Optogenetic regulation of embryo implantation in mice using photoactivatable CRISPR-Cas9.
Embryo implantation is achieved upon successful interaction between a fertilized egg and receptive endometrium and is mediated by spatiotemporal expression of implantation-associated molecules including leukemia inhibitory factor (LIF). Here we demonstrate, in mice, that LIF knockdown via a photoactivatable CRISPR-Cas9 gene editing system and illumination with a light-emitting diode can spatiotemporally disrupt fertility. This system enables dissection of spatiotemporal molecular mechanisms associated with embryo implantation and provides a therapeutic strategy for temporal control of reproductive functions in vivo.


Class II (B) general transcription factor (TFIIB) that binds to the template-committed preinitiation complex is different from general transcription factor BTF3.
A class II (B) general transcription factor of 34 kDa has been purified from HeLa cells to apparent homogeneity. This factor appears to be transcription factor IIB (TFIIB), since it binds in vitro to template-committed preinitiation complexes formed between a template containing the TATA box/cap-site elements of the adenovirus type 2 major late promoter (Ad2MLP) and recombinant human or yeast TFIID (previously called BTF1) expressed in Escherichia coli. DNase I footprint studies show an extended pattern of protection of Ad2MLP TATA box/cap-site sequences when TFIIB is bound to template-committed complexes, even though TFIIB does not bind on its own to the template in the absence of TFIID. We also show that TFIIB is different from BTF3 by a number of criteria.


Stability of coagulation factors in feline fresh frozen plasma intended for transfusion after 1 year of storage.
The most common use of plasma transfusion is for haemostatic purposes, but coagulation factor activities in stored feline plasma are unknown. The concentration and stability of coagulation factors I (fibrinogen), II, V, VII, VIII, IX, X, XI and XII in feline fresh frozen plasma (fFFP) stored for 1 year were studied. Fifty-five units of fFFP were produced from 55 fresh whole-blood donations obtained from indoor healthy blood donor cats. Twenty-one units were stored for <2 weeks (T0) and 34 were stored for 1 year (T1). After the completion of storage, specific coagulation factor activities for factors II, V, VII, VIII, IX, X, XI and XII were tested using modified one-stage activated partial thromboplastin or prothrombin time assays. Fibrinogen was determined using the Clauss method. Significantly decreased activities were observed for factors II (T0: 101.94% ± 19.06%; T1: 73.23% ± 39.06% [P = 0.001]), VII (T0: 102.78% ± 24.69%; T1: 60.08% ± 38.17% [P <0.001]), VIII (T0: 77.52% ± 30.39%; T1: 50.32% ± 23.8% [P = 0.001]), XI (T0: 88.76% ± 22.73%; T1: 66.28% ± 22.2% [P = 0.001]) and XII (T0: 89.50% ± 21.85%; T1: 55.46% ± 23.18% [P <0.001]) when comparing units at time 0 and after 1 year of storage. No significant difference was observed for factors IX (T0: 84.86% ± 29.35%; T1: 71.37% ± 22.23% [P = 0.064]) and X (T0: 96.24% ± 25.1%; T1: 83.91% ± 49.54% [P = 0.236]). Unexpectedly, a significant increase was observed for factor V (T0: 71.94% ± 24.14%; T1: 97.89% ± 62.33%; P = 0.046). Fibrinogen was 2.76 ± 1.09 g/l at T1. Factors VIII, XII and VII had the lowest mean activities after 1 year. Although a decrease in most coagulation factors activities was noted with storage, 1-year-old fFFP was haemostatically active in vitro. The most suitable factors for quality control assessment of fFFP are factors VII and VIII. Approximately 13-20 ml/kg of fFFP is required to administer a minimum of 10 IU/kg coagulation factor activity.


Elimination of schistosomiasis transmission in Zanzibar: baseline findings before the onset of a randomized intervention trial.
Gaining and sustaining control of schistosomiasis and, whenever feasible, achieving local elimination are the year 2020 targets set by the World Health Organization. In Zanzibar, various institutions and stakeholders have joined forces to eliminate urogenital schistosomiasis within 5 years. We report baseline findings before the onset of a randomized intervention trial designed to assess the differential impact of community-based praziquantel administration, snail control, and behavior change interventions. In early 2012, a baseline parasitological survey was conducted in ~20,000 people from 90 communities in Unguja and Pemba. Risk factors for schistosomiasis were assessed by administering a questionnaire to adults. In selected communities, local knowledge about schistosomiasis transmission and prevention was determined in focus group discussions and in-depths interviews. Intermediate host snails were collected and examined for shedding of cercariae. The baseline Schistosoma haematobium prevalence in school children and adults was 4.3% (range: 0-19.7%) and 2.7% (range: 0-26.5%) in Unguja, and 8.9% (range: 0-31.8%) and 5.5% (range: 0-23.4%) in Pemba, respectively. Heavy infections were detected in 15.1% and 35.6% of the positive school children in Unguja and Pemba, respectively. Males were at higher risk than females (odds ratio (OR): 1.45; 95% confidence interval (CI): 1.03-2.03). Decreasing adult age (OR: 1.04; CI: 1.02-1.06), being born in Pemba (OR: 1.48; CI: 1.02-2.13) or Tanzania (OR: 2.36; CI: 1.16-4.78), and use of freshwater (OR: 2.15; CI: 1.53-3.03) showed higher odds of infection. Community knowledge about schistosomiasis was low. Only few infected Bulinus snails were found. The relatively low S. haematobium prevalence in Zanzibar is a promising starting point for elimination. However, there is a need to improve community knowledge about disease transmission and prevention. Control measures tailored to the local context, placing particular attention to hot-spot areas, high-risk groups, and individuals, will be necessary if elimination is to be achieved.


Optimizing MSLT Specificity in Narcolepsy With Cataplexy.
Multiple sleep onset rapid eye movement (R) periods (SOREMPs) and a mean sleep latency of ≤8 minutes on the multiple sleep latency test (MSLT) are diagnostic criteria of narcolepsy (NC), but also occur in other conditions with increased sleep pressure, including insufficient sleep syndrome (ISS), sleep-disordered breathing (SDB), or Parkinson's disease (PD). These false positives are common, may create diagnostic uncertainty, and highlight the need for complementary MSLT measures with high specificity for NC. Detailed analysis of MSLT findings in 56 NC, 83 PD, 89 SDB, and 23 ISS patients, using receiver operating characteristic curves. A positive MSLT (mean sleep latency ≤ 8.0 minutes and ≥2 SOREMPs) was found in 53 NC (95%), 1 PD (1%), 8 SDB (9%), and 12 ISS patients (52%). MSLT-based differentiation between NC and non-NC patients was best when applying a mean R latency of ≤5 minutes (sensitivity/specificity/positive predictive value [PPV]: 49%/95%/96%) or a mean percentage of sleep stage R ≥ 40% (sensitivity/specificity/PPV: 60%/100%/100%) as cutoffs. When analyzing all 252 naps with SOREMPs in isolation, the combination of both R latency of ≤5 minutes and R percentage of ≥50% yielded a sensitivity/specificity/PPV of 50%/99%/99%. In addition, a sleep stage sequence with R occurring prior to N2 was more common in NC than in non-NC (71% vs. 32%, p < .001), and in combination with R percentage of ≥50% yielded a sensitivity/specificity/PPV of 53%/96%/97%. A better characterization of R sleep by latency, duration, and sleep stage sequence facilitates detection of false positives and, hence, contributes to a higher MSLT specificity in NC.


A Neurologist's View of Lyme Disease and Other Tick-Borne Infections.
Tick-borne infections-including tick-borne encephalitis viruses, represented in the United States by rare infections with Powassan and deer tick viruses, and more often Lyme disease-are of increasing importance to neurologists. Lyme neuroborreliosis (LNB) causes all or part of a triad including meningitis, radiculoneuritis, and cranial neuritis. Rarely, parenchymal brain and spinal cord involvement occur, with focal findings on examination and magnetic resonance imaging (MRI). LNB diagnosis requires plausible exposure, objective evidence of nervous system involvement, and, generally, positive two-tier serology. Central nervous system (CNS) LNB is almost always accompanied by abnormal cerebrospinal fluid (CSF) (cells, protein), often with intrathecal antibody production, which is determined by concentration-adjusted comparison of serum and CSF antibody. Measuring CSF antibody in isolation and nucleic acid-based testing of CSF are not useful in LNB and should be avoided. LNB treatment is highly effective with a 2- to 3-week course of antibiotics. Increasing evidence suggests that LNB not involving the CNS parenchyma can be treated successfully with oral doxycycline.


Different Infectivity of Mediterranean and Southern Asian Tomato Leaf Curl New Delhi Virus Isolates in Cucurbit Crops.
Tomato leaf curl New Delhi virus (ToLCNDV) became an alerting virus in Europe from 2017 to 2020 because of its significant damage to Cucurbitaceae cultivation. Until now, just some cucurbit crops including sponge gourd, melon, pumpkin, and cucumber were reported to be resistant to ToLCNDV, but no commercial cultivars are available. In this study, a new isolate of ToLCNDV was identified in Pakistan and analyzed together with ToLCNDV-ES which was previously isolated in Italy. Furthermore, infectious clones of two ToLCNDV isolates were constructed and agroinoculated into different cucurbit crops to verify their infectivity. Results showed that both isolates exhibited severe infection on all tested cucurbit (>70%) except watermelon. Thus, those cultivars may be good candidates in the first step of screening genetic resources for resistance on both Southeast Asian and Mediterranean ToLCNDV isolates. Additional, comparison pathogenicity of different geographical ToLCNDV isolates will be aided to understand viral characterization as such knowledge could facilitate breeding resistance to this virus.


Burden of hospitalization in clinically diagnosed peripheral artery disease: A community-based study.
The burden and predictors of hospitalization over time in community-based patients with peripheral artery disease (PAD) have not been established. This study evaluates the frequency, reasons and predictors of hospitalization over time in community-based patients with PAD. We assembled an inception cohort of 1798 PAD cases from Olmsted County, MN, USA (mean age 71.2 years, 44% female) from 1 January 1998 through 31 December 2011 who were followed until 2014. Two age- and sex-matched controls ( n = 3596) were identified for each case. ICD-9 codes were used to ascertain the primary reasons for hospitalization. Patients were censored at death or last follow-up. The most frequent reasons for hospitalization were non-cardiovascular: 68% of 8706 hospitalizations in cases and 78% of 8005 hospitalizations in controls. A total of 1533 (85%) cases and 2286 (64%) controls ( p < 0.001) were hospitalized at least once; 1262 (70%) cases and 1588 (44%) controls ( p < 0.001) ≥ two times. In adjusted models, age, prior hospitalization and comorbid conditions were independently associated with increased risk of recurrent hospitalizations in both groups. In cases, severe PAD (ankle-brachial index < 0.5) (HR: 1.25; 95% CI: 1.15, 1.36) and poorly compressible arteries (HR: 1.26; 95% CI: 1.16, 1.38) were each associated with increased risk for recurrent hospitalization. We demonstrate an increased rate of hospitalization in community-based patients with PAD and identify predictors of recurrent hospitalizations. These observations may inform strategies to reduce the burden of hospitalization of PAD patients.


Two-Step Approach Using Degradable Magnesium to Inhibit Surface Biofilm and Subsequently Kill Planktonic Bacteria.
Bacterial infection remains a great risk in medical implantation surgery. In this paper, we found that degradable metals may be a feasible alternative option of antibacterial implantation materials. It is known that the spalling mechanism of magnesium (Mg) during degradation leads to Mg ions-induced alkaline environment, which is harmful to planktonic bacteria. In this study, we showed that alkaline pH environment is almost harmless to those adhesive bacteria protected in well-formed biofilms. Moreover, experimental results demonstrated that the biofilm formed in the place where Mg spalls are destroyed, releasing the covered bacteria to be planktonic in the alkaline environment. As a result, the colonization of biofilms continues to shrink during the degradation of Mg. It implies that if degradable metal is employed as implantation material, even if bacterial infection occurs, it may be possibly cured without second surgery.


[A Survey on the Care of Patients with Cancer during the COVID-19 Pandemic].
The COVID-19 pandemic is expected to significantly impact cancer treatment due to the shortage of medical supply, including the anxiety of patients and their families-specifically those associated with the risk of infection. We conducted a questionnaire survey using the internet service formrun between March 1 and 31, 2021. A total of 126 hospitals and 65 patients responded to the survey. Telemedicine perception differed between hospitals and patients. Hospitals found telemedicine to be useful, whereas patients did not; the main reasons for this were inadequate examinations and consultations and difficulty in communicating symptoms. Although telemedicine is regarded as one of the most effective tools during the COVID-19 pandemic, there are still issues with its widespread application. Notably, it would be essential to collaborate with local hospitals to improve cancer treatment during this pandemic.


Characterization of neo red cells (NRCs), their function and safety in vivo tests.
The Neo Red Cells (NRCs), a new type of artificial oxygen carriers have been developed and investigated for oxygen transport capacity, hemodynamics and safety in experimental animals. Stroma free hemoglobin (Hb) prepared from outdated human red blood cells was encapsulated together with inositol hexaphosphate (as an allosteric effector), coenzyme and substrates for reducing metHb back to Hb in liposomes under an aseptic condition. The NRCs were subsequently coated with polyethylene glycol bound to phosphatidylethanolamine as surface modifier to prevent aggregation of NRCs in plasma. The metHb formation was reduced from 1%/hr to 0.37%/hr by the addition of metHb reduction system. The blood pressure increased transiently during injection, and then immediately returned to pre-injection levels. The efficiency of the NRCs in tissue oxygenation and recovery from anemia was studied in rabbits which had been made severely anemic by drawing 85% of their blood and replacing it with NRCs. All of the animals infused with NRCs recovered to pre-anemic conditions within 6-8 hr and survived until they were sacrificed, 6 months after the exchange transfusion. Our observations suggest that the NRCs are pathogen free, efficient oxygen carriers without causing serious adverse reactions, with controllable metHb formation in vivo and during storage.


'Just talking about it opens your heart': meaning-making among Black African migrants and refugees living with HIV.
Meaning-making has emerged as a core construct in addressing trauma, loss or crisis. This paper considers how diasporic Black Africans living with HIV, who come from interdependent collectivist cultures where the norm is one of implicit support, extend their meaning-making strategies when faced with a diagnosis of HIV. In this qualitative study, 13 Black African migrants and refugees living with HIV in New Zealand were interviewed and the transcripts analysed. After their diagnosis, participants began a journey of reconceptualising situational and global meaning. They extended their meaning-making strategies to include a community of like others to gain explicit support. Caregivers in host countries must understand the meaning-making processes of HIV-positive Black African migrants in order to provide competent services that lead to good social and health outcomes. All healthcare and social services workers should regularly assess Black African migrants and refugees living with HIV for positive social connectedness as well as medication adherence and more specific health concerns.


Electrospun Cellulose Nanocrystals/Chitosan/Polyvinyl Alcohol Nanofibrous Films and their Exploration to Metal Ions Adsorption.
Cellulose nanocrystals/chitosan/polyvinyl alcohol (CNC/CS/PVA) composite nanofibrous films were prepared while using an electrospinning technique and successfully thiol-functionalized. Then, the modified films were used for the sorption-desorption of Cu(II) and Pb(II) ions. Subsequently, the adsorption capacity of the films was investigated by changing the CNC loading level, solution pH, and adsorption time. Results showed that the adsorption of metal ions by the films was the best with CNC loading level of 5 wt %, pH of 6, and adsorption time of 4 h. The adsorption behavior of the films was agreed with the Freundlich model. The adsorption equation of metal ions could be described while using a pseudo-second order model. Based on the Langmuir model, the maximum adsorption capacities of Cu(II) and Pb(II) ions were estimated to be 484.06 and 323.49 mg/g, respectively. The Cu(II) and Pb(II) ions adsorption efficiencies of the films after 4 adsorption-desorption cycles were 90.58% and 90.21%, respectively. This study may provide a feasible approach for the application of functional CNC/CS/PVA nanofibrous films in the treatment of water.


Application of near-infrared reflectance spectroscopy (NIRS) to the evaluation of carotenoids content in maize.
The importance of including antioxidant compounds in the diet is well recognized. These compounds remediate the detrimental activity on animal cells of the so-called reactive oxygen substances (ROS). Many papers have reported on the determination of both hydrophilic and hydrophobic antioxidant compounds present in a large number of vegetables, and all methods involve the extraction from the matrix of the compounds to be determined. Because some problems may arise, such as the completeness of the extraction and the stability of the extracted compound during the extraction steps, the possibility of analyzing these compounds in the native matrix would be useful. Here is reported the application of near-infrared spectroscopy (NIRS) to the determination of the content of carotenoids in maize, comparing the obtained data with those derived from high-performance liquid chromatography (HPLC) determination of the extract obtained from the same samples. Equations for predicting carotenoid content in maize were derived using scores from modified partial least-squares (MPLS) as independent variables. Cross-validation procedures indicated good correlations between HPLC values and NIRS estimates. The results show that NIRS, a well-established and widely applied technique, can be applied to determine the maize carotenoids and that samples are readily analyzed in minutes, the only required step being their grinding.


Strategies for Optimizing the Production of Proteins and Peptides with Multiple Disulfide Bonds.
Bacteria can produce recombinant proteins quickly and cost effectively. However, their physiological properties limit their use for the production of proteins in their native form, especially polypeptides that are subjected to major post-translational modifications. Proteins that rely on disulfide bridges for their stability are difficult to produce in Escherichia coli. The bacterium offers the least costly, simplest, and fastest method for protein production. However, it is difficult to produce proteins with a very large size. Saccharomyces cerevisiae and Pichia pastoris are the most commonly used yeast species for protein production. At a low expense, yeasts can offer high protein yields, generate proteins with a molecular weight greater than 50 kDa, extract signal sequences, and glycosylate proteins. Both eukaryotic and prokaryotic species maintain reducing conditions in the cytoplasm. Hence, the formation of disulfide bonds is inhibited. These bonds are formed in eukaryotic cells during the export cycle, under the oxidizing conditions of the endoplasmic reticulum. Bacteria do not have an advanced subcellular space, but in the oxidizing periplasm, they exhibit both export systems and enzymatic activities directed at the formation and quality of disulfide bonds. Here, we discuss current techniques used to target eukaryotic and prokaryotic species for the generation of correctly folded proteins with disulfide bonds.


Non-neoplastic salivary gland diseases.
A wide range of non neoplastic disorders can affect the salivary glands, although the more common are: mumps, acute suppurative sialadenitis, Sjögren's syndrome and drug-induced xerostomia. Salivary dysfunction is not a normal consequence of old age, and can be due to systemic diseases, medications or head and neck radiotherapy. Diagnosis of salivary disorders begins with a careful medical history, followed by a cautious examination. While complaints of xerostomia may be indicative of a salivary gland disorder, salivary diseases can present without symptoms. Therefore, routine examination of salivary function must be part of any head, neck, and oral examination. Health-care professionals can play a vital role in identifying patients at risk for developing salivary dysfunction, and should provide appropriate preventive and interventive techniques that will help to preserving a person's health, function, and quality of life. The present work provides an overview of most of the non neoplastic disorders of the salivary glands, in which the general presentation, pathology, and treatments are discussed.


Development and validation of the Moral Sensitivity Inventory for people with intellectual disabilities.
This article presents the Moral Sensitivity Inventory, a unique reading-free tool for evaluating the moral sensitivity of people with intellectual disability. Moral sensitivity, one of the four components of Rest's Four Component Model of Morality (1994), is thought to influence moral behavior. The Moral Sensitivity Inventory is intended for people aged 16-30 years with mild or moderate intellectual disabilities. The Moral Sensitivity Inventory is comprised of 10 stories with pictures illustrating the aspects of morality, which are grouped into six categories: responsibility; respect for the common good and the property of other people; harming other people; seeking and seeing the good in others; conformance to principles and norms; understanding. The Moral Sensitivity Inventory identifies competences and gaps in moral sensitivity, which makes it a helpful tool for educating and social rehabilitation of people with intellectual disabilities. The overall reliability of the tool was .89 and the reliability of individual stories ranged from .87 to .90.


The combination of glycolic acid peels with a topical regimen in the treatment of melasma in dark-skinned patients: a comparative study.
Melasma continues to be a difficult condition to treat, especially in dark-skinned patients, although various topical modalities including hydroquinone, tretinoin, and/or topical steroids have been used singly or in combination with variable results. To determine if serial glycolic acid peels provide additional improvement when combined with a time-tested topical regimen, a modification of Kligman's formula (hydroquinone 5%, tretinoin 0.05%, hydrocortisone acetate 1% in a cream base). All cases had epidermal melasma as detected by Wood's light examination. Forty Indian melasma patients were divided into two groups of 20 each. One group received serial glycolic acid peel combined with a topical regimen, modified Kligman's formula. The other, a control group, received only modified Kligman's formula. The results were evaluated by a clinical investigator both subjectively and with photographs taken at baseline, 12 (before the fourth peel), and 21 (3 weeks after the sixth peel) weeks. For clinical evaluation, the Melasma Area and Severity Index (MASI) was used. A significant decrease in the MASI score from baseline to 21 weeks was observed in both groups (P <.001). The group receiving the glycolic acid peels showed a trend toward more rapid and greater improvement, with statistically significant results (P <.001). Only a few side effects were observed in the peel group. This study demonstrates that serial glycolic acid peels provide an additional effect to a topical regimen which is a modification of the time-tested Kligman's regimen for treating melasma in dark-complexioned individuals if used judiciously and under supervision. It demonstrates that superficial chemical peels are beneficial in the treatment of melasma.


Sex of parent and offspring in the transmission of alcoholism. A meta-analysis.
Familial studies of alcoholism were reviewed to evaluate the role of sex of parent and offspring in alcoholism transmission. Data from 32 familial alcoholism studies were evaluated by meta-analysis. The results indicated that both male and female alcoholic patients more frequently come from homes in which their father, rather than their mother, is alcoholic, even when sex differences in alcoholism prevalence rates are taken into account. Although female offspring of alcoholic mothers show alcoholism rates that are elevated relative to those expected in the general population, male offspring of alcoholic mothers do not.


ASPICov: An automated pipeline for identification of SARS-Cov2 nucleotidic variants.
ASPICov was developed to provide a rapid, reliable and complete analysis of NGS SARS-Cov2 samples to the biologist. This broad application tool allows to process samples from either capture or amplicon strategy and Illumina or Ion Torrent technology. To ensure FAIR data analysis, this Nextflow pipeline follows nf-core guidelines and use Singularity containers. Pipeline is implemented and available at https://gitlab.com/vtilloy/aspicov.


[Antibiotic use in the Hungarian hospitals in the last two decades (1996-2015)].
Growing bacterial resistance threatens public health, which can be tempered by prudent antibiotic use. To quantify systemic antibacterial use in Hungarian hospitals. Consumption data were analysed using the Anatomical-Therapeutic-Chemical - Defined Daily Dose (ATC/DDD) methodology. Data were standardized for patient turnover and also for population to enable international benchmarking. Hospital antibiotic use was quite constant (22.4 ± 1.5 DDD/100 patient-days), but its composition changed substantially. The use of parenteral products rose gradually (in 1996 26.4% and in 2015 41.6%). The pattern of use was homogenised due to the headway of co-amoxiclav use. A substantial increase of fluoroquinolone (2.3 vs. 4.2 DDD/100 patient-days) and third generation cephalosporin (1.0 vs. 2.9 DDD/100 patient-days) use was detected. In parallel the use of narrow spectra penicillins diminished. Hungarian hospital antibiotic use is low. The causes and the justification of this low use together with the internationally outstanding use of certain antibacterials should be addressed in future studies. Orv. Hetil., 2016, 157(46), 1839-1846.


Folding on the assembly line.
Deciphering the mechanism of folding of newly synthesized proteins in the cell is a major challenge because of the large size and multiplicity of molecular components involved and the asynchrony of biosynthesis. Fluorescently labeled ribosome-bound nascent chains of a defined length were prepared and subjected to dynamic fluorescence depolarization spectroscopy measurements. Nanosecond anisotropy decay correlation times of proteins' nascent chains at different stages of polypeptide elongation were determined for the first time. Striking dependence of the chain dynamics on the stages of elongation was observed and revealed chain length dependence of folding on the ribosome.


Update: Complications and care related to pregnancy, labor and delivery, active component, U.S. Armed Forces, 2001-2010.
Diagnoses of "complications of pregnancy, childbirth, and the puerperium" (ICD-9-CM: 630-679) include both morbid complications and indications for routine care of pregnant women. During 2001-2010, U.S. service members with live births (n=156,314) had 1.16 million medical encounters with primary diagnoses of "pregnancy complications". The most frequent diagnoses were "other current conditions", "other threatened labor" (e.g., false labor), "previous cesarean delivery" and "normal delivery". The number of complication-related medical encounters per pregnancy more than doubled during 2001-2010. Diagnoses of gestational diabetes, previous cesarean delivery and advanced maternal age contributed to the largest increases in pregnancy-related medical encounters during the period. The complications that accounted for the largest increases in the percents of pregnancies affected by them were urinary tract infections, fetal heart rate/rhythm abnormalities and back pain in pregnancy.


Caffeine effects on behavioral thermoregulation.
Caffeine has a wide range of behaviorally active properties. Varying doses of caffeine solutions were administered (ip) prior to fixed-interval 2-min. schedules of microwave reinforcement in rats tested in a cold environment. Four Sprague-Dawley rats were conditioned to regulate their thermal environment with 5-sec. exposures of microwave reinforcement. Graphic descriptions showed that small doses of caffeine produced higher response rates for microwave heat than high doses of caffeine and saline controls, yet Friedman's nonparametric test showed no significant differences between dose levels. Synergism between thermogenic and discriminative properties of caffeine is proposed.


Extrapolation, interpolation, and identification of spots in Hartmann patterns.
Based on Fourier analysis, we propose a simple method to extrapolate spots outside an aperture that bounds a Hartmann pattern, and interpolate spots in those regions where some spots are lost or damaged. It allows us to smoothen or remove the discontinuities of the wavefront slopes at the border of the aperture or in those inner regions where some spots are lost. The method changes the spot pattern by a fringe pattern. The extrapolated or interpolated spots are obtained from the intersection of the fringe maxima. The fringe pattern is also used to identify the spots, and it is particularly useful in highly distorted Hartmann patterns. Experimental results show how the method works.


[Assessment of respiratory function in patients with chronic obstructive pulmonary diseases treated with budesonide].
The aim of the study was to carry out assessment of the respiratory function of patients with COPD undergoing a three month (600 microgram) treatment with an inhaled steroid--Budesonide. This was a double blind study carried out on 57 patients. During treatment with Budesonide a clinical improvement was seen. Respiratory function parameters did not alter throughout the 3 month treatment period, although bronchial reactivity diminished. Side effects were seen only in 3 patients--in all it was dysphonia. It seems that Budesonide can be added to other treatment protocols in patients with exacerbations of chronic bronchitis.


"Yin and Yang" tuned fluorescence sensing behavior of branched 1,4-bis(phenylethynyl)benzene.
Achieving high sensing performance and good photostability of fluorescent films based on adlayer construction represents a significant challenge in the area of functional fluorescent film research. A solution may be offered by "Yin and Yang", a balance idea from Chinese philosophy, for the design of a fluorophore and the relevant assembly. Accordingly, a 1,4-bis(phenylethynyl)benzene (BPEB) derivative (C2) with two cholesteryl residues in the side chains and two glucono units in the head and tail positions was designed and synthesized. As a control, compound C1 was also prepared. The only difference between C1 and C2 is that the hydroxyl groups in the glucono residues of C1 are fully acetylated. Studies of the fluorescence behaviors of the two compounds in solution revealed that both the profile and the intensity of the fluorescence emission of the compounds, in particular C2, are dependent on their concentration and on the nature of solvents employed. Presence of HCl also alters the emission of the compounds in solution. On the basis of the studies, three fluorescent films were prepared, and their sensing performances to HCl in vapor state were studied. Specifically, Film 1 and Film 3 were fabricated via physical coating, separately, of C2 and C1 on glass plate surfaces. As another comparison, Film 2 was also fabricated with C2 as a fluorophore but at a much lower concentration if compared to that for the preparation of Film 1. As revealed by SEM and fluorescent microscopy studies, Film 1 and Film 2 exhibit well-defined microstructures, which are spherical particles and spherical pores, respectively, while Film 3 is characterized by irregular aggregates of C1. Fluorescence measurements demonstrated that Film 1 and Film 3 both display an aggregation emission, of which the emission from Film 1 is supersensitive to the presence of HCl vapor (detection limit: 0.4 ppb, a lowest value reported in the literatures). For Film 3, however, its emission is insensitive to the presence of the vapor. Similarly, the emission from the nonaggregated state of C2, a characteristic emission of Film 2, is also insensitive to the presence of the vapor. Furthermore, the sensing process of Film 1 to the vapor is highly selective and fully reversible, which lays foundation for its real-life uses. As for C2, the results from solution studies and those from film studies demonstrate clearly that introduction of auxiliary structures with opposite properties onto a typical fluorophore is a good strategy to develop fluorescent supramolecular motifs with rich assembly properties and great potential of applications.


Zinc Oxide and Copper Chitosan Composite Films with Antimicrobial Activity.
The role of the oral microbiome and its effect on dental diseases is gaining interest. Therefore, it has been sought to decrease the bacterial load to fight oral cavity diseases. In this study, composite materials based on chitosan, chitosan crosslinked with glutaraldehyde, chitosan with zinc oxide particles, and chitosan with copper nanoparticles were prepared in the form of thin films, to evaluate a new alternative with a more significant impact on the oral cavity bacteria. The chemical structures and physical properties of the films were characterized using by Fourier transform infrared spectroscopy (FTIR,) Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), elemental analysis (EDX), thermogravimetric analysis (TGA), X-ray diffraction (XRD), scanning electron microscopy (SEM), and contact angle measurements. Subsequently, the antimicrobial activity of each material was evaluated by agar diffusion tests. No differences were found in the hydrophilicity of the films with the incorporation of ZnO or copper particles. Antimicrobial activity was found against S. aureus in the chitosan film crosslinked with glutaraldehyde, but not in the other compositions. In contrast antimicrobial activity against S. typhimurium was found in all films. Based on the data of present investigation, chitosan composite films could be an option for the control of microorganisms with potential applications in various fields, such as medical and food industry.


Radiotherapy-centered multimodal treatment of unresectable pancreatic carcinoma.
Multimodal treatment procedures, including intraoperative and external beam radiotherapy, chemotherapy and hyperthermotherapy, used for treatment of unresectable pancreatic carcinoma for the past two years have been described. Among the ten progressive cases where multimodal treatment was applied, marked reduction in tumor mass was observed in three cases. The cases receiving such treatment reported prolonged survival, the median survival period being 250 days as compared with 85.4 days in the non-multimodal group. The conclusion is that optimal palliative effects can be achieved by sustained application of both intraoperative and postoperative radiotherapy, hyperthermia and other techniques of multimodal therapy in cases of unresectable pancreatic carcinoma.


Protective effects of simultaneous splenectomy on small-for-size liver graft injury in rat liver transplantation.
Splenectomy is an effective technique in living donor liver transplantation (LDLT) with small-for-size (SFS) liver grafts for overcoming SFS liver graft injury. However, the protective mechanism of splenectomy is still unclear. The aim of this study was to investigate how splenectomy could attenuate SFS graft injury through the measurement of biochemical factors, particularly the expression of endothelin (ET)-1, which is a key molecule of microcirculatory disorders by mediating sinusoidal vasoconstriction. We performed rat orthotopic liver transplantation using SFS liver grafts with or without splenectomy. We investigated intragraft expression of ET-1 mRNA and hepatic protein levels of ET-1. In addition, portal pressure, hepatic injury and morphological changes, and survival rate were evaluated. In result, intragraft ET-1 mRNA expression after SFS liver transplantation was significantly downregulated by splenectomy, and hepatic expression of ET-1 in SFS grafts was rarely observed. Splenectomy inhibited the increase in portal pressure, ameliorated SFS liver graft injury and improved the graft survival rate after SFS liver transplantation. In conclusion, splenectomy improved the SFS liver injury and decreased the expression of ET-1 by attenuating portal hypertension on SFS liver transplantation. Downregulation of intragraft ET-1 expression plays important roles in the protective mechanism of splenectomy in SFS liver transplantation.


Transposable element profiles reveal cell line identity and loss of heterozygosity in Drosophila cell culture.
Cell culture systems allow key insights into biological mechanisms yet suffer from irreproducible outcomes in part because of cross-contamination or mislabeling of cell lines. Cell line misidentification can be mitigated by the use of genotyping protocols, which have been developed for human cell lines but are lacking for many important model species. Here, we leverage the classical observation that transposable elements (TEs) proliferate in cultured Drosophila cells to demonstrate that genome-wide TE insertion profiles can reveal the identity and provenance of Drosophila cell lines. We identify multiple cases where TE profiles clarify the origin of Drosophila cell lines (Sg4, mbn2, and OSS_E) relative to published reports, and also provide evidence that insertions from only a subset of long-terminal repeat retrotransposon families are necessary to mark Drosophila cell line identity. We also develop a new bioinformatics approach to detect TE insertions and estimate intra-sample allele frequencies in legacy whole-genome sequencing data (called ngs_te_mapper2), which revealed loss of heterozygosity as a mechanism shaping the unique TE profiles that identify Drosophila cell lines. Our work contributes to the general understanding of the forces impacting metazoan genomes as they evolve in cell culture and paves the way for high-throughput protocols that use TE insertions to authenticate cell lines in Drosophila and other organisms.


The Iterative Reweighted Mixed-Norm Estimate for Spatio-Temporal MEG/EEG Source Reconstruction.
Source imaging based on magnetoencephalography (MEG) and electroencephalography (EEG) allows for the non-invasive analysis of brain activity with high temporal and good spatial resolution. As the bioelectromagnetic inverse problem is ill-posed, constraints are required. For the analysis of evoked brain activity, spatial sparsity of the neuronal activation is a common assumption. It is often taken into account using convex constraints based on the l1-norm. The resulting source estimates are however biased in amplitude and often suboptimal in terms of source selection due to high correlations in the forward model. In this work, we demonstrate that an inverse solver based on a block-separable penalty with a Frobenius norm per block and a l0.5-quasinorm over blocks addresses both of these issues. For solving the resulting non-convex optimization problem, we propose the iterative reweighted Mixed Norm Estimate (irMxNE), an optimization scheme based on iterative reweighted convex surrogate optimization problems, which are solved efficiently using a block coordinate descent scheme and an active set strategy. We compare the proposed sparse imaging method to the dSPM and the RAP-MUSIC approach based on two MEG data sets. We provide empirical evidence based on simulations and analysis of MEG data that the proposed method improves on the standard Mixed Norm Estimate (MxNE) in terms of amplitude bias, support recovery, and stability.


Linking shifts in bacterial community with changes in dissolved organic matter pool in a tropical lake.
Bacterioplankton communities have a pivotal role in the global carbon cycle. Still the interaction between microbial community and dissolved organic matter (DOM) in freshwater ecosystems remains poorly understood. Here, we report results from a 12-day mesocosm study performed in the epilimnion of a tropical lake, in which inorganic nutrients and allochthonous DOM were supplemented under full light and shading. Although the production of autochthonous DOM triggered by nutrient addition was the dominant driver of changes in bacterial community structure, temporal covariations between DOM optical proxies and bacterial community structure revealed a strong influence of community shifts on DOM fate. Community shifts were coupled to a successional stepwise alteration of the DOM pool, with different fractions being selectively consumed by specific taxa. Typical freshwater clades as Limnohabitans and Sporichthyaceae were associated with consumption of low molecular weight carbon, whereas Gammaproteobacteria and Flavobacteria utilized higher molecular weight carbon, indicating differences in DOM preference among clades. Importantly, Verrucomicrobiaceae were important in the turnover of freshly produced autochthonous DOM, ultimately affecting light availability and dissolved organic carbon concentrations. Our findings suggest that taxonomically defined bacterial assemblages play definite roles when influencing DOM fate, either by changing specific fractions of the DOM pool or by regulating light availability and DOC levels.


Promotion of children's early psychosocial development through primary health care services.
The paper presents the study design of the development and organisation of an EU/WHO multicentre study on the promotion of children's early psychosocial development through primary health care services. The aim of the programme is to develop training for primary health care workers (PHCWs) that improves their ability to assess factors relevant to psychosocial development and gives them preventive approaches to foster that development, building on families' strengths. The focus is on the period from conception to the end of the second year of the child's' life. Two main components are the training of the PHCWs and the semi-structured interviews that guide the workers in practice. The training has content and structure that is feasible within primary health care resources working in collaboration with mental health services. The programme is evaluated in terms of (a) the impact of training on both the PHCWs' knowledge and attitudes with regard to infant behaviour and development, and on practice as assessed by the delivery of the semi-structured interview; and (b) the effect of the intervention on maternal well-being, infant language development and behavior, home environment and mother-infant interaction. The programme is taking place in Cyprus, Greece, the Federal Republic of Yugoslavia, Portugal, Slovenia and Turkey.


Design, fabrication, and performance of enhanced magneto-optic quadrilayers with controllable ellipticity.
An outline is given of a technique for designing quadrilayers that enhance the magneto-optic polar Kerr effect by maximizing the signal-to-noise ratio associated with readout systems utilizing differential detection. The method is particularly novel in that it produces maximized Kerr rotations with controllable ellipticities by using a systematic design procedure that does not involve computer-driven iterative techniques that search for optimum film structures. A number of quadrilayers, based on the ZnS-TbFeCo-ZnS-Al system, have been prepared using rf and dc sputtering and conventional evaporation techniques. The performance of the devices in terms of the basic optical and magneto-optical functions is good compared with theoretical predictions, both at the design wavelength and at all other regions of the spectrum, in the range 400-800 nm.


A Comparison of Pigment-Protein Complexes among Normal, Chlorophyll-Deficient and Senescent Soybean Genotypes.
Pigment-protein complexes were isolated from chloroplasts of normal green and several types of chlorophyll-deficient soybeans. The complexes were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and comparisons were made between normal and chlorophyll-deficient genotypes of the relative amounts of chlorophyll associated with Photosystem I (PSI), Photosystem II (PSII), light-harvesting, and free pigment complexes.Chlorophyll-deficient genotypes, compared to normal green genotypes, have fewer light-harvesting complexes and a higher ratio of PSII to PSI complexes. Chlorophyll associated with PSII in yellow genotypes is in relatively higher amounts in spite of the fact that these genotypes have much less grana stacking than normal green genotypes. Although PSII activity has been associated with appressed regions of grana in normal plants, our work shows that the association does not always hold true.


Syk kinase-coupled C-type lectin receptors engage protein kinase C-δ to elicit Card9 adaptor-mediated innate immunity.
C-type lectin receptors (CLRs) that couple with the kinase Syk are major pattern recognition receptors for the activation of innate immunity and host defense. CLRs recognize fungi and other forms of microbial or sterile danger, and they induce inflammatory responses through the adaptor protein Card9. The mechanisms relaying CLR proximal signals to the core Card9 module are unknown. Here we demonstrated that protein kinase C-δ (PKCδ) was activated upon Dectin-1-Syk signaling, mediated phosphorylation of Card9 at Thr231, and was responsible for Card9-Bcl10 complex assembly and canonical NF-κB control. Prkcd(-/-) dendritic cells, but not those lacking PKCα, PKCβ, or PKCθ, were defective in innate responses to Dectin-1, Dectin-2, or Mincle stimulation. Moreover, Candida albicans-induced cytokine production was blocked in Prkcd(-/-) cells, and Prkcd(-/-) mice were highly susceptible to fungal infection. Thus, PKCδ is an essential link between Syk activation and Card9 signaling for CLR-mediated innate immunity and host protection.


Beneficial effect of phosphodiesterase-5 inhibitor in experimental inflammatory bowel disease; molecular evidence for involvement of oxidative stress.
ABSTRACT Inflammatory bowel disease (IBD) is a common and chronic gastrointestinal disorder characterized by intestinal inflammation and mucosal tissue damage. Reactive oxygen metabolites (ROMs) play a pathogenic role in IBD. We aimed to examine the protective effect of sildenafil, a cGMP phosphodiesterase inhibitor, in the experimental mouse model of IBD. Intrarectal instillation of acetic acid was used to induce IBD. Prednisolone was used as the standard drug for comparison. Sildenafil was used at doses of 0.75, 1.5, and 3 mg/kg. Biochemicals and macroscopic and microscopic examinations of colonic tissue were performed. Results indicated that activity of myeloperoxidase (MPO) and lipid peroxidation product (TBARS) markers of oxidative stress are increased in acetic acid-treated groups and are recovered by sildenafil pretreatment and prednisolone. Sildenafil- (1.5 and 3 mg/kg) and prednisolone-treated groups showed significantly lower score values of macroscopic and microscopic characters when compared to the acetic acid-treated group. The beneficial effect of sildenafil (3 mg/kg) was comparable to that of prednisolone. It is concluded that sildenafil is helpful in the management of IBD, which is presumably related to its strong antioxidative stress potential mediated through enhanced cGMP. Results of proper clinical trials will determine the possible efficacy of phosphodiesterase-5 inhibitors in human IBD.


Simultaneous detection of multiple DNA targets by integrating dual-color graphene quantum dot nanoprobes and carbon nanotubes.
Simultaneous detection of multiple DNA targets was achieved based on a biocompatible graphene quantum dots (GQDs) and carbon nanotubes (CNTs) platform through spontaneous assembly between dual-color GQD-based probes and CNTs and subsequently self-recognition between DNA probes and targets.


Sex differences in material-specific cognitive functions related to language dominance: an intracarotid amobarbital study in left temporal lobe epilepsy.
Population-based studies of material-specific cognitive functions yield evidence of sex differences: women are superior on verbal tasks whereas men are superior on figural and visuo-spatial tasks. Although there is still no direct evidence, these sex differences have been assumed to be related to a different cerebral organisation of language and nonlanguage functions, i.e. language in women is suggested to be more bilaterally organised than in men. We investigated this issue with respect to verbal/nonverbal memory functions in 42 men and 43 women with left temporal lobe epilepsies who all underwent intracarotid amobarbital testing for language dominance. The results indicate that atypical language dominance is not significantly more frequent in women than in men (44% vs 33%). Atypical dominance was related to an onset of epilepsy before puberty and appeared to be a consequence of the left hemisphere pathology rather than being naturally predisposed. Furthermore, women indeed showed the expected advantage with respect to verbal memory, and men showed an advantage in figural memory. With consideration of language dominance, however, the sex difference in verbal memory was observed particularly with atypical dominance, whereas the sex difference in figural memory was observed with left dominance.


Role of the endogenous nitric oxide inhibitor asymmetric dimethylarginine (ADMA) and brain-derived neurotrophic factor (BDNF) in depression and behavioural changes: clinical and preclinical data in chronic kidney disease.
Patients with chronic kidney disease (CKD) exhibit a high prevalence of neuropsychiatric alterations, including depression and behavioural changes. CKD is also associated with decreased physical activity not fully explained by co-morbidities. In patients without CKD, the brain-derived neurotropic factor (BDNF) as well as the endogenous NOS inhibitor asymmetric dimethylarginine (ADMA) had been suspected to be involved in major depression. The aim of our study was to examine the role of ADMA and BDNF in the behaviour of haemodialysis patients (CKD5D) as well as in a rat model of 5/6 nephrectomy and chronic ADMA infusion alone. Eleven (5F/6M) CKD5D patients underwent Beck Depression Inventory (BDI) testing along with analysis of ADMA and BDNF. Male Sprague-Dawley rats were randomly assigned to four groups: (i) saline infusion; (ii) ADMA (250 µg/kg/day) infusion via osmotic mini pumps; (iii) 5/6 nephrectomy; (iv) untreated controls. After 28 days, the animals underwent behavioural tests measuring anxiety, locomotion and investigative behaviour. Animals were sacrificed, blood samples were drawn and analysed and hippocampal immunohistology for BDNF was performed. In CKD5D patients, decreased BDNF levels correlated with higher scores of depression (Pearson r = -0.8156, P = 0.002). ADMA infusion led to a significant decrease of BDNF while the decrease of BDNF in 5/6 nephrectomy was not significant. However, an attenuated hippocampal BDNF expression could be detected in 5/6 nephrecomized animals. Decreased spontaneous locomotor activity was shown in ADMA-infused rats [15.9 (13.5-26.1) lines crossed/min] and 5/6 nephrectomy [14.6 (6.1-20.2) lines crossed/min] when compared with controls [32.5 (15.3-42.4) lines crossed/min]. Anxiety-like behaviour tested by hole investigation time was significantly more pronounced in 5/6 nephrectomy [24 (6-44) s] when compared with ADMA infusion [64 (28-93) s] and controls [33 (26-65) s]. Progressive renal failure in rats is accompanied by a marked increase of ADMA and a decrease in BDNF. 5/6 nephrectomy leads to significantly decreased exploratory behaviour and locomotion. Both behaviours could be reproduced by ADMA infusion alone. Indicators of anxiety were more pronounced in ADMA-infused animals when compared with 5/6 nephrectomized rats. Furthermore, an inverse relationship of BDNF and BDI in 11 CKD5D patients was shown.


Down-regulation of c-Myc following MEK/ERK inhibition halts the expression of malignant phenotype in rhabdomyosarcoma and in non muscle-derived human tumors.
Expression of c-myc proto-oncogene is inappropriate in a wide range of human tumors, and is a downstream target of Ras/Raf/ERK pathway, which promotes c-Myc stability by enhancing c-Myc expression and activity. The aim of this study was to investigate whether the oncogenic phenotype in the human muscle-derived Rhabdomyosarcoma (RD) cell line and in non muscle-derived human tumor cell lines (SW403, IGR39 and PC3) can be blocked by disrupting the c-Myc pathway either by means of pharmacological MEK/ERK inhibition or by direct inactivation of the c-Myc protein. We demonstrate that, in all the tumor cell lines used, the MEK/ERK inhibitor U0126 rapidly induces c-Myc de-phosphorylation, which is followed by a marked reduction in its expression level, by inhibition of proliferation and by reversion of anchorage-independent growth. These data suggest that the targeting of pathways controlling c-Myc expression or stability reverses deregulated growth of different tumor-derived cell lines. Indeed, in RD cells, we found a marked down-regulation of cyclins E2, A and B and CDK2, all of which are known to be targets of c-Myc. Moreover, ectopic MadMyc chimera, a c-Myc function antagonist, causes dramatic growth arrest, CDK and cyclin modulation as well as inhibition of anchorage-independent growth in RD cells, as occurs in U0126-treated cells. In particular, we found that the mere inhibition of c-Myc by MadMyc chimera rescues the myogenic program, MHC expression and the acquisition of the myogenic-like phenotype in RD cells. Our data provide evidence of the key role played by the MEK/ERK pathway in the growth arrest and transformation phenotype of Rhabdomyosarcoma and of non muscle-derived tumor cell lines. In fact, MEK/ERK inhibitor, U0126, induces growth arrest, anchorage-dependent growth of these cell lines. In addition, the results of this study demonstrate that the direct inactivation of c-Myc by Mad/Myc chimera rescues myogenic program and leads to the reversal of the Rhabdomyosarcoma phenotype. In conclusion these data strongly suggest that the targeting of c-Myc by means of the MEK inhibitor can be tested as a promising strategy in anti-cancer therapy.


Accurate and Robust Calibration of the Uniform Affine Transformation Between Scan-Camera Coordinates for Atom-Resolved In-Focus 4D-STEM Datasets.
Accurate geometrical calibration between the scan coordinates and the camera coordinates is critical in four-dimensional scanning transmission electron microscopy (4D-STEM) for both quantitative imaging and ptychographic reconstructions. For atomic-resolved, in-focus 4D-STEM datasets, we propose a hybrid method incorporating two sub-routines, namely a J-matrix method and a Fourier method, which can calibrate the uniform affine transformation between the scan-camera coordinates using raw data, without a priori knowledge of the crystal structure of the specimen. The hybrid method is found robust against scan distortions and residual probe aberrations. It is also effective even when defects are present in the specimen, or the specimen becomes relatively thick. We will demonstrate that a successful geometrical calibration with the hybrid method will lead to a more reliable recovery of both the specimen and the electron probe in a ptychographic reconstruction. We will also show that, although the elimination of local scan position errors still requires an iterative approach, the rate of convergence can be improved, and the residual errors can be further reduced if the hybrid method can be firstly applied for initial calibration. The code is made available as a simple-to-use tool to correct affine transformations of the scan-camera coordinates in 4D-STEM experiments.


[Comparison of restoring and maintaining the cervical curvature and height by using three different anterior cervical plate systems].
To investigate and compare the effects of restoring and maintaining the cervical curvature and height of the fused segment by using three different anterior cervical plate systems. From January 2002 to June 2004, 122 patients underwent anterior cervical decompression, autogenous iliac bone graft and plate fixation. Of the 122 patients (85 males, 37 females, aged 14-70), 37 underwent surgery involving the fixation with the Orion plate system, 39 with the Zephir plate system, and 46 with the Codman plate system. The cervical curvature and height of the fused segment were measured on the lateral X-ray films so as to compare the changes of the conditions preoperatively, 1 week after surgery, and during the follow-up, and also to compare the difference among the three groups. The follow-up of the patients for 6-35 months (average 17.3 months) showed that all the patients developed the bone fusion 6 months after operation. There was a significant improvement in the cervical curvature and height of the fused segment before operation versus 1 week after operation (P < 0.05); however, there were no significant changes 1 week after operation versus during the follow-up in each group(P > 0.05); there was no significant difference among the three groups (P > 0.05). The three plate systems can effectively reconstruct and maintain the cervical curvature and height of the fused segment, with a satisfactory effect in a short term.


NASBA isothermal enzymatic in vitro nucleic acid amplification optimized for the diagnosis of HIV-1 infection.
Isothermal nucleic acid amplification of target RNA or DNA sequences is accomplished by the simultaneous enzymatic activity of AMV reverse transcriptase, T7 RNA polymerase and RNase H. Amplification factors of the nucleic acid sequence based amplification (NASBA) method range from 2 x 10(6) to 5 x 10(7) after 2.5 h incubation at 41 degrees C. During NASBA there is a major accumulation of specific single stranded RNA. RNA:DNA hybrid and double stranded DNA are also synthesized, although to a minor extent. The system is optimized for the detection of HIV-1 sequences in in vitro infected cells, blood and plasma. Detection levels are 10 molecules of HIV-1 in a model system with in vitro generated HIV-1 RNA as input and 5 infected cells on a background of 5 x 10(4) non-infected cells. Blood and plasma can also be used as the source of nucleic acid for detection of HIV-1 sequences using a specifically developed sample preparation method. Using NASBA it is possible to amplify specifically RNA or DNA from a pool of total nucleic acid, which permits the investigation of the expression of specific genes involved in pathogenesis of infectious agents. The combination of NASBA with a rapid and user-friendly nucleic acid extraction method makes the whole procedure suitable for large scale diagnosis of infectious agents (e.g. HIV-1).


Anticoagulation, Bleeding, Mortality, and Pathology in Hospitalized Patients With COVID-19.
Thromboembolic disease is common in coronavirus disease-2019 (COVID-19). There is limited evidence on the association of in-hospital anticoagulation (AC) with outcomes and postmortem findings. The purpose of this study was to examine association of AC with in-hospital outcomes and describe thromboembolic findings on autopsies. This retrospective analysis examined the association of AC with mortality, intubation, and major bleeding. Subanalyses were also conducted on the association of therapeutic versus prophylactic AC initiated ≤48 h from admission. Thromboembolic disease was contextualized by premortem AC among consecutive autopsies. Among 4,389 patients, median age was 65 years with 44% women. Compared with no AC (n = 1,530; 34.9%), therapeutic AC (n = 900; 20.5%) and prophylactic AC (n = 1,959; 44.6%) were associated with lower in-hospital mortality (adjusted hazard ratio [aHR]: 0.53; 95% confidence interval [CI]: 0.45 to 0.62 and aHR: 0.50; 95% CI: 0.45 to 0.57, respectively), and intubation (aHR: 0.69; 95% CI: 0.51 to 0.94 and aHR: 0.72; 95% CI: 0.58 to 0.89, respectively). When initiated ≤48 h from admission, there was no statistically significant difference between therapeutic (n = 766) versus prophylactic AC (n = 1,860) (aHR: 0.86; 95% CI: 0.73 to 1.02; p = 0.08). Overall, 89 patients (2%) had major bleeding adjudicated by clinician review, with 27 of 900 (3.0%) on therapeutic, 33 of 1,959 (1.7%) on prophylactic, and 29 of 1,530 (1.9%) on no AC. Of 26 autopsies, 11 (42%) had thromboembolic disease not clinically suspected and 3 of 11 (27%) were on therapeutic AC. AC was associated with lower mortality and intubation among hospitalized COVID-19 patients. Compared with prophylactic AC, therapeutic AC was associated with lower mortality, although not statistically significant. Autopsies revealed frequent thromboembolic disease. These data may inform trials to determine optimal AC regimens.


A multi-center clinical trial of a new chairside test in distinguishing between diseased and healthy periodontal sites. II. Association between site type and test outcome before and after therapy.
The aim of the present study was to evaluate the association between the outcome of a chairside test measuring gingival crevicular fluid (GCF) levels of the enzyme aspartate aminotransferase (AST) and other clinical measures of disease including probing depth, severity of inflammation, and GCF flow before and after therapy. We studied 91 patients with moderate to severe periodontitis. Eight sites with probing depths between 5 mm and 8 mm and obvious signs of inflammation were selected and designated diseased sites. Four sites with probing depth < or = 3 mm with no or minimal signs of inflammation were selected and designated non-diseased sites in patients. Thirty healthy individuals were enrolled and four sites in each were selected and designated healthy controls. Patients were treated with scaling and root planing and control subjects with supragingival prophylaxis. Measurements including GCF volume, gingival inflammation, and probing depth were performed at screening baseline, 1 week later at pretreatment baseline, and at weeks 2 and 4 after treatment. AST content of GCF was measured using a chairside colorometric test. It was concluded that the outcome of the test is an effective objective measure distinguishing between diseased sites and non-diseased sites in patients and control subjects when evaluated both prior to and following application of therapy. Use of this simple chairside test, when combined with other standard diagnostic procedures, provides an objective measurement permitting improved capacity to distinguish between diseased and non-diseased periodontal sites, and to better assess and monitor the outcome of therapy.


Multiple fistulas and tracheobronchial stenoses require extensive stenting of the central airways and esophagus in squamous-cell carcinoma.
We report here on two cases of double airway and esophageal stenting in patients with multiple esophagotracheal fistulas and stenoses of the airways and esophagus due to squamous-cell carcinoma. Dumon stents and a Strecker device were used for tracheobronchial stenting. Covered Gianturco Z-stents were implanted into the esophagus. In one case, extrinsic compression of the trachea and tumor progression required recanalization by Nd:YAG laser. Clinical improvement led to discharge of the patients within two weeks after the procedure. The results show that simultaneous implantation of stents in the central airways and covered Gianturco Z-stents in the esophagus is an effective therapeutic strategy in patients with tracheal and esophageal obstructions and esophagorespiratory fistulas. Further systematic studies evaluating double stenting are warranted.


Requirement of ONE-HELIX PROTEIN 1 (OHP1) in early Arabidopsis seedling development and under high light intensity.
Plant ONE-HELIX PROTEINS (OHPs) are part of the light-harvesting complex superfamily whose members are involved in various processes related to sensing and capturing light as well as light protection. We recently showed the requirement of a functional OHP1-OHP2 heterodimer for efficient D1 synthesis. Interestingly, while the ohp1 knockout mutant showed a strong defect in accumulation of the photosystem II and is hardly viable, virus-induced gene silencing of OHP1 had no detectable impact on plant growth and performance under standard growth conditions. However, in vivo labeling assays with 35S-methionine indicate a reduced D1 synthesis rate. Here, we show that VIGS-OHP1 plants are more susceptible towards elevated light intensities than control plants. This underlines an obligatory function of OHP1 for light acclimation.


Nanocrystalline diamond films grown in nitrogen rich atmosphere: structural and field emission properties.
The field emission behaviour of a series of nanocrystalline N-doped diamond films has been investigated and interpreted on the basis of the structural and compositional characteristics of the layers. The diamond films, formed by crystallites with grain size in the range 20-100 nm were produced from CH4/H2 mixtures using a HF-CVD apparatus. Nitrogen was added to the gaseous reactants in form of both N2 and of Urea. Micro-Raman spectroscopy and cathodoluminescence have been used to define the structure of the deposits on a nanometric scale. The field emission measurements have been carried out under a pressure of 10(-6) mbar using a sphere-to-plane anode-cathode configuration. The characteristics of the emission from the various nanodiamond samples and from different regions of the same sample are discussed in terms of field threshold, current density, current stability.


Bombesin potentiates taurocholic acid-induced neurotensin release in rats.
The hypothesis of synergistic effects between luminal stimulants and the intramural neural network in the control of intestinal neurotensin (NT) release was investigated with an isolated vascularly perfused rat jejuno-ileum. Luminal administration of low doses of taurocholic acid (TC; 1, 5, and 10 mM) provoked only a small increase in NT-like immunoreactivity (NT-LI) in the portal effluent (50%, 100%, and 130%, respectively, above basal). Increasing the concentration of TC to 20 mM induced a strong and sustained release of NT-LI (700% above basal). The arterial infusion of bombesin (BOM; 10(-9) M) induced only a transient rise in NT-LI levels (200% above basal), with a rapid return to basal values. In contrast, 10(-9) M BOM synergistically enhanced NT-LI responses induced by 10 mM TC; the integrated response of NT-LI release was 3.2-fold higher than the sum of responses to 10(-9) M BOM and 10 mM TC alone. These cooperative effects were apparent with 10(-10) M BOM and TC concentrations over the physiological range 1-10 mM. Neither tetrodotoxin (10(-6) M) nor atropine (10(-5) M) was able to modify this synergy. Substance-P and methacholine, two other NT secretagogues, did not potentiate the TC-induced NT responses. In conclusion, BOM potentiated the release of NT induced by TC, thus suggesting that bombesinergic neurons, among other stimulatory neurons of the enteric nervous system, may modulate the sensitivity of N-cells to luminal stimulants.


Dual-wavelength photoacoustic atlas method to estimate fractional methylene blue and hemoglobin contents.
Methylene blue (MB) is an exogenous contrast agent that has the potential to assist with visualization and penetration challenges in photoacoustic imaging. However, monitoring the local concentration between MB and endogenous chromophores is critical for avoiding unnecessary MB accumulations that could lead to adverse effects such as hemolysis when exposed to increased dose and photodamage when exposed to high laser energies. We developed a modified version of a previously proposed acoustic-based atlas method to estimate concentration levels from a mixture of two photoacoustic-sensitive materials after two laser wavelength emissions. Photoacoustic data were acquired from mixtures of 100-μM MB and either human or porcine blood (Hb) injected in a plastisol phantom, using laser wavelengths of 710 and 870 nm. An algorithm to perform linear regression of the acoustic frequency response from an atlas composed of pure concentrations was designed to assess the concentration levels from photoacoustic samples obtained from 11 known MB/Hb volume mixtures. The mean absolute error (MAE), coefficient of determination (i.e., R2), and Spearman's correlation coefficient (i.e., ρ) between the estimated results and ground-truth labels were calculated to assess the algorithm performance, linearity, and monotonicity, respectively. The overall MAE, R2, and ρ were 12.68%, 0.80, and 0.89, respectively, for the human Hb dataset and 9.92%, 0.86, and 0.93, respectively, for the porcine Hb dataset. In addition, a similarly linear relationship was observed between the acoustic frequency response at 2.3 MHz and 870-nm laser wavelength and the ground-truth concentrations, with R2 and | ρ | values of 0.76 and 0.88, respectively. Contrast agent concentration monitoring is feasible with the proposed approach. The potential for minimal data acquisition times with only two wavelength emissions is advantageous toward real-time implementation in the operating room.


Relationship between the goniometric alignment and articular cartilage damage in knee osteoarthritis
Osteoarthritis of the knee is common among the elderly. The alignment of the knee plays an important part in disease progression. The aim was to determine the relationship between goniometric measurement of malalignment and gross and histological damage to articular cartilages of osteoarthritic knees. Anterior plane knee alignment was measured using a handheld metal goniometer in 84 patients awaiting knee replacement. Articular cartilages removed at surgery were stained and examined for macroscopic and microscopic damage which were quantified with macroscopic visual assessment score and Mankins histochemical score, respectively. The relationship between the type of knee angulation and macroscopic and microscopic damage to medial and lateral compartments was analyzed using analysis of variance (ANOVA), and the relationship between the degree of angulation and the degree of damage was determined using multiple linear regression models. Varus knees had markedly higher macroscopic damage scores in the medial than the lateralcompartment (p<0.0001), while valgus knees showed the opposite (p<0.0001). The knee angulation, together with age and BMI, predicted 54% of the macroscopic (p<0.0001) and 60% of the microscopic damage (p<0.0001) in varus knees. Anterior plane goniometric knee alignment is a significant predictor of the articular cartilage damage present in an osteoarthritic knee. Goniometric measurement is a valuable tool in assessing the disease severity as it is a simple, safe and cost-effective method.


[Pharmacologic treatment of hypoxemia in adult respiratory distress syndrome].
New drugs that improve arterial oxygenation (nitric oxide, almitrine, inhaled prostacyclin and cyclooxygenase inhibitors) are useful in the treatment of severe hypoxemia unresponsive to conventional treatment that is mainly seen in patients with acute respiratory distress syndrome (ARDS). By acting selectively on pulmonary blood flow and redistributing it, these drugs achieve effects unattainable until now. Thus, they decrease perfusion in non-ventilated zones (V/Q = O) responsible for shunt, or increase perfusion in well ventilated zones, guaranteeing adequate oxygenation. To apply these drugs the physician must understand their mechanism of action, guidelines for dosing, constraints on their use and side effects.


Variable selection and interpretation in structure-affinity correlation modeling of estrogen receptor binders.
A computational approach for the identification and investigation of correlations between a chemical structure and a selected biological property is described. It is based on a set of 132 compounds of known chemical structures, which were tested for their binding affinities to the estrogen receptor. Different multivariate modeling methods, i.e., partial least-squares regression, counterpropagation neural network, and error-back-propagation neural network, were applied, and the prediction ability of each model was tested in order to compare the results of the obtained models. To reduce the extensive set of calculated structural descriptors, two types of variable selection methods were applied, depending on the modeling approach used. In particular, the final partial least-squares regression model was built using the "variable importance in projection" variable selection method, while genetic algorithms were applied in neural network modeling to select the optimal set of descriptors. A thorough statistical study of the variables selected by genetic algorithms is shown. The results were assessed with the aim to get insight to the mechanisms involved in the binding of estrogenic compounds to the receptor. The variable selection on the basis of genetic algorithm was controlled with the test set of compounds, extracted from the data set available. To compare the predictive ability of all the optimized models, a leave-one-out cross-validation procedure was applied, the best model being the nonlinear neural network model based on error back-propagation algorithm, which resulted in R2= 92.2% and Q2= 70.8%.


New approach to the reduction of recombination in dye-sensitised solar cells via complexation of oxidised species.
Although dye-sensitised solar cells (DSSCs) have received great attention as low-cost and clean energy conversion devices, their conversion efficiency still lags behind that of inorganic solar cells. One of the reasons is due to the recombination of injected electrons with the oxidised species of the redox couple that are present in the electrolyte near the surface of the bare TiO2 particles. While most research has focused on blocking the bare surface of the TiO2 nanoparticles, we have introduced a new approach that can directly reduce the concentration of the oxidised species of the redox couple present in the electrolytes through complex formation. Recombination was reduced by the addition of cyclodextrins (CDs) to a polyethyleneglycol dimethyl ether (PEGDME) electrolyte containing iodide/triiodide redox couples that can form a complex with the triiodide. Experimental and theoretical investigation of the complex formation between triiodide and the CD in PEGDME matrix was performed. Increase in total power conversion efficiency was achieved using the alpha-CD as an additive in a low volatile PEGDME based electrolyte. Electrochemical impedance spectra and intensity modulated photovoltage spectroscopy measurements showed that the increase in the short-circuit current density is due to the suppression of surface recombination by the complex formation between the CDs and the triiodide ions.


Wild Boars as Reservoir of Highly Virulent Clone of Hybrid Shiga Toxigenic and Enterotoxigenic Escherichia coli Responsible for Edema Disease, France.
Edema disease is an often fatal enterotoxemia caused by specific strains of Shiga toxin-producing Escherichia coli (STEC) that affect primarily healthy, rapidly growing nursery pigs. Recently, outbreaks of edema disease have also emerged in France in wild boars. Analysis of STEC strains isolated from wild boars during 2013-2019 showed that they belonged to the serotype O139:H1 and were positive for both Stx2e and F18 fimbriae. However, in contrast to classical STEC O139:H1 strains circulating in pigs, they also possessed enterotoxin genes sta1 and stb, typical of enterotoxigenic E. coli. In addition, the strains contained a unique accessory genome composition and did not harbor antimicrobial-resistance genes, in contrast to domestic pig isolates. These data thus reveal that the emergence of edema disease in wild boars was caused by atypical hybrid of STEC and enterotoxigenic E. coli O139:H1, which so far has been restricted to the wildlife environment.


[Sleep disorders and panic attacks].
The aim of this work is to describe the clinical and polisomnographical characteristics of patients suffering prevailing night panic attacks. The most usual therapeutics are also briefly mentioned. The Polisomnography and especially the EGG are mentioned as the methods which allow the most accurate diagnosis since the night panic attack produces patognomonical disturbances.


Effects of month and age on prolactin concentrations in stallion serum.
Prolactin concentrations in stallion serum were measured by a newly developed radioimmunoassay based on anti-dog prolactin serum and radiolabelled horse prolactin. Samples of serum from a total of 444 stallions were obtained at a commercial abattoir monthly from April to the following March. Ages of stallions were estimated from eruption and wear patterns of incisors. In the analysis of variance, both month (P less than 0.01) and age (P less than 0.05) were significant sources of variation whereas there was no interaction between these factors. Monthly means for prolactin concentrations were greatest between May and August with peak concentrations occurring in July (7.1 +/- 0.5 ng/ml). Lowest mean concentrations of prolactin (1.2 ng/ml) were observed during the winter months (December and January). Concentrations of prolactin were lowest in stallions less than or equal to 5 years old and were highest in stallions greater than 10 years old. It appears that prolactin secretion in the stallion is strongly influenced by season, as it is in the mare and in other seasonally breeding animals. Moreover, prolactin secretion in the stallion tends to increase as the stallion ages.


Influence of Temperature on the Mechanical Properties and Reactive Behavior of Al-PTFE under Quasi-Static Compression.
Al-PTFE (aluminum-polytetrafluoroethylene) is a typical kind of Reactive Material (RM), which has a variety of potential applications in weapon systems. In this paper, quasi-static compression experiments were carried out for a pressed and sintered mixture of Al and PTFE powders using a microcomputer-controlled electronic universal testing machine. The results show that both the mechanical property and reactive behavior of Al-PTFE are strongly temperature-dependent. The material undergoes a brittle-ductile transition associated with a temperature-induced crystalline phase transformation of the PTFE matrix. At low temperatures (-18, 0, and 16 °C), samples of Al-PTFE failed with shear crack and no reaction was observed. As the temperature increased (22, 35, and 80 °C), Al-PTFE exhibited a high toughness and violent reaction occurred in all of the tested samples. Scanning electron microscope observations showed different fracture mechanisms of the PTFE matrix and the increase in toughness was due to the formation of PTFE fibrils which could dissipate energy and bridge crack plane during plastic deformation.


Carotid artery: prospective blinded comparison of two-dimensional time-of-flight MR angiography with conventional angiography and duplex US.
A prospective blinded comparison of two-dimensional (2D) time-of-flight (TF) magnetic resonance (MR) angiography and color duplex flow ultrasound (US) with conventional angiography as a standard of reference was performed in 50 patients with hemispheric ischemic symptoms. The guidelines of the North American Symptomatic Carotid Endarterectomy Trial for measuring stenosis of the internal carotid artery were utilized for maximum-intensity-projection (MIP) images and conventional angiograms. While the 2D TF MIP technique overrepresented the degree of stenosis in the internal carotid arteries, it matched the performance of color duplex flow US as measured by means of receiver operating characteristic curves. A signal void on the MIP images corresponded to a 70% or greater internal carotid artery stenosis in 17 of 20 arteries. At its current state of development, the accuracy of 2D TF MR angiography equals that of US in characterizing the degree of carotid stenosis but cannot be considered a replacement for conventional angiography.


Magnetic resonance imaging of hepatocellular carcinoma.
Hepatocellular carcinoma (HCC), the most common primary hepatic malignancy, usually develops in patients with cirrhosis, growing sequentially from low-grade dysplastic nodules to frank malignant HCC. Its recognition is critical because curative treatment and prognosis require early diagnosis. Survival in patients with HCC relates directly to the number, size, and extent of lesions at diagnosis. Imaging of HCC is complicated because the tumor has a varied imaging appearance and frequently coexists with other cirrhotic nodules. Magnetic resonance imaging (MRI), the best available diagnostic technique, offers good contrast resolution and diagnostic sensitivity ranging from 33% to 77%. The main difficulty is not in diagnosing large tumors, but rather small tumors (<2 cm), because of considerable overlap on imaging between benign (regenerative), borderline (dysplastic), and malignant nodules. Increasing degrees of histological malignancy are associated with increasing arterialization and loss of portal blood supply; therefore, recognition of HCC requires dynamic imaging with gadolinium-enhanced T1-weighted sequence. Typically, HCC is a focal lesion with high signal intensity on T2-weighted images, variable signal intensity on T1-weighted images, intense arterial phase enhancement after gadolinium injection, and isointensity or hypointensity at the portal venous phase. The sensitivity of MRI for detecting small lesions is low, and improvement is still needed. Newer contrast agents, higher field strength (3 Tesla) imaging, and perfusion and diffusion MRI techniques possibly will provide greater sensitivity and specificity for detecting small HCCs in the future.


Ethnicity and Health in Colombia: What Do Self-Perceived Health Indicators Tell Us?
To compare self-perceived health indicators between ethnic groups in Colombia. Cross-sectional study with data from the 2007 National Public Health Survey (ENSP-2007). Data from 57,617 people ≥18 years were used. Variables included: belonging to an ethnic group (exposure); self-rated health; mental health problems, injuries for accidents/violence (outcomes); sex, age, education level and occupation (explicative/control). A descriptive study was carried out of the explicative variables, and the prevalence of the outcomes was calculated according to ethnicity, education level and occupation. The association between the exposure variable and the outcomes was estimated by means of adjusted odds ratios (OR) with 95% CI using logistic regression. Analyses were conducted separately for men and women. The prevalence of outcomes was higher in people reporting to belong to an ethnic group and differences were found by sex, ethnic groups and health outcomes. Women from the Palenquero group were more likely to report poor self-rated health (aOR 7.04; 95%CI 2.50-19.88) and injuries from accidents/violence (aOR 7.99; 95%CI 2.89-22.07). Indigenous men were more likely to report mental health problems (aOR 1.75; 95%CI 1.41-2.17). Gradients according to ethnicity, education, occupation and sex were found. Minority ethnic groups are vulnerable to reporting poor health outcomes. Political actions are required to diminish health inequalities in these groups.


Simultaneous mapping of membrane voltage and calcium in zebrafish heart in vivo reveals chamber-specific developmental transitions in ionic currents.
The cardiac action potential (AP) and the consequent cytosolic Ca(2+) transient are key indicators of cardiac function. Natural developmental processes, as well as many drugs and pathologies change the waveform, propagation, or variability (between cells or over time) of these parameters. Here we apply a genetically encoded dual-function calcium and voltage reporter (CaViar) to study the development of the zebrafish heart in vivo between 1.5 and 4 days post fertilization (dpf). We developed a high-sensitivity spinning disk confocal microscope and associated software for simultaneous three-dimensional optical mapping of voltage and calcium. We produced a transgenic zebrafish line expressing CaViar under control of the heart-specific cmlc2 promoter, and applied ion channel blockers at a series of developmental stages to map the maturation of the action potential in vivo. Early in development, the AP initiated via a calcium current through L-type calcium channels. Between 90 and 102 h post fertilization (hpf), the ventricular AP switched to a sodium-driven upswing, while the atrial AP remained calcium driven. In the adult zebrafish heart, a sodium current drives the AP in both the atrium and ventricle. Simultaneous voltage and calcium imaging with genetically encoded reporters provides a new approach for monitoring cardiac development, and the effects of drugs on cardiac function.


Defective in vitro growth of primitive hematopoietic cells from pediatric patients with acute myeloid leukemia.
Acute myeloid leukemia (AML) is a neoplastic hematologic disorder that arises at the level of a primitive stem/progenitor cell. Most studies on the biology of the hematopoietic system in AML have focused on cells from adult patients; much less is known about hematopoietic cells from childhood AML. By using a negative immunoselection system, we have obtained a primitive cell population (enriched for CD34(+) Lin(-) cells) from the bone marrow (BM) of 17 pediatric AML patients and characterized its proliferation, expansion, and differentiation potentials in liquid cultures supplemented with a mixture of 8 different recombinant stimulatory cytokines. The proportion of CD34(+) cells in AML patients was extremely heterogeneous, ranging from 0% to 74%. Regardless of their CD34(+) cell content, and in contrast to normal cells, AML cells showed a deficient capacity to proliferate even in the presence of the stimulatory cytokines. AML progenitors were unable to generate new progenitor cells, indicating their inability to expand. Interestingly, AML cells were able to differentiate in culture, giving rise to morphologically recognizable precursors. A major difference, however, as compared to hematopoietic progenitors from normal subjects, was the fact that whereas in cultures of normal cells both myeloid and erythroid precursors were produced, in AML cultures the vast majority of the cells generated corresponded to myeloid cells, mostly mature macrophages. As compared to their normal counterparts, primitive hematopoietic cells from pediatric patients with AML possess impaired proliferation, expansion, and differentiation potentials in vitro.


Overexpression of HIV-1 proteins in Escherichia coli by a modified expression vector and their one-step purification.
A synthetic expression system, pMH, was constructed for high-level expression and rapid purification of HIV-1 proteins in Escherichia coli, by introducing two synthetic sequences. The first sequence permitted a high-level expression via a second ribosome-binding site plus an A/T-rich region. This minicistron between the 5' untranslated region of the mRNA and the protein-coding region minimized the formation of local secondary structures and favored ribosome binding. The second region coded for six histidines, which allowed for easy purification of the proteins. Purification was based on interaction of the protein with metal ions: immobilized metal affinity chromatography. The p17 and p24 structural genes of HIV-1 were cloned into this modified expression vector. Expression of the structural proteins represented up to 30% of the total protein in E. coli, i.e., 40 mg of highly purified protein could be obtained per liter of culture.


In Vitro Model of Human Skeletal Muscle Tissues with Contractility Fabricated by Immortalized Human Myogenic Cells.
In the development process for drugs used to treat skeletal muscle, cell-based contractile force assays have been considered as a useful in vitro test. Immortalized human myogenic cells are promising as cell sources for reproducible and well-characterized in vitro models. In this study, it is investigated whether immortalized human myogenic cells, Hu5/KD3, have suitable contractile ability and the potential to be used as cell sources for contractile force assays. Muscle tissues are fabricated using Hu5/KD3 cells on the microfabricated devices used to measure contractile force. The tissues generate a tetanic force of ≈30 µN in response to the electrical pulse stimulation (EPS). Gene expression analysis of the myosin heavy chain (MYH) isoform indicates that the tissues mostly consisted of muscle fibers expressing MYH7 or/and MYH8. The addition of dexamethasone or lovastatin decreases the contractile force of the tissues, indicating that the tissues have the potential to evaluate drug candidates designed to treat muscle atrophy or statin-induced myopathy. It is also demonstrated that the contractile force of tissues increased when EPS is applied as an artificial exercise. These results indicate that the Hu5/KD3 tissues can be employed for contractile force assays and would be useful for in vitro human skeletal muscle models.


Cloned animal products in the human food chain: FDA should protect American consumers.
Animal cloning is "complex process that lets one exactly copy the genetic, or inherited, traits of an animal." In 1997, Dolly the sheep was the first animal cloned and since then "scientists have used animal cloning to breed dairy cows, beef cattle, poultry, hogs and other species of livestock." Cloned animals are highly attractive to livestock breeders because "cloning essentially produces an identical copy of an animal with superior traits." The main purpose of cloning livestock is "more focused on efficiency and economic benefits of the producer rather than the overall effect of cloning on an animal's physical and mental welfare." The focus of this article is threefold. First, the science behind animal cloning is explained and some potential uses and risks of this technology are explored. Second, FDA's historical evolution, current regulatory authority, and limitations of that authority, is described. Lastly, a new regulatory vision recognizes the realities of 21st century global markets and the dynamic evolution of scientific discovery and technology.


The effect of nordihydroguaiaretic acid, an inhibitor of prostaglandin and leukotriene biosynthesis, on hematopoiesis of gamma-irradiated mice.
The effects of the inhibition of the cyclooxygenase and lipoxygenase metabolic pathways of arachidonic acid on the postirradiation recovery of hematopoietic functions in mice were investigated. Nordihydroguaiaretic acid (NDGA), an inhibitor of prostaglandin (PG) and leukotriene (LT) production, was given to animals in single doses (0.015 to 0.75 mg/mouse) 1 hour before 5 Gy of total-body gamma-irradiation. Enhanced hematopoietic recovery in terms of exogenous and endogenous spleen colonies, femoral granulocyte-macrophage colony-forming cells and peripheral blood granulocyte levels was observed at higher doses of NDGA. The treatment used influenced neither lymphocyte nor erythrocyte postirradiational levels or hemoglobin concentration. A comparison of the effects induced by a high dose of NDGA (0.3 mg per mouse) with those observed after an isomolar dose of indomethacin (an inhibitor of PG production) indicated only slight differences between these two drugs. An isomolar dose of esculetin (an inhibitor of LT production) had no effect on the postirradiation behavior of hematopoiesis. The results suggest that the inhibition of PG production plays the main role in the mechanism of NDGA action. Inhibition of LT production seems to be of less importance for hematopoiesis in these in vivo conditions.


Beta-blockade benefits patients following a subarachnoid haemorrhage.
Previous studies have shown that ECG changes following a subarachnoid haemorrhage are associated with increased catecholamine levels, necrotic myocardial lesions, and a poor prognosis. Furthermore, beta-blockade using propranolol reverses some of the ECG changes and prevents necrotic myocardial lesions. This study was established to assess the affects of adrenergic blockade on morbidity and mortality following subarachnoid haemorrhage. Patients were admitted to the randomized double-blind between-patients study if they presented at the neurosurgical unit within 48 hours of a subarachnoid haemorrhage confirmed by lumbar puncture. Of 224 patients, the first 118 received an alpha-blocker, phentolamine 20 mg three-hourly, and either the beta-blocker propranolol 80 mg eight-hourly, or placebo. The last 106 patients received either propranolol or placebo. Treatment was continued for three weeks. Assessment at four weeks revealed significant improvements in the treated group for neurological deficit (p = 0.003) and death (p = 0.02). More treated patients underwent operation and those that did had a better outcome (p = 0.01). Assessment at one year showed that although patients had improved in both groups, patients in the treated group had significantly fewer neurological deficits (p = 0.003). There were fewer deaths in the treated group but this difference was not significant (p = 0.09). Possible mechanisms for this protective effect of propranolol may include a reduction in plasma renin activity, a reduction in pulmonary oedema, prevention of myocardial infarcts, and a reduction in cerebral oxygen requirements. It is concluded that early beta-blockade benefits patients with subarachnoid haemorrhage, in terms of fewer neurological deficits, for up to one year.


Support groups as open systems: a model for practice and research.
Despite the increasing of use of support groups to respond to the needs of people dealing with crises, life transitions, and chronic conditions, there is no conceptual framework to guide practice and research. On the basis of a review of the literature, this article distinguishes support groups from self-help and treatment groups. It then proposes an open systems model for understanding the factors that affect support groups, for guiding interventions, and for evaluating their outcomes. Four conceptual dimensions are described: (1) environmental conditions (resources and constraints), (2) participant group characteristics (size, composition, and technology), (3) group conditions (goals, structural form, and development), and (4) outcomes (positive and negative effects for members, leaders, and organizations; group problems; and ethical and legal issues). Findings from a pilot study of support group practice based on this framework are presented and discussed.


Molecular markers for corneal epithelial cells in larval vs. adult Xenopus frogs.
Corneal Epithelial Stem Cells (CESCs) and their proliferative progeny, the Transit Amplifying Cells (TACs), are responsible for maintaining the integrity and transparency of the cornea. These stem cells (SCs) are widely used in corneal transplants and ocular surface reconstruction. Molecular markers are essential to identify, isolate and enrich for these cells, yet no definitive CESC marker has been established. An extensive literature survey shows variability in the expression of putative CESC markers among vertebrates; being attributed to species-specific variations, or other differences in developmental stages of these animals, approaches used in these studies and marker specificity. Here, we expanded the search for CESC markers using the amphibian model Xenopus laevis. In previous studies we found that long-term label retaining cells (suggestive of CESCs and TACs) are present throughout the larval basal corneal epithelium. In adult frogs, these cells become concentrated in the peripheral cornea (limbal region). Here, we used immunofluorescence to characterize the expression of nine proteins in the corneas of both Xenopus larvae and adults (post-metamorphic). We found that localization of some markers change between larval and adult stages. Markers such as p63, Keratin 19, and β1-integrin are restricted to basal corneal epithelial cells of the larvae. After metamorphosis their expression is found in basal and intermediate layer cells of the adult frog corneal epithelium. Another protein, Pax6 was expressed in the larval corneas, but surprisingly it was not detected in the adult corneal epithelium. For the first time we report that Tcf7l2 can be used as a marker to differentiate cornea vs. skin in frogs. Tcf7l2 is present only in the frog skin, which differs from reports indicating that the protein is expressed in the human cornea. Furthermore, we identified the transition between the inner, and the outer surface of the adult frog eyelid as a key boundary in terms of marker expression. Although these markers are useful to identify different regions and cellular layers of the frog corneal epithelium, none is unique to CESCs or TACs. Our results confirm that there is no single conserved CESC marker in vertebrates. This molecular characterization of the Xenopus cornea facilitates its use as a vertebrate model to understand the functions of key proteins in corneal homeostasis and wound repair.