HIV treatment as prevention: population effect vs. individual protection?
There are compelling data suggesting that antiretroviral (ARV) drugs can prevent the transmission of HIV: 1) ARV therapy reduces the intrinsic risk of transmission by cutting down the viral load in infected people; 2) ARVs may be used to prevent transmission in uninfected subjects who experienced high risk exposures. Despite these encouraging preliminary results, there are several questions that need to be answered before recommending the use of ARV among uninfected subjects: 1) the efficacy of these strategies amongst women is not yet established; 2) the cost-effectiveness of pre-exposure prophylaxis (PrEP) in the long term is undefined since PrEP efficacy may depend on adherence; 3) treating uninfected individuals may be counter-effective, as it may increase risky behavior due to the belief that ARV drugs have unlimited protective power. In conclusion, the public-health use of ARV drugs represents a new paradigm in the field of HIV prevention. However, ARV treatment cannot be considered as a substitute for other measures for preventing transmission but a complement to individual protection. In particular, post-exposure prophylaxis may be used in selected cases, whereas PrEP appears to be a highly costly method with limited public-health potential and possible safety concerns.


Lateral extension of π conjugation along the bay regions of bisanthene through a Diels-Alder cycloaddition reaction.
Diels-Alder cycloaddition reactions at the bay regions of bisanthene (1) with dienophiles such as 1,4-naphthoquinone have been investigated. The products were submitted to nucleophilic addition followed by reductive aromatization reactions to afford the laterally extended bisanthene derivatives 2 and 3. Attempted synthesis of a larger expanded bisanthene 4 revealed an unexpected hydrogenation reaction at the last reductive aromatization step. Unusual Michael addition was observed on quinone 14, which was obtained by Diels-Alder reaction between 1 and 1,4-anthraquinone. Compounds 1-3 exhibited near-infrared (NIR) absorption and emission with high-to-moderate fluorescent quantum yields. Their structures and absorption spectra were studied by density function theory and non-planar twisted structures were calculated for 2 and 3. All compounds showed amphoteric redox behavior with multiple oxidation/reduction waves. Oxidative titration with SbCl(5) gave stable radical cations, and the process was followed by UV/Vis/NIR spectroscopic measurements. Their photostability was measured and correlated to their different geometries and electronic structures.


The psychiatric physical examination - Part I: methodology.
The frequency of medical findings in psychiatric patients is quite high. Regardless, none of the responding psychiatrists in a recent survey do routine physical examinations. In response to this survey, the authors describe a method of rapid physical examination specifically designed for the psychiatric patient. The average duration of the examination in 75 patients was 16.9 minutes. There was no evidence of negative transference or countertransference. The level of competency required was that of a medical student, yet there were 8.2 physical findings per patient with 5.3 physical findings per patient being unknown previously.


Topical application of 17beta-estradiol increases extracellular matrix protein synthesis by stimulating tgf-Beta signaling in aged human skin in vivo.
To investigate the effects of topically applied 17beta-estradiol on the expression of extracellular matrix proteins in aged human skin, 17beta-estradiol (0.01%) and its vehicle (70% propylene glycol, 30% ethanol) were applied to aged (68-82 y, eight females and five males) human buttock skin under occlusion for 2 wk (three times per week). Topical 17beta-estradiol was found to increase the expression of type 1 procollagen mRNA and protein significantly in human aged skin in vivo. In addition, metalloproteinase (MMP-1 protein levels were reduced by topical 17beta-estradiol. The expressions of TGF-beta1, TGF-beta type II receptor, and Sma and Mad related (Smad)3 were increased by topical 17 beta-estradiol in aged human skin, and TGF-beta1 neutralizing antibody inhibited 17beta-estradiol-induced procollagen synthesis in cultured fibroblasts. We also found that the expressions of tropoelastin and fibrillin-1 mRNA and protein, and elastic fibers in aged skin were also increased by topical 17beta-estradiol. Topical 17beta-estradiol also increased keratinocyte proliferation and the epidermal thickness in aged human skin. We also observed the same effects of topical 17beta-estradiol in young skin. In conclusion, our results suggest that topical 17beta-estradiol treatment may improve the cutaneous function of aged human skin by improving the connective tissue and increasing epidermal thickness.


Plain radiography does not add any clinically significant advantage to multidetector row computed tomography in diagnosing cervical spine injuries in blunt trauma patients.
Cervical spine (c-spine) injuries (CSI) in trauma patients are common and potentially catastrophic. Numerous guidelines involving clinical and radiologic criteria have been devised to diagnose such injuries. It is not clear whether using plain X-ray films in addition to helical computed tomography (CT) provides any additional benefit in trauma patients who require radiologic clearance of their c-spine. We hypothesized that three standard X-ray views (anteroposterior, lateral, and odontoid) of the c-spine do not provide clinically significant advantage to Multidetector row CT in diagnosing CSI in trauma patients. We reviewed the charts of consecutive adult patients with CSI who were admitted to the Trauma Service at a Level I Trauma Center between January 2001 and December 2004. Patients who had CT plus X-ray at admission were entered into the study. Exclusion criteria were age < or = 16 years, incomplete radiology reports, inadequate plain films, or no CSI identified. A total of 121 patients with diagnosed CSI were included in the study. CT picked up 100% of patients who had a CSI diagnosed on plain films and also detected 47 additional CSI that were missed by plain films. The sensitivity for CT was 100%, whereas that of plain films was 61%. Nine patients with CSI (19.1%) who had false-negative plain films required operative intervention. Three standard X-ray views of the c-spine provided no clinically significant advantage to Multidetector row CT in diagnosing CSI. Revision of current clinical guidelines on c-spine clearance is recommended.


Management of corneal epithelial defects in a population of mature chuck-will's-widows (Antrostomus carolinensis) in South Florida.
To describe ocular clinical findings, gross/histopathologic findings, and treatment regimens in a series of migratory chuck-will's-widows (Antrostomus carolinensis) (CWW) with corneal epithelial defects. Seven CWW were presented to the South Florida Wildlife Center (SFWC). Four presented with bilateral (OU) corneal ulceration; two developed corneal ulceration OU; one had no ocular lesions. Treatment protocols for patients with corneal ulcers included the following: medical therapy only or medical therapy combined with an additional procedure. Four patients including the bird with no ocular lesions were euthanized, and one patient died. Their globes were submitted for histopathology. Two patients were released. Clinical findings prior to enucleation included superficial corneal ulceration with redundant epithelium persisting weeks to >1 month. On histopathology, epithelium in nonulcerated globes was remarkably thin; this was considered normal. Common histopathologic findings of ulcerated globes revealed epithelial and conjunctival attenuation with an acellular superficial stromal layer and hypercellular mid-stromal layer. One globe healed with medical therapy and cotton tip applicator debridement. Four globes healed by combination of medical therapy, equine amnion, nictitating membrane (NM) flap, and temporary tarsorrhaphy. No globes healed with diamond burr debridement or grid keratotomy. Factors that may be contributing to these corneal epithelial defects include, but are not limited to, normally thin epithelium, exposure keratopathy, neurotrophic disease, epithelial turnover and inadequate stem cell recruitment, inherited/genetic causes, and unidentified infectious agents (eg, viral etiologies). Of the 12 eyes treated, one healed with medical therapy/cotton tip applicator debridement, and four healed with medical therapy/equine amnion/nictitating membrane flap/temporary tarsorrhaphy.


The Multi-Strain Probiotic OMNi-BiOTiC® Active Reduces the Duration of Acute Upper Respiratory Disease in Older People: A Double-Blind, Randomised, Controlled Clinical Trial.
Immunosenescence is the adverse change in the human immune function during aging, leaving older people more prone to an increased risk of infections and morbidity. Acute upper respiratory tract infections (URTIs) are very common among older people, often resulting in continued morbidity and mortality. Therefore, approaches, such as consuming probiotics, that shorten the duration or even reduce the incidence of URTIs in older people are being studied. The aim of this study was to determine the effects of a multi-strain probiotic OMNi-BiOTiC® Active, which contains 11 live probiotic strains, on the incidence, duration, and severity of URTIs in older people. In this randomized double-blinded placebo-controlled study, 95 participants, with an average age of 70.9 years in the probiotic group and 69.6 years in the placebo group, were randomly allocated to two groups: 1010 cfu per day of the multi-strain probiotic intervention OMNi-BiOTiC® Active (49) or placebo (46). The incidence of URTIs in older people after 12 weeks supplementation with OMNi-BiOTiC® showed no statistically significant difference between the two groups (p = 0.5244). However, the duration of the URTI infections was statistically significantly different between the groups (p = 0.011). The participants that consumed the probiotic had an average duration of illness of 3.1 ± 1.6 days, whilst participants that received the placebo had symptoms for an average of 6.0 ± 3.8 days (p = 0.011). Statistically significant differences in lymphocyte counts in both groups after supplementation (p = 0.035 for the probiotic group and p = 0.029 for the placebo group) and between both groups were found (p = 0.009). Statistically significant differences in eosinophil (p = 0.002) and basophil counts (p = 0.001) in the probiotic groups before and after supplementation with probiotics were also found. Supplementation with the multi-strain probiotic OMNi-BiOTiC® Active may benefit older people with URTIs. Larger randomised controlled clinical trials are warranted. Clinical Trial Registration; identifier NCT05879393.


Well-Being and Loneliness in Swiss Older Adults During the COVID-19 Pandemic: The Role of Social Relationships.
The current coronavirus disease 2019 (COVID-19) pandemic and social distancing measures are an extreme stressor that might result in negative emotional experiences and feelings of loneliness. However, it is possible that social relationships might have a protective effect. In the present study, we examine how the COVID-19 pandemic affected older adults' well-being and loneliness, and the role of structural and functional characteristics of social relationships. We use data from 99 older adults in Switzerland who participated (a) in a 3-week microlongitudinal study on social relationships and well-being in 2019 and (b) in a weekly online survey during 4 weeks of the COVID-19 lockdown. Our findings show that the global pandemic had substantial adverse effects on older adults' emotional well-being and loneliness. In addition, aspects of social relationships were related to loneliness both before and during the pandemic. Only one functional feature of social relationships (satisfaction with communication during the pandemic) buffered adverse effects of the major stressful event. Although the social distancing measures during COVID-19 presented a major stressor for older adults' well-being and loneliness, being able to maintain social communication to a satisfactory level during that time reduced this effect. Therefore, enabling older adults to stay in touch with their social circle based on their personal preferences might reduce the impact that any future lockdown might have on their well-being.


Robotic Hartmann's reversal-feasibility and technical aspects.
Hartmann's reversal as a staged procedure after emergency surgery is a major abdominal operation with undeniable skill-demanding steps. The robotic approach and its advantages seem to be a safe and feasible technique and could overcome necessary technical compromises of laparoscopy, ensuring a significant reduction in restoration failures or conversion.


An electronic family health history tool to identify and manage patients at increased risk for colorectal cancer: protocol for a randomized controlled trial.
Colorectal cancer is the fourth most commonly diagnosed cancer in the United States. Approximately 3-10% of the population has an increased risk for colorectal cancer due to family history and warrants more frequent or intensive screening. Yet, < 50% of that high-risk population receives guideline-concordant care. Systematic collection of family health history and decision support may improve guideline-concordant screening for patients at increased risk of colorectal cancer. We seek to test the effectiveness of a web-based, systematic family health history collection tool and decision support platform (MeTree) to improve risk assessment and appropriate management of colorectal cancer risk among patients in the Department of Veterans Affairs primary care practices. In this ongoing randomized controlled trial, primary care providers at the Durham Veterans Affairs Health Care System and the Madison VA Medical Center are randomized to immediate intervention or wait-list control. Veterans are eligible if assigned to enrolled providers, have an upcoming primary care appointment, and have no conditions that would place them at increased risk for colorectal cancer (such as personal history, adenomatous polyps, or inflammatory bowel disease). Those with a recent lower endoscopy (e.g. colonoscopy, sigmoidoscopy) are excluded. Immediate intervention patients put their family health history information into a web-based platform, MeTree, which provides both patient- and provider-facing decision support reports. Wait-list control patients access MeTree 12 months post-consent. The primary outcome is the risk-concordant colorectal cancer screening referral rate obtained via chart review. Secondary outcomes include patient completion of risk management recommendations (e.g. colonoscopy) and referral for genetic consultation. We will also conduct an economic analysis and an assessment of providers' experience with MeTree clinical decision support recommendations to inform future implementation efforts if the intervention is found to be effective. This trial will assess the feasibility and effectiveness of patient-collected family health history linked to decision support to promote risk-appropriate screening in a large healthcare system such as the Department of Veterans Affairs. ClinicalTrials.gov, NCT02247336 . Registered on 25 September 2014.


Cholinergic modulation of microglial activation by alpha 7 nicotinic receptors.
Almost all degenerative diseases of the CNS are associated with chronic inflammation. A central step in this process is the activation of brain mononuclear phagocyte cells, called microglia. While it is recognized that healthy neurons and astrocytes regulate the magnitude of microglia-mediated innate immune responses and limit excessive CNS inflammation, the endogenous signals governing this process are not fully understood. In the peripheral nervous system, recent studies suggest that an endogenous 'cholinergic anti-inflammatory pathway' regulates systemic inflammatory responses via alpha 7 nicotinic acetylcholinergic receptors (nAChR) found on blood-borne macrophages. These data led us to investigate whether a similar cholinergic pathway exists in the brain that could regulate microglial activation. Here we report for the first time that cultured microglial cells express alpha 7 nAChR subunit as determined by RT-PCR, western blot, immunofluorescent, and immunohistochemistry analyses. Acetylcholine and nicotine pre-treatment inhibit lipopolysaccharide (LPS)-induced TNF-alpha release in murine-derived microglial cells, an effect attenuated by alpha 7 selective nicotinic antagonist, alpha-bungarotoxin. Furthermore, this inhibition appears to be mediated by a reduction in phosphorylation of p44/42 and p38 mitogen-activated protein kinase (MAPK). Though preliminary, our findings suggest the existence of a brain cholinergic pathway that regulates microglial activation through alpha 7 nicotinic receptors. Negative regulation of microglia activation may also represent additional mechanism underlying nicotine's reported neuroprotective properties.


Coccidioidomycosis in patients infected with human immunodeficiency virus: review of 91 cases at a single institution.
We retrospectively evaluated the clinical manifestations, diagnosis, treatment, and outcome of coccidioidomycosis in 91 patients infected with human immunodeficiency virus (HIV) at a single institution. Coccidioidomycosis was the AIDS-defining illness in 37 patients. Fever and chills, weight loss, and night sweats were the most frequent symptoms. The lung was the most frequently involved organ (80%), followed by the meninges (15%). A diffuse reticulonodular infiltrate was seen in 59 patients (65%), and 13 (14%) had focal pulmonary disease; for 15 patients (16%), the chest radiograph was normal. Coccidioidal serologies were positive for 60 patients (68%), while for 23% with proven coccidioidomycosis such tests were negative Most patients were treated with systemic amphotericin B and then an oral azole. The mortality for the whole group was 60%. Patients with diffuse pulmonary disease had the highest mortality (68%), with a median duration of survival of 54 days (P < .05; 95% confidence interval, 147-175 days). The presence of diffuse pulmonary disease and a CD4 lymphocyte count of < 50/microL were independent predictors of death. In our experience, coccidioidomycosis is an important opportunistic infection that causes substantial morbidity and mortality among HIV-infected patients living in an area of endemicity.


Influence of equine growth hormone, insulin-like growth factor-I and its interaction with gonadotropins on in vitro maturation and cytoskeleton morphology in equine oocytes.
In horses, successful in vitro fertilization procedures are limited by our inability to consistently mature equine oocytes by in vitro methods. Growth hormone (GH) is an important regulator of female reproduction in mammals, playing an important role in ovarian function, follicular growth and steroidogenesis. The objectives of this research were to investigate: the effects of equine growth hormone (eGH) and insulin-like growth factor-I (IGF-I) on the in vitro maturation (IVM) of equine oocytes, and the effects of eGH in addition to estradiol (E2), gonadotropins (FSH and LH) and fetal calf serum (FCS) on IVM. We also evaluated the cytoskeleton organization of equine oocytes after IVM with eGH. Equine oocytes were aspirated from follicles <30 mm in diameter and matured for 30 h at 38.5°C in air with 5% CO2. In experiment 1, selected cumulus-oocyte complexes (COCs) were randomly allocated as follows: (a) control (no additives); (b) 400 ng/ml eGH; (c) 200 ng/ml IGF-I; (d) eGH + IGF-I; and (e) eGH + IGF-I + 200 ng/ml anti-IGF-I. In addition to these treatment groups, we also added 1 μg/ml E2, 5 IU/ml FSH, 10 IU/ml LH and 10% FCS in vitro (experiment 2). Oocytes were stained with markers for microtubules (anti-α-tubulin antibody), microfilaments (AlexaFluor 488 Phalloidin) and chromatin (TO-PRO3-iodide) and assessed via confocal microscopy. No difference was observed when eGH and IGF-I was added into our IVM system. However, following incubation with eGH alone (40%) and eGH, E2, gonadotropins and FCS (36.6%) oocytes were classified as mature v. 17.6% of oocytes in the control group (P < 0.05). Matured equine oocytes showed that a thin network of filaments concentrated within the oocyte cortex and microtubules at the metaphase spindle showed a symmetrical barrel-shaped structure, with chromosomes aligned along its midline. We conclude that the use of E2, gonadotropins and FCS in the presence of eGH increases the number of oocytes reaching oocyte competence.


Gamma-tubulin complexes and microtubule nucleation.
Microtubules are dynamic cytoskeletal polymers that assemble from alpha/beta-tubulin and are vital for the establishment of cell polarity, vesicle trafficking and formation of the mitotic/meiotic spindle. gamma-Tubulin, a protein related to alpha/beta-tubulin, is required for initiating the polymerization of microtubules in vivo. gamma-Tubulin has been found in two main protein complexes: the gamma-tubulin ring complex and its subunit, the gamma-tubulin small complex. The latter is analogous to the yeast Tub4 complex. In the past year, important advances have been made in understanding the structure and function of the gamma-tubulin ring complex and how it interacts with microtubules.


Bayesian meta-analysis of the effect of fasting, transport and lairage times on four attributes of pork meat quality.
Technological meat quality is a significant economic factor in pork production, and numerous publications have shown that it is strongly influenced both by genetic status and by rearing and slaughter conditions. The quality of meat is often described by meat pH at different times postmortem, as well as by color and drip loss. A meta-analysis based on a database built from 27 studies corresponding to a total of 6526 animals classified was carried out. The purpose of this meta-analysis was to study the effect of fasting, lairage and transport durations on four main attributes of the technological pork meat quality. A Bayesian hierarchical meta-regression approach was adopted. The results of our meta-analysis showed that fasting time had a significant effect on pH measured 24h post-mortem (pHu) and drip loss (DL) measured in longissimus muscle. While, lairage affected only the pHu in semimembranosus muscle. Interestingly, we found that DL was the lone attribute that was affected by transport time and its interaction with fasting time.


Lyme borreliosis: from infection to autoimmunity.
Lyme borreliosis in humans is an inflammatory disease affecting multiple organ systems, including the nervous system, cardiovascular system, joints and muscles. The causative agent, the spirochaete Borrelia burgdorferi, is transmitted to the host by a tick bite. The pathogenesis of the disease in its early stages is associated largely with the presence of viable bacteria at the site of inflammation, whereas in the later stages of disease, autoimmune features seem to contribute significantly. In addition, it has been suggested that chronic persistence of B. burgdorferi in affected tissues is of pathogenic relevance. Long-term exposure of the host immune system to spirochaetes and/or borrelial compounds may induce chronic autoimmune disease. The study of bacterium-host interactions has revealed a variety of proinflammatory and also immunomodulatory-immunosuppressive features caused by the pathogen. Therapeutic strategies using antibiotics are generally successful, but chronic disease may require immunosuppressive treatment. Effective and safe vaccines using recombinant outer surface protein A have been developed, but have not been propagated because of fears that autoimmunity might be induced. Nevertheless, new insights into the modes of transmission of B. burgdorferi to the warm-blooded host have been generated by studying the action of these vaccines.


Characterization of Free and Porous Silicon-Encapsulated Superparamagnetic Iron Oxide Nanoparticles as Platforms for the Development of Theranostic Vaccines.
Tracking vaccine components from the site of injection to their destination in lymphatic tissue, and simultaneously monitoring immune effects, sheds light on the influence of vaccine components on particle and immune cell trafficking and therapeutic efficacy. In this study, we create a hybrid particle vaccine platform comprised of porous silicon (pSi) and superparamagnetic iron oxide nanoparticles (SPIONs). The impact of nanoparticle size and mode of presentation on magnetic resonance contrast enhancement are examined. SPION-enhanced relaxivity increased as the core diameter of the nanoparticle increased, while encapsulation of SPIONs within a pSi matrix had only minor effects on T2 and no significant effect on T2* relaxation. Following intravenous injection of single and hybrid particles, there was an increase in negative contrast in the spleen, with changes in contrast being slightly greater for free compared to silicon encapsulated SPIONs. Incubation of bone marrow-derived dendritic cells (BMDC) with pSi microparticles loaded with SPIONs, SIINFEKL peptide, and lipopolysaccharide stimulated immune cell interactions and interferon gamma production in OT-1 TCR transgenic CD8+ T cells. Overall, the hybrid particle platform enabled presentation of a complex payload that was traceable, stimulated functional T cell and BMDC interactions, and resolved in cellular activation of T cells in response to a specific antigen.


Distribution of putative male sex pheromones among Lutzomyia sandflies (Diptera: Psychodidae).
Male Lutzomyia longipalpis produce terpene sex pheromones in glandular tissue underlying the cuticle. The pheromones are transmitted to the surface via cuticle-lined ducts (measuring 0.25 microm in diameter), each of which reaches the surface in the centre of a papule (measuring 3-3.5 microm in diameter). Similar papules, in a range of shapes but all characterized by the presence of a central pore and absence of macroserae, occur in some other species of sandfly. The aim of the present study was to determine the distribution of sex pheromones in sandflies of the genus Lutzomyia that do and do not have the papules. The results indicate that sex pheromones are not widely distributed amongst male Lutzomyia spp. Male members of the genus can be subdivided into three groups: those that produce terpenes and have cuticular papules; those that do not produce terpenes but still have the associated papules; and those that have neither terpenes nor papules. The papules seen in the species that do not synthesise sex pheromones are presumably vestigial, non-functional structures. Such species may have stopped producing pheromone as the result of changes in the way in which the females found and selected mates or changing feeding preferences. A similar event has occurred in the Lepidoptera, where vestigial pheromone-secreting structures remain in some species which no longer produce pheromone. Lutzomyia lenti collected in southern Brazil produced a novel diterpene whereas male L. lenti from north-eastern Brazil did not, supporting suggestions by others that L. lenti is, like L. longipalpis, a species complex.


Metrological sharp shooting for plasma proteins and peptides: The need for reference materials for accurate measurements in clinical proteomics and in vitro diagnostics to generate reliable results.
Reliable study results are necessary for the assessment of discoveries, including those from proteomics. Reliable study results are also crucial to increase the likelihood of making a successful choice of biomarker candidates for verification and subsequent validation studies, a current bottleneck for the transition to in vitro diagnostic (IVD). In this respect, a major need for improvement in proteomics appears to be accuracy of measurements, including both trueness and precision of measurement. Standardization and total quality management systems (TQMS) help to provide accurate measurements and reliable results. Reference materials are an essential part of standardization and TQMS in IVD and are crucial to provide metrological correct measurements and for the overall quality assurance process. In this article we give an overview on how reference materials are defined, prepared and what role they play in standardization and TQMS to support the generation of reliable results. We discuss how proteomics can support the establishment of reference materials and biomarker tests for IVD applications, how current reference materials used in IVD may be beneficially applied in proteomics, and we provide considerations on the establishment of reference materials specific for proteomics. For clarity, we solely focus on reference materials related to serum and plasma.


[The clinical importance of the radioimmunological determination of the beta 2-microglobulin in acute leukemia patients].
The concentration of beta 2-microglobulin in the blood serum was examined in 141 patients with acute leukemia. There were 63 men and 78 women aged 14 to 78 years. Studies performed before chemotherapy was instituted revealed a statistically significant increase in serum beta 2-microglobulin in all varieties of acute leukemia as applicable to the control values. There was a considerable scattering of individual indicators of protein concentration in the serum, which did not allow the use of measurements of beta 2-microglobulin in the blood serum as a criterion for differential diagnosis in identification of certain clinicomorphological varieties of acute leukemia. A direct correlation was established between the concentration of serum beta 2-microglobulin and the phase of the disease. The data obtained indicate that the beta 2-microglobulin test is of great help for the control of remission completeness in acute leukemias and for early diagnosis of relapses.


Modeling the links between young swimmers' performance: energetic and biomechanic profiles.
The aim was to develop a path-flow analysis model for young swimmers' performance based on biomechanical and energetic parameters, using structural equation modeling. Thirty-eight male young swimmers served as subjects. Performance was assessed by the 200-m freestyle event. For biomechanical assessment the stroke length, the stroke frequency and the swimming velocity were analyzed. Energetics assessment included the critical velocity, the stroke index and the propulsive efficiency. The confirmatory model explained 79% of swimming performance after deleting the stroke index-performance path, which was nonsignificant (SRMR = 0.06). As a conclusion, the model is appropriate to explain performance in young swimmers.


The metabolism of beta-phenylpropionic acid by an Achromobacter.
1. When a species of Achromobacter grew with beta-phenylpropionate as carbon source, 2-hydroxy-beta-phenylpropionate and 2,3-dihydroxy-beta-phenylpropionate appeared in the growth medium. The concentrations of these compounds were maximal during exponential growth. 2. The cells contained an oxygenase that required Fe(2+) ions and cleaved the benzene nucleus between the adjacent carbon atoms that bear the side chain and one hydroxyl group of 2,3-dihydroxy-beta-phenylpropionate. 3. The ring-fission product, formed with the consumption of 1mol. of oxygen/mol. of substrate, was isolated and a chemical structure assigned. Sephadex-treated cell extracts converted 1mol. of this compound into 1mol. of 4-hydroxy-2-oxovalerate without oxygen consumption; succinic acid was also formed. 4. When Mn(2+) ions or Mg(2+) ions were added, dialysed extracts converted 4-hydroxy-2-oxovalerate into pyruvate and acetaldehyde, but the reaction did not proceed to completion.


Axonal plasticity underpins the functional recovery following surgical decompression in a rat model of cervical spondylotic myelopathy.
Cervical spondylotic myelopathy (CSM) is the most common spinal cord disorder and a major cause of disability in adults. Improvements following surgical decompression are limited and patients often remain severely disabled. Post mortem studies indicate that CSM is associated with profound axonal loss. However, our understanding of the pathophysiology of CSM remains limited.To investigate the hypothesis that axonal plasticity plays a role in the recovery following surgical decompression, we adopted a novel preclinical model of mild to moderate CSM. Spinal cord compression resulted in significant locomotor deterioration, increased expression of the axonal injury marker APP, and loss of serotonergic fibres. Surgical decompression partially reversed the deficits and attenuated APP expression. Decompression was also associated with axonal sprouting, reflected in the restoration of serotonergic fibres and an increase of GAP43 expression. The re-expression of synaptophysin indicated the restoration of functional synapses following decompression. Promoting axonal plasticity may therefore be a therapeutic strategy for promoting neurological recovery in CSM.


Modulation of the norfloxacin resistance in Staphylococcus aureus by Cordia verbenaceae DC.
Several chemical compounds isolated from natural sources have antibacterial activity and some enhance the antibacterial activity of antibiotics reversing the natural resistance of bacteria to certain antibiotics. In this study, the hexane and methanol extract of Cordia verbenaceae were assessed for antibacterial activity alone and combinated with norfloxacin against the Staphylococcus aureus strain SA1199B. The minimum inhibitory concentration (MIC) of extracts was assayed using microdilution assay and the modulatory activity was evaluated using plate diffusion assay. The MIC observed varied between 256 to >1024 μg/ml. However, the antibiotic activity of norfloxacin was enhanced in the presence of subinhibitory concentrations of hexane extract of C. verbenaceae (HECV). INTERPRETATIONS & CONCLUSIONS: Our results indicate that Cordia verbenaceae DC. can be a source of plant derived products with antibiotic modifying activity.


Molecular and immunopathological studies of borna disease virus infection in rats.
Borna disease virus is an agent distinct from all known viruses. Pathogenesis of its infection is also unique. This review highlights several aspects of the biology of this viral infection and the preliminary characterization of the agent. BDV can be used to answer important questions in neurobiology. These include neuroinvasiveness and neurotropism of viral agents, CD4+ T cell-mediated immunopathology and tolerance in newborn animals to a persistent viral infection in the CNS and behavioral diseases and eating disorders induced by neurotropic viruses. This review is dedicated to Prof. Dr. Rott on occasion of his 65th birthday in recognition of his immense contributions to studies on Borna disease and also for his success focusing the attention of the scientific community to this still evolving unique viral disease.


Domain separation and characterization of PriC, a replication restart primosome factor in Escherichia coli.
In Escherichia coli the oriC-independent primosome plays an essential role in replication restart after dissociation of the replication DNA-protein complex by DNA damage. Primosome is thought to form via two pathways: one PriA dependent and the other PriA independent. PriC is a key protein in the replication restart of the PriA-independent pathway. In this study, we determined that PriC was divided into two domains. Then, we obtained information that: (i) the C-terminal domain preferentially binds to single-stranded DNA (ssDNA); (ii) the binding of PriC to ssDNA depends on salt concentration; and (iii) the binding site size of PriC is approximately 7-9 nucleotides. The protease digestion of PriC suggested that a possible DNA-binding site is the N-terminus of the C-terminal domain where basic amino acid residues are concentrated. Interestingly, α-helical induction of the C-terminal domain of PriC occurred after the addition of DNAs. Also, we examined the role of heptad repeat of leucine or valine residues in the C-terminal domain and PriC oligomerization. This study describes the structure and function analysis of PriC which forms the primosome complex in replication restart.


Helping or punishing strangers: neural correlates of altruistic decisions as third-party and of its relation to empathic concern.
Social norms are a cornerstone of human society. When social norms are violated (e.g., fairness) people can either help the victim or punish the violator in order to restore justice. Recent research has shown that empathic concern influences this decision to help or punish. Using functional magnetic resonance imaging (fMRI) we investigated the neural underpinnings of third-party help and punishment and the involvement of empathic concern. Participants saw a person violating a social norm, i.e., proposing unfair offers in a dictator game, at the expense of another person. The participants could then decide to either punish the violator or help the victim. Our results revealed that both third-party helping as well as third-party punishing activated the bilateral striatum, a region strongly related with reward processing, indicating that both altruistic decisions share a common neuronal basis. In addition, also different networks were involved in the two processes compared with control conditions; bilateral striatum and the right lateral prefrontal cortex (lPFC) during helping and bilateral striatum as well as left lPFC and ventral medial prefrontal cortex (vmPFC) during punishment. Further we found that individual differences in empathic concern influenced whether people prefer to help or to punish. People with high empathic concern helped more frequently, were faster in their decision and showed higher activation in frontoparietal regions during helping compared with punishing. Our findings provide insights into the neuronal basis of human altruistic behavior and social norm enforcement mechanism.


Anticancer activity of a broccoli derivative, sulforaphane, in barrett adenocarcinoma: potential use in chemoprevention and as adjuvant in chemotherapy.
The incidence of Barrett esophageal adenocarcinoma (BEAC) has been increasing at an alarming rate in western countries. In this study, we have evaluated the therapeutic potential of sulforaphane (SFN), an antioxidant derived from broccoli, in BEAC. BEAC cells were treated with SFN, alone or in combination with chemotherapeutic, paclitaxel, or telomerase-inhibiting agents (MST-312, GRN163L), and live cell number determined at various time points. The effect on drug resistance/chemosensitivity was evaluated by rhodamine efflux assay. Apoptosis was detected by annexin V labeling and Western blot analysis of poly(ADP-ribose) polymerase cleavage. Effects on genes implicated in cell cycle and apoptosis were determined by Western blot analyses. To evaluate the efficacy in vivo, BEAC cells were injected subcutaneously in severe combined immunodeficient mice, and after the appearance of palpable tumors, mice were treated with SFN. SFN induced both time- and dose-dependent decline in cell survival, cell cycle arrest, and apoptosis. The treatment with SFN also suppressed the expression of multidrug resistance protein, reduced drug efflux, and increased anticancer activity of other antiproliferative agents including paclitaxel. A significant reduction in tumor volume was also observed by SFN in a subcutaneous tumor model of BEAC. Anticancer activity could be attributed to the induction of caspase 8 and p21 and down-regulation of hsp90, a molecular chaperon required for activity of several proliferation-associated proteins. These data indicate that a natural product with antioxidant properties from broccoli has great potential to be used in chemoprevention and treatment of BEAC.


Hand function with touch screen technology in children with normal hand formation, congenital differences, and neuromuscular disease.
To measure and compare hand function for children with normal hand development, congenital hand differences (CHD), and neuromuscular disease (NMD) using a function test with touch screen technology designed as an iPhone application. We measured touch screen hand function in 201 children including 113 with normal hand formation, 43 with CHD, and 45 with NMD. The touch screen test was developed on the iOS platform using an Apple iPhone 4. We measured 4 tasks: touching dots on a 3 × 4 grid, dragging shapes, use of the touch screen camera, and typing a line of text. The test takes 60 to 120 seconds and includes a pretest to familiarize the subject with the format. Each task is timed independently and the overall time is recorded. Children with normal hand development took less time to complete all 4 subtests with increasing age. When comparing children with normal hand development with those with CHD or NMD, in children aged less than 5 years we saw minimal differences; those aged 5 to 6 years with CHD took significantly longer total time; those aged 7 to 8 years with NMD took significantly longer total time; those aged 9 to 11 years with CHD took significantly longer total time; and those aged 12 years and older with NMD took significantly longer total time. Touch screen technology has becoming increasingly relevant to hand function in modern society. This study provides standardized age norms and shows that our test discriminates between normal hand development and that in children with CHD or NMD. Diagnostic III.


75 W 40% efficiency single-mode all-fiber erbium-doped laser cladding pumped at 976 nm.
Optimization of Yb-free Er-doped fiber for lasers and amplifiers cladding pumped at 976 nm was performed in this Letter. The single-mode fiber design includes an increased core diameter of 34 μm and properly chosen erbium and co-dopant concentrations. We demonstrate an all-fiber high power laser and power amplifier based on this fiber with the record slope efficiency of 40%. To the best of our knowledge, the achieved output power of 75 W is the highest power reported for such lasers.


Organocatalyzed Photo-Controlled Synthesis of Ultrahigh-Molecular-Weight Fluorinated Alternating Copolymers.
Ultrahigh-molecular-weight (UHMW) polymers with tailored structures are highly desirable for the outstanding properties. In this work, we developed a novel photoorganocatalyzed controlled radical alternating copolymerizations of fluoroalkyl maleimide and diverse vinyl comonomers, enabling efficient preparation of fluorinated copolymers of predetermined UHMWs and well-defined structures at high conversions. Versatility of this method was demonstrated by expanding to controlled terpolymerization, which allows facial access toward fluorinated terpolymers of UHMWs and functional pendants. The obtained copolymers exhibited attractive physical properties and furnished thermoplastic, anticorrosive and (super)hydrophobic attributes as coatings on different substrates. Molecular simulations provided insights into the coating morphology, which unveiled a fluorous protective layer on the top surface with polar groups attached to the bottom substrate, resulting in good adhesion and hydrophobicity, simultaneously. This synthetic method and customized copolymers shed light on the design of high-performance coatings by macromolecular engineering.


Representation of Women in Academic Orthopaedic Leadership: Where Are We Now?
Women have long been underrepresented in orthopaedic surgery; however, there is a lack of quantitative data on the representation of women in orthopaedic academic program leadership. (1) What is the proportion of women in leadership roles in orthopaedic surgery departments and residency programs in the United States (specifically, chairs, vice chairs, program directors, assistant program directors, and subspecialty division chiefs)? (2) How do women and men leaders compare in terms of years in position in those roles, years in practice, academic rank, research productivity as represented by publications, and subspecialty breakdown? (3) Is there a difference between men and women in the chair or program director role in terms of whether they are working in that role at institutions where they attended medical school or completed their residency or fellowship? We identified 161 academic orthopaedic residency programs from the Accreditation Council for Graduate Medical Education (ACGME) website. Data (gender, length of time in position, length of time in practice, professorship appointment, research productivity as indirectly measured via PubMed publications, and subspecialty) were collected for chairs, vice chairs, program directors, assistant program directors, and subspecialty division chiefs in July 2020 to control for changes in leadership. Information not provided by the ACGME and PubMed was found using orthopaedic program websites and the specific leader's curriculum vitae. Complete data were obtained for chairs and program directors, but there were missing data points for vice chairs, assistant program directors, and division chiefs. All statistical analysis was performed using SPSS using independent t-tests for continuous variables and the Pearson chi-square test for categorical variables, with p < 0.05 considered significant. Three percent (4 of 153) of chairs, 8% (5 of 61) of vice chairs, 11% (18 of 161) of program directors, 27% (20 of 75) of assistant program directors, and 9% (45 of 514) of division chiefs were women. There were varying degrees of missing data points for vice chairs, assistant program directors, and division chiefs as not all programs reported or have those positions. Women chairs had fewer years in their position than men (2 ± 1 versus 9 ± 7 [95% confidence interval -9.3 to -5.9]; p < 0.001). Women vice chairs more commonly specialized in hand or tumor compared with men (40% [2 of 5] and 40% [2 of 5] versus 11% [6 of 56] and 4% [2 of 56], respectively; X2(9) = 16; p = 0.04). Women program directors more commonly specialized in tumor or hand compared with men (33% [6 of 18] and 17% [3 of 18] versus 6% [9 of 143] and 11% [16 of 143], respectively; X2(9) = 20; p = 0.02). Women assistant program directors had fewer years in practice (9 ± 4 years versus 14 ± 11 years [95% CI -10.5 to 1.6]; p = 0.045) and fewer publications (11 ± 7 versus 30 ± 48 [95% CI -32.9 to -5.8]; p = 0.01) than men. Women division chiefs had fewer years in practice and publications than men and were most prevalent in tumor and pediatrics (21% [10 of 48] and 16% [9 of 55], respectively) and least prevalent in spine and adult reconstruction (2% [1 of 60] and 1% [1 of 70], respectively) (X2(9) = 26; p = 0.001). Women program directors were more likely than men to stay at the same institution they studied at for medical school (39% [7 of 18] versus 14% [20 of 143]; odds ratio 3.9 [95% CI 1.4 to 11.3]; p = 0.02) and trained at for residency (61% [11 of 18] versus 42% [60 of 143]; OR 2.2 [95% CI 0.8 to 5.9]; p = 0.01). The higher percentage of women in junior leadership positions in orthopaedic surgery, with the data available, is a promising finding. Hand, tumor, and pediatrics appear to be orthopaedic subspecialties with a higher percentage of women. However, more improvement is needed to achieve gender parity in orthopaedics overall, and more information is needed in terms of publicly available information on gender representation in orthopaedic leadership. Proportional representation of women in orthopaedics is essential for quality musculoskeletal care, and proportional representation in leadership may help encourage women to apply to the specialty. Our findings suggest movement in an improving direction in this regard, though more progress is needed.


Clinical and laboratory characteristics of hepatitis d in Republic of Sakha (Yakutia).
Inrtroduction: The epidemiological situation for hepatitis D has changed significantly. Reduced population authors infection due to a sharp decline in hospitalizations from Central Asia regions, the Caucasus and Moldova, which are known to be endemic for hepatitis D. Currently, the incidence of chronic hepatitis D (HGD) in Russia is 1%, while in the countries of Central Asia, and in particular in Turkmenistan, the share of HGD among chronic viral hepatitis is 8%. The aim of research was to establish the clinical features, depending on the activity of the replication of hepatitis viruses B and D. Materials and Methods: We studied 26 patients with viral hepatitis D with a determined activity replicative virus by PCR (polymerase chain reaction). The age of patients ranged from 28 to 78 years. The patients performed the ELISA (enzyme-linked immunosorbent assay) study for the presence of markers of parenteral viral hepatitis (HBsAg, a-HCV and a-HDV), the standard general clinical biochemical blood tests. of the instrumental methods survey used ultrasonography (ultrasound), EGD (fibrogastroduodenoscopy). Grading the severity of liver cirrhosis established by Child-Pugh (eng. Child-Pugh, Child-Turcotte, Child-Turcotte-Pugh, sometimes Child-Paquet) is designed to assess the severity of cirrhosis. The severity of liver cirrhosis is assessed on a point system, which are calculated from 5 or 6 parameters. Results: It is established that most HGD more prevalent among young people bodied (69%) and occurs mainly in severe symptoms and portal hypertension leading to the rapid development of liver cirrhosis (53%). It showed that hyperenzymemia reaches high levels of ALT to 1715 U / L. with a high viral DNA load virus (HBV) 2648226,0 ± 953892,7 copies / ml in the presence of an RNA virus D (HDV +). Conclusion: Thus, the main feature of chronic hepatitis D is its predominant tsirrogennost.


Specificity of electrical coupling among neurons innervating forelimb muscles of the adult bullfrog.
1. The specificity of electrical connections among sensory fibers and motoneurons in the bullfrog's spinal cord was studied by recording intracellularly from brachial motoneurons. Synaptic potentials evoked by stimulation of individual muscle nerves were recorded in normal and reduced-calcium solutions and after acute section of dorsal or ventral roots. 2. Homonymous motoneurons are electrically coupled. After the dorsal roots were cut to abolish sensory input, short-latency potentials were almost always evoked anti-dromically in a motoneuron by stimulation of its own muscle nerve but rarely by stimulation of nerves innervating other muscles. These potentials differed from chemically mediated synaptic potentials in this preparation; they had a shorter latency and remained after perfusion with reduced-calcium solutions. This evidence suggests that they are mediated electrically. 3. Some motoneurons that innervate functionally equivalent muscles are electrically coupled. Approximately two-thirds of the motoneurons innervating the internal or external heads of the triceps muscles received coupling potentials on stimulation of the other nerve, although never on stimulation of the heteronymous, medial, triceps nerve. 4. The monosynaptic potentials evoked by muscle sensory afferents in motoneurons often have both electrical and chemical components. The electrical component occurred with short delay and persisted in reduced-calcium solutions. The chemical component occurred 1.5-2.0 ms later, at 14 degrees C, and was abolished by reducing calcium in the bathing solution. Muscle sensory afferents make these mixed synapses on homonymous, heteronymous, and other motoneurons.


Osteopathic Medicine and Primary Care looks forward to 2009.
Osteopathic Medicine and Primary Care, which enters its third year of operation in 2009 under the umbrella of BioMed Central, continues to promote and advance open access publishing through universal online access without charge, indexing in PubMed and archiving in PubMed Central, retention of authors' copyright, and expeditious peer review. Notable accomplishments during 2008 included a median lag time of four months from initial manuscript submission to publication, designation of eight articles as "highly accessed," and achievement of a balanced proportion of publications in our core topic areas of osteopathic medicine and primary care. In October 2008, Springer Science+Business Media, a major publisher of journals in science, technology, and medicine, acquired the BioMed Central Group. Our 2009 Editorial Board is presented herein, as well as a new mechanism for posting book reviews on the Osteopathic Medicine and Primary Care website. We continue to encourage manuscript submissions and reader comments on our articles. Waivers or discounts of article processing charges are available via several mechanisms for eligible authors who submit qualified manuscripts.


Actin remodeling, the synaptic tag and the maintenance of synaptic plasticity.
Activity-dependent plasticity of synaptic connections is a hallmark of the mammalian brain and represents a key mechanism for rewiring neural circuits during development, experience-dependent plasticity, and brain disorders. Cellular models of memory, such as long-term potentiation and long-term depression, share common principles to memory consolidation. As for memory, the maintenance of synaptic plasticity is dependent on the synthesis of de novo protein synthesis. The synaptic-tagging and capture hypothesis states that the maintenance of synaptic plasticity is dependent on the interplay between input-specific synaptic tags and the allocation or capture of plasticity-related proteins (PRPs) at activated synapses. The setting of the synaptic tag and the capture of PRPs are independent processes that can occur separated in time and different groups of activated synapses. How are these two processes orchestrated in time and space? Here, we discuss the synaptic-tagging and capture hypothesis in the light of neuronal compartmentalization models and address the role of actin as a putative synaptic tag. If different groups of synapses interact by synaptic-tagging and capture mechanisms, understanding the spatial rules of such interaction is key to define the relevant neuronal compartment. We also discuss how actin modulation can allow an input-specific capture of PRPs and try to conciliate the temporal dynamics of synaptic actin with the maintenance of plasticity. Understanding how multiple synapses interact in time and space is fundamental to predict how neurons integrate information and ultimately how memory is acquired.


Mammalian blastocyst: transport functions in a developing epithelium.
This article reviews the current state of knowledge concerning morphological and physiological mechanisms important to growth and differentiation of the mammalian blastocyst between compaction and implantation. Morphological processes occur in conjunction with major changes in transport systems that control the movement of substances into and out of the embryo. Compaction is a morphological development that is associated with the formation of an outer squamous epithelium, the trophectoderm, which regulates the composition of the medium bathing the presumptive embryo (the inner cell mass). Implantation involves the interaction of two epithelia, the adhesion between the trophectoderm and the maternal endometrium. Before adhesion, the blastocyst lies free in the uterine fluid and exchanges occur between this fluid and the embryo. Apposition of these epithelia is brought about in part by expansion of the blastocyst and removal of the uterine fluid. Blastocyst physiology is an inherently important field because vectorial transport system development and the genes that regulate it can be studied.


Silver-mediated temperature-controlled selective deposition of Pt on hexoctahedral Au nanoparticles and the high performance of Au@AgPt NPs in catalysis and SERS.
Nanomaterials with high catalytic activity and good SERS properties can be used for sensitive and real-time in situ tracking of a catalytic process via SERS, which can be a powerful tool for investigating the products and mechanisms of the catalytic reaction. In the present work, Au@AgPt NPs with a {431}-faceted hexoctahedral Au core and an AgPt alloy shell exhibiting enhanced catalysis and good SERS activity were prepared by a facile silver-mediated temperature-controlled selective deposition of Pt. The complex hexoctahedral Au nanoparticles were synthesized first as nano-templates, followed by coating with a thin layer of Ag. Then, a temperature-controlled synthesis method for preferably depositing Pt on the hexoctahedral Au NPs was proposed to prepare Au@AgPt NPs. With the increase of the synthesis temperature, the Pt atoms were controlled to selectively deposit on the tips, edges or the entire surface of the nano-templates. By systematically investigating the effects of temperature, precursor consumption and synthesis time on the morphology, composition, optical properties, catalysis and SERS properties of the Au@AgPt NPs, the kinetic and thermodynamic mechanisms of the deposition of Pt on hexoctahedral Au nanoparticles were explored. The performance of the Au@AgPt NPs in SERS-based real-time in situ monitoring of the catalytic reaction was also investigated and verified. Besides, it is easy to regulate and control their SERS and catalytic performances through the selective deposition of Pt, according to the demand of the catalytic reaction and SERS monitoring. This work not only presents a new Au@AgPt nanostructure with good catalytic and SERS properties, but also develops a facile, universal and controllable method for selective deposition of Pt on Au nano-templates with a variety of morphologies.


Circulating miRNA profiles in COVID-19 patients and meta-analysis: implications for disease progression and prognosis.
We compared circulating miRNA profiles of hospitalized COVID-positive patients (n = 104), 27 with acute respiratory distress syndrome (ARDS) and age- and sex-matched healthy controls (n = 18) to identify miRNA signatures associated with COVID and COVID-induced ARDS. Meta-analysis incorporating data from published studies and our data was performed to identify a set of differentially expressed miRNAs in (1) COVID-positive patients versus healthy controls as well as (2) severe (ARDS+) COVID vs moderate COVID. Gene ontology enrichment analysis of the genes these miRNAs interact with identified terms associated with immune response, such as interferon and interleukin signaling, as well as viral genome activities associated with COVID disease and severity. Additionally, we observed downregulation of a cluster of miRNAs located on chromosome 14 (14q32) among all COVID patients. To predict COVID disease and severity, we developed machine learning models that achieved AUC scores between 0.81-0.93 for predicting disease, and between 0.71-0.81 for predicting severity, even across diverse studies with different sample types (plasma versus serum), collection methods, and library preparations. Our findings provide network and top miRNA feature insights into COVID disease progression and contribute to the development of tools for disease prognosis and management.


Emerging drugs for diabetic neuropathy.
Diabetic neuropathy (DN) is a very common and disabling diabetes-related complication. DN is associated with significant morbidity and mortality. Diabetic peripheral neuropathy (DPN) can be painful in the earlier stages of the disease before becoming painless. Most of the currently available therapies are symptomatic (focusing on pain relief) rather than disease-modifying. With the exception of good glycemic control, there is currently no effective treatment to slow the progression of or reverse DPN. In this article, we review the epidemiology, pathogenesis, currently available and future treatments for DPN, and the potential development issues/challenges related to such new therapies. Literature search was performed using PubMed, Medline and Pharmaprojects from 1950 onwards. Search terms include a combination of terms such as diabetic neuropathy, pathogenesis, pathophysiology, mechanisms, treatment, therapy, oxidative/nitrosative stress, anti-oxidants, serotonin, nitrotyrosine, protein kinase C, aldose reductase, sodium channels, taurine, lipoic acid and poly (ADP-ribose) polymerase. The reader will gain an overview of the epidemiology, clinical features and risk factors of DN. In addition, the reader will have a better understanding of the mechanisms that underpin the development of DPN and their relationships to the current and future therapies. The reader will also develop an insight into the limitations of the current approach to DPN treatment and the potential avenues for future research. DN is a very common and disabling complication that currently has no effective treatments other than diabetes control. The pathogenesis of DPN is complex and multi-factorial. Several disease-modifying and symptomatic treatments are currently under development. Oxidative and nitrosative stress have been identified as key pathogenic factors in the development of DPN and new treatments target these pathways and/or their downstream consequences. Gene therapy and growth factors have also emerged as potential new therapies that target particular cellular compartments as opposed to being delivered systemically. The recognition of the difficulty in reversing established DN has focused efforts on slowing its progression.


A minimalist approach to gene mapping: locating the gene for acheiropodia, by homozygosity analysis.
Acheiropodia is an autosomal recessive disease that results in hemimelia (lack of formation of the distal extremities). We performed a complete genome screen of seven members of an extended pedigree that included three siblings with acheiropodia. Homozygosity mapping was used to identify regions most likely to harbor the gene for acheiropodia in this pedigree. In these two key regions (14p and 7q), further genotyping of one additional affected member of this pedigree plus seven additional unaffected siblings provided evidence, through linkage analysis, that the 7q36 region contains the acheiropodia gene. In this region, a maximum two-point LOD score of 3.81 (4.2 with multipoint analysis) was achieved, and a homozygous haplotype spanning a region of 11.7 cM was seen in all affected in this pedigree. Finally, genotypic analysis of two additional cases of acheiropodia with no known relation to the other samples revealed homozygous sharing of a portion of the same haplotype on 7q36, which reduces the chromosomal location of the acheiropodia gene to an 8.6-cM region. Localization of this gene, at the screening level, by use of data from only three affected subjects, provides an example of how certain genes may be mapped by use of a minimal number of affected cases.


Reactive Carbonyl Species Mediate Methyl Jasmonate-Induced Stomatal Closure.
Production of reactive oxygen species (ROS) is a key signal event for methyl jasmonate (MeJA)- and abscisic acid (ABA)-induced stomatal closure. We recently showed that reactive carbonyl species (RCS) stimulates stomatal closure as an intermediate downstream of hydrogen peroxide (H2O2) production in the ABA signaling pathway in guard cells of Nicotiana tabacum and Arabidopsis thaliana. In this study, we examined whether RCS functions as an intermediate downstream of H2O2 production in MeJA signaling in guard cells using transgenic tobacco plants overexpressing A. thaliana 2-alkenal reductase (n-alkanal + NAD(P)+ ⇌ 2-alkenal + NAD(P)H + H+) (AER-OE tobacco) and Arabidopsis plants. The stomatal closure induced by MeJA was impaired in the AER-OE tobacco and was inhibited by RCS scavengers, carnosine and pyridoxamine, in the wild-type (WT) tobacco plants and Arabidopsis plants. Application of MeJA significantly induced the accumulation of RCS, including acrolein and 4-hydroxy-(E)-2-nonenal, in the WT tobacco but not in the AER-OE plants. Application of MeJA induced H2O2 production in the WT tobacco and the AER-OE plants and the H2O2 production was not inhibited by the RCS scavengers. These results suggest that RCS functions as an intermediate downstream of ROS production in MeJA signaling and in ABA signaling in guard cells.


Effects of intra-session exercise sequence during water-based concurrent training.
The aim was to investigate the effects of the intra-session exercise order during water-based concurrent training on the neuromuscular adaptations in young women. 26 women (25.1±2.9 years) were placed into 2 groups: resistance prior to (RA) or after (AR) aerobic training. Subjects performed resistance (sets at maximal effort) and aerobic training (exercises at heart rate corresponding to the second ventilatory threshold) twice a week over 12 weeks, performing both exercise types in the same training session. Upper (elbow flexion) and lower-body (knee extension) one-repetition maximum test (1RM) and peak torque (PT) were evaluated. The muscle thickness (MT) of upper (sum of MT of biceps brachii and brachialis) and lower-body (sum of MT of vastus lateralis, vastus medialis, vastus intermedius, and rectus femoris) was determined by ultrasonography. Moreover, the maximal electromyographic activity (EMG) of upper (biceps brachii) and lower-body (sum of EMG of vastus lateralis and rectus femoris) was measured. Both RA and AR groups increased the upper and lower-body 1RM and PT, while the lower-body 1RM increases observed in the RA was greater than AR (43.58±14.00 vs. 27.01±18.05%). RA and AR showed MT increases in all muscles evaluated, while the lower-body MT increases observed in the RA were also greater than AR (10.24±3.11 vs. 5.76±1.88%). There were increases in the maximal EMG of upper and lower-body in both RA and AR, with no differences between groups. Performing resistance prior to aerobic exercise during water-based concurrent training seems to optimize the lower-body strength and hypertrophy.


The oral splicing modifier RG7800 increases full length survival of motor neuron 2 mRNA and survival of motor neuron protein: Results from trials in healthy adults and patients with spinal muscular atrophy.
Spinal muscular atrophy (SMA) is a rare genetic and progressively debilitating neuromuscular disease. It is the leading genetic cause of death among infants. In SMA, low levels of survival of motor neuron (SMN) protein lead to motor neuron death and muscle atrophy as the SMN protein is critical to motor neuron survival. SMA is caused by mutations in, or deletion of, the SMN1 gene. A second SMN gene, SMN2, produces only low levels of functional SMN protein due to alternative splicing which excludes exon 7 from most transcripts, generating truncated, rapidly degraded SMN protein. Patients with SMA rely on limited expression of functional SMN full-length protein from the SMN2 gene, but insufficient levels are generated. RG7800 is an oral, selective SMN2 splicing modifier designed to modulate alternative splicing of SMN2 to increase the levels of functional SMN protein. In two trials, oral administration of RG7800 increased in blood full-length SMN2 mRNA expression in healthy adults and SMN protein levels in SMA patients by up to two-fold, which is expected to provide clinical benefit.


W values and radial dose distributions for protons in TE-gas and air at energies up to 500 MeV.
The dosimetry of charged-particle beams is frequently performed by measuring the ionization yield produced in gas-filled ionization chambers and converting it into absorbed dose using the mean energy W expended in a gas per ion pair formed, or its differential value omega for the filling gas. Because of the increasing importance of proton therapy and the lack of accurate values of W and omega at high proton energies in particular, we investigated the ionization yield formation by protons in air and methane-based tissue-equivalent gas in the energy range up to 500 MeV. For this purpose, we analysed the available experimental values of W and omega and, in a second step, investigated the ionization-yield formation by protons as a function of their initial energy using an analytical method based on the Spencer-Fano equation in the framework of the continuous-slowing-down approximation. To estimate possible ionization losses which can be expected if the ionometric method is applied in the case of an incomplete secondary-electron equilibrium, a new analytical model has been developed which can be used to calculate the radial distribution of the ionization yield produced by secondary electrons around high-energy proton tracks.


Knowledge, attitudes and acceptability to provider-initiated HIV testing and counseling: patients' perspectives in Moshi and Rombo Districts, Tanzania.
Provider-initiated HIV testing and counseling (PITC) is referred to as routine testing in a clinical setting as part of a standard programme of medical services. PITC is initiated in order to avoid missed opportunities for people to get tested for HIV. While advocated as a strategy, there is dearth of information on patients' views on PITC in a number of districts in Tanzania. The objective of this study was to assess the knowledge, attitude and acceptability to PITC services among patients attending health care facilities in rural and urban settings in Kilimanjaro region A total of 12 focus group discussions (FGDs) were conducted with 99 (73 female and 26 male) patients enrolled into out-patient clinics in 8 (2 hospitals and 6 primary care centers) health facilities in Moshi Urban and Rombo districts in northern Tanzania. The study explored on knowledge, attitudes and acceptability of PITC, perceived benefits and barriers of PITC, and ethical issues related to PITC. Interviews were audio taped, transcribed, translated, and analyzed using Non-numerical Unstructured Data Indexing and Theorizing (NUDIST) software. Knowledge about PITC services was generally low. Compared to men, women had a more positive attitude towards PITC services, because of its ability to identify and treat undiagnosed HIV cases. HIV stigma was regarded as a major barrier to patients' uptake of PITC. Institutional factors such as lack of supplies and human resources were identified as barriers to successful provision of PITC. In conclusion, the findings highlight both opportunities and potential barriers in the successful uptake of PITC, and underscore the importance of informed consent, counseling and confidentiality and the need for specific strategies on advocacy for the service.


In-vitro and in-vivo antibacterial activity of quinupristin/dalfopristin.
Quinupristin/dalfopristin is a new water-soluble streptogramin antimicrobial agent comprising quinupristin and dalfopristin in a ratio of 30:70. The in-vitro spectrum of activity includes most multi-resistant Gram-positive aerobes, important Gram-negative aerobes, Gram-positive anaerobes and intracellular bacteria that are causal agents of respiratory, blood and cutaneous infections. Of particular note, quinupristin/dalfopristin is active against multidrug-resistant isolates of Staphylococcus aureus, Staphylococcus epidermidis and Enterococcus faecium, and against penicillin-resistant and/or erythromycin-resistant Streptococcus pneumoniae. The combination is also active against staphylococci showing both constitutive and inducible erythromycin resistance. Bactericidal activity and a prolonged post-antibiotic effect have also been noted for quinupristin/dalfopristin against Gram-positive cocci. Gram-negative bacteria susceptible to quinupristin/dalfopristin include Moraxella catarrhalis, Legionella spp. and Mycoplasma spp. Overall, the spectrum of antibacterial activity indicates a potential role for this combination in the treatment of difficult-to-treat Gram-positive infections, including those caused by multidrug-resistant organisms. Since this activity extends to Gram-negative respiratory bacteria, quinupristin/dalfopristin may also find a role in the treatment of atypical, as well as typical, pneumonia.


Effects of the long-term (17-19 months) feeding of high-iodine eggs on lipid metabolism and thyroid function in rats.
The present paper describes the effects of long-term (17-19 months) feeding of high-iodine eggs on lipid metabolism and thyroid function of rats, and also the effects of inorganic iodine on lipid metabolism. Rats were meal-fed on a diet containing 1% (w/w) of ordinary egg powder (OE diet as control: 35 micrograms I/100 g) or high-iodine egg powder (IE diet: 392 micrograms I/100 g). After the 19-month dietary treatment, rats fed on the IE diet, compared with the controls, showed a higher tissue lipoprotein lipase activity, a lower lipid peroxide level in the brain and a trend toward lower serum triacylglycerol levels and body fat storage without alterations in serum levels of thyroid-related hormones (TSH, T3 and T4). From the results of cold exposure and anti-thyroid drug-treatment conducted on rats fed on the OE and IE diets for 17 months, high-iodine eggs seemed to improve the age-related defects in thermogenic and thyroid hormone responses to cold, and also to result in a resistance to the anti-thyroid drug. The effects of the IE diet on lipid metabolism of rats were partly exhibited by feeding of the OE diet with an equivalent amount of iodine added as KI or KIO3. Thus, it is suggested that iodine ingestion through high-iodine eggs modulates both lipid metabolism and thyroid function in rats.


Using Concept Mapping within a Community-Academic Partnership to Examine Obesity among Mexican Immigrants.
Weight gain is common following migration to a new country. Mexican immigrants have a disparate prevalence of overweight/obesity and food insecurity. Social stressors, such as unemployment, discrimination, and the threat of deportation, may fuel both food insecurity and weight gain in this population. We sought to (1) examine community-defined causes and correlates of obesity among Mexican-Americans; (2) determine how current social stressors, policies, and programs impact food insecurity and obesity; and (3) identify community-defined priorities for preventive interventions. Group concept mapping (GCM) was used in a community-academic partnership (CAP) to describe the factors contributing to weight gain and obesity among Mexican immigrant families. Activities included community brainstorming, sorting and rating, multivariate statistical analysis, and community interpretation of results. Eighty statements were generated in the brainstorming sessions. These statements were sorted into nine clusters, which were organized into three regions: (1) intrapersonal factors; (2) community-level factors; and (3) social policy-related barriers. Statements reflecting the impact of immigration-related stressors were found in all three regions, addressing participants' fears of deportation, and the prioritization of resources away from healthy eating, resulting in food insecurity. Community members identified five priority areas for intervention planning: (1) lack of exercise; (2) lack of knowledge of a healthy diet; (3) expense of healthy foods; (4) "junk" food; and (5) stress management. Results suggest high levels of social stress are contributing to food insecurity and obesity among Mexican immigrant families. Areas identified for intervention planning reflect the need for a multifaceted approach toward obesity prevention.


Evaluation of a hospital work redesign: patient-focused care.
The 1990s have seen a frenzy of work redesign in acute care hospitals/health systems. Much of the redesign work in acute care hospitals has centered on changing how nurses organize and deliver care to patients. But have we really determined that redesigning nursing care delivery does what we want? The author describes the introduction of patient-focused care at one tertiary care university teaching hospital and answers the following question: Is there a change in provider or patient outcomes when the nursing care delivery system is changed from primary/total patient care to patient-focused care?


Cloning and characterization of a gene required for the secretion of extracellular enzymes across the outer membrane by Xanthomonas campestris pv. campestris.
Nonpathogenic mutants of Xanthomonas campestris pv. campestris, generated from transposon mutagenesis, accumulated extracellular polygalacturonate lyase, alpha-amylase, and endoglucanase in the periplasm. The transposon Tn5 was introduced by a mobilizable, suicidal plasmid, pSUP2021 or pEYDG1. Genomic banks of wild-type X. campestris pv. campestris, constructed on the broad-host-range, mobilizable cosmid pLAFR1 or pLAFR3, were conjugated with one of the mutants, designated XC1708. Recombinant plasmids isolated by their ability to complement XC1708 can be classified into two categories. One, represented by pLASC3, can complement some mutants, whereas the other, represented by a single plasmid, pLAHH2, can complement all of the other mutants. Restriction mapping showed that the two recombinant plasmids shared an EcoRI fragment of 8.9 kb. Results from subcloning, deletion mapping, and mini-Mu insertional mutation of the 8.9-kb EcoRI fragment suggested that a 4.2-kb fragment was sufficient to complement the mutant XC1708. Sequence analysis of this 4.2-kb fragment revealed three consecutive open reading frames (ORFs), ORF1, ORF2, and ORF3. Hybridization experiments showed that Tn5 in the genome of XC1708 and other mutants complemented by pLASC3 was located in ORF3, which could code for a protein of 83.5 kDa. A signal peptidase II processing site was identified at the N terminus of the predicted amino acid sequence. Sequence homology of 51% was observed between the amino acid sequences predicted from ORF3 and the pulD gene of Klebsiella species.


Comparing differences in support needs as perceived by parents of adult offspring with down syndrome, autism spectrum disorder and cerebral palsy.
Parents often face many barriers when taking care of their offspring with disabilities. In childhood, support needs vary with families of children with Down syndrome often reporting less caregiving challenges. However, it is unclear whether support needs vary in adulthood. This study compared parents of adults with Down syndrome (DS), autism spectrum disorder (ASD) and cerebral palsy (CP) regarding support needs of their offspring with intellectual and developmental disabilities (IDD) and their families. Data were collected via a national survey in the United States with 189 parents of adults with IDD. Across the quantitative and qualitative analyses, parents of adults with DS (versus CP and ASD) reported significantly greater recreational, natural supports, more formal services and less future planning barriers. The results indicate that the DS advantage may persist in adulthood regarding support needs. More research is needed to understand different types of support needs.


Decontamination of equipment and the environment.
This article explains the various ways decontamination of equipment and the environment can be carried out and highlights the importance of nurses being familiar with such methods so to reduce the possibility of infection.


Effects of dexamphetamine with and without alcohol on simulated driving.
In party circuits dexamphetamine is frequently used in combination with alcohol. It is hypothesized that co-administration of dexamphetamine to alcohol might reduce the sedative effects of alcohol, but may potentiate risk-taking behaviour. The study was aimed at assessing the effects of alcohol, dexamphetamine and the combination of both on simulated driving and cognitive performance. Eighteen subjects participated in a randomized, crossover, placebo-controlled study employing four conditions: 10 mg dexamphetamine, 0.8 g/kg alcohol, 10 mg dexamphetamine + 0.8 g/kg alcohol, and placebo. Fundamental driving skills and risk-taking behaviour were assessed in a driving simulator. Subjects also completed vigilance and divided attention tasks, and subjective ratings. Mean BAC levels during simulated driving were between 0.91‰ and 0.64‰. Subjects using alcohol showed a significantly larger mean standard deviation of lateral position and shorter accepted gap time and distance. Use of alcohol or dexamphetamine + alcohol was associated with a higher frequency of red light running and collisions than the dexamphetamine or placebo conditions. Performance of vigilance and divided attention tasks was significantly impaired in the alcohol condition and, to a lesser degree, in the dexamphetamine + alcohol condition. Single doses of 0.8 g/kg alcohol increased risk-taking behaviours and impaired tracking, attention and reaction time during a 3-h period after drinking when BACs declined from 0.9 to 0.2 mg/ml. The stimulatory effects of co-administration of dexamphetamine 10 mg were not sufficient to overcome the impairing effects of alcohol on skills related to driving.


Randomized prospective evaluation of nephrostomy tube configuration: impact on postoperative pain.
Conduct a prospective randomized single-blind comparison of two nephrostomy catheter designs, evaluating specifically intraoperative placement and postoperative comfort. The single-blind, prospective randomized trial was conducted with institutional review board approval. All patients undergoing percutaneous nephrolithotomy who gave informed consent were randomized to placement of either a Boston Scientific Flexima 8.3F pigtail nephrostomy tube #27-180 (PIG) or a Boston Scientific 8.2F nephroureteral stent #410-126 (NUS). Randomization was concealed from the surgeon until time of placement. Subjective intraoperative placement characteristics were rated by the surgeon on a scale of 1 = excellent, 2 = fair, 3 = good and 4 = poor. The patient's postoperative pain intensity was evaluated with a Visual Analog Pain Score (0 = no pain to 10 = worst pain). Nine patients were randomized to each group. The PIG group was rated significantly better than the NUS group with regards to ease of placement (p = 0.007) and radiopacity of the tube (p = 0.007) by surgeon. Visual analog pain scores on postoperative day one, was significantly lower in the PIG group (mean = 2+/-2) than the NUS group (mean = 5+/-1) (p =0.004). The mean amount of intra-venous morphine equivalent given in the PIG group (mean = 1+/4 Eq morphine) was less on average compared to the NUS group (mean = 6+/13 Eq morphine), but the differences did not reach statistical significance (p = 0.06). Following percutaneous nephrolithotomy, use of a small pig-tail nephrostomy tube results in greater ease of placement and less postoperative pain than a nephroureteral catheter.


High-throughput sperm assay using label-free microscopy: morphometric comparison between different sperm structures of boar and stallion spermatozoa.
The capacity for microscopic evaluation of sperm is useful for assisted reproductive technologies (ART), because this can allow for specific selection of sperm cells for in vitro fertilization (IVF). The objective of this study was to analyze the same sperm samples using two high-resolution methods: spatial light interference microscopy (SLIM) and atomic force microscopy (AFM) to determine if with one method there was more timely and different information obtained than the other. To address this objective, there was evaluation of sperm populations from boars and stallions. To the best of our knowledge, this is the first reported comparison when using AFM and high-sensitivity interferometric microscopy (such as SLIM) to evaluate spermatozoa. Results indicate that with the use of SLIM microscopy there is similar nanoscale sensitivity as with use of AFM while there is approximately 1,000 times greater throughput with use of SLIM. With SLIM, there is also allowace for the measurement of the dry mass (non-aqueous content) of spermatozoa, which may be a new label-free marker for sperm viability. In the second part of this study, there was analysis of two sperm populations. There were interesting correlations between the different compartments of the sperm and the dry mass in both boars and stallions. Furthermore, there was a correlation between the dry mass of the sperm head and the length and width of the acrosome in both boars and stallions. This correlation is positive in boars while it is negative in stallions.


Individual and community level factors associated with modern contraceptive utilization among married women in the emerging region of Ethiopia: a multilevel mixed effects analysis of the 2019 Ethiopia Mini-Demographic and health survey.
A demonstrated technique to enhance reproductive health and economic progress is through ensuring that family planning services are accessible universally. Those studies that used Ethiopia Demographic and Health Survey (EDHS) data did not assess individual and community-level factors in contraceptive utilization. Thus, the study employs a multilevel mixed effects analysis approach, which allows for the examination of individual and community-level factors that influence contraceptive utilization. This study analyzed the 2019 Ethiopia Min Demographic and Health Survey datasets. A total of 1916 married women in the 2019 surveys were included in the analysis. The data were analyzed using Stata version 17.0. The data were analyzed using Multi-level mixed-effect logistic regression to identify the individual and community-level factors associated with modern contraceptive utilization. An adjusted odds ratio with a 95% confidence interval was used to. Show the strength and direction of the association and statistical significance was declared at a P value less than 0.05. Factors significantly associated with modern contraceptive utilization were; Muslim and protestant followers [AOR = 0.31, 95% CI: (0.134, 0.714)] and [AOR = 0.35, 95% CI: (0.173, 0.691)] respectively, women with no education (OR = 0.46; 95% CI: 0.293, 0.710), those women who belong to the poor and middle wealth of household [AOR = 0.35, 95% CI: (0.237, 0.527)] and [AOR = 0.56, 95% CI: (0.347, 0.919)] respectively, women who had one to five and greater than or equal to six living children [AOR = 11.36, 95% CI:(2.119, 60.918)] and [AOR = 7.44, 95% CI:(1.437, 38.547)]respectively, Women in clusters poor wealth status [AOR = 0.40, 95% CI: (0.183, 0.875)] and women who belong to the Somali region [AOR = 0.20, 95% CI: (0.0.070, 0.506)]. The study revealed that both individual and community-level factors determined modern contraceptive utilization. At the individual level, the religion of women, educational status, the wealth of the household, and the total number of living children were significantly associated with modern contraceptive utilization. At community-level factors, community wealth status and belonging to the Somali region were significantly associated with modern contraceptive utilization. The findings suggest that interventions aimed at increasing modern contraceptive utilization should target women with lower levels of education, those living in households with lower wealth, and those with larger families. Additionally, efforts should be made to improve access to modern contraceptives in communities with lower wealth status and in regions where traditional beliefs may hinder their use.


Glutathione redox dynamics and expression of glutathione-related genes in the developing embryo.
Embryonic development involves dramatic changes in cell proliferation and differentiation that must be highly coordinated and tightly regulated. Cellular redox balance is critical for cell fate decisions, but it is susceptible to disruption by endogenous and exogenous sources of oxidative stress. The most abundant endogenous nonprotein antioxidant defense molecule is the tripeptide glutathione (γ-glutamylcysteinylglycine, GSH), but the ontogeny of GSH concentration and redox state during early life stages is poorly understood. Here, we describe the GSH redox dynamics during embryonic and early larval development (0-5 days postfertilization) in the zebrafish (Danio rerio), a model vertebrate embryo. We measured reduced and oxidized glutathione using HPLC and calculated the whole embryo total glutathione (GSHT) concentrations and redox potentials (Eh) over 0-120 h of zebrafish development (including mature oocytes, fertilization, midblastula transition, gastrulation, somitogenesis, pharyngula, prehatch embryos, and hatched eleutheroembryos). GSHT concentration doubled between 12h postfertilization (hpf) and hatching. The GSH Eh increased, becoming more oxidizing during the first 12h, and then oscillated around -190 mV through organogenesis, followed by a rapid change, associated with hatching, to a more negative (more reducing) Eh (-220 mV). After hatching, Eh stabilized and remained steady through 120 hpf. The dynamic changes in GSH redox status and concentration defined discrete windows of development: primary organogenesis, organ differentiation, and larval growth. We identified the set of zebrafish genes involved in the synthesis, utilization, and recycling of GSH, including several novel paralogs, and measured how expression of these genes changes during development. Ontogenic changes in the expression of GSH-related genes support the hypothesis that GSH redox state is tightly regulated early in development. This study provides a foundation for understanding the redox regulation of developmental signaling and investigating the effects of oxidative stress during embryogenesis.


Wall Shear Stress and Pressure Fluctuations under Oscillating Stimulation in Helical Square Ducts with Cochlea-like Geometrical Curvature and Torsion.
Our study aims to provide basic insights on the impact of the spiral shape of the cochlea, i.e., of geometric torsion and curvature, on wall pressure and wall shear stress. We employed computational fluid dynamics in square duct models with curvature and torsion similar to those found in human cochleae. The results include wall pressures and wall shear stresses within the ducts under oscillating axial flow. Our findings indicate that the helical shape generates higher transverse wall shear stresses compared to exclusively curved or twisted ducts. The wall pressures and transverse wall shear stresses we found rise to amounts that may be physiologically relevant in the cochlea.Clinical relevance- The role of the spiral shape of the cochlea in hearing physiology remains, for a large part, elusive. For a better apprehension of hearing and its disorders, it is important to investigate the influence of geometric properties on biofluids motion and emerging phenomena in the cochlea.


Nested cytochrome B polymerase chain reaction diagnostics detect sporozoites of hemosporidian parasites in peripheral blood of naturally infected birds.
Some discrepancies between microscopy and PCR-based methods have been recently recorded in the diagnosis of Leucocytozoon spp. infection in naturally infected birds. To clarify this issue, blood samples from 109 yellow-whiskered greenbuls Andropadus latirostris were investigated using both the microscopic examination of blood films and a nested mitochondrial cytochrome b PCR. The overall prevalence of Leucocytozoon spp. infection was 4% after the standard microscopic examination and 17% using the PCR diagnostics. Samples from 9 randomly chosen birds that were microscopy negative, but PCR positive, were then examined microscopically by screening 2 entire blood films from each individual bird. Sporozoites of Leucocytozoon spp. were observed in 4 birds, and 1 gametocyte of the parasite was seen in each of 2 birds. We conclude that sensitive PCR-based diagnostics are able to detect extremely light parasitemias of circulating sporozoites and gametocytes of hemosporidian parasites. Because of the PCR detection of sporozoites of unknown fate in the peripheral circulation, conclusions regarding the distribution of hemosporidian lineages in wildlife should be made with caution. To be accepted as the lineages of successfully developing species of hemosporidians, such PCR-based information should be supported with the detection of blood stages of the parasites. The present study emphasizes the crucial need for a synthesis of information provided by the tools of traditional parasitology and molecular biology, particularly in field studies of blood parasites.


Drosophila RB proteins repress differentiation-specific genes via two different mechanisms.
The RB and E2F proteins play important roles in the regulation of cell division, cell death, and development by controlling the expression of genes involved in these processes. The mechanisms of repression by the retinoblastoma protein (pRB) have been extensively studied at cell cycle-regulated promoters. However, little is known about developmentally regulated E2F/RB genes. Here, we have taken advantage of the simplicity of the E2F/RB pathway in flies to inspect the regulation of differentiation-specific target genes. These genes are repressed by dE2F2/RBF and a recently identified RB-containing complex, dREAM/MMB, in a cell type- and cell cycle-independent manner. Our studies indicate that the mechanism of repression differs from that of cell cycle-regulated genes. We find that two different activities are involved in their regulation and that in proliferating cells, both are required to maintain repression. First, dE2F2/RBF and dREAM/MMB employ histone deacetylase (HDAC) activities at promoter regions. Remarkably, we have also uncovered an unconventional mechanism of repression by the Polycomb group (PcG) protein Enhancer of zeste [E(Z)], which is involved in silencing of these genes through the dimethylation of histone H3 Lys27 at nucleosomes located downstream of the transcription start sites (TSS).


[Validity of carbohydrate-deficient transferrine (CDT) in assessing chronic abuse of ethyl alcohol in urban public transport workers].
The aim of this study was to verify the ability of some chemical-clinical parameters, with particular emphasis on carbohydrate-deficient transferrin (CDT), in assessing chronic abuse of ethanol in a group of urban public transport workers. In the 512 subjects, public transport tram drivers, all males, who entered the study, information on the intake of alcoholic beverages was collected during the periodical health surveillance controls performed according to Italian legislation (DM88/99). In the study subjects the following clinical-chemical parameters were measured: CDT gamma-glutamyltransferase (GGT), mean corpuscular volume of erythrocytes (MCV), aspartate aminotransferase (AST), alanine aminotransferase (ALT). The subjects were divided into five groups according to different levels of alcohol intake: non-drinkers, occasional drinkers, moderate drinkers, habitual drinkers and heavy drinkers. The median values of CDT GGT and MCV were higher in drinkers than in non-drinkers, with an increasing trend in proportion to the amount of ethanol ingested. The validity of each parameter in determining chronic abuse of ethyl alcohol was calculated taking as true the statement on alcohol intake made spontaneously by the subject. CDT was confirmed as the parameter with the best sensitivity and specificity: 90% and 98%, respectively, the negative predictive value was 99%, while the positive predictive power was 45%. The combination of CDT with GGT or MCV led to small improvements in the positive predictive ability, which reached 50% for CDT and MCV and 60% for CDT and GGT. Our results confirmed that, also in the workplace, CDT is the most important parameter for the diagnosis of chronic abuse of alcohol intake, but also showed that the positivity of this marker cannot be taken as certainty of abuse. The adoption of further diagnostic tools is therefore proposed, such as a specific questionnaire to collect information on alcohol intake, and in case of positive CDT a second-level test with a high specificity.


[Leukaemia inhibitory factor (LIF): structure and biological activity].
Leukaemia inhibitory factor (LIF) is a multifunctional cytokine, which plays a role in growth-promotion and differentiation, regulates calcium and bone metabolism, induces acute phase proteins and causes cachexia in organisms with neoplastic disorders. Moreover, LIF participates in the induction of inflammation, and therefore represents an important pathogenic factor of many disorders. The multifunctional properties of LIF have become of special interest to investigators from different disciplines of medicine.


Rational design of human metapneumovirus live attenuated vaccine candidates by inhibiting viral mRNA cap methyltransferase.
The paramyxoviruses human respiratory syncytial virus (hRSV), human metapneumovirus (hMPV), and human parainfluenza virus type 3 (hPIV3) are responsible for the majority of pediatric respiratory diseases and inflict significant economic loss, health care costs, and emotional burdens. Despite major efforts, there are no vaccines available for these viruses. The conserved region VI (CR VI) of the large (L) polymerase proteins of paramyxoviruses catalyzes methyltransferase (MTase) activities that typically methylate viral mRNAs at positions guanine N-7 (G-N-7) and ribose 2'-O. In this study, we generated a panel of recombinant hMPVs carrying mutations in the S-adenosylmethionine (SAM) binding site in CR VI of L protein. These recombinant viruses were specifically defective in ribose 2'-O methylation but not G-N-7 methylation and were genetically stable and highly attenuated in cell culture and viral replication in the upper and lower respiratory tracts of cotton rats. Importantly, vaccination of cotton rats with these recombinant hMPVs (rhMPVs) with defective MTases triggered a high level of neutralizing antibody, and the rats were completely protected from challenge with wild-type rhMPV. Collectively, our results indicate that (i) amino acid residues in the SAM binding site in the hMPV L protein are essential for 2'-O methylation and (ii) inhibition of mRNA cap MTase can serve as a novel target to rationally design live attenuated vaccines for hMPV and perhaps other paramyxoviruses, such as hRSV and hPIV3. Human paramyxoviruses, including hRSV, hMPV, and hPIV3, cause the majority of acute upper and lower respiratory tract infections in humans, particularly in infants, children, the elderly, and immunocompromised individuals. Currently, there is no licensed vaccine available. A formalin-inactivated vaccine is not suitable for these viruses because it causes enhanced lung damage upon reinfection with the same virus. A live attenuated vaccine is the most promising vaccine strategy for human paramyxoviruses. However, it remains a challenge to identify an attenuated virus strain that has an optimal balance between attenuation and immunogenicity. Using reverse genetics, we generated a panel of recombinant hMPVs that were specifically defective in ribose 2'-O methyltransferase (MTase) but not G-N-7 MTase. These MTase-defective hMPVs were genetically stable and sufficiently attenuated but retained high immunogenicity. This work highlights a critical role of 2'-O MTase in paramyxovirus replication and pathogenesis and a new avenue for the development of safe and efficacious live attenuated vaccines for hMPV and other human paramyxoviruses.