate decarboxylase (GAD), and glutaminase activities, whereas glutamine synthetase (GS) and 2',3'-cyclic nucleotide-3'-phosphohydrolase (CNPase) activities were used to judge the maturation of astroglial and oligodendroglial cells. Treatment with either thyroid hormone or NGF from birth significantly increased the expression of ChAT activity in the basal forebrain of neonatal rats. When both agents were administered to the same animal, in agreement with our earlier in vitro findings, the stimulation in ChAT activity was much greater than the sum of the individual effects. In hypothyroid rats, significant effects of NGF at the low doses used were not detectable, although the increase of ChAT activity induced by thyroxine was potentiated by NGF in these animals. Under the present experimental conditions neither thyroxine nor NGF treatment had an appreciable effect on the activities of glutaminase, GS, and lactate dehydrogenase. However, the administration of thyroxine markedly increased CNPase activity in normal rats, whereas in hypothyroid rats the effect on both CNPase and GAD was also significant. Similar elevations in CNPase and GAD activities were not observed after NGF treatment, suggesting that the effect of NGF was specific to the cholinergic cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of irradiated medium with or without cells on bystander cell responses.
Recent studies have indicated that extranuclear or extracellular targets are important in mediating the bystander genotoxic effects of alpha-particles. In the present study, human-hamster hybrid (A(L)) cells were plated on either one or both sides of double-mylar dishes 2-4 days before irradiation, depending on the density requirement of experiments. One side (with or without cells) was irradiated with alpha-particles (from 0.1 to 100 Gy) using the track segment mode of a 4 MeV Van de Graaff accelerator. After irradiation, cells were kept in the dishes for either 1 or 48 h. The non-irradiated cells were then collected and assayed for both survival and mutation. When one side with cells was irradiated by alpha-particles (1, 10 and 100 Gy), the surviving fraction among the non-irradiated cells was significantly lower than that of control after 48 h co-culture. However, such a change was not detected after 1h co-culture or when medium alone was irradiated. Furthermore, co-cultivation with irradiated cells had no significant effect on the spontaneous mutagenic yield of non-irradiated cells collected from the other half of the double-mylar dishes. These results suggested that irradiated cells released certain cytotoxic factor(s) into the culture medium that killed the non-irradiated cells. However, such factor(s) had little effect on mutation induction. Our results suggest that different bystander end points may involve different mechanisms with different cell types.


Radiation reduces carboplatin sensitivity and enhances nedaplatin sensitivity in cervical squamous cell carcinoma in vitro.
The study was performed to examine how the platinum anticancer drugs other than cisplatin, such as carboplatin (CBDCA) and nedaplatin (NEP) can be effectively used in chemoradiotherapy for cervical squamous cell carcinoma patients. The radiosensitive human cervical squamous cell carcinoma cell line ME180 was examined to investigate the radiation effects on CBDCA and NEP sensitivities of the cells. Irradiation significantly reduced cellular CBDCA sensitivity. There were no significant changes in CBDCA sensitivity between the cells concurrently irradiated and those treated with CBDCA 8 h before or 8 h after irradiation. However NEP sensitivity of the cells treated 8 h before or 8 h after irradiation was significantly higher than that in cells concurrently irradiated. Although CBDCA sensitivity in the concurrently irradiated cells is reduced, NEP sensitivity is enhanced by irradiation. NEP, but not CBDCA, therefore, may be a candidate anticancer drug for concurrent chemoradiotherapy for cervical cancer. For the greatest efficacy, NEP should be administered to patients several hours before or after irradiation.


Immunoexpression status and prognostic value of mammalian target of rapamycin and hypoxia-induced pathway members in papillary cell renal cell carcinomas.
Dysregulation of the mammalian target of rapamycin and hypoxia-induced pathways has been consistently identified in clear cell renal cell carcinomas. However, experience with non-clear cell renal cell carcinoma subtypes is scant. In this study, we evaluated the immunohistochemical expression of upstream (PTEN and phosphorylated AKT) and downstream (phosphorylated S6 and 4EBP1) effectors of the mammalian target of rapamycin pathway, as well as related cell-cycle proteins (p27 and c-MYC), and a member of the hypoxia-induced pathway (HIF-1α) in 54 patients with papillary renal cell carcinoma treated by nephrectomy. PTEN was lower in tumor than in normal kidney, and loss of PTEN expression was found in 48% of the patients. In tumor tissues, phosphorylated S6, 4EBP1, and HIF-1α were higher than in normal kidney. Conversely, scores of p27 were lower in tumor than in normal kidney. Finally, scores of c-MYC and phosphorylated AKT were similar in tumor and in normal kidney. Overall mortality and cancer-specific mortality were 24% and 11%, respectively. Tumor progression was observed in 17% of the patients. None of the tested biomarkers predicted cancer-specific mortality or tumor progression. As expected, patients with high T-stage tumors had higher hazard ratios for cancer-specific mortality (hazard ratio, 6.9) and tumor progression (hazard ratio, 6.7). Patients with higher Fuhrman grades also had higher risks for cancer-specific mortality (hazard ratio, 11.4) and tumor progression (hazard ratio, 4.5). In summary, our study provides evidence of dysregulation of the mammalian target of rapamycin and hypoxia-induced pathways in papillary renal cell carcinoma. Immunohistochemistry for members of the mammalian target of rapamycin pathway and for HIF-1α lacked prognostic significance in our cohort.


Color Rendering in Medical Extended-Reality Applications.
Cross-platform development of medical applications in extended-reality (XR) head-mounted displays (HMDs) often relies on game engines with rendering capabilities currently not standardized in the context of medical visualizations. Many aspects of the visualization pipeline including the characterization of color have yet to be consistently defined across rendering models and platforms. We examined the transfer of color properties from digital objects, through the rendering and image processing steps, to the RGB values sent to the display device. Five rendering pipeline configurations within the Unity engine were evaluated using 24 digital color patches. In the second experiment, the same configurations were evaluated with a tissue slide sample image. Measurements of the change in color associated with each configuration were characterized using the CIE 1976 color difference ([Formula: see text]). We found that the distribution of [Formula: see text] for the first experiment ranges from zero, as in the case using an Unlit Shader, to 25.97, as in the case using default configurations. The default Unity configuration consistently returned the highest [Formula: see text] across all 24 colors and also the largest range of color differences. In the second experiment, [Formula: see text]E ranged from 7.49 to 34.18. The Unlit configuration resulted in the highest [Formula: see text] in three of four selected pixels in the tissue sample image. Changes in color image properties associated with texture import settings were then evaluated in a third experiment using the TG18-QC test pattern. Differences in pixel values were found in all nine of the investigated texture import settings. The findings provide an initial characterization of color transfer and a basis for future work on standardization, consistency, and optimization of color in medical XR applications.


Sensitivity of the Western blot detection of prion protein PrPres in natural sheep scrapie.
The sensitivity of Western blot detection of PrPres using two different extraction procedures on brain material of 30 scrapie-affected sheep was compared. Whereas PrPres could be detected in all sheep after extraction with the first method, 30% did not give any signal after extraction with the second method. However, the second method, when positive, permitted the detection of PrPres from smaller amounts of infected brain tissue. When used with the ruminant specific monoclonal antibody p4, the second method gave positive signals corresponding to less than 12.5 microg of scrapie-infected brain, that, up to now, is the highest sensitivity described for PrPres detection from naturally infected ruminant brains. The overall results showed highly variable levels of PrPres between sheep and are presented in relation to breed, survival time and animal genotype data. Further progress can thus be expected for PrPres detection in prion diseases, if more efficient extraction procedures and more sensitive immunological reagents are used. Such technical improvements could contribute to more accurate diagnosis in animals affected naturally.