Molecular characteristics and virulence analysis of eight Aeromonas hydrophila isolates obtained from diseased Amur sturgeon Acipenser schrenckii Brandt, 1869.
Aeromonas hydrophila is an opportunistic pathogen of a variety of aquatic animals that displays extreme diversity in drug resistance, phenotypes, virulence genes, and virulence. In this study, eight pathogenic A. hydrophila strains were isolated from diseased Amur sturgeons and investigated for their sensitivity to select antibiotics, their phenotype, virulence genes, and virulence. According to the phylogenetic analysis of the DNA gyrase subunit B protein, the eight isolates formed a single branch in the A. hydrophila group. The antibiotics ceftazidime, cefuroxime, cefoperazone, cefotaxime, ceftriaxone, aztreonam, and cefepime appeared effective against them. All of the isolates possessed the virulence genes for aerolysin, flagellin, heat-stable cytotonic enterotoxin, heat-labile cytotonic enterotoxin, hemolysin, and elastase, while only one isolate, HZ8, possessed the gene for lateral flagella. The cytolytic enterotoxin and lipase genes were present in all isolates, except in ZJ10 and ZJ12. Enterobacterial repetitive intergenic consensus sequence PCR indicated that the eight A. hydrophila isolates could be divided into four types. Isolates YW2, TR3, HZ8 and ZJ10, each representing a different type, were selected for challenge experiments. The challenge tests revealed that isolate HZ8 had the lowest lethal dose, causing 50% mortality at 2.30 × 104 colony forming units (cfu)/ml. The isolate ZJ10 had the highest LD50, 1.25 × 106 cfu/ml. Knowledge of the characteristics of the A. hydrophila isolates obtained from Amur sturgeon will be beneficial in developing potential disease control strategies.


Microencapsulation and characterization of liposomal vesicles using a supercritical fluid process coupled with vacuum-driven cargo loading.
A new technique of liposomal microencapsulation, consisting of supercritical fluid extraction followed by rapid expansion of the supercritical solution and vacuum-driven cargo loading, was successfully developed. It is a continuous flow-through process without usage of any toxic organic solvent. For use as a coating material, the solubility of soy phospholipids in supercritical carbon dioxide was first determined using a dynamic equilibrium system and the data was correlated with the Chrastil model with good agreement. Liposomes were made with D-(+)-glucose as a cargo and their properties were characterized as functions of expansion pressure, temperature, and cargo loading rates. The highest encapsulation efficiency attained was 31.7% at the middle expansion pressure of 12.41MPa, highest expansion temperature of 90°C, and lowest cargo loading rate of 0.25mL/s. The large unilamellar vesicles and multivesicular vesicles were observed to be a majority of the liposomes produced using this eco-friendly process.


Rising Rates and Widening Socioeconomic Disparities in Diabetic Ketoacidosis in Type 1 Diabetes in Scotland: A Nationwide Retrospective Cohort Observational Study.
Whether advances in the management of type 1 diabetes are reducing rates of diabetic ketoacidosis (DKA) is unclear. We investigated time trends in DKA rates in a national cohort of individuals with type 1 diabetes monitored for 14 years, overall and by socioeconomic characteristics. All individuals in Scotland with type 1 diabetes who were alive and at least 1 year old between 1 January 2004 and 31 December 2018 were identified using the national register (N = 37,939). DKA deaths and hospital admissions were obtained through linkage to Scottish national death and morbidity records. Bayesian regression was used to test for DKA time trends and association with risk markers, including socioeconomic deprivation. There were 30,427 DKA admissions and 472 DKA deaths observed over 393,223 person-years at risk. DKA event rates increased over the study period (incidence rate ratio [IRR] per year 1.058 [95% credibility interval 1.054-1.061]). Males had lower rates than females (IRR male-to-female 0.814 [0.776-0.855]). DKA incidence rose in all age-groups other than 10- to 19-year-olds, in whom rates were the highest, but fell over the study. There was a large socioeconomic differential (IRR least-to-most deprived quintile 0.446 [0.406-0.490]), which increased during follow-up. Insulin pump use or completion of structured education were associated with lower DKA rates, and antidepressant and methadone prescription were associated with higher DKA rates. DKA incidence has risen since 2004, except in 10- to 19-year-olds. Of particular concern are the strong and widening socioeconomic disparities in DKA outcomes. Efforts to prevent DKA, especially in vulnerable groups, require strengthening.


Cytokine and hormonal profile in blood serum and follicular fluids during ovarian stimulation with the multidose antagonist or the long agonist protocol.
The aim of our study was to explore cytokine and hormonal profiles in blood and follicular fluids from normal women stimulated with either the multidose antagonist or the long agonist protocol. Fifty-six patients were stimulated with the multidose antagonist protocol and 12 with the long agonist protocol. Interleukin (IL)-1beta, IL-6, tumour necrosis factor-alpha (TNFalpha), leptin, vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), estradiol (E(2)), progesterone and testosterone levels were measured in serum and follicular fluids by immunoassays. The two treatment groups had similar cytokine concentrations in serum. The intrafollicular concentrations of IL-1beta, IL-6, VEGF and leptin were also similar in the two groups. The concentrations of bFGF in follicular fluids from the antagonist group (169.5 +/- 113.2 ng/ml) were lower than those from the agonist group (249.7 +/- 119.8 ng/ml). bFGF concentrations were correlated with the amount of administered gonadotrophins (R = 0.364, P < 0.01) which was significantly lower in the antagonist group (antagonist group: 2037.7 +/- 725.8 IU; agonist group: 2836.4 +/- 1163.5 IU). Normal women stimulated with either the multidose antagonist or the long agonist protocol generally have similar cytokine profiles in serum and follicular fluids. The intrafollicular levels of bFGF tend to be lower in antagonist cycles because of the lower amount of administered gonadotrophins.


Tarantula toxin ProTx-I differentiates between human T-type voltage-gated Ca2+ Channels Cav3.1 and Cav3.2.
ProTx-I peptide, a venom toxin of the tarantula Thrixopelma pruriens, has been reported to interact with voltage-gated ion channels. ProTx-I reduced Ba(2+) currents through recombinant human T-type voltage-gated Ca(2+) channels, Ca(v)3.1 (hCa(v)3.1), with roughly 160-fold more potency than through hCa(v)3.2 channels. Chimeric channel proteins (hCa(v)3.1/S3S4 and hCa(v)3.2/S3S4) were produced by exchanging fourteen amino acids in the hCa(v)3.1 domain IV S3-S4 linker region and the corresponding region of hCa(v)3.2 between each other. The ProTx-I sensitivity was markedly reduced in the hCa(v)3.1/S3S4 chimera as compared to the original hCa(v)3.1 channel, while the hCa(v)3.2/S3S4 chimera exhibited greater ProTx-I sensitivity than the original hCa(v)3.2 channel. These results suggest that the domain IV S3-S4 linker in the hCa(v)3.1 channel may contain residues involved in the interaction of ProTx-I with T-type Ca(2+) channels.


TTF-1 regulates α5 nicotinic acetylcholine receptor (nAChR) subunits in proximal and distal lung epithelium.
Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels comprised of five similar subunits that influence signal transduction and cell turnover. α5 is a structural subunit detected in many non-neuronal tissues; however, its function during pulmonary development is unknown. α5 was assessed by immunohistochemistry and RT-PCR in mouse lungs from embryonic day (E)13.5 to post-natal day (PN)20. From E13.5 to E18.5, α5 expression was primarily observed in primitive airway epithelial cells while mesenchymal expression was faint and sporadic. α5 expression was detected throughout the proximal lung at PN1 and extensively expressed in the peripheral lung at PN4, an early stage of murine alveologenesis. An interesting shift occurred wherein α5 expression was almost undetectable in the proximal lung from PN4-PN10, but significant localization was again observed at PN20. Transcriptional control of α5 was determined by assessing the activity of reporters containing 2.0-kb and 850-bp of the mouse α5 promoter. Because perinatal expression of α5 was abundant in bronchiolar and alveolar epithelium, we assessed transcriptional control of α5 in Beas2B cells, a human bronchiolar epithelial cell line, and A-549 cells, an alveolar type II cell-like human epithelial cell line. Thyroid Transcription Factor-1 (TTF-1), a key transcription regulator of pulmonary morphogenesis, significantly increased α5 transcription by acting on both the 2.0-kb and 850-bp α5 promoters. Site-directed mutagenesis revealed that TTF-1 activated α5 transcription by binding specific TTF-1 response elements. Exogenous TTF-1 also significantly induced α5 transcription. These data demonstrate that α5 is specifically controlled in a temporal and spatial manner during pulmonary morphogenesis. Ongoing research may demonstrate that precise regulation of α5 is important during normal organogenesis and misexpression correlates with tobacco related lung disease.


Modeling impacts on populations: fathead minnow (Pimephales promelas) exposure to the endocrine disruptor 17beta-trenbolone as a case study.
Evaluation of population-level impacts is critical to credible ecological risk assessments. In this study, a predictive model was developed to translate changes in fecundity of the fathead minnow (Pimephales promelas) in a short-term laboratory toxicity test to alterations in population growth rate. The model uniquely combines a Leslie population projection matrix and the logistic equation. Application of the model requires only a life table for the organism of interest, a measure of carrying capacity for the given population, and an estimation of the effect of a stressor on vital rates. The model was applied to investigate population dynamics for fathead minnow exposed to the androgen receptor agonist 17beta-trenbolone. Organismal-level responses for fathead minnows exposed to varying levels of 17beta-trenbolone were used to determine projected alterations in a population existing in a small body of water containing varying concentrations of the androgen. Fathead minnow populations occurring at carrying capacity and subsequently exposed to 0.027 microg/L of 17beta-trenbolone exhibited a 51% projected decrease in average population size after 2 years of exposure. Populations at carrying capacity exposed to concentrations of 17beta-trenbolone > or = 0.266 microg/L exhibited a 93% projected decrease in average population size after 2 years of exposure. Overall, fathead minnow populations exposed to continued concentrations of 17beta-trenbolone equal to or greater than 0.027 microg/L were projected to have average equilibrium population sizes that approached zero.


A Community-Partnered, Participatory, Cluster-Randomized Study of Depression Care Quality Improvement: Three-Year Outcomes.
Community Partners in Care, a community-partnered, cluster-randomized trial with depressed clients from 93 Los Angeles health and community programs, examined the added value of a community coalition approach (Community Engagement and Planning [CEP]) versus individual program technical assistance (Resources for Services [RS]) for implementing depression quality improvement in underserved communities. CEP was more effective than RS in improving mental health-related quality of life, reducing behavioral health hospitalizations, and shifting services toward community-based programs at six months. At 12 months, continued evidence of improvement was found. This study examined three-year outcomes. Among 1,004 participants with depression who were eligible for three-year follow-up, 600 participants from 89 programs completed surveys. Multiple regression analyses estimated intervention effects on poor mental health-related quality of life and depression, physical health-related quality of life, behavioral health hospital nights, and use of services. At three years, no differences were found in the effects of CEP versus RS on depression or mental health-related quality of life, but CEP had modest effects in improving physical health-related quality of life and reducing behavioral health hospital nights, and CEP participants had more social- and community-sector depression visits and greater use of mood stabilizers. Sensitivity analyses with longitudinal modeling reproduced these findings but found no significant differences between groups in change from baseline to three years. At three years, CEP and RS did not have differential effects on primary mental health outcomes, but CEP participants had modest improvements in physical health and fewer behavioral health hospital nights.


What we would like to know about the antihypertensive mechanisms of angiotensin converting enzyme inhibition.
Angiotensin converting enzyme (ACE) inhibitors induce haemodynamic, humoral and metabolic actions which have been well characterized during the last decade. The precise mechanisms involved in these actions still need clarification. Among the many questions we need answered are the following: under steady-state conditions, does plasma ACE activity reflect angiotensin I conversion in tissues? Do the low levels of angiotensin II observed after ACE inhibition still contribute to vascular tone? What is the precise role of extrarenal tissue renin in the regulation of vascular tone? What is the role of bradykinin, prostaglandins and the interaction with the sympathetic nervous system in the antihypertensive effect of ACE inhibition? We hope that definite answers to these questions will be found during the second decade of the use of ACE inhibitors.


Prediction of Hemodynamic Response to Epinephrine via Model-Based System Identification.
In this study, we present a system identification approach to the mathematical modeling of hemodynamic responses to vasopressor-inotrope agents. We developed a hybrid model called the latency-dose-response-cardiovascular (LDC) model that incorporated 1) a low-order lumped latency model to reproduce the delay associated with the transport of vasopressor-inotrope agent and the onset of physiological effect, 2) phenomenological dose-response models to dictate the steady-state inotropic, chronotropic, and vasoactive responses as a function of vasopressor-inotrope dose, and 3) a physiological cardiovascular model to translate the agent's actions into the ultimate response of blood pressure. We assessed the validity of the LDC model to fit vasopressor-inotrope dose-response data using data collected from five piglet subjects during variable epinephrine infusion rates. The results suggested that the LDC model was viable in modeling the subjects' dynamic responses: After tuning the model to each subject, the r (2) values for measured versus model-predicted mean arterial pressure were consistently higher than 0.73. The results also suggested that intersubject variability in the dose-response models, rather than the latency models, had significantly more impact on the model's predictive capability: Fixing the latency model to population-averaged parameter values resulted in r(2) values higher than 0.57 between measured versus model-predicted mean arterial pressure, while fixing the dose-response model to population-averaged parameter values yielded nonphysiological predictions of mean arterial pressure. We conclude that the dose-response relationship must be individualized, whereas a population-averaged latency-model may be acceptable with minimal loss of model fidelity.


Crohn's disease and schistosomiasis: a rare association.
Schistosomiasis is a chronic enteropathogenic disease caused by blood flukes of the genus Schistosoma. Coexistence of schistosomiasis with Crohn's disease is very rare. To the best of our knowledge, this association has been described in literature only once. A 20-year-old male patient with a past medical history of appendectomy and ileocecal Crohn's disease, presented with abdominal pain and vomiting. Ileocolonoscopy showed an ulcerated and congested appearance of the upper rectum and sigmoid. Computed tomography scan revealed a circumferential thickening of the terminal ileum with luminal stenosis. Histopathological examination of the biopsy specimens revealed a focally ulcerated colonic epithelium. The lamina propria was fibrous harbouring a polymorphic inflammatory infiltrate including lymphocytes and plasma cells organized in lymphoid follicles admixed with eosinophils and neutrophils. In the submucosa, there were two well-preserved schistosoma eggs surrounded by a thick shell with a barely visible terminal spine. The final pathological diagnosis was colonic schistosomiasis associated with Crohn's disease. The patient underwent an ileocecal resection for stenosis of the terminal ileum complicated with enterocutaneous fistula. The postoperative course was uneventful. A stool examination and serology tests were planned for this patient who was lost to follow-up.


Improving signal-to-noise performance for DNA translocation in solid-state nanopores at MHz bandwidths.
DNA sequencing using solid-state nanopores is, in part, impeded by the relatively high noise and low bandwidth of the current state-of-the-art translocation measurements. In this Letter, we measure the ion current noise through sub 10 nm thick Si3N4 nanopores at bandwidths up to 1 MHz. At these bandwidths, the input-referred current noise is dominated by the amplifier's voltage noise acting across the total capacitance at the amplifier input. By reducing the nanopore chip capacitance to the 1-5 pF range by adding thick insulating layers to the chip surface, we are able to transition to a regime in which input-referred current noise (∼ 117-150 pArms at 1 MHz in 1 M KCl solution) is dominated by the effects of the input capacitance of the amplifier itself. The signal-to-noise ratios (SNRs) reported here range from 15 to 20 at 1 MHz for dsDNA translocations through nanopores with diameters from 4 to 8 nm with applied voltages from 200 to 800 mV. Further advances in bandwidth and SNR will require new amplifier designs that reduce both input capacitance and input-referred amplifier noise.


Enhanced gene transfection efficiency by polyamidoamine (PAMAM) dendrimers modified with ornithine residues.
Aim of the study was to prepare and to evaluate gene transfection efficiency and cytotoxicity of the ornithine-conjugated PAMAMG4 dendrimers. Ornithine-conjugated PAMAMG4 dendrimers were prepared by Fmoc synthesis. A comparative gene transfection study between PAMAMG4 dendrimers and the surface modified dendrimers was conducted in HEK 293T, GM7373 and NCI H157G cell lines. Effect of excess of ornithine (100muM) on transfection efficiency of the ornithine-conjugated PAMAMG4 dendrimers was investigated in separate experiment. Cytotoxicity of the dendriplexes was tested in HEK 293T cells by MTT assay. (1)H NMR and MALDI-TOF spectral analysis showed that about 60 molecules of ornithine (PAMAMG4-ORN60) were conjugated to a PAMAMG4 dendrimer. Preliminary studies indicated that dendriplexes at charge ratio (N/P 10) show higher transfection efficiency and presence of serum does not affect the transfection efficiency of the dendriplexes. Transfection efficiency of PAMAMG4-ORN60 dendriplexes was slightly higher in cancer cells (NCI H157G) as compared to HEK 293T cells. Transfection efficiency of the PAMAMG4-ORN60 dendrimers decreased in presence of excess of ornithine while there was no effect on the parent PAMAMG4 dendrimers. Cytotoxicity assay has shown that PAMAMG4-ORN60 dendriplexes at N/P 10 were safe at concentrations <or=50 microg/mL. It may be concluded that the ornithine-conjugated dendrimers possess the potential to be novel gene carrier.


Interaction between meteorological parameters and COVID-19: an ecological study on 406 authorities of the UK.
Understanding the factors affecting COVID-19 transmission is critical in assessing and mitigating the spread of the pandemic. This study investigated the transmissibility and death distribution of COVID-19 and its association with meteorological parameters to study the propagation pattern of COVID-19 in UK regions. We used the reported case and death per capita rate (as of November 13, 2020; before mass vaccination) and long-term meteorological data (temperature, humidity, precipitation, wind speed, and visibility) in 406 UK local authority levels based on publicity available secondary data. We performed correlation and regression analysis between COVID-19 variables and meteorological parameters to find the association between COVID-19 and independent variables. Student's T and Mann-Whitney's tests were used to analyze data. The correlation and regression analyses revealed that temperature, dew point, wind speed, and humidity were the most important factors associated with spread and death of COVID-19 (P <0.05). COVID-19 cases negatively correlated with humidity in areas with high population density, but the inverse in low population density areas. Wind speeds in low visibility areas, which are considered polluted air, may increase the spread of disease (r=0.42, P <0.05) and decrease the spread in high visibility areas (r=-0.16, P <0.05). Among low (T <10°C) and high (T >10°C) temperature areas, the average incidence rates were 2056.86 (95% confidence interval (CI): 1909.49-2204.23) and 1446.76 (95% CI: 1296.71-1596.81). Also, COVID-19 death per capita rates were 81.55 (95% CI: 77.40-85.70) and 69.78 (95% CI: 64.39-75.16) respectively. According to the comprehensive analysis, the spread of disease will be suppressed as the weather warms and humidity and wind speed decrease. Different environmental conditions can increase or decrease spread of the disease due to affecting spread of disease vectors and by altering people's behavior.


First Report of Pea as a Natural Host of tomato mottle mosaic virus in China.
Tomato mottle mosaic virus, a new species in the genus Tobamovirus and family Virgaviridae, was first reported on tomato in Mexico in 2013 (Li et al. 2013). Subsequently, tomato mottle mosaic virus (ToMMV) was found infecting pepper and eggplant in China (Li et al. 2014; Chai et al. 2018). ToMMV was thought to pose a serious risk to solanaceous crops due to its potential to break resistance and numerous transmission ways (Li et al. 2020), meanwhile, some plant species in the families of Amaranthaceae, Brassicaceae, Cucurbitaceae, and Verbenaceae, were found as the hosts of ToMMV under experimental condition (Sui et al. 2017; Li et al. 2020). To clarify the occurrence of ToMMV in Yunnan province of China, 395 plant samples, including tomato (Solanum lycopersicum L.), pepper (Capsicum annuum L.) and pea (Pisum sativum L.), exhibiting viral-like symptoms were collected in major crop plantations in Yunnan province in 2020. Total nucleic acids were extracted from the diseased samples using a CTAB method (Li et al. 2008), and tested by reverse transcription polymerase chain reaction (RT-PCR) with ToMMV specific primers, ToMMVMPF (5'-ATGGCTCTAACTGTTAGTGGT-3') and ToMMVMPR (5'-TTAATACGAATCAGATCCCGCG-3'), which were designed based on the movement protein gene sequence of ToMMV YYMLJ isolate (KR824950). ToMMV was detected in 13 symptomatic samples (11 cherry tomato and 2 green pea plants) with a total detection rate of 3.29%. An 807-bp fragment was amplified from 2 out of 86 pea samples showing foliar chlorosis, mosaic, malformation and necrosis symptoms, and the amplicons were cloned and sequenced. Sequences obtained from the two pea samples were identical. Therefore, one sequence was deposited in the GenBank (accession no. MW561348). BLAST search result showed that the nucleotide sequence had the highest identity of 99.88% with the ToMMV TiLhaLJ isolate (KR824951). The ToMMV infection on the two pea samples was also verified by dot-enzyme immunoassay (Dot-ELISA) using ToMMV monoclonal antibody (kindly provided by Dr. Jianxiang Wu, Zhejiang University, China). To determine the pathogenicity of ToMMV on pea, ToMMV infectious cDNA clone was used to inoculate into 3-week-old healthy pea plants via an Agrobacterium-mediated method (Tu et al. 2021). Mottle or slight mosaic, chlorosis and malformation symptoms were observed on the upper leaves of the 8 out of 12 inoculated plants 8 days postinoculation, and ToMMV could be detected by RT-PCR from the 8 symptomatic plants but not from the asymptomatic and healthy control pea plants with the above described ToMMV specific primer pair. ToMMV has now only been detected in Mexico, China, Spain, USA, Israel and Australia on tomato, pepper and eggplant (Webster et al. 2014; Turina et al. 2016; Ambrós et al. 2017; Lovelock et al. 2020). To the best of our knowledge, this is the first report of natural infection of ToMMV in pea as well as the natural infection of ToMMV on plants outside of the family Solanaceae. The result also implies that ToMMV has a potential risk to more crops in the field. Since pea, pepper and tomato are economically important cash crops in China, proper virus management strategies for the cultivation of these crops should be adopted.


Packed cell volume and heart rate to predict medical and surgical cases and their short-term survival in horses with gastrointestinal-induced colic.
The objective of this prospective case-control study of 125 horses with gastrointestinal tract-induced colic was to determine whether heart rate (HR) and packed cell volume (PCV) can predict surgical versus medical treatment and its short-term survival (time of discharge of the patient). Sixty-four horses were treated medically and 61 surgically (29 small intestinal and 32 large intestinal cases). At admission, both PCV and HR were higher in horses treated surgically than in horses treated medically; however, with longer duration of colic before presentation, the PCV was higher in the small intestinal surgical group only. In addition, both PCV and HR were higher, and the duration of colic was longer in non-survivors compared to survivors. Binary logistic regression demonstrated a significant association between HR and type of treatment, and both HR and PCV were predictive of survival. Simple parameters such as HR and PCV provide useful information for management of colic cases.


Cerebral malaria in Indian adults: a prospective study of 441 patients from Bikaner, north-west India.
As per WHO (1993) the assessment and analysis of local problems and an appropriate epidemiological information system is an essential part of a control programme before embarking any control activity. Four hundred and fourty one (441) adults of strictly defined admitted cerebral malaria patients were studied. Detailed clinical/neurological examination was done at the time of admission, daily thereafter, at the time of regaining consciousness, at the time of discharge and at weekly intervals in those having neurological sequelae. All patients were treated by i.v./oral quinine and specific syndromes were managed according to WHO guidelines. Apart from fever and unconsciousness in all the patients, other features were convulsion (21.31%), neck rigidity (19%), psychosis (5.21%), conjugate deviation of eyes (2.26%), extrapyramidal rigidity (2.25%), trismus (1.31%), decorticate rigidity (1.13%) and decerebrate rigidity (0.90%). One hundred forty five (32.87%) patients expired and mortality was highest in pregnant ladies (39.28%). The important neurological sequelae in survivors were psychosis in 15 (5.06%), cerebellar ataxia in 14 (4.72%), hemiplegia in five (1.68%), extrapyramidal rigidity (EPR) in four (1.35%), peripheral neuropathy in three (1.01%), EPR with trismus in one (0.33%) and isolated sixth nerve palsy in one (0.33%) patients and all showed complete recovery in further follow up. The important observations of this study were stormy presentation, increased incidence of haemoglobinuria and jaundice, presence of neck rigidity, no prognostic relation to fundus abnormalities and high incidence of cerebellar ataxia and psychosis as neurological sequelae in survivors. Knowledge of self-limiting course of neurological sequelae may be helpful in reducing economic strain of expensive investigations and treatment.


Glucocorticoids inhibit neurogenic plasma extravasation and prevent virus-potentiated extravasation in the rat trachea.
Capsaicin increases the permeability of blood vessels in the rat tracheal mucosa through a mechanism involving the release of tachykinins from sensory nerves. This capsaicin-induced increase in vascular permeability is potentiated by viral infections of the respiratory tract. The present study was done to determine whether this "neurogenic plasma extravasation" can be inhibited by glucocorticoids, to learn the time course of this inhibition, and to determine whether glucocorticoids can prevent the potentiating effect of viral respiratory infections on neurogenic plasma extravasation. Groups of pathogen-free F344 rats were treated with dexamethasone for 2 or 8 h (4 mg/kg i.p.) or 48 or 120 h (0.5-4 mg/kg per d i.p.). Another group of rats was treated with dexamethasone for 120 h following the intranasal inoculation of Sendai virus. The magnitude of plasma extravasation produced by capsaicin or substance P was assessed after this treatment by using Monastral blue pigment and Evans blue dye as intravascular tracers. We found that dexamethasone reduced, in a dose-dependent fashion, the magnitude of plasma extravasation produced in the rat trachea by capsaicin and substance P. Significant inhibition was produced by a dose of dexamethasone as small as 0.5 mg/kg i.p. The effect of dexamethasone had a latency of several hours and reached a maximum after 2 d of treatment. Furthermore, dexamethasone prevented the potentiation of neurogenic plasma extravasation usually present after 5 d of Sendai virus respiratory infection.


Genome-wide association study of pre-harvest sprouting resistance in Chinese wheat founder parents.
Pre-harvest sprouting (PHS) is a major abiotic factor affecting grain weight and quality, and is caused by an early break in seed dormancy. Association mapping (AM) is used to detect correlations between phenotypes and genotypes based on linkage disequilibrium (LD) in wheat breeding programs. We evaluated seed dormancy in 80 Chinese wheat founder parents in five environments and performed a genome-wide association study using 6,057 markers, including 93 simple sequence repeat (SSR), 1,472 diversity array technology (DArT), and 4,492 single nucleotide polymorphism (SNP) markers. The general linear model (GLM) and the mixed linear model (MLM) were used in this study, and two significant markers (tPt-7980 and wPt-6457) were identified. Both markers were located on Chromosome 1B, with wPt-6457 having been identified in a previously reported chromosomal position. The significantly associated loci contain essential information for cloning genes related to resistance to PHS and can be used in wheat breeding programs.


Identification of bacteria and bacteria-associated chemical cues that mediate oviposition site preferences by Aedes aegypti.
The yellow fever mosquito, Aedes aegypti, the global vector of dengue and yellow fever, is inexorably linked to water-filled human-made containers for egg laying and production of progeny. Oviposition is stimulated by cues from water containers, but the nature and origin of these cues have not been elucidated. We showed that mosquito females directed most of their eggs to bamboo and white-oak leaf infusions, and only a small fraction of the eggs were laid in plain water containers. In binary choice assays, we demonstrated that microorganisms in leaf infusions produced oviposition-stimulating kairomones, and using a combination of bacterial culturing approaches, bioassay-guided fractionation of bacterial extracts, and chemical analyses, we now demonstrate that specific bacteria-associated carboxylic acids and methyl esters serve as potent oviposition stimulants for gravid Ae. aegypti. Elucidation of these compounds will improve understanding of the chemical basis of egg laying behavior of Ae. aegypti, and the kairomones will likely enhance the efficacy of surveillance and control programs for this disease vector of substantial global public health importance.


Occupational health problems in processing of X-ray photographic films.
Hazards to health from exposure to chemicals in automatic processing of photographic film is well documented by ILO and others. In the early 1980s increasing reports of miscellaneous health problems including respiratory and skin effects in radiographers processing X-ray films were published. In some cases asthmatic type responses were reported to be so severe as to exclude the radiographer from further work. It has been suggested that the increased number of such cases coincided with process changes, in particular the increased addition of glutaraldehyde as a hardening agent in the developer to offset the softening of the film especially those of low silver content. Attention must also be focused however on other miscellaneous chemicals present and of the possibility of synergistic reactions. The major problems appear to be associated with heavy workloads, poorly designed and ill-ventilated darkrooms or processing rooms, and lack of appropriate safe handling techniques for the chemicals used. Several studies illustrate improvements by attention to good occupational hygiene practice. The necessity for a holistic approach to control of such a complex micro-environment is stressed. It is unlikely that a single causal agent can be identified.


Spatial Transcriptomic Analysis of Pituitary Corticotroph Tumors Unveils Intratumor Heterogeneity.
Spatial transcriptomic (ST) analysis of tumors provides a novel approach on studying gene expression along with the localization of tumor cells in their environment to uncover spatial interactions. Herein, we present ST analysis of corticotroph pituitary neuroendocrine tumors (PitNETs) from formalin-fixed, paraffin-embedded (FFPE) tissues. We report that the in situ annotation of tumor tissue can be inferred from the gene expression profiles and is in concordance with the annotation made by a pathologist. Furthermore, relative gene expression in the tumor corresponds to common protein staining used in the evaluation of PitNETs, such as reticulin and Ki-67 index. Finally, we identify intratumor heterogeneity; clusters within the same tumor may present with different secretory capacity and transcriptomic profiles, unveiling potential intratumor cell variability with possible therapeutic interest. Together, our results provide the first attempt to clarify the spatial cell profile in PitNETs.


Deep pericardial sling versus lateral pericardial sutures in off-pump coronary artery bypass surgery.
The aim of this study was to evaluate the safety and efficacy of deep pericardial sling (DPS) versus lateral pericardial sutures (LPSs) for heart stabilization and adequate coronary artery exposure during off-pump coronary artery bypass surgery (OPCAB). One surgeon employed in 101 consecutive patients a series of four to six 2-0 polyglactin sutures placed laterally between the left phrenic nerve and the left pulmonary veins (LPS). Two other surgeons used in 104 consecutive patients a single 0-0 braided silk suture with moistened gauze placed in the oblique sinus of the posterior pericardium, between the inferior vena cava and the right lower pulmonary vein (DPS). One conversion to beating heart surgery with cardiopulmonary bypass support occurred in each study group. No patient in the LPS group was converted to DPS technique. The use of LPSs allowed a number of distal anastomoses somewhat higher than the DPS technique (4.1+/-1.1 vs. 3.7+/-1.1, P=0.02). Postoperative results were similar in both study groups. A lower incidence of postoperative low-cardiac output syndrome and of prolonged need of inotropes has been observed in the LPS group, but the difference failed to reach statistical significance. One patient in the LPS group had postoperative left phrenic nerve palsy. One patient in the DPS group suffered of intraoperative bleeding secondary to rupture of the inferior vena cava likely related to placement of DPS, which was successfully repaired. LPS technique is as effective as DPS technique and allows complete revascularization with a postoperative outcome similar to the latter technique.


The role of the chemical burns caused by hydroxide ion in the toxicity of dermal exposure to tetramethylammonium ion in a rat model.
To evaluate the role of the chemical burns caused by hydroxide ion in the fatal effects of tetramethylammonium ion (TMA) in dermal exposure to tetramethylammonium hydroxide (TMAH), we conducted a rat study consisting of two-step treatments with dermal exposure to NaOH and tetramethylammonium chloride (TMACl). In the first step, NaOH or saline was administered in the gauze on the shaved skin for 5 min, and in the second step, TMAH, TMACl, or saline was administered in the same way. The mean blood pressure (MBP), heart rate (HR), and survival in rats were compared among seven groups. Dermal exposure to saline and then 2.75 M TMACl introduced limited and temporary non-fatal effects. Exposure to 2.75 M NaOH and then saline had almost no effects and caused no deaths. Treatments with more concentrated NaOH or TMACl resulted in suppressions of MBP and HR, and deaths were observed after the dosing of TMACl. The toxicity of dermal exposure to TMA alone is limited, but fatal effects can be introduced by pre-treatment with hydroxide ion. Therefore, the chemical burn caused by hydroxide ion plays an essential role in the toxicity, implicating that effective neutralizing may help decreasing the fatality rate.


Automated selection of aptamers against cholangiocarcinoma cells on an integrated microfluidic platform.
An integrated microfluidic system capable of automatically identifying aptamers specific to cholangiocarcinoma (CCA) cells was developed herein. The developed system was capable of performing cell-based systematic evolution of ligands via an exponential enrichment (Cell-SELEX) process on-chip, and only six rounds of Cell-SELEX were required to identify high specificity aptamers; this represents a significant improvement in speed over conventional SELEX, in which 15-20 rounds are typically required. Using the microfluidic chip developed, three aptamers specific to CCA cells (one for SNU-478 cells and two for HuCCT-1 cells) were successfully screened. This automated system could be modified to uncover aptamer probes against other cancer cells, thereby allowing for earlier diagnosis and consequently a potentially improved prognosis.


Psychosis associated with HIV infection.
Five cases of psychiatric illness, presenting as functional psychosis, occurring in male homosexuals with human immunodeficiency virus (HIV) infection are described and compared with similar cases in the literature. The association between psychosis and infection with HIV is discussed with particular emphasis on the significance of functional versus organic presentation.


Brain surface ependymoma with repeated episodes of intratumoral hemorrhage--case report.
A 70-year-old woman presented with a rare brain surface ependymoma with repeated intratumoral hemorrhage. She was admitted with progressive dementia. Two years earlier, a diagnosis of subcortical hematoma in the right frontal lobe had been made following a fall. On admission, magnetic resonance imaging showed a huge right frontal mass lesion with multiple hemorrhagic cysts. She underwent gross total resection. The tumor was located on the surface of the frontal lobe, and was sharply demarcated from the surrounding brain tissue with no attachment to the ventricular wall. The histological features were consistent with an ependymoma forming perivascular pseudorosettes. Immunohistochemistry showed positive staining for glial fibrillary acidic protein. Electron microscopy showed microvilli and zonula adherens. This case demonstrates the natural course of malignant progression of ectopic ependymomas. Ependymoma should be included in the differential diagnosis of tumors associated with repeated subcortical hematomas, even if located on the brain surface and distant from ventricles.


MHC class I in dopaminergic neurons suppresses relapse to reward seeking.
Major histocompatibility complex class I (MHCI) is an important immune protein that is expressed in various brain regions, with its deficiency leading to extensive synaptic transmission that results in learning and memory deficits. Although MHCI is highly expressed in dopaminergic neurons, its role in these neurons has not been examined. We show that MHCI expressed in dopaminergic neurons plays a key role in suppressing reward-seeking behavior. In wild-type mice, cocaine self-administration caused persistent reduction of MHCI specifically in dopaminergic neurons, which was accompanied by enhanced glutamatergic synaptic transmission and relapse to cocaine seeking. Functional MHCI knockout promoted this addictive phenotype for cocaine and a natural reward, namely, sucrose. In contrast, wild-type mice overexpressing a major MHCI gene (H2D) in dopaminergic neurons showed suppressed cocaine seeking. These results show that persistent cocaine-induced reduction of MHCI in dopaminergic neurons is necessary for relapse to cocaine seeking.


Indirect hypertensive actions of long-term intracarotid angiotensin II infusion during ovine pregnancy.
Angiotensin II (AngII) influences the regulation of mean arterial pressure (MAP) through numerous mechanisms, including an action of circulating AngII on the brain to alter autonomic activity. We have obtained evidence that the relative importance of this effect is increased during pregnancy. Consequently, these studies were undertaken to assess the effects of bilateral infusion of AngII (0.35 ng.kg-1.min-1.artery-1) into the internal carotid arteries (ica) of sheep for 13 days. Six non-pregnant (NP) and six 105- to 125-day pregnant (PG) ewes were maintained in large metabolism cages, where MAP was continuously monitored. By day 10 of ica AngII infusion in NP ewes, MAP was increased from 83.9 +/- 1.6 to 92.9 +/- 2.8 mmHg (1 mmHg = 133.3 Pa) (p = 0.001). Twenty-four hour urine volume (UV, 2664 +/- 341 to 1583 +/- 228 mL; p = 0.005) and sodium excretion (UNaV, 190 +/- 5 to 113 +/- 19 mmol/day; p = 0.005) were decreased. 51Cr-tagged blood volume (BV) was increased on day 13 (3643 +/- 187 to 4379 +/- 446 mL; p = 0.05). In contrast, by only day 6 of ica AngII infusion in PG ewes, MAP increased from 79.1 +/- 1.9 to 84.1 +/- 1.4 mmHg (p = 0.03) in association with a BV expansion from 3999 +/- 274 to 4207 +/- 275 mL. These changes were preceded by decreases in UV (2813 +/- 413 to 2198 +/- 362 mL; p = 0.01) and UNaV (190 +/- 15 to 118 +/- 26 mmol/day; p = 0.01). By day 13, MAP had plateaued at 93.0 +/- 1.2 mmHg.(ABSTRACT TRUNCATED AT 250 WORDS)


Molecular epidemiology is becoming complex under the dynamic HIV prevalence: The perspective from Harbin, China.
Unlike most areas of China, HIV transmission via men who have sex with men (MSM) is increasing rapidly, and has become the main route of HIV transmission in Harbin city. The purpose of the current study was to elaborate the molecular epidemiologic characteristics of the new HIV epidemic. Eighty-one HIV-1 gag gene sequences (HXB2:806-1861) from local HIV infections were isolated; CRF01_AE predominated among HIV infections (71.6%), followed by subtype B (16.5%), CRF07_BC (6.2%), and unique recombinant strains (URFs; 6.2%). URFs were most often identified in the MSM population, which consisted of a recombination of CRF01_AE with subtype B or CRF07_BC. Six clusters were formed in this analysis; clusters I and II mainly circulated in southwest China. Clusters III and IV mainly circulated in southwest, southeast, and central China. Clusters V and VI mainly circulated in north and northeast China. Clusters III and IV may facilitate the transmission of the CRF01_AE strain from the southwest to the north and northeast regions of China. HIV subtypes are becoming diverse with the persistent epidemic in this geographic region. In brief, our results indicate that the molecular epidemiology of HIV is trending to be more complex. Thus, timely molecular epidemiologic supervision of HIV is necessary, especially for the MSM population.


Increased neutrophil-to-lymphocyte ratio is a novel marker for nutrition, inflammation and chemotherapy outcome in patients with locally advanced and metastatic esophageal squamous cell carcinoma.
Esophageal squamous cell carcinoma (ESCC) is one of the most common types of cancer, and its progression is strongly influenced by the presence of inflammation. Recently, there has been growing interest in the host inflammatory response, and increasing evidence has indicated that the neutrophil-to-lymphocyte ratio (NLR), a useful marker of systemic inflammation, may be an effective prognostic indicator in various types of malignant diseases. In the present study, 260 patients with ESCC were enrolled, including 110 who received chemoradiation therapy (CRT) involving irradiation and chemotherapy of 5-fluorouracil and cisplatin, and 150 received chemotherapy using 5-fluorouracil and cisplatin (FP). The patients of each group were both divided into two groups according to their NLR: High NLR (NLR>3.0) and low NLR (NLR≤3.0). Serum levels of prealbumin and retinol binding protein, which are nutritional parameters, were both significantly inversely correlated with NLR in patients treated with CRT, and patients treated with FP. Levels of CRP, a marker of inflammation, were significantly correlated with NLR, and stimulation indices, markers of immune reactions, were inversely correlated with NLR in both of CRT patients and FP patients. In patients treated with CRT, a partial response was significantly higher in patients with a low NLR and with progressive disease compared to those with a high NLR. In patients treated with FP, a partial response was also significantly higher in patients with a low NLR and with progressive disease compared to those with a high NLR. The overall survival of patients with CRT and FP were both significantly worse in patients with a high NLR than in those with a low NLR. NLR may serve as a useful marker of the tumor response, immune suppression, malnutrition and prognosis upon CRT or FP in patients with locally advanced or metastatic ESCC.


Intermittent drug techniques for schizophrenia.
Antipsychotic medication is considered the mainstay of treatment for schizophrenia and is generally regarded as highly effective, especially in controlling positive symptoms. However, long-term antipsychotic exposure has been associated with a range of adverse effects, including extra-pyramidal symptoms (EPS), neuroleptic malignant syndrome (NMS), tardive dyskinesia and death. Intermittent drug techniques refers to the 'use of medication only during periods of incipient relapse or symptom exacerbation rather than continuously'. The aim is to reduce the risk of typical adverse effects of antipsychotics by 'reducing long-term medication exposure for patients who are receiving maintenance treatment while limiting the risk of relapse', with a further goal of improving social functioning resulting from the reduction of antipsychotic-induced side effects To review the effects of different intermittent drug techniques compared with maintenance treatment in people with schizophrenia or related disorders. We searched The Cochrane Schizophrenia Group Trials Register (April 2012) and supplemented this by contacting relevant study authors, handsearching relevant intermittent drug treatment articles and manually searching reference lists. All randomised controlled trials (RCTs) that compared intermittent drug techniques with standard maintenance therapy for people with schizophrenia. Primary outcomes of interest were relapse and hospitalisation. At least two review authors selected trials, assessed quality and extracted data. We calculated risk ratios (RR) and 95% confidence intervals (CI) of homogeneous dichotomous data and estimated the 95% confidence interval (CI) around this. For non-skewed continuous endpoint data extracted from valid scales, we estimated mean difference (MD) between groups with a 95% CI. Where data displayed heterogeneity, these were analysed using a random-effects model. Skewed data are presented in tables. We assessed overall quality for clinically important outcomes using the GRADE approach. Of 241 records retrieved by the search, 17 trials conducted between 1961 and 2011, involving 2252 participants with follow-up from six weeks to two years, were included. Homogenous data demonstrated that instances of relapse were significantly higher in people receiving any intermittent drug treatment in the long term (n = 436, 7 RCTs, RR 2.46, 95% CI 1.70 to 3.54, moderate quality evidence). Intermittent treatment was shown to be more effective than placebo, however, and demonstrated that significantly less people receiving intermittent antipsychotics experienced full relapse by medium term (n = 290, 2 RCTs, RR 0.37, 95% CI 0.24 to 0.58, very low quality evidence). Hospitalisation rates were higher for people receiving any intermittent drug treatment by long term (n = 626, 5 RCTs, RR 1.65, 95% CI 1.33 to 2.06, moderate quality evidence). Results demonstrated little difference in instances of tardive dyskinesia in groups with any intermittent drug technique versus maintenance therapy, with equivocal results (displaying slight heterogeneity) at long term (n = 165, 4 RCTs, RR 1.15, 95% CI 0.58 to 2.30, low quality evidence). Results of this review support the existing evidence that intermittent antipsychotic treatment is not as effective as continuous, maintained antipsychotic therapy in preventing relapse in people with schizophrenia. More research is needed to assess any potential benefits or harm of intermittent treatment regarding adverse effects typically associated with maintained antipsychotic treatment, as well as any cost-effectiveness of this experimental treatment.


Extinction, reacquisition, and rapid forgetting of eyeblink conditioning in developing rats.
Eyeblink conditioning is a well-established model for studying the developmental neurobiology of associative learning and memory. However, age differences in extinction and subsequent reacquisition have yet to be studied using this model. The present study examined extinction and reacquisition of eyeblink conditioning in developing rats. In Experiment 1, post-natal day (P) 17 and 24 rats were trained to a criterion of 80% conditioned responses (CRs) using stimulation of the middle cerebellar peduncle (MCP) as a conditioned stimulus (CS). Stimulation CS-alone extinction training commenced 24 h later, followed by reacquisition training after the fourth extinction session. Contrary to expected results, rats trained starting on P17 showed significantly fewer CRs to stimulation CS-alone presentations relative to P24s, including fewer CRs as early as the first block of extinction session 1. Furthermore, the P17 group was slower to reacquire following extinction. Experiment 2 was run to determine the extent to which the low CR percentage observed in P17s early in extinction reflected rapid forgetting versus rapid extinction. Twenty-four hours after reaching criterion, subjects were trained in a session split into 50 stimulation CS-unconditioned stimulus paired trials followed immediately by 50 stimulation CS-alone trials. With this "immediate" extinction protocol, CR percentages during the first block of stimulation CS-alone presentations were equivalent to terminal acquisition levels at both ages but extinction was more rapid in the P17 group. These findings indicate that forgetting is observed in P17 relative to P24 rats 24 h following acquisition. The forgetting in P17 rats has important implications for the neurobiological mechanisms of memory in the developing cerebellum.


Antifouling Slippery Surface against Marine Biofouling.
Titanium and its alloys have become the most excellent structure materials for naval seawater pipelines due to their high strength and good corrosion resistance. However, marine biofouling poses a serious threat to titanium alloy piping systems because of their good biocompatibility. Recently, the biomimetic antifouling coating, a novel antifouling method, has received great attention. Here, based on this biomimetic idea, we develop a nontoxic antifouling slippery surface (AFSS) using silicone oil, silane coupling agent, nanosilica, nanoceramic coating, epoxy resin, and capsaicin. The developed AFSS has excellent slippery performance for various droplets, good durability, and a superior self-cleaning property. Additionally, the antifouling performance of the AFSS was significantly enhanced, as confirmed by the reduced adhesion of proteins (70.7%), bacteria (97.2%), and algae (97.7%) compared to the ordinary titanium alloy. With these excellent properties, the AFSS was expected to be a promising candidate for protecting titanium alloy piping systems from marine biofouling.


Algorithmic Approach to Virtual Machine Migration in Cloud Computing with Updated SESA Algorithm.
Cloud computing plays an important role in every IT sector. Many tech giants such as Google, Microsoft, and Facebook as deploying their data centres around the world to provide computation and storage services. The customers either submit their job directly or they take the help of the brokers for the submission of the jobs to the cloud centres. The preliminary aim is to reduce the overall power consumption which was ignored in the early days of cloud development. This was due to the performance expectations from cloud servers as they were supposed to provide all the services through their services layers IaaS, PaaS, and SaaS. As time passed and researchers came up with new terminologies and algorithmic architecture for the reduction of power consumption and sustainability, other algorithmic anarchies were also introduced, such as statistical oriented learning and bioinspired algorithms. In this paper, an indepth focus has been done on multiple approaches for migration among virtual machines and find out various issues among existing approaches. The proposed work utilizes elastic scheduling inspired by the smart elastic scheduling algorithm (SESA) to develop a more energy-efficient VM allocation and migration algorithm. The proposed work uses cosine similarity and bandwidth utilization as additional utilities to improve the current performance in terms of QoS. The proposed work is evaluated for overall power consumption and service level agreement violation (SLA-V) and is compared with related state of art techniques. A proposed algorithm is also presented in order to solve problems found during the survey.


Electrohydrodynamic flow through a 1 mm(2) cross-section pore placed in an ion-exchange membrane.
In recent years, the control of ionic currents has come to be recognized as one of the most important issues related to the efficient transport of single molecules and microparticles in aqueous solutions. However, the complicated liquid flows that are usually induced by applying electric potentials have made it difficult to address a number of unsolved problems in this area. In particular, the nonequilibrium phenomena that occur in electrically non-neutral fields must be more thoroughly understood. Herein, we report on the development of a theoretical model of liquid flows resulting from ion interactions while focusing on the so-called electrohydrodynamic (EHD) flow. We also discuss the development of an experimental system to optically and electrically observe EHD flows using a 1 mm(2) cross-section pore placed in an ion-exchange membrane where cation and anion flows can be separated without the use of a charged environment. Although micro/nanosized flow channels are usually applied to induce electric double layer overlaps to utilize strong electroosmotic effects, our system does not require such laborious fabrication processes. Instead, we visualize EHD flows by using a millimeter size pore immersed in an alkaline aqueous solution. In this setup, liquid flows passing through the pore along the direction of ion flow, whose velocity reaches on the order of 1 mm/s, can be clearly observed by applying a few volts of electric potential. Furthermore, the transient phenomena associated with ionic responses are theoretically elucidated.


Context-dependent catalepsy intensification is due to classical conditioning and sensitization.
Haloperidol-induced catalepsy represents a model of neuroleptic-induced Parkinsonism. Daily administration of haloperidol, followed by testing for catalepsy on a bar and grid, results in a day-to-day increase in catalepsy that is completely context dependent, resulting in a strong placebo effect and in a failure of expression after a change in context. The aim of this study was to analyse the associative learning process that underlies context dependency. Catalepsy intensification was induced by a daily threshold dose of 0.25 mg/kg haloperidol. Extinction training and retesting under haloperidol revealed that sensitization was composed of two components: a context-conditioning component, which can be extinguished, and a context-dependent sensitization component, which cannot be extinguished. Context dependency of catalepsy thus follows precisely the same rules as context dependency of psychostimulant-induced sensitization. Catalepsy sensitization is therefore due to conditioning and sensitization.


A guide to nucleic acid detection by single-molecule kinetic fingerprinting.
Conventional methods for detecting small quantities of nucleic acids require amplification by the polymerase chain reaction (PCR), which necessitates prior purification and introduces copying errors. While amplification-free methods do not have these shortcomings, they are generally orders of magnitude less sensitive and specific than PCR-based methods. In this review, we provide a practical guide to a novel amplification-free method, single-molecule recognition through equilibrium Poisson sampling (SiMREPS), that provides both single-molecule sensitivity and single-base selectivity by monitoring the repetitive interactions of fluorescent probes to immobilized targets. We demonstrate how this kinetic fingerprinting filters out background arising from the inevitable nonspecific binding of probes, yielding virtually zero background signal. As practical applications of this digital detection methodology, we present the quantification of microRNA miR-16 and the detection of the mutation EGFR L858R with an apparent single-base discrimination factor of over 3 million.


Quality of life in patients with antiphospholipid syndrome is related to disease burden and anticoagulant therapy.
To evaluate health-related quality of life (HRQoL) in primary antiphospholipid syndrome (PAPS) and correlate it with a crude estimate of accrual organ damage, comorbidity (diabetes mellitus, hypertension and dyslipidemia) and treatment (oral anticoagulation, immunosuppressors and prednisone). We assessed HRQoL with the Short-Form 36 (SF-36) and the Lupus Quality of Life instrument (LupusQoL) and the disease burden with a modified Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index (SLICC/ACR SDI). As controls we used SF-36 data from a Mexican general population within the same age range. We included 50 PAPS patients (86% women), mean age 47.6 ± 14.5 years, median disease duration 9.4 years, median SLICC/ACR score of 1 point and 80% had thrombotic events. PAPS patients had lower HRQoL than controls. We found a positive correlation between SF-36 and LupusQoL (r = 0.85, P < 0.0001). The SLICC/ACR SDI correlated negatively with both LupusQoL and SF-36, specifically the peripheral vascular domain (r = -0.29, P = 0.03, for both). Patients on oral anticoagulants (n = 37) had lower LupusQoL, physical functioning, intimate relationships, burden to others and pain scores as well as a lower SF-36 physical functioning score. We did not find differences in HRQoL regarding comorbidities and other treatments. HRQoL in PAPS was related to burden of the disease specifically at the vascular peripheral area and use of anticoagulants.


Effects of algal bloom (AB) on sediment microorganisms with special functions at different AB stages in Chaohu Lake.
Algal bloom (AB) is a serious water environment problem in China, but it is still unclear how AB affects microorganisms with special functions (such as nitrogen-, phosphorus-, sulfur- and iron-related bacteria) at different AB stages. Samples of water and sediment were taken in the AB region and free algal bloom (FAB) region of Chaohu Lake at different AB stages, and the numbers of N-, P-, S- and Fe-related bacteria were determined by culture-dependent methods. Results showed that during the whole experiment, numbers of N-, P-, S-, and Fe-related bacteria in the AB region were significantly higher than those in the FAB region, especially at the AB outbreak stage. In the FAB region, numbers of above bacteria showed little variation with time. In the AB region, however, numbers of N-, S-, and Fe-related bacteria evidently changed as 'Λ'-type with AB process. They presented an upward trend at the AB formation stage and the initial stage of AB outbreak, and then showed a downward trend. Numbers of P-related bacteria also showed a similar trend as the above bacteria, but they kept high level for long time at the AB outbreak stage. This study indicated that AB produced different effects on microorganisms with special functions at different AB stages.


"[I] don't wanna just be like a cog in the machine": Narratives of autism and skilled employment.
Autistic people are less likely to be employed than the general population. Autistic people with skilled training (e.g. training for jobs in acting, plumbing, science, or social work) might be even less likely to get a good job in their field. Little is known about the experiences of autistic people in skilled employment or what employment success means to them. We interviewed 45 autistic people with skilled training in a wide range of fields, 11 job supervisors, and 8 topic experts. We asked them about their experiences, what they felt helped them to be successful at work, and what employment success means to them. Participants talked about the high stakes of disclosure, taking unconventional pathways to careers, disconnects with service and support systems, mental health challenges from trauma and burnout, the autistic advantages in the workplace, and complex dimensions of discrimination. Participants said success meant opportunities for growth, good work/life balance, financial independence, sense of community, and feeling valued, accepted, and like their work had meaning. Things that helped them be successful included flexible, accepting workplaces, supportive and respectful supervisors, and direct communication. What we learned suggests that an individualized, wholistic approach to autism employment intervention that considers both employers and employees and employee mental health could be useful. We also recommend more research into disclosure and destigmatizing disability at work.


Target-initiated fluorescent aptasensor based on multisite strand displacement amplification for label-free detection of ochratoxin A.
We developed a fluorescent aptasensor for label-free detection of ochratoxin A (OTA) based on TdT- and DNA polymerase-assisted multisite strand displacement amplification. This aptasensor exhibits good specificity and high sensitivity with a limit of detection (LOD) of 0.18 ng mL-1, and it can be further applied for the accurate quantification of OTA in complex real samples, holding promising applications in the field of food safety.


Steering Photocatalytic CO2 Conversion over CsPbBr3 Perovskite Nanocrystals by Coupling with Transition-Metal Chalcogenides.
Transition-metal chalcogenides (TMCs) have received enormous attention by virtue of their large light absorption coefficient, abundant catalytically active sites, and markedly reduced spatially vectorial charge-transfer distance originating from generic structural merits. However, the controllable construction of TMC-based heterostructured photosystems for photocatalytic carbon dioxide (CO2) reduction is retarded by the ultrashort charge lifetime, sluggish charge-transfer kinetics, and low target product selectivity. Herein, we present the rational design of two-dimensional (2D)/zero-dimensional (0D) heterostructured CO2 reduction photosystems by an electrostatic self-assembly strategy, which is enabled by precisely anchoring CsPbBr3 quantum dots (QDs) on the 2D TMC (CdIn2S4, ZnIn2S4, In2S3) frameworks. The peculiar 2D/0D integration mode and suitable energy-level alignment between these two assembly units afford maximal interfacial contact and applicable potential for CO2 photoreduction, thus endowing the self-assembled TMCs/CsPbBr3 nanocomposites with considerably improved visible-light-driven photocatalytic performances toward CO2 reduction to carbon monoxide with high selectivity. The enhanced photocatalytic performances of TMCs/CsPbBr3 heterostructures are attributed to the abundant active sites on the TMC frameworks, excellent light absorption of CsPbBr3 QDs, and well-defined 2D/0D heterostructures of TMCs/CsPbBr3 QDs photosystems, which synergistically boosts the directional charge transport from CsPbBr3 QDs to TMCs, enhancing the interfacial charge migration/separation. Our work would inspire the construction of novel TMCs-involved photosystems for solar-to-fuel conversion.


Septic arthritis and Reiter's syndrome in sickle cell disorders: case reports and implications for management.
Two patients with sickle cell disorders developed gram-negative septic arthritis and responded poorly to conventional managment. Unfavorable factors included (1) confusion with the nonseptic arthropathy of a sickle crisis; (2) impaired articular perfusion, perhaps related to local sickling; (3) conversion of an infecting organism to an L-form in the presence of cell-wall inhibitors; and (4) interference with cultures and antibacterial titrations of synovial fluid caused by intro-articular administration of antibiotics. Prolonged treatment with very high doses of antibiotics was required to eradicate infection in both patients. Because sickling and local hypoxia apparently interfere with defenses against infection, anemia should be corrected by transfusion. One patient had had Reiter's syndrome; the dysenteric form of this disease may be a cause of arthritis in some sicklemic patients.


Toward the design of a nursing ontology system.
The unambiguous and consistent representation of the healthcare domain is the foundation of modernized healthcare (eg, electronic medical records). However, the nursing domain often fails to meet this requirement. In this article, we address this challenge by presenting a three-stage methodological approach that can be used to (1) capture knowledge in a nursing domain; (2) design a nursing ontology, composed of data models linked with terminology concepts in a multirelational property graph; and (3) implement and (4) evaluate the ontology. Through the feasibility, development, and evaluation phases of our methodological approach, we modeled a nursing domain (ontology) and the indices that partition the domain into an efficient, searchable space, where the solution to a nursing problem is seen as abstractly defined traversals through its graph vertices and edges. Thus, the use of the three-phase ontology development process and multirelational property graph was sufficiently comprehensive for achieving the representation of a nursing domain ontology and its instantiation.


Permeabilization of Beauveria bassiana blastospores for in situ enzymatic assays.
Vigorous agitation of an aqueous suspension of blastospores (BS) of Beauveria bassiana mixed with nine volumes of a 1:4 (v/v) mixture of toluene:ethanol (95%) for 10 min permits blastospore permeabilization. Agitation results in greater membrane permeabilization than heating blastospores in the presence of toluene:ethanol or the detergents Triton X-100, sodium dodecyl sulfate, hexadecyltrimethylammonium bromide and Brij-35. The β-galactosidase activity in permeabilized blastospores was determined with these methods. The effectiveness of permeabilization in detecting enzyme activity was assessed by comparison to whole BS lysates prepared by mechanical disruption and pressurized disruption of BS biomass. The toluene:ethanol method was applied to study the incorporation of (3)H-thymidine triphosphate into blastospore DNA. Whole BS permeabilization allows the examination of enzyme activity and DNA synthesis at a cellular level in this important mycoinsecticide.


The solar UV radiation level needed for cutaneous production of vitamin D3 in the face. A study conducted among subjects living at a high latitude (68 degrees N).
Populations at high latitudes experience several winter months with insufficient UV solar radiation to induce a significant cutaneous production of vitamin D. This unique study was designed to pursue an in vivo threshold of UV radiation needed for cutaneous production of vitamin D to take place if only the face was exposed to UV radiation. The vitamin D status were measured by analyzing blood samples weekly from a study group of 15 subjects over a period of 2 months during late winter, when UV radiation can be expected to increase substantially from rising solar elevations. Statistical analysis showed no significant positive association between the mean UV radiation dose and the mean 25(OH)D (25-hydroxy vitamin D) for the group. On an individual basis, however, we found indications that subjects with very low initial concentration of 25(OH)D (<30 nmol l(-1)) seemed to respond to UV radiation as early as in the beginning of March. For other individuals diet seemed to be the dominant controlling factor for 25(OH)D levels.


The stimulating effect of a cytosol extract from regenerating liver on isolated hepatocytes and the positive role of insulin.
Several kinds of damage induce a regenerative response in the liver. Partial hepatectomy is a well known method, suitable for studying liver regeneration and factor/s involved in it. The presence of the so called Hepatic Stimulator Substance has been demonstrated in the cytosol obtained from regenerating hepatocytes, and it has been partially characterized, from a molecular point of view. Mainly, it stimulates liver regeneration and Tritiated Thymidine incorporation in cultured hepatocytes as demonstrated in this paper (Control Hepatocytes: 8447 +/- 660.4 cpm, (mean +/- sd), Test Hepatocytes: 16974 +/- 2720.9 cpm; Index of Stimulation: 2.01). Furthermore, the stimulating activity of HSS is increased by the presence, in the culture media, of insulin which has been proved to have a positive role in liver regeneration (Control Hepatocytes: 8889 +/- 367.3 cpm; Test Hepatocytes: 21588.5 +/- 1232.4 cpm; Index of Stimulation: 2.42, p less than 0.05). These data seem to suggest that liver is probably the site of production/activity and action of factor/s that stimulate/s liver regeneration; but other substances exist, such as insulin, which play a permissive role.


A new species of Neossos Malloch (Diptera: Heleomyzidae) from the Yukon Territory, Canada, and a revised key to the Nearctic species.
The rarely collected genus Neossos Malloch contains three Nearctic and one western European species. Most known specimens have been collected from bird nests. Two specimens of an undescribed species of Neossos were collected by sweeping in subarctic tundra and a mesic meadow in the Yukon Territory, Canada. This represents a significant northward extension of the known Nearctic range of the genus. Neossos tombstonensis sp. n. is described from the Yukon Territory. This represents the fourth described Nearctic species of Neossos. Although the type specimens were collected by sweeping, the species is predicted to be associated with bird nests, based on habits of other members of the genus. A revised key to the Nearctic species of Neossos is provided.


Human Adenomyosis Endometrium Stromal Cells Secreting More Nerve Growth Factor: Impact and Effect.
Abnormal expression of nerve growth factor (NGF) was found in adenomyosis (AM). We collected AM foci from patients and eutopic endometrium from non-AM controls. Endometrium stromal cells (ESCs) were cultured. Different levels of 17β-estradiol, tumor necrosis factor (TNF), CoCl2, and H2O2 were added to the culture system separately, then the expression level of NGF in ESCs was detected. After adding different levels of NGF, the proliferation and apoptosis of ESCs and aromatase expression were detected. We found that 17β-estradiol promoted NGF production in AM ESCs but not in control ESCs; TNF promoted NGF production in both AM and control ESCs; and CoCl2 inhibited NGF production in control ESCs, but had no effect in AM ESCs. Nerve growth factor promoted the proliferation and synthesis of aromatase in AM ESCs. In conclusion, locally increased estrogen levels and inflammation may cause increased NGF production in the uterus of patients with AM. Nerve growth factor stimulated the proliferation and increased aromatase expression of ESCs from AM foci, suggesting NGF might contribute to the pathology and etiology of AM.


Variability of central conduction in the course of multiple sclerosis. Serial recordings of evoked potentials in the evaluation of therapy.
In 27 normal subjects evoked potentials (EPs) were serially measured across an interval of 3.5 months. The variation in latencies of the P100 component of the VEP and of the components N13 and N20 of the SEP did not exceed 12% of the absolute values of these latencies on the first recording. The VEP and SEP were also serially recorded over the same interval in 35 patients with 'clinically definite' multiple sclerosis. Changes of the latencies exceeding those of normal subjects were noted in 30% of the cases. The changes in the EPs showed no correlation with the variations in the overall clinical disability. It is argued that EPs are probably more sensitive to changes in function of parts of the white matter than the clinical examination and are useful in the quantification of these changes. As a model for future studies, EPs were used in the 'evaluation' of ACTH therapy. It appeared, that therapy did not have effect on the EPs.


Experimental and ab initio study of a new D 1Deltag state of the C3 radical.
We report here the first observation of the D (1)Delta(g) state of the C(3) radical, which provides the first comprehensively analyzed example of the dynamic Renner-Teller splitting in Delta symmetry. Two color double resonance spectroscopy via the A (1)Pi(u) state was employed to experimentally probe an extensive range of vibronic levels in this D (1)Delta(g) state, covering all three modes of vibration of C(3). The analysis was supported by ab initio potential energy surface calculations on the C(3) radical to outline the lowest eight singlet electronic states. Two methods were used to analyze the Renner-Teller effect. The first method is an empirical Hamiltonian based on normal modes, using harmonic oscillator functions as a basis, with Renner-Teller and other terms added as required, which allows conventional vibrational parameters to be determined. The second is a much larger program that uses the exact kinetic energy operator for a triatomic molecule to calculate vibronic energy levels directly from the Renner-Teller pair of potential energy surfaces. Both methods give a good fit to the experimental results, with only a small adjustment to the ab initio surfaces required for the latter. One of the overall conclusions is that the Renner-Teller effect is rather smaller in the D (1)Delta(g) state than in the A (1)Pi(u) state.


Action pattern of human pancreatic alpha-amylase on maltoheptaose, a substrate for determining alpha-amylase in serum.
An enzymatic assay for the determination of alpha-amylase in serum was developed which employed a soluble substrate, maltoheptaose, and a coupled enzymatic indicator reaction consisting of alpha-glucosidase and the hexokinase-glucose-6-phosphate dehydrogenase system. We used high-performance liquid chromatography (HPLC) to establish the action pattern of maltoheptaose under the test conditions: (A) the action pattern of alpha-amylase, (B) that of the combined action of alpha-amylase and alpha-glucosidase. Conductive to this effect was: the availability of pure maltoheptaose and human pancreatic alpha-amylase; the development of an adequate procedure for sample pretreatment (partition chromatography on a mixed-bed ion exchange) and of an HPLC system for separation of substrate and reaction products without interference from by products of the assay (partition chromatography on a cation-exchange column with acetonitrile-water); and the use of a new, very sensitive refractometric detector revealing sugar amounts as low as 40 ng. We derived the following stoichiometric equations: (see formula index).


Electrochemical Behavior and Redox-Dependent Disassembly of Gallic Acid/FeIII Metal-Phenolic Networks.
Metal-phenolic networks (MPNs) are a versatile class of organic-inorganic hybrid systems that are generating interest for applications in catalysis, bioimaging, and drug delivery. These self-assembled MPNs possess metal-coordinated structures and may potentially serve as redox-responsive platforms for triggered disassembly or drug release. Therefore, a comprehensive study of the reduction and oxidation behavior of MPNs for evaluating their redox responsiveness, specific conditions required for their disassembly, and the kinetics of metal ion release, is necessary. Using a representative MPN gallic acid-iron (GA/FeIII) system, we conducted electrochemical studies to provide fundamental insights into the redox behavior of these MPNs. We demonstrate that GA/FeIII is redox active, and evaluate its electrochemical reversibility, identify the oxidation state of the redox-active species, and provide information regarding the stability of the networks toward reductive stimuli and specific redox conditions required for the "on-off" or continuous release of FeIII. Overall, through studying the redox properties of GA/FeIII films, we advance the understanding of multifunctional iron-containing MPN platforms that may have practical significance for biologically relevant applications.


Serological Screening of Immunoglobulin G against SARS-CoV-2 Nucleocapsid and Spike Protein before and after Two Vaccine Doses among Healthcare Workers in Japan.
This study sought to evaluate the effects of two vaccine doses and the extent of SARS-CoV-2 infection among healthcare workers. We measured immunoglobulin G antibody titers against SARS-CoV-2 nucleocapsid and spike protein among healthcare workers at Gunma University Hospital. In March 2021, prior to BNT-162b2 vaccination, two of 771 participants were seropositive for nucleocapsid and spike protein, whereas 768 were seronegative. The remaining one participant was seropositive for nucleocapsid protein but seronegative for spike protein. A total of 769 participants were seropositive for spike protein after two vaccination doses. The two seropositive participants prior to vaccination showed the highest antibody titers after the second vaccination. They were probably infected with SARS-CoV-2 without clinical symptoms before March 2021. Four weeks after the second vaccination, a younger age was associated with higher antibody titers against SARS-CoV-2 spike protein. Thirty-two weeks after the second vaccination, blood samples were collected from 342 of 769 participants. Antibody titers at 32 weeks after the second vaccination significantly decreased compared with those at 4 weeks after the second vaccination among all age groups. The rate of decrease in antibody titers between 4 and 32 weeks after the second vaccination was greater in the female participants. No sex differences were observed in the antibody titers within each age group. BNT-162b2 vaccination thus induced seroconversion in an age-dependent manner. Serological screening could further establish the likelihood of subclinical SARS-CoV-2 infection.


The sensitivity of commercial kits in detecting the genes of pathogenic bacteria in venison.
The expansion of the wild deer population is a major problem for the Japanese farm and forestry industries because their damage to farm products and vegetation results in huge economic loss. To promote game meat consumption, hygiene inspections should be performed to detect main bacterial pathogens before products are shipped. In this study, we aimed to evaluate the ability of commercial test kits to genetically detect EHEC, Salmonella and Listeria monocytogenes in venison. Our results demonstrated that the kits for three pathogens could be useful for venison as well as other domestic meat products. Our comparative study showed that the LAMP kits were more sensitive than the RT-qPCR kits in the detection of all of these pathogens.


Smoking habits and smoking cessation among North Carolina nurses.
A 1987 questionnaire survey of a 1% random sample (n = 356) of registered nurses in North Carolina provided data on the smoking habits and smoking cessation. Fifty-six percent were never smokers; 19% were current smokers. Among the ever smokers, 31% had quit smoking for at least one year. Twenty-two percent of the former smokers had smoked less than 5 years and 39% less than 10 years before quitting. Anecdotal notes from never smokers suggested that their major deterrent to smoking was their own parents smoking. Concerns about the addictive smoking behavior and health effects of smoking observed in their parents as well as concerns about potential health risks to themselves deterred them from smoking. Concerns about the adverse consequences of smoking was the most influential factor influencing smoking cessation and reduction of cigarette smoking. Friends' and family's encouragement to stop smoking was the most influential external factor motivating nurses to quit or reduce cigarette consumption. Fifty-seven percent of the former smokers quit smoking after one or two attempts while 53 of the current smokers had tried to quit 3 or more times - 90% had tried at least once to quit smoking; however, only 18% of the current smokers had abstained for more than one year during any of their attempts to quit. Implications of the results include: (1) smoking cessation programs for nurses in the workplace may have considerable impact since the majority of nurses who smoke are tying to quit; (2) relapse prevention strategies need to be an integral part of such smoking cessation programs including involvement of family and friends to support the smokers in their cessation efforts.


Nonsurgical correction of a severe anterior deep overbite accompanied by a gummy smile and posterior scissor bite using a miniscrew-assisted straight-wire technique in an adult high-angle case.
In the present report, we describe the successful use of miniscrews to achieve vertical control in combination with the conventional sliding MBT™ straight-wire technique for the treatment of a 26-year-old Chinese woman with a very high mandibular plane angle, deep overbite, retrognathic mandible with backward rotation, prognathic maxilla, and gummy smile. The patient exhibited skeletal Class II malocclusion. Orthodontic miniscrews were placed in the maxillary anterior and posterior segments to provide rigid anchorage and vertical control through intrusion of the incisors and molars. Intrusion and torque control of the maxillary incisors relieved the deep overbite and corrected the gummy smile, while intrusion of the maxillary molars aided in counterclockwise rotation of the mandibular plane, which consequently resulted in an improved facial profile. After 3.5 years of retention, we observed a stable, well-aligned dentition with ideal intercuspation and more harmonious facial contours. Thus, we were able to achieve a satisfactory occlusion, a significantly improved facial profile, and an attractive smile for this patient. The findings from this case suggest that nonsurgical correction using miniscrew anchorage is an effective approach for camouflage treatment of high-angle cases with skeletal Class II malocclusion.


[The history of the discovery of the Helicobacter pylori].
Helicobacter pylori is a spiral-shaped gram-negative bacterium that colonizes the stomach lining. The presence of a microorganism in humans was described more than a century ago, but from detection to recognition of its role in the etiology and pathogenesis of diseases of the stomach, researchers had to overcome a long path of criticism and mistrust. Coiled bacteria have been mentioned several times in the medical literature, but these bacteria were thought to be contaminants, and any evidence of the bacteria in the stomach was ignored by the medical community. The discovery of H. pylori led to a revolutionary rethinking of the mechanisms of development of a number of diseases: the role of bacteria in the development of chronic gastritis, peptic ulcer disease, stomach cancer and MALT lymphoma was proved. The principles of their prevention and treatment have changed. For this discovery in 2005, Barry Marshall and Robin Warren were awarded the Nobel Prize in Medicine and Physiology.


Myosin 1b and F-actin are involved in the control of secretory granule biogenesis.
Hormone secretion relies on secretory granules which store hormones in endocrine cells and release them upon cell stimulation. The molecular events leading to hormone sorting and secretory granule formation at the level of the TGN are still elusive. Our proteomic analysis of purified whole secretory granules or secretory granule membranes uncovered their association with the actomyosin components myosin 1b, actin and the actin nucleation complex Arp2/3. We found that myosin 1b controls the formation of secretory granules and the associated regulated secretion in both neuroendocrine cells and chromogranin A-expressing COS7 cells used as a simplified model of induced secretion. We show that F-actin is also involved in secretory granule biogenesis and that myosin 1b cooperates with Arp2/3 to recruit F-actin to the Golgi region where secretory granules bud. These results provide the first evidence that components of the actomyosin complex promote the biogenesis of secretory granules and thereby regulate hormone sorting and secretion.


Adhesion and signaling in axonal fasciculation.
Numerous in vitro assays and in vivo perturbation studies have led to a model of neural development in which selective fasciculation helps to define accurate axonal projections. Genetic analysis in vivo confirms the hypothesis that axonal fasciculation and defasciculation are controlled by adhesion mechanisms, but also suggests that, in many cases, adhesion and guidance are separable phenomena. In addition, receptors that control the level of tyrosine phosphorylation may play an important role in fasciculation, suggesting that complex intracellular pathways lie just beneath the surface.


Mitogen-activated protein kinases and calcium-dependent protein kinases are involved in wounding-induced ethylene biosynthesis in Arabidopsis thaliana.
Ethylene, an important hormone in plant growth, development and response to environmental stimuli, is rapidly induced by mechanical injury or wounding. Although induction of ACS (1-aminocyclopropane-1-carboxylic acid synthase) gene expression has been associated with this process, the detailed regulatory mechanism is unclear. Here, we report that the wounding-induced ethylene production is modulated by both mitogen-activated protein kinase (MAPK) pathway and calcium-dependent protein kinase (CPK) pathway. Study using acs mutants demonstrated that four ACS isoforms, including ACS2, ACS6, ACS7 and ACS8, contribute to ethylene production in response to wounding. Loss-of-function analysis defines the role of MPK3 and MPK6, and their upstream MKK4 and MKK5, in wounding-induced ethylene production. They play an important role in the wounding-induced up-regulation of all four ACS genes expression. Independent of MAPK pathway, CPK5 and CPK6 are also involved in the wounding-induced ethylene production by regulating the expression of ACS2, ACS6 and ACS8 genes. Taken together, we demonstrate that two independent kinase signalling pathways, MPK3/MPK6 cascade and CPK5/CPK6, are involved in the wounding-induced ethylene biosynthesis via differential regulation of ACS genes at transcriptional level.


[A new species of wild mice on the Island of Cyprus].
A mitochondrial and nuclear gene analysis allowed us to precise the taxonomical position of the two sympatric species of mice known to be present on Cyprus. One of them is the commensal house mouse M. m. domesticus, and the other revealed to be a new taxon that is a sister species of M. spicilegus and M. macedonicus. The new species is equidistant from each of these, the divergence dating around 0.5-1 Myr. Its origin either results from an ancient accidental colonisation of the island or from a recent transportation by the first epipalaeolithic settlers. In this last eventuality, the new species would also exist somewhere else in Asia Minor.


Neutral endopeptidase and natriuretic peptide receptors participate in the regulation of C-type natriuretic peptide expression in renal interstitial fibrosis.
The initiation and progression of renal interstitial fibrosis (RIF) is a complicated process in which many factors may play an activate role. Among these factors, C-type natriuretic peptide (CNP) is an endothelium-derived hormone and acts in a local, paracrine fashion to regulate vascular smooth muscle tone and proliferation. In this study, we established a rat model of unilateral ureteral obstruction (UUO). CNP expression tends to be higher immediately after ligation and declined at later time points, occurring predominantly in tubular epithelial cells. A high-level CNP may contribute to the elevated expression of natriuretic peptide receptor (NPR)-B in the early phase of UUO. However, the sustained expression of NPR-C and neutral endopeptidase (NEP) observed throughout the study period (that is up to 3 months) helps to, at least partly, explain the subsequent decline of CNP. Thus, NEP and NPRs participate in the regulation of CNP expression in RIF.


Inter-protein dependences during assembly of the 50S subunit from Escherichia coli ribosomes.
Recently, we published an assembly map of the 50S ribosomal subunit from E. coli, incorporating 23S and 5S RNA and 26 out of the 32 L-proteins (Röhl and Nierhaus (1982) Proc. Natl. Acad. Sci. U.S.A., in press). In this paper we present the experimental details which led to the compilation of this 50S assembly map. The following experimental strategy was pursued: In order to elucidate binding of a protein to 23S RNA or a protein-protein interdependence, 23S rRNA and a set of purified ribosomal proteins were incubated under reconstitution conditions, the RNA-protein complex was separated from the non-bound proteins by sucrose gradient centrifugation and the proteins contained in the complex were determined by means of gel electrophoresis.


Ventilation-perfusion inequality during endotoxin-induced pulmonary vasoconstriction in conscious sheep: mechanisms of hypoxia.
In sheep low-dose endotoxin infusion causes transient thromboxane-mediated pulmonary vaso- and bronchoconstriction. These changes are paralleled by a decrease of arterial PO2. To elucidate the mechanisms of hypoxia we analysed ventilation-perfusion (VA/Q) distributions by the multiple inert gas elimination technique before and after endotoxin infusion in six conscious sheep. We found that hypoxia after endotoxin could be explained by both a VA/Q inequality and a simultaneous decrease in the mixed venous oxygen tension. The latter was due to a decrease of cardiac output. True shunt did not contribute to hypoxia as it was unchanged.


The role of antipsychotics in treating delirium.
The mainstay of the pharmacologic management of delirium remains typical antipsychotics, primarily haloperidol. Typical antipsychotics are associated with significant side effects, particularly in the elderly. This article reviews the literature on the use of both typical and atypical antipsychotics in the management of acute delirium, with a focus on the elderly. In this population, typical antipsychotics are associated with substantially more drug induce side effects--either extrapyramidal side effects or anticholinergic effects from the antipsychotics alone or in combination with benztropine or trihexyphenidyl. Anticholinergic toxicity is especially problematic in delirious, demented patients, because most dementias are associated with pre-existing deficiencies in cholinergic neurotransmission. These issues will be reviewed for typical antipsychotics as well as the emerging literature on the use of atypical antipsychotics-- risperidone, olanzapine, and quetiapine--for pharmacologic management of acute delirium. Data from two studies conducted at the Wesley Woods Center at Emory University will be briefly reviewed as they constitute the largest series to date investigating the pharmacologic management of delirious demented patients.


[Eighty cases of minimally invasive direct coronary artery bypass grafting].
Between March 1996 and November 1997, 80 patients with a mean age of 70 years (45-89) have undergone minimally invasive direct coronary artery bypass grafting via anterior minithoracotomy or subxiphoid incision with left internal thoracic artery and right epigastric artery using local coronary occlusion on a beating heart. Cardiac-related hospital mortality was 2.5% (2/80). Routine angiographic assessment of anastomotic patency showed an overall patency. rate of 94.6%, but demonstrated the severe stenosis at the anastomotic site in 8 patients. Further study is required to establish the efficacy of minimally invasive direct coronary artery bypass grafting and combination therapy with PTCA.


Sulphasalazine and Delagil--a comparative study in patients with juvenile chronic arthritis.
Thirty nine consecutive patients with pauciarticular and polyarticular juvenile chronic arthritis were randomised to treatment with either sulphasalazine or Delagil (chlorochinum diphosphoricum) in a parallel 6 month clinical trial. We compared clinical and laboratory signs of activity before and after the treatment. Of 21 patients with Sulphasalazine 10 were improved, 7 unchanged and in 4 patients had the therapy to be withdrawn. Of 18 patients with Delagil 5 were improved, 12 without effect and withdrawal was necessary in 1.


Live imaging of the Drosophila spermatogonial stem cell niche reveals novel mechanisms regulating germline stem cell output.
Adult stem cells modulate their output by varying between symmetric and asymmetric divisions, but have rarely been observed in living intact tissues. Germline stem cells (GSCs) in the Drosophila testis are anchored to somatic hub cells and were thought to exclusively undergo oriented asymmetric divisions, producing one stem cell that remains hub-anchored and one daughter cell displaced out of the stem cell-maintaining micro-environment (niche). We developed extended live imaging of the Drosophila testis niche, allowing us to track individual germline cells. Surprisingly, new wild-type GSCs are generated in the niche during steady-state tissue maintenance by a previously undetected event we term 'symmetric renewal', where interconnected GSC-daughter cell pairs swivel such that both cells contact the hub. We also captured GSCs undergoing direct differentiation by detaching from the hub. Following starvation-induced GSC loss, GSC numbers are restored by symmetric renewals. Furthermore, upon more severe (genetically induced) GSC loss, both symmetric renewal and de-differentiation (where interconnected spermatogonia fragment into pairs while moving towards then establishing contact with the hub) occur simultaneously to replenish the GSC pool. Thus, stereotypically oriented stem cell divisions are not always correlated with an asymmetric outcome in cell fate, and changes in stem cell output are governed by altered signals in response to tissue requirements.


[Venous blood flow in ocular vessels of patients with active retinopathy of prematurity].
To study the characteristics of venous circulation in the eyes with retinopathy of prematurity (ROP) observed in examination of blood flow in the central retinal vein and superior orbital vein in patients with different forms, stages and state of the disease. Color duplex scanning of the central retinal vein and superior orbital vein was performed using color Doppler imaging and pulsed Doppler sonography in 55 premature babies (55 eyes) with active ROP and 8 premature babies (8 eyes) with no signs of ROP. All babies included in the study were born at 25-32 weeks of gestation, their birth weight was 680-1760 g. A characteristic feature of hemodynamic parameters in the central retinal vein at stages 1-3 of active ROP was a decrease in both the maximum and minimum blood flow velocities (Vmax and Vmin), indicating a serious disorder of ocular hemodynamics in this category of patients. An increase in Vmax in the central retinal vein was recorded for patients with aggressive posterior ROP, which occurs in response to venous stasis that is common in this form of the disease. The performed statistical assessment of the prognostic significance of hemodynamic parameters of the central retinal vein in relation to the type of active ROP revealed a high information content of Vmin, which is promising for early detection of unfavorable course of the disease helping to ensure timely treatment. The blood flow in the superior orbital vein showed distinctive absence of any correlations with the course of the disease, disallowing any immediate conclusions on the informativeness and predictive value of its parameters. The revealed features of venous blood flow in patients with active ROP help expand the understanding of vascular changes in this pathology, and can also be applied in clinical practice to improve the accuracy of predicting the course of the disease.