Measurement of the kinetics of protein unfolding in viscous systems and implications for protein stability in freeze-drying.
The aim of the study is to determine the degree of coupling between protein unfolding rate and system viscosity at low temperatures in systems relevant to freeze-drying. The cold denaturation of both phosphoglycerate kinase (PGK) and beta-lactoglobulin were chosen as models for the protein unfolding kinetics study. The system viscosity was enhanced by adding stabilizers (such as sucrose), and denaturant (guanidine hydrochloride or urea) was added to balance the stabilizing effect of sucrose to maintain the cold denaturation temperature roughly constant. The protein unfolding kinetics were studied by both temperature-controlled tryptophan emission fluorescence spectroscopy and isothermal high-sensitivity modulated differential scanning calorimetry (MDSC) (Tzero). Viscometers were used to determine the system viscosity. To verify the predictions of structure based on protein unfolding dynamics, protein formulations were freeze-dried above the glass transition temperatures, and the protein structures in dry products were determined by fluorescence spectroscopy of reconstituted solids by extrapolation of the solution data to the time of reconstitution. Empirical equations describing the effect of sucrose and denaturant (urea and guanidine hydrochloride) on protein cold denaturation were developed based on DSC observations [X. C. Tang and M. J. Pikal. The Effects of Stabilizers and Denaturants on the Cold Denaturation Temperature of Proteins and Implications for Freeze-Drying. Pharm. Res. Submitted (2004)]. It was found that protein cold denaturation temperature can be maintained constant in system of increasing sucrose concentration by simultaneous addition of denaturants (urea and guanidine hydrochloride) using the empirical equations as a guide. System viscosities were found to increase dramatically with increasing sucrose concentration and decreasing temperature. The rate constants of protein unfolding (or the half-life of unfolding) below the cold denaturation temperature were determined by fitting the time dependence of either fluorescence spectroscopy peak position shift or DSC heat capacity increase to a first-order reversible kinetic model. The half-life of unfolding did slow considerably as system viscosity increased. The half-life of PGK unfolding, which was only 3.5 min in dilute buffer solution at -10 degrees C, was found to be about 200 min in 37% sucrose at the same temperature. Kinetics of protein unfolding are identical as measured by tryptophan fluorescence emission spectroscopy and by high-sensitivity modulated DSC. The coupling between protein unfolding kinetics and system viscosity for both proteins was significant with a stronger coupling with PGK than with beta-lactoglobulin. The half-lives of PGK and beta-lactoglobulin unfolding are estimated to be 5.5 x 10(11) and 2.2 years, respectively, even when they are freeze-dried in sucrose formulations 20 degrees C above Tg'. Thus, freeze-drying below Tg' should not be necessary to preserve the native conformation. In support of this conclusion, native PGK was obtained after the freeze-drying of PGK at a temperature more than 60 degrees C above the system Tg' in a thermodynamically unstable system during freeze-drying. Protein unfolding kinetics is highly coupled with system viscosity in high viscosity systems, and the coupling coefficients are protein dependent. Protein unfolding is very slow on the time scale of freeze-drying, even when the system is freeze-dried well above Tg'. Thus, it is not always necessary to freeze-dry protein formulations at temperature below Tg' to avoid protein unfolding. That is, protein formulations could be freeze-dried at product temperature far above the Tg', thereby allowing much shorter freeze-drying cycle times, with dry cake structure being maintained by the simultaneous use of a bulking agent and a disaccharide stabilizer.


Analogs of C-type natriuretic peptide as a potential new non-surgical treatment strategy in knee osteoarthritis.
Osteoarthritis (OA) is a common, chronic, progressive, and multifactorial musculoskeletal system disease affecting millions of people around the world. Despite the use of several treatment modalities, the search for a disease modifying drug continuous. Recent evidence suggest involvement of C-type natriuretic peptide (CNP) signaling in induction of chondroprotective pathways. A CNP analog (BMN 111) with an extended plasma half-life due to its neutral-endopeptidase resistance has shown to be pharmacologically active in achondroplasia enabling to hypothesize that BMN 111 may also be used as a treatment strategy in OA, in which CNP signaling has been suggested to be protective and/or reparative.


Narcolepsy presenting as REM sleep behaviour disorder.
REM sleep behaviour disorder (RBD) is a neurological condition well known to be associated with the synucleinopathies in middle-aged patients. However, there is much less data concerning its development, evolution, and association with other disorders in younger patients. We report two patients aged less than 33 years who presented with clinical and polysomnographical features of RBD, both of whom proved to have previously undiagnosed narcolepsy. Whilst the association of narcolepsy with RBD has been previously recognised, this is the first report of narcoleptic patients presenting with RBD. Narcolepsy should be included in the differential diagnosis of young patients presenting with abnormal behaviour during sleep compatible with RBD.


Can vibrotactile working memory store multiple items?
Vibrotactile working memory is increasing in popularity as a model system to test theories of working memory. Notably, however, we know little about vibrotactile working memory capacity. While most other domains of working memory are able to store multiple items (for example, the seven-plus-or-minus-two capacity of verbal memory [17]), previous examinations of vibrotactile working memory suggest that stored items may suffer from high levels of interference in the form of overwriting or representation-based interference [2,4], potentially limiting capacity and also limiting our ability to draw comparisons between vibrotactile working memory and other forms of working memory. In the present study, we use a two-item delayed match-to-sample paradigm to demonstrate that subjects are able to store multiple items in vibrotactile working memory, suggesting that interference does not catastrophically limit capacity, and strengthening our ability to compare vibrotactile working memory to other working memory tasks.


Jaggery: an avoidable cause of severe, deadly pediatric burns.
Jaggery is the non-industrial refinement of sugar cane into a sugar product. Sugar cane cultivation, harvest and refinement are central aspects of rural Indian life. We present a retrospective review of pediatric burns at a single institution in Southern India, drawing special attention to scald burns incurred when young children fall into the cauldron of boiling jaggery. Descriptive statistics comparing children burned by jaggery and children burned by other mechanisms were performed. Multivariable logistic regression including burn size and mechanism of burn (jaggery and non-jaggery) was performed to determine the increased risk of death when burned by jaggery. Children burned by jaggery immersions are older, more likely male, and have larger burns. They have longer hospital stays, more operations, and are more likely to die. When controlling for age, gender, size of burn, and mechanism, jaggery exposure was associated with a higher mortality. Jaggery burns are deadly, devastating burns which could be prevented. While jaggery and sugar cane production can lead to economic independence for rural Indian villages, the cost it exacts from burns and death to the youngest and most vulnerable children must be addressed and prevented.


Coil-like structure of the inner core of chordae tendineae.
Scanning electron microscopy was used to examine the chordae tendineae in young (approximately 20 years old) and old (approximately 85 years old) cadavers. A network of collagen fibrils was observed in both the outer and intersurface layers of the chordae tendineae. In young subjects, a regularly arranged series of disconnected collagenous ring-like structures surrounded the longitudinal collagen bundles forming the inner core of the chordae tendineae. In old subjects, the ring-like structures of the chordae tendineae were few or absent. The ring-like structures may be related to reducing mechanical stress during the tightening, twisting, and slackening of the chordae tendineae.


Improving blood donor recruitment and retention: integrating theoretical advances from social and behavioral science research agendas.
Increasing blood donor recruitment and retention is of key importance to transfusion services. Research within the social and behavioral science traditions has adopted separate but complementary approaches to addressing these issues. This article aims to review both of these types of literature, examine theoretical developments, identify commonalities, and offer a means to integrate these within a single intervention approach. The social and behavioral science literature on blood donor recruitment and retention focusing on theory, interventions, and integration is reviewed. The role of emotional regulation (anticipated anxiety and vasovagal reactions) is central to both the behavioral and the social science approaches to enhancing donor motivation, yet although intentions are the best predictor of donor behavior, interventions targeting enactment of intentions have not been used to increase donation. Implementation intentions (that is, if-then plans formed in advance of acting) provide a useful technique to integrate findings from social and behavioral sciences to increase donor recruitment and retention. After reviewing the literature, implementation intention formation is proposed as a technique to integrate the key findings and theories from the behavioral and social science literature on blood donor recruitment and retention.


Effect of two graded doses of whole-body X-irradiation and radioprotection by the use of S-phenethyl formamidino 4(N-ethyl isothioamide) morpholine dihydrochloride.
The protection offered by a newly synthesized compound (S-phenethyl-formamidino-4(N-ethyl isothioamide) morpholine dihydrochloride) against radiation effects on DNA, RNA and protein biosynthetic processes in the brain, and on metabolites of 5-HT and nor-adrenalin, i.e., 5-HIAA and VMA, in the urine, including the radiobiological damage to thyroid and testes, was evaluated. The use of the compound prior to irradiation prevented radiation-induced changes in the thyroid and testes. The radiation-induced alterations in the pattern of DNA, RNA, protein in the brain, and in 5-HIAA and VMA in urine could be averted by treatment with this compound prior to each dose of X-irradiation.


Reinduction of remission of chronic myeloid leukemia by donor leukocyte transfusion following relapse after bone marrow transplantation: recovery complicated by initial pancytopenia and late dermatomyositis.
A significant proportion of patients relapse after allogeneic BMT for CML. These relapses have been treated by induction of a graft-versus-leukemia effect by transfusing donor leukocytes. We have treated a 27-year-old woman with interferon and donor leukocyte transfusion and a complete haematological and cytogenetic remission was obtained coincident with the onset of GVHD. Her course was complicated by prolonged and profound pancytopenia which was fully reversed by the administration of rGM-CSF. She remains in CR with mild dermatomyositis due to chronic GVHD 17 months after the procedure.


What is usual about "treatment-as-usual"? Data from two multisite effectiveness trials.
Despite increased emphasis on broadening the implementation of empirically supported therapies (ESTs) to improve standard clinical practice and patient outcomes, objective descriptions of what actually constitutes standard practice in community-based drug abuse treatment do not exist. We present data from independent ratings of 379 audiotapes drawn from the "treatment-as-usual" arm of two multisite randomized effectiveness trials in the National Institute on Drug Abuse Clinical Trials Network. As expected, the most frequently occurring strategies involved assessing the participant's substance use and social functioning, asking open-ended questions, discussing problems and feedback, and giving advice and direction. However, a number of interventions associated with ESTs were very rarely implemented in these early sessions. These data suggest missed opportunities for optimally engaging patients in the early stages of treatment and enhancing substance use outcomes and only moderate success to date of efforts to bridge the gap between research and practice.


Radiation dose optimization potential of deep learning-based reconstruction for multiphase hepatic CT: A clinical and phantom study.
This clinical and phantom study aimed to evaluate the impact of deep learning-based reconstruction (DLR) on image quality and its radiation dose optimization capability for multiphase hepatic CT relative to hybrid iterative reconstruction (HIR). Task-based image quality was assessed with a physical evaluation phantom; the high- and low-contrast detectability of HIR and DLR images were computed from the noise power spectrum and task-based transfer function at five different size-specific dose estimate (SSDE) values in the range 5.3 to 18.0-mGy. For the clinical study, images of 73 patients who had undergone multiphase hepatic CT under both standard-dose (STD) and lower-dose (LD) examination protocols within a time interval of about four-months on average, were retrospectively examined. STD images were reconstructed with HIR, while LD with HIR (LD-HIR) and DLR (LD-DLR). SSDE, quantitative image noise, and contrast-to-noise ratio (CNR) were compared between protocols. The noise magnitude, noise texture, streak artifact, image sharpness, interface smoothness, and overall image quality were subjectively rated by two independent radiologists. In phantom study, the high- and low-contrast detectability of DLR images obtained at 5.3-mGy and 7.3-mGy, respectively, were slightly higher than those obtained with HIR at the STD protocol dose (18.0-mGy). In clinical study, LD-DLR yielded lower image noise, higher CNR, and higher subjective scores for all evaluation criteria than STD (all, p ≤ 0.05), despite having 52.8% lower SSDE (8.0 ± 2.5 vs. 16.8 ± 3.4-mGy). DLR improved the subjective and objective image quality of multiphase hepatic CT compared with HIR techniques, even at approximately half the radiation dose.


Case reports and hypothesis: a neglect-like syndrome may be responsible for the motor disturbance in reflex sympathetic dystrophy (Complex Regional Pain Syndrome-1).
Reflex sympathetic dystrophy (RSD, Complex Regional Pain Syndrome-1, CRPS-1) is a chronic pain disorder associated with autonomic dysregulation that most commonly involves a limb. In addition to pain, motor dysfunction in the involved extremity may be a significant cause of disability. To spur reassessment of underlying mechanisms and development of novel rehabilitative approaches for RSD, we propose a new hypothesis: a neurologic neglect-like syndrome may explain the motor dysfunction that occurs in a subgroup of RSD patients. This hypothesis is based on a selected series of 11 patients who underwent specific neglect testing. The etiology of neglect in RSD is not clear, but we hypothesize that changes within central nervous system (CNS) structures may occur following persistent abnormal activation of the peripheral and autonomic nervous systems, which then may result in a neglect-like syndrome. Further study is needed to verify our clinical observations and test this hypothesis.


Genetic variation and intestinal cholesterol absorption in humans: A systematic review and a gene network analysis.
Intestinal cholesterol absorption varies widely between individuals, which may translate into differences in responsiveness to cholesterol-lowering drugs or diets. Therefore, understanding the importance of genetic variation on cholesterol absorption rates and the complex intestinal cholesterol network is important. Based on a systematic review, genetic variants in seven genes (ABCG5, ABCG8, ABO, APOE, MTTP, NPC1L1, and LDLR) were identified that were associated with intestinal cholesterol absorption. No clear associations were found for variants in APOA4, APOB, CETP, CYP7A1, HMGCR, SCARB1, SLCO1B1, and SREBF1. The seven genes were used to construct an intestinal cholesterol absorption network. Finally, a network with fifteen additional genes (APOA1, APOA4, APOB, APOC2, APOC3, CETP, HSPG2, LCAT, LDLRAP1, LIPC, LRP1, OLR1, P4HB, SAR1B, and SDC1) was generated. The constructed network shows that cholesterol absorption is complex. Further studies are needed to validate and improve this network, which may ultimately lead to a better understanding of the wide inter-individual variability in intestinal cholesterol absorption and the development of personalized interventions.


Can urinary excretion rate of malondialdehyde, uric acid and protein predict the severity and impending death in perinatal asphyxia?
Perinatal asphyxia (PA) associated with multi-organ damage is a leading cause of neonatal mortality and morbidity. We evaluated if urinary malondialdehyde:creatinine (UMDA:Cr), uric acid:creatinine (UUA:Cr) and protein:creatinine (UP:Cr) vary with the severity of PA and if these parameters can predict the impending death in PA. Study included 20 asphyxiated and 20 healthy newborn males. Hypoxic-ischemic encephalopathy (HIE) staging, APGAR (activity, pulse, grimace, appearance and respiration) score and urinary protein, uric acid, creatinine and MDA were evaluated. UMDA:Cr, UUA:Cr and UP:Cr were significantly higher and correlated with APGAR and HIE in PA. By regression analysis also, urinary parameters were found to have significant association with HIE stage and APGAR in PA. Receiver operating characteristics (ROC) curve of UP:Cr, UUA:Cr and UMDA:Cr showed area under curve of 0.896 (p=0.003), 0.859 (p=0.008) and 0.849 (p=0.010) with cut-off value of 9.04 mg, 2.34 mg and 3.49 microg/mg of creatinine respectively that can optimally predict the impending death in PA. SDS-PAGE of unconcentrated urine detected both high (73 kDa and 68 kDa) and low molecular weight proteins (52 kDa, 47 kDa, 25 kDa and 20 kDa) in PA but not in controls. Urinary excretion rate of uric acid, MDA and proteins is higher and has potential to act as biochemical markers for severity evaluation and death prediction in PA.


Subcutaneous Infliximab, CT-P13 SC: A Profile of Its Use in the EU.
CT-P13 (Remsima®; Inflectra®), a biosimilar of reference infliximab (Remicade®), provides a useful alternative for patients requiring infliximab therapy and, as with other biosimilar agents, has the potential to reduce treatment costs. Furthermore, the availability of CT-P13 in a subcutaneous formulation (CT-P13 SC), with the possibility (after adequate training) of self-administration at home, has the potential to both improve patient convenience and reduce the burden on the healthcare system. The initial approval of CT-P13 SC, for use in the treatment of rheumatoid arthritis in adults, was based on the findings of a randomised, double-blind phase I/III trial which demonstrated the non-inferiority of CT-P13 SC administered once every 2 weeks to intravenous CT-P13 (CT-P13 IV) administered once every 8 weeks in reducing disease activity in patients with active rheumatoid arthritis. Subsequently, based on pharmacokinetic data in patients with inflammatory bowel disease, CT-P13 SC has also been approved in the EU for use in the treatment of Crohn's disease, ulcerative colitis and, by extrapolation, ankylosing spondylitis, psoriatic arthritis and psoriasis, in adults.


Control of Aβ release from human neurons by differentiation status and RET signaling.
Few studies have compared the processing of endogenous human amyloid precursor protein (APP) in younger and older neurons. Here, we characterized LUHMES cells as a human model to study Alzheimer's disease-related processes during neuronal maturation and aging. Differentiated LUHMES expressed and spontaneously processed APP via the secretase pathways, and they secreted amyloid β (Aβ) peptide. This was inhibited by cholesterol depletion or secretase inhibition, but not by block of tau phosphorylation. In vitro aged cells increased Aβ secretion without upregulation of APP or secretases. We identified the medium constituent glial cell line-derived neurotrophic factor (GDNF) as responsible for this effect. GDNF-triggered Aβ release was associated with rapid upregulation of the GDNF coreceptor "rearranged during transfection" (RET). Other direct (neurturin) or indirect (nerve growth factor) RET activators also increased Aβ, whereas different neurotrophins were ineffective. Downstream of RET, we found activation of protein kinase B (AKT) to be involved. Accordingly, inhibitors of the AKT regulator phosphatidylinositol-3-kinase completely blocked GDNF-triggered AKT phosphorylation and Aβ increase. This suggests that RET signaling affects Aβ release from aging neurons.


The hepatocellular transport of sulfobromophthalein-glutathione by clofibrate treated, perfused rat liver.
The hypolipidemic drug clofibrate is known to affect the hepatic transport of various organic anions including bilirubin, fatty acids and sulfobromophthalein. Changes in the rate of metabolism and/or intracellular transport have been claimed responsible for the effect. To evaluate these possibilities, the transport of sulfobromophthalein-glutathione, a model compound that does not require metabolism for biliary excretion, was studied in perfused livers isolated from clofibrate-treated and control rats. Cytosolic fatty acid binding protein and glutathione S-transferase activity were also measured. Clofibrate treatment significantly increased liver weight; as a result glutathione S-transferase activity (toward 1-chloro-2,4-dinitrobenzene) fell if expressed per gram of liver (4560 +/- 420 (SE) vs 7010 +/- 260 nmoles/min for clofibrate treated and controls respectively, p less than 0.002), but was unchanged when expressed per total liver (60.8 +/- 6.5 vs 64.6 +/- 3.5 mumoles/min for clofibrate and controls p greater than 0.5). Irrespective of how it was expressed fatty acid binding protein was significantly increased by the drug treatment. Steady state sulfobromophthalein-glutathione removal velocity was saturable with increasing concentrations of sulfobromophthalein-glutathione in both control and clofibrate-treated livers. Steady state extraction ratio, as well as Vmax and Km for removal, did not differ between the two groups. In keeping with other observations, these data collectively indicate that the hepatic steady state removal of nonmetabolized compounds is not affected by clofibrate.(ABSTRACT TRUNCATED AT 250 WORDS)


Inter-animal radiation as potential heat stressor in lying animals.
A model for predicting inter-animal radiant heat exchange in shaded animals is presented, with emphasis on mature cattle. When a cow's surface temperature is 35 °C, as is common in warmer climates, it loses ∼510 Watt m(-2) as radiant heat. Net radiant heat balance depends on radiation coming from bodies in the vicinity. In the 30 °C radiant temperature shaded environment typical of warm climates, net radiant loss from a lactating cow is ∼60 Watt m(-2), i.e., 30 % of its ∼173 Watt m(-2) heat production. Cows rest for 8-14 h day(-1). The heat exchange of a lying cow differs from that of a standing one: the body center is low and 20-30 % of its surface contacts a surface of relatively low heat conductance. Lying reduces the impact of the surrounding shaded area on heat exchange but increases that of heat radiating from neighboring cows. When a cow rests adjacent to other cows, with 1.25 m between body centers when in stalls, it occupies about 140° of the horizontal plane of view. Heat emitted from the animal's surface reduces the net radiant heat loss of a resting cow by ∼30 Watt m(-2). In contrast, the presence of cows at 5 and 10 m distance, e.g., cows resting on straw in loose yard housing, reduces the net radiant heat loss of the resting cow by 9 and 5 Watt m(-2), respectively. Radiant heat input increases with animal density, which is beneficial in cooler climates, but acts as a stressor in warm climates.


Medial arterial calcification and its association with mortality and complications of diabetes.
Medical arterial calcification was studied among 4,553 subjects in a 20-year, longitudinal study of Pima Indians. The prevalence and incidence of medial arterial calcification were highest among men, the elderly, and patients with Type 2 (non-insulin-dependent) diabetes mellitus. Medial arterial calcification was most commonly observed in the feet and appeared to progress proximally. Proportional hazards analysis was used to evaluate risk factors for medial arterial calcification in the feet and to evaluate medial arterial calcification as a risk factor for death and for complications of diabetes. Among diabetic patients, risk factors for medial arterial calcification were impaired vibration perception, long duration of diabetes, and high plasma glucose concentration (p less than 0.01 for each). Among nondiabetic subjects, age, male gender (p less than 0.01 for each), and high serum cholesterol concentration (p = 0.02) were risk factors for medial arterial calcification. Nondiabetic subjects with medial arterial calcification did not have higher mortality rates than subjects without medial arterial calcification (rate ratio = 0.95, 95% confidence interval = 0.7-1.3). Diabetic patients with medial arterial calcification, compared with diabetic patients without medial arterial calcification, had 1.5-fold the mortality rate (95% confidence interval = 1.0-2.1), 5.5-fold the rate of amputations (95% confidence interval = 2.1-14.1), 2.4-fold the rate of proteinuria (95% confidence interval = 1.3-4.5), 1.7-fold the rate of retinopathy (95% confidence interval = 0.98-2.8), and 1.6-fold the rate of coronary artery disease (95% confidence interval = 0.48-5.4).


Interpretive policy analysis: Marshallese COFA migrants and the Affordable Care Act.
Since the enactment of the Affordable Care Act (ACA), the rate of uninsured in the United States has declined significantly. However, not all legal residents have benefited equally. As part of a community-based participatory research (CBPR) partnership with the Marshallese community, an interpretative policy analysis research project was conducted to document Marshallese Compact of Free Association (COFA) migrants' understanding and experiences regarding the ACA and related health policies. This article is structured to allow the voice of Marshallese COFA migrants to explain their understanding and interpretation of the ACA and related polices on their health in their own words. Qualitative data was collected from 48 participants in five focus groups conducted at the local community center and three individual interviews for those unable to attend the focus groups. Marshallese community co-investigators participated throughout the research and writing process to ensure that cultural context and nuances in meaning were accurately captured and presented. Community co-investigators assisted with the development of the semi-structured interview guide, facilitated focus groups, and participated in qualitative data analysis. Content analysis revealed six consistent themes across all focus groups and individual interviews that include: understanding, experiences, effect on health, relational/historical lenses, economic contribution, and pleas. Working with Marshallese community co-investigators, we selected quotations that most represented the participants' collective experiences. The Marshallese view the ACA and their lack of coverage as part of the broader relationship between the Republic of the Marshall Islands (RMI) and the United States. The Marshallese state that they have honored the COFA relationship, and they believe the United States is failing to meet its obligations of care and support outlined in the COFA. While the ACA and Medicaid Expansion have reduced the national uninsured rate, Marshallese COFA migrants have not benefited equally from this policy. The lack of healthcare coverage for the Marshallese COFA migrants exacerbates the health disparities this underserved population faces. This article is an important contribution to researchers because it presents the Marshallese's interpretation of the policy, which will help inform policy makers that are working to improve Marshallese COFA migrant health.


Can the alcohol withdrawal scale be applied to post-operative patients?
Assessment scales are commonly used to diagnose and treat alcohol withdrawal syndrome (AWS) in acute hospitals, although they have only been validated for use in detoxification facilities. There is a significant overlap between the symptoms and signs of AWS and other clinical presentations, including systemic inflammatory response syndrome (SIRS) and the physiological response to surgery. This may lead to both over-diagnosis and inappropriate treatment of AWS. This study sought to determine the false-positive rate for the commonly used Clinical Institute Withdrawal Assessment for Alcohol-Revised (CIWA-Ar) among post-operative patients. This was a prospective study of patients undergoing major abdominal surgery at University Hospital Geelong. Patients were recruited who were NOT at risk of alcohol dependency (using the World Health Organisation Alcohol Use Disorders Identification Test). Patients were assessed for AWS using the CIWA-Ar day one post-operatively with a false positive measured as a CIWA-Ar > 7. A total of 67 patients were included in the study. There were 31 (46%) men and 36 women. Their median age was 52 years (range 27-85). Thirty-six (52%) of patients underwent elective procedures, and 32 were emergencies. Twelve of the 67 patients (18%) had CIWA-Ar scores >seven. In the early post-operative period, the CIWA-Ar tool over-diagnoses AWS in 18% of patients. These false-positives could lead to delayed treatment of serious underlying conditions. We call for caution in the use of alcohol withdrawal scales in the acute hospital setting.


Acid-induced esophagitis in cats is prevented by sucralfate but not synthetic prostaglandin E.
The cytoprotective effects of liquid sucralfate and a synthetic analog of prostaglandin E1 (PGE1) on acid-induced esophagitis in cats were studied. Esophagitis was induced in adult cats using a constant infusion of 0.1 N HCl at 1 ml/min for 20 min. Animals were infused for either one or three days. Mucosal lesions were evaluated by blinded investigators using both fiberoptic endoscopy and light microscopy. Histologic changes included basal cell hyperplasia, intraepithelial leukocytosis, and subepithelial leukocytosis. Liquid sucralfate given prior to acid infusion consistently prevented acid-induced lesions in both one- and three-day infusions, demonstrated by both endoscopy and quantitative histologic scoring. Indomethacin (200 micrograms/kg) given prior to sucralfate and acid did not affect sucralfate cytoprotection. Synthetic PGE1, given in doses of 5 micrograms/kg and 100 micrograms/kg, afforded no esophageal cytoprotection. These studies indicate that sucralfate is cytoprotective against acid-induced esophageal injury in cats, an effect that does not appear to be mediated by prostaglandin. In addition, synthetic PGE1 does not confer protection in this animal model.


Photo-acclimatory thresholds anticipate sudden shifts in seagrass ecosystem state under reduced light conditions.
Seagrass ecosystems usually respond in a nonlinear fashion to increasing pressures and environmental changes. Feedback mechanisms operating at the ecosystem level and involving multiple interactions among the seagrass meadow, its associated community and the physical environment are known to play a major role in such nonlinear responses. Phenotypic plasticity may also be important for buffering these ecological thresholds (i.e., regime shifts) as many physiological processes show nonlinear responses to gradual environmental changes, conferring the appearance of resistance before the effects at the organism and population levels are visible. However, the potential involvement of plant plasticity in driving catastrophic shifts in seagrass ecosystems has not yet been assessed. In this study, we conducted a manipulative 6-month light-gradient experiment in the field to capture nonlinearities of the physiological and population responses of the seagrass Cymodocea nodosa to gradual light reduction. The aim was to explore if and how the photo-acclimatory responses of shaded plants are translated to the population level and, hence, to the ecosystem level. Results showed that the seagrass population was rather stable under increasing shading levels through the activation of multilevel photo-acclimative responses, which are initiated with light reduction and modulated in proportion to shading intensity. The activation of photo-physiological and metabolic compensatory responses allowed shaded plants to sustain nearly constant plant productivity (metabolic carbon balance) along a range of shading levels before losing linearity and starting to decline. The species then activated plant- and meadow-scale photo-acclimative responses and drew on its energy reserves (rhizome carbohydrates) to confer additional population resilience. However, when the integration of all these buffering mechanisms failed to counterbalance the effects of extreme light limitation, the population collapsed, giving place to a phase shift from vegetated to bare sediments with catastrophic ecosystem outcomes. Our findings evidence that ecological thresholds in seagrass ecosystems under light limitation can be explained by the role of species' compensatory responses in modulating population-level responses. The thresholds of these plastic responses anticipate the sudden loss of seagrass meadows with the potential to be used as early warning indicators signalling the imminent collapse of the ecosystem, which is of great value for the real-world management of seagrass ecosystems.


Segregation studies in CHO hybrid cells: I. Spontaneous and mutagen-induced segregation events of two recessive drug-resistant loci.
The process of segreation or phenotypic expression of two recessive drug-resistant loci from heterozygous Chinese hamster ovary hybrid lines is examined. The spontaneous segregation rates of phytohaemagglutinin resistance (Phar) and a temperature-dependent 8-azaguanine-resistant locus (Azarts) from heterozygous quasitetraploid lines using Luria-Delbruck fluctuation analysis were 5 X 10(-5) and 10(-5) events/cell/generation, respectively. In quasihexaploid lines, the latter rates increased 40- and 200-fold, respectively, and were dependent on the number of presumptive drug-sensitive allelel. The mutagens EMS, MNNG, ICR-170, ICR-191, and gamma rays significantly increased the frequency of segregation events. The mutagen-induced frequency of dominant mutations to ouabain (Ouar) and alpha-amanitin (Amar) rsistance in the same hybrid line was much lower in comparison to segregation events and was mutagen specific. The chromosome number per metaphase cell was more variable than DNA content in quasitetraploid lines. These properties of marker segregation are consistent with mechanisms of either restricted chromosome loss, rearrangement, or mutation.


Flexible cystoscopy in surgical repair of traumatic urethral strictures in children.
Flexible cystoscopy via a suprapubic cystostomy tract was utilized to locate the proximal segment of a complete traumatic disruption of the bulbar urethra secondary to a pelvic fracture. With the fiberoptic light of the instrument visible in the perineum, the proximal urethra was easily and safely dissected free from the enveloping fibrotic scar tissue and a simple one-stage urethroplasty performed. The patient is voiding normally eight months postoperatively.


Kinetics of methane production during anaerobic fermentation of chicken manure with sawdust and fungi pre-treated wheat straw.
The goal of this study was to determine the kinetic parameters of methane production during anaerobic fermentation of mixtures of chicken manure with sawdust and wheat straw overgrown with fungi. Pre-treatment of wheat straw was carried out with Pleurotus ostreatus and Trametes versicolor white-rot fungi. Mixtures of chicken manure with sawdust and wheat straw overgrown with fungi at different mass ratios (50:50, 60:40 and 80:20) were used as a substrate for anaerobic fermentation. For the control, ordinary wheat straw was used. Anaerobic fermentations were performed at (35, 40 and 45) °C. An individual process of anaerobic fermentation was maintained at constant temperature for 21 days. During the process, the volume and concentration of biogas produced were monitored. The most biogas produced was recorded for the straw overgrown with Pleurotus ostreatus fungi (ratio 50:50) at 45 °C, and the least in the case of straw overgrown with Trametes versicolor fungi (ratio 80:20) at 35 °C. At the beginning of anaerobic fermentation, the methane concentration increased faster at a higher temperature, while after 21 days, it was between 53 and 56% regardless of temperature.


Brain activity changes in a monkey model of central post-stroke pain.
Central post-stroke pain (CPSP) can occur after stroke in the somatosensory pathway that includes the posterolateral region of the thalamus. Tactile allodynia, in which innocuous tactile stimuli are perceived as painful, is common in patients with CPSP. Previous brain imaging studies have reported plastic changes in brain activity in patients with tactile allodynia after stroke, but a causal relationship between such changes and the symptoms has not been established. We recently developed a non-human primate (macaque) model of CPSP based on thalamic lesions, in which the animals show behavioral changes consistent with the occurrence of tactile allodynia. Here we performed functional magnetic resonance imaging under propofol anesthesia to investigate the changes in brain activation associated with the allodynia in this CPSP model. Before the lesion, innocuous tactile stimuli significantly activated the contralateral sensorimotor cortex. When behavioral changes were observed after the thalamic lesion, equivalent stimuli significantly activated pain-related brain areas, including the posterior insular cortex (PIC), secondary somatosensory cortex (SII), anterior cingulate cortex (ACC), and amygdala. Moreover, when either PIC/SII or ACC was pharmacologically inactivated, the signs of tactile allodynia were dampened. Our results show that increased cortical activity plays a role in CPSP-induced allodynia.


Grip strength test and infrared thermometry as non-invasive methods to complement acute toxicity data in mice.
Acute toxicity testing in-vivo seeks a new approach minimising the number of animals used and increasing the number of quantitative and qualitative observations. The measurement of grip strength of mice (forelimbs), a classical tool for the evaluation of muscular force and the emitted heat measurement, as a tool for evaluation of basic metabolism, have been used. The aim of acute toxicity was, up to now, to determine LD50 level or eventually a minimal lethal dose. These non-invasive tests give an idea of general health condition and allow the determination of the minimal toxic dose while in the search for the minimal lethal dose. A computer-assisted grip strength test and an infrared thermometry test have been developed and calibrated with Na+ pentobarbital, theophylline and diazepam given once orally or intraperitoneally to 10 male NMRI (MAI) mice/dose. The tests were performed within one hour after administration and daily until the values of the surviving animals went back to normal. Dose-response curves were obtained and plotted against control animals (n = 10). Oral and intraperitoneal administration in ascending doses of Na+-pentobarbital, theophylline and diazepam (i.p. only) up to the lethal level have a parallel influence on grip strength and heat emission; the correlation with mortality is clear for Na+-pentobarbital and diazepam but not for theophylline.


Two cycles of the PS-341/bortezomib, adriamycin, and dexamethasone combination followed by autologous hematopoietic cell transplantation in newly diagnosed multiple myeloma patients.
Treatment with 3-6 cycles of PS-341/bortezomib, adriamycin, and dexamethasone (PAD) has been explored in terms of induction therapy prior to autologous stem cell transplantation (ASCT) in patients with multiple myeloma (MM). We evaluated the effects of two cycles of PAD given before ASCT. Patients received two 21-d cycles of PAD (bortezomib 1.3 mg/m(2) × 4 d, adriamycin 9 mg/m(2) × 4 d, and dexamethasone 40 mg × 4 d × 2). Starting on day 12 of cycle 2, patients were given subcutaneous granulocyte-colony stimulating factor to mobilize peripheral blood stem cells (PBSCs). Following PBSC harvesting, ASCT was performed using high-dose melphalan, followed by thalidomide. A total of 32 patients were enrolled. Of 31 who completed two cycles of PAD, 25 (81%) achieved a partial response (PR) or better. Major adverse events were cytopenia, with grade I/II neurotoxicity evident during 4.8% of PAD cycles. Two patients were withdrawn from the study prior to PBSC collection. Thirty patients showed successful mobilization of PBSCs and underwent ASCT, with all 30 showing adequate neutrophil and platelet recovery. Following ASCT, 14 patients (47%) achieved a complete response (CR), 8 (27%) a very good partial response (VGPR), and 6 (20%) PR. Thalidomide was given to 25 patients after ASCT, as maintenance therapy. Twelve patients showed better responses after administration of thalidomide, and a total of 21 patients (70%) achieved CR. The 5-yr probabilities of overall and progression-free survival were 71.1% and 23.5%, respectively. A short course of PAD was effective as an induction treatment before ASCT in patients newly diagnosed with MM. Prospective comparisons with longer courses of such treatment are needed.


MEN1 and pituitary adenomas.
MEN1 gene mutations predispose carriers to pituitary tumors. Molecular pathways involved in the development of these tumors seem different to what is known in sporadic tumors. Clinical studies showed that all types of adenomas can be found with a predominance of prolactinoma and macroadenoma compared to a control population. These MEN1 tumors seem more aggressive, invasive and resistant to treatment requiring a very careful long-life follow-up. Occurrence of these tumors can be described in the pediatric population and it can be the first and only manifestation of MEN1 for some years asking the question of the systematic screening for MEN1 gene mutation in pediatric population with pituitary adenoma.


Should NS5A inhibitors serve as the scaffold for all-oral anti-HCV combination therapies?
Chronic hepatitis C virus (HCV) infection represents a global health problem that affects up to 130-150 million people worldwide. The HCV treatment landscape has been transformed recently by the introduction of direct-acting antiviral (DAA) agents that target viral proteins, including the NS3 protease, the NS5B polymerase, and the NS5A protein. Treatment with multiple DAAs in combination has been shown to result in high rates of sustained virologic response, without the need for pegylated interferon, and a shorter duration of therapy compared with interferon-based regimens; however, the optimal combination of DAAs has yet to be determined. The class of NS5A inhibitors has picomolar potency with pangenotypic activity, and recent clinical studies have shown these inhibitors to be an important component of DAA combination regimens. This review discusses the rational design of an optimal anti-HCV DAA cocktail, with a focus on the role of NS5A in the HCV life cycle, the attributes of the NS5A class of inhibitors, and the potential for NS5A inhibitors to act as a scaffold for DAA-only treatment regimens.


N-Methyl-2-pyridone-5-carboxamide is 1-methylnicotinamide metabolite of low cyclooxygenase-dependent vasodilating activity.
1-Methylnicotinamide (MNA) is a primary metabolite of nicotinamide recently proven to cause systemic increase in PGI(2) plasma levels in an unknown mechanism. Our present study was aimed at verifying whether the increased production of PGI(2), a vasodilating prostanoid, in response to MNA, its metabolite N-methyl-2-pyridone-5-carboxamide (Met2PY), and nicotinamide may be reproduced under in vitro conditions. Since prostacyclin is a vasodilating prostanoid, we also performed the functional tests in the ex vivo model of coronary vascular bed perfusion to evaluate the vasoactive properties of those compounds. We did not observe any significant effect of the tested drugs on either PGI(2) or PGE(2) secretion in our in vitro model. Nicotinamide at the concentrations of 10 and 100 μmol/l and 100 μmol/l Met2PY slightly but significantly increased coronary flow in rat heart. These increases, however, remained very low when compared to that induced by the reference compound, bradykinin (100 nmol/l). Perfusion of rat hearts with Met2PY in the presence of 50 μmol/l indomethacin resulted in decreased coronary flow, which proves that the effect is cyclooxygenase dependent. We conclude that MNA metabolites should be more carefully addressed in reference to pro-prostacyclin activity and that systemic mechanism of MNA-induced PGI(2) production needs further clarification.


A removal experiment with sympatric populations of Apodemus sylvaticus (L.) and A. flavicollis (Melchior) (Rodentia: Muridae).
This study examines the role of competition in the relationship between sympatric populations of Apodemus sylvaticus and A. flavicollis at Woodchester Park, Gloucestershire, England. Three grids were trapped for twelve months to monitor changes in population size, survival, reproduction, body weight, spatial segregation and microhabitat associations. The A. flavicollis was removed from one grid, A. sylvaticus from another while the rodent community on the third was left intact as a control. Subsequent changes in ecological parameters were observed over a further period of 7 months. Isolated populations of both species entered reproductive condition earlier than on the control grid. This resulted in a rapid increase of the A. flavicollis population but the dynamics of A. sylvaticus were unaffected. In the absence of their congener both species made greater use of areas of woodland once frequented by the absent species. However specific microhabitat associations observed in two species communities remained after experimental manipulation. It is concluded that although A. sylvaticus and A. flavicollis do compete for space, this is a comparatively weak interaction which does not lead to habitat exclusion. Competition is probably ameliorated by intrinsic differences in the biology of these Apodemus species.


Effects of sulfasalazine on biopsy mucosal pathologies and histological grading of patients with active ulcerative colitis.
To investigate the mechanisms of sulfasalazine (SASP) in the treatment of ulcerative colitis (UC). Changes of pathological signs and histological grading of 106 patients with active UC were observed before and after the treatment with SASP, 1 g, thrice daily for 6 wk. The effect of SASP on the vasculitis in lamina propria was 48.2% and 17.4% in the mild active UC (P<0.001) and 68% and 26.7% in the moderate active UC (P<0.001) before and after treatment. Fibroid necrosis of vessel wall was found in one case of mild UC and two cases of moderate UC before treatment and was not found after treatment. No thrombosis was found in mild UC before and after treatment, while thrombosis was found in one case of moderate UC before treatment. The effect on mucosal glandular abnormality was 30.4% and 13.0% in mild UC (P<0.05), and 42% and 40% in moderate UC (P>0.05) before and after treatment. The rate of eosinophil infiltration was 98.2% and 80.4% in mild UC (P<0.01), and 100% and 91.1% in moderate UC (P<0.05) before and after treatment. The effect on crypt abscess was 21.4% and 4.4% in mild UC (P<0.05), and 48% and 13.3% in moderate UC (P<0.001) before and after treatment. The effect on mucosal pathohistological grading was 2.00+/-0.84 and 0.91+/-0.46 in mild UC (P<0.001), and 2.49+/-0.84 and 1.31+/-0.75 in moderate UC (P<0.001) before and after treatment. SASP can improve small vessel lesions and crypt abscesses and reduce neutrophilic and eosinophilic leukocyte infiltration in inflammatory mucosa of UC.


Panax notoginseng saponins improve the erectile dysfunction in diabetic rats by protecting the endothelial function of the penile corpus cavernosum.
Diabetes mellitus (DM)-associated ED is predominantly due to neurovascular dysfunction mediated by nitric oxide (NO) suppression. Panax notoginseng saponins (PNS) are widely used for treating cardiovascular disease in China. The aim of this study was to evaluate the effects of PNS on penile erection and corpus cavernosum tissues in rats with diabetes-associated ED. Four weeks after PNS treatment, erectile function was assessed by intracavernous pressure (ICP) and mean arterial pressure (MAP) measurements. The level of NO, cyclic guanosine monophosphate (cGMP) and advanced glycation end products (AGEs) in cavernous tissue were assessed. Immunohistochemical staining and TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) were performed for detecting endothelial NO synthase (eNOS) and apoptosis, respectively. The results show that ICP/MAP ratio was significantly increased in high-dose (150 mg kg(-1) per day) PNS-treated group compared with the diabetic ED untreated group (DM group). Compared with the untreated group, the expression of eNOS and the levels of NO and cGMP were increased in the PNS-treated groups. Moreover, apoptosis was markedly decreased in the group that received 150 mg kg(-1) per day of PNS. These results suggest that PNS may be used for improving the ED in diabetic rats via the NO/cGMP pathway and restores the function of endothelium in corpus cavernosum.


A novel reductive amination method with isotopic formaldehydes for the preparation of internal standard and standards for determining organosulfur compounds in garlic.
Garlic (Allium sativum) is a long-cultivated plant that is widely utilized in cooking and has been employed as a medicine for over 4000 years. In this study, we fabricated standards and internal standards (ISs) for absolute quantification via reductive amination with isotopic formaldehydes. Garlic has four abundant organosulfur compounds (OSCs): S-allylcysteine, S-allylcysteinine sulfoxide, S-methylcysteine, and S-ethylcysteine are abundant in garlic. OSCs with primary amine groups were reacted with isotopic formaldehydes to synthesize ISs and standards. Cooked and uncooked garlic samples were compared, and we utilized tandem mass spectrometry equipped with a selective reaction monitoring technique to absolutely quantify the four organosulfur compounds.


Intraventricular neuroendoscopy: complication avoidance and management.
In the modern era, neuroendoscopy has had an increasingly prominent role in neurosurgery. As attention has focused the development of minimally invasive surgical methods, neuroendoscopy has advanced both as an independent treatment modality for various neurologic disorders and as an adjunct to microneurosurgery. Neuroendoscopy is distinct from traditional surgery, and a thorough understanding of its unique attributes is required to attain maximal benefit. In addition to its advantages, neuroendoscopy is associated with unique obstacles that must be anticipated, appreciated, and accounted for to prevent complications. This article outlines techniques to reduce and manage complications during more common intraventricular neuroendoscopic procedures including endoscopic third ventriculostomy (ETV), colloid cyst resection, tumor biopsy and resection, and treatment of loculated hydrocephalus.


Neural correlates and gait characteristics for hypoxic-ischemic brain injury induced freezing of gait.
To investigate gait characteristics in patients with freezing of gait (FOG) after hypoxic-ischemic brain injury (HIBI) and to elucidate neural correlates for FOG using F-18 fluoro-2-deoxy-d-glucose positron emission tomography. We enrolled 12 patients with FOG after HIBI and 17 patients without FOG after HIBI. We performed three-dimensional gait analyses and compared each parameter and gait variability. Brain metabolism was measured, and we compared regional brain metabolism using a voxel-by-voxel-based statistical mapping analysis. The FOG group revealed a significantly decreased joint range of motion (ROM), particularly in the sagittal plane for three-joint summated ROM (p < 0.0025). Spatiotemporal results demonstrated that stride length and step length were decreased in the with FOG group (p < 0.005). FOG severity was negatively correlated with brain metabolism in the left thalamus, and three-joint summated ROM in the sagittal plane was positively associated with brain metabolism in the left thalamus and midbrain (p < 0.05). Central organizational level amplitude disorder may play an important role in the pathophysiology, and disturbance in the cholinergic pathway might contribute to the development of FOG in patients with HIBI. These findings contribute to understanding FOG in HIBI.


Early growth response transcription factors are required for development of CD4(-)CD8(-) thymocytes to the CD4(+)CD8(+) stage.
Progression of immature CD4(-)CD8(-) thymocytes beyond the beta-selection checkpoint to the CD4(+)CD8(+) stage requires activation of the pre-TCR complex; however, few of the DNA-binding proteins that serve as molecular effectors of those pre-TCR signals have been identified. We demonstrate in this study that members of the early growth response (Egr) family of transcription factors are critical effectors of the signals that promote this developmental transition. Specifically, the induction of three Egr family members (Egr1, 2, and 3) correlates with pre-TCR activation and development of CD4(-)CD8(-) thymocytes beyond the beta-selection checkpoint. Enforced expression of each of these Egr factors is able to bypass the block in thymocyte development associated with defective pre-TCR function. However, Egr family members may play somewhat distinct roles in promoting thymocyte development, because there are differences in the genes modulated by enforced expression of particular Egr factors. Finally, interfering with Egr function using dominant-negative proteins disrupts thymocyte development from the CD4(-)CD8(-) to the CD4(+)CD8(+) stage. Taken together, these data demonstrate that the Egr proteins play an essential role in executing the differentiation program initiated by pre-TCR signaling.


Association of genetic variants in GABRA3 gene and thyrotoxic hypokalaemic periodic paralysis in Thai population.
Genetic predisposition has been suggested to play role in the pathogenesis of thyrotoxic hypokalaemic periodic paralysis (THPP). In this study, we assessed the differences of single-nucleotide polymorphisms (SNP) allelic frequency between THPP patients and well-characterized controls in order to find the susceptibility genetic variants related to THPP using microarray-based assessments on pooled DNA. Fifty cases of THPP and 50 male hyperthyroid patients without hypokalaemia as controls were recruited. Equal amounts of individual genomic DNA were pooled from each group. Estimated allele frequencies of SNPs were derived by averaging relative allele signal score obtained by Affymetrix GeneChip(R) Mapping 10K Arrays. Sixty-nine loci that display robust allele frequency differences between THPP and controls were identified. SNP rs750841 (A > T) in intron 3 of the gamma-aminobutyric acid (GABA) receptor alpha3 subunit (GABRA3) gene possessed the most significant difference in allele frequency (27% in THPP case and 5% in controls, P = 0.007). Actual allele frequencies obtained from genotyping in each individual were very similar to the estimated frequency from the pools (28% in THPP and 2% in controls, and P = 0.0002). Nearby DNA sequences of GABRA3 were sequenced and an additional two SNPs were found (A > C at exon 1 and G > T of rs12688128). Allele A of rs750841 and allele G of rs12688128 in intron 3 were predominantly found in THPP with significant genetic relative risk of 19 (P < 0.0002; 95%CI 2.4-151.6). Whole-genome scanning on pooled DNA provides an accurate, useful screening tool for elucidating genetic underpinnings of THPP. SNPs at intron 3 of GABRA3 are found to be associated with THPP.


Early and late hematologic toxicity following CD19 CAR-T cells.
Autologous T cells transduced with CD19-directed chimeric antigen receptors have recently been approved by several regulatory agencies for the treatment of relapsed and refractory leukemia and lymphoma, after demonstrating remarkable remission rate in advanced patients. The most common adverse events reported are cytokine-release syndrome (CRS), neurotoxicity, and hematologic toxicity. Here, we focus on early and late cytopenia occurring after CD19 CAR-T cells in 38 patients treated with CD19 CAR-T cells. Neutropenia, thrombocytopenia, and anemia occur frequently (94, 80, and 51%, respectively) after CAR-T cell infusion, and are associated with a biphasic nature, as in 93% of patients hematologic toxicity occurs after 21 days from cell infusion. Late hematologic toxicity was more common in patients with high grade CRS and in patients treated after a recent stem cell transplantation. Interestingly, since these events occur late after the lymphodepleting chemotherapy and after resolution of CRS, we found perturbations in SDF-1 levels to correlate with events of late neutropenia, likely associated with B-cell recovery.


Characterisation of genes encoding for extended spectrum β-lactamase in Gram-negative bacteria causing healthcare-associated infections in Mwanza, Tanzania.
Healthcare-associated infections (HCAIs) caused by extended spectrum β-lactamase-producing Gram-negative bacteria (ESBL-GNB) increase morbidity and mortality. This cross-sectional study characterised ESBL genes (bla CTX-M, bla TEM and bla SHV) among 30 ceftriaxone-resistant GNB causing HCAIs between January 2022 and July 2022 by multiplex polymerase chain reaction assay at the zonal referral hospital in Mwanza, Tanzania. Twenty-five (83.3%) had at least one ESBL gene, of which 23/25 (92.0%) carried the bla CTX-M gene. Seventy-two percent (18/25) of the GNB-ESBL isolates carried more than one ESBL gene, of which the majority (88.8%; n = 16/25) carried the bla CTX-M and bla TEM genes. Extended spectrum β-lactamase genes, particularly bla CTX-M, are common among ceftriaxone-resistant GNB causing HCAIs. This study revealed the distribution of genes (bla CTX-M, bla TEM and bla SHV) coding for ESBL production among ceftriaxone resistant GNB causing HCAIs However, all ESBL producing GNB were susceptible towards ceftriaxone-sulbactam indicating that ceftriaxone-sulbactam may be empirically prescribed for treating patients with HCAIs.


Analysis of cortical thickness in narcolepsy patients with cataplexy.
To investigate differences in cortical thickness in narcolepsy patients with cataplexy and control subjects. Cortical thickness was measured using a 3-D surface-based method that enables more accurate measurement in deep sulci and localized regional mapping. University hospital. We enrolled 28 patients with narcolepsy and cataplexy and 33 age-and sex-matched control subjects. Cortical thickness was measured using a direct method for calculating the distance between corresponding vertices from inner and outer cortical surfaces. We normalized cortical surfaces using 2-D surface registration and performed diffusion smoothing to reduce the variability of folding patterns and to increase the power of the statistical analysis. Localized cortical thinning in narcolepsy patients with cataplexy was found in orbitofrontal gyri, dorsolateral and medial prefrontal cortexes, insula, cingulate gyri, middle and inferior temporal gyri, and inferior parietal lobule of the right and left hemispheres at the level of a false discovery rate P<0.05. No significant local increases in cortical thickness were observed in narcolepsy patients. A significant negative correlation was observed between the narcolepsy patients' scores on the Epworth Sleepiness Scale and the cortical thickness of the left supramarginal gyrus. Cortical thinning in narcolepsy patients with cataplexy in localized anatomic brain regions may serve as a possible neuroanatomic mechanism of the disturbances in attention, memory, emotion, and sleepiness.


Choosing the best method to utilise single positron emission computed tomography (SPECT) scans in the management of unilateral condylar hyperplasia.
Facial asymmetry secondary to unilateral condylar hyperplasia can be a diagnostic challenge to oral and maxillofacial surgeons. Single positron emission computed tomography (SPECT) scans provide a useful adjunct. We report a brief summary of the evidence describing the effectiveness of different methods of analysing SPECT scans and compare it with the results of a 10-year study at Sunderland Royal Hospital. Overall, both the evidence base and our study strongly favour use of the condyle:condyle ratio over the condyle:reference bone ratio, suggesting that no further comparisons are needed.


Discovery of novel (1S)-(-)-verbenone derivatives with anti-oxidant and anti-ischemic effects.
A series of novel (1S)-(-)-verbenone derivatives was synthesized bearing a 4-styryl scaffold. The synthesized compounds were tested for their anti-oxidant, anti-excitotoxic, and anti-ischemic activities. These derivatives significantly reduced oxygen-glucose deprivation-induced neuronal injury and N-methyl-D-aspartic acid-evoked excitotoxicity in cortical neurons. Furthermore, compound 3f was identified as a potent anti-ischemic agent in an in vitro ischemic model, potentially due to the inhibition of N-methyl-D-aspartic acid-evoked excitotoxicity and oxidative/nitrosative stress.


[Reform and practice of Biopharmaceutical Technology Comprehensive Experiments in higher vocational colleges in the context of skills competition].
Comprehensive experiments course is a bridge for higher vocational students to integrate theoretical knowledge with production practice. The article introduces that our biological pharmacy department is committed to the principles of "promotion of teaching, learning and construction through skills competition so as to integrate education and training". By taking penicillin fermentation process as an example, reform has been made in several aspects including teaching objectives, teaching content and teaching methods. We integrate the practical operation of fermentation equipment with virtual simulation software to develop a two-way interactive course. By reducing the subjective dependence, the quantitative management and evaluation of fermentation process parameter control were put into place, which efficiently integrated the skills competition with practical teaching. Improved teaching performance has been achieved over recent years, which may facilitate the reform and practice of similar courses based on skills competition.


Health Care Professionals' Perspectives on Teleneonatology Through the Lens of Normalization Process Theory.
Little research has been done on tele-intensive care unit (ICU) implementation across different types of ICUs, and there exist few studies that have used qualitative research methods to analyze the human and organizational factors influencing optimization of telemedicine for newborn resuscitation. The objective of this study was to understand health care professionals' acceptance, utilization, and integration of video telemedicine for newborn resuscitation (termed teleneonatology) in community hospital settings. Focus group and individual interviews were conducted with 49 health care professionals at six affiliated health system hospitals. Data were gathered from physicians (n = 18), nurses (n = 30), and a nurse practitioner. Data were inductively analyzed using a thematic approach, and then constructs from normalization process theory (NPT) were deductively applied. NPT rendered a general framework to describe and assess how care teams perceive the implementation of teleneonatology and how they interact with this telemedicine service in their local setting. Local health care professionals accepted teleneonatology as an important, helpful service, yet its implementation was perceived as both valuable and a threat to professional traditions. Utilization may depend on perceived benefit, mutual understanding of the guidelines, and expectations of use, and other relational, human, contextual, and system factors. Participants in this study agreed on the need for collective work to successfully integrate teleneonatology into the local practice. NPT uncovered that successful implementation of a teleneonatology program may be facilitated by strong interpersonal relationships among care teams, continuous programmatic training and education, and communicating the clinical value of teleneonatology, including its opportunities and benefits.


Assay of reducing end-groups in oligosaccharide homologues with 2,2'-bicinchoninate.
For the first time, reducing values of homologous series of oligosaccharides have been determined by the 2,2'-bicinchoninate assay. The extent of Cu2+ reduction was monitored by spectrophotometric measurement of Cu+/2,2'-bicinchoninate complexes. Conditions of the assay were optimized so that relatively uniform reducing values were obtained with oligosaccharides derived from starch, polygalacturonic acid, and chitin, regardless of degree of polymerization. The uniform values resulted because members of each oligosaccharide series were oxidized to similar levels beyond the aldonic acid stage, while oxidation was limited to the reducing-terminal residue. This conclusion was based on direct measurements of quantitative loss of paramagnetic copper (Cu2+) by electron paramagnetic resonance spectroscopy.


Dental pedagogy in the 'new normal' COVID-19 era: a transition template of teaching protocols.
Delivery of clinical dental education, as opposed to clinical medicine, is particularly challenging due to the obligatory aerosol-generating procedures (AGPs) used in dentistry, which are known to facilitate the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Hence, using AGPs and working in close proximity to patients for extended periods in dental hospital/university settings with multiple teaching clinics have been a formidable prospect for all stake holders. Therefore, several professional and governmental organizations have promulgated variations of infection control guidelines for general practice dentistry in the pandemic era to mitigate SARS-CoV-2 transmission. In the absence of unified guidelines for modified infection control/clinical procedures for dental education. We implemented a clinical protocol template and modified operating procedures (MOP) for teaching clinical dentistry to fit the infection control requirements during the pandemic/post-pandemic period at the Sharjah University, College of Dentistry, UAE. MOPs ranged from various engineering control measures (e.g., negative-pressure ventilation systems in operatories) to administrative control measures featuring post-procedure fallow periods of treatment-abeyance between patient sessions. The new MOPs for clinical dentistry in the COVID-19 pandemic era, trialled in a UAE dental teaching hospital, have successfully eliminated infection transmission amongst the students, clinicians, ancillary staff, or attending patients, thus far. The proposed MOPs that complement the standard operating protocols in clinical dentistry were an attempt to mitigate nosocomial infection transmission and protect four different groups of stakeholders, i) the patients, ii) the dental students, iii) the clinical academics, and iv) the para-dental personnel/assistants. Due to the endemicity of the COVID-19 in many regions of the World, the suggested MOPs need periodic review and revision, to fit the emerging data on the disease. Finally, as there are no studies to date comparing the relative efficacy of the MOPs in various dental academic institutions, there is an urgent need for future workers to address this issue.


Thermal Bridges on Building Rooftops.
Thermal Bridges on Building Rooftops (TBBR) is a multi-channel remote sensing dataset. It was recorded during six separate UAV fly-overs of the city center of Karlsruhe, Germany, and comprises a total of 926 high-resolution images with 6927 manually-provided thermal bridge annotations. Each image provides five channels: three color, one thermographic, and one computationally derived height map channel. The data is pre-split into training and test data subsets suitable for object detection and instance segmentation tasks. All data is organized and structured to comply with FAIR principles, i.e. being findable, accessible, interoperable, and reusable. It is publicly available and can be downloaded from the Zenodo data repository. This work provides a comprehensive data descriptor for the TBBR dataset to facilitate broad community uptake.


Lacunar infarction of the basal ganglia as a complication of hemolytic-uremic syndrome. MRI and clinical correlations.
Central nervous system (CNS) complications of hemolytic-uremic syndrome (HUS) commonly consist of alterations in mental status, seizures, and rarely hemiparesis. The authors report the clinical evolution of left hemiparesis and later choreo-athetoid movements in a patient who sustained a right lacunar infarction as a complication of HUS. The infarction is demonstrated on magnetic resonance imaging (MRI).


Rapid Infusion of Hydroxyethyl Starch 70/0.5 but not Acetate Ringer's Solution Decreases the Plasma Concentration of Propofol during Target-controlled Infusion.
Rapid fluid infusion resulting in increased hepatic blood flow may decrease the propofol plasma concentration (Cp) because propofol is a high hepatic extraction drug. The authors investigated the effects of rapid colloid and crystalloid infusions on the propofol Cp during target-controlled infusion. Thirty-six patients were randomly assigned to 1 of 3 interventions (12 patients per group). At least 30 min after the start of propofol infusion, patients received either a 6% hydroxyethyl starch (HES) solution at 24 ml·kg·h or acetated Ringer's solution at 24 or 2 ml·kg·h during the first 20 min. In all groups, acetated Ringer's solution was infused at 2 ml·kg·h during the next 20 min. The propofol Cp was measured every 2.5 min as the primary outcome. Cardiac output, blood volume, and indocyanine green disappearance rate were determined using a pulse dye densitogram analyzer before and after the start of fluid administration. Effective hepatic blood flow was calculated as the blood volume multiplied by the indocyanine green disappearance rate. The rapid HES infusion significantly decreased the propofol Cp by 22 to 37%, compared to the Cp at 0 min, whereas the rapid or maintenance infusion of acetate Ringer's solution did not decrease the propofol Cp. Rapid HES infusion, but not acetate Ringer's solution infusion, increased the effective hepatic blood flow. Rapid HES infusion increased the effective hepatic blood flow, resulting in a decreased propofol Cp during target-controlled infusion. Rapid HES infusion should be used cautiously as it may decrease the depth of anesthesia.


[The elderly population in Rio de Janeiro (Brazil): a pilot-study of confiability and validity of the mental health segment of the BOAS questionnaire].
A first step in the pilot-study undertaken as part of a research project on psychiatric morbidity--depression and organic brain syndrome--in the elderly population of Rio de Janeiro is described. The main objective is to "fine-tune" the instrument used (check list of DSM-III) and testing validity of the BOAS questionnaire (Brazilian version of Short-Care). The methodological aspects, and conceptual definitions are focused as well some procedures for the next stage of the project are suggested.


Cerebral white matter hyperintensity in Parkinson's disease: a major risk factor for mild cognitive impairment.
Mild cognitive impairment (MCI) and dementia contribute to a poor quality of life among patients with PD. The influence of cerebral ischemia as a risk factor for MCI in PD has not been adequately investigated. To address this issue, we examined the influence of the volume and distribution of white matter hyperintensity (WMH) as a risk factor for MCI in early PD. Prospective study of patients with early idiopathic PD. All patients had baseline MRI-FLAIR, clinical assessment and detailed neuropsychological evaluation. Data on demographics, vascular risk factors, cognitive performance and WMH volumes were analyzed. 91 patients; mean age 64.9 years, mean education of 10.5 years. 24 patients fulfilled the Movement Disorder Society criteria for MCI and were classified as PD-MCI while the rest were classified as PD with no cognitive impairment (PD-NCI). Patients with PD-MCI and PD-NCI did not differ in Hoehn & Yahr staging. PD-MCI patients had a higher prevalence of diabetes mellitus, hypertension and hyperlipidemia. PD-MCI patients had significantly greater volume of periventricular (6.04 ml vs. 2.66 ml, p = 0.001) and deep subcortical WMH (2.16 vs.1.44, p = 0.002). Regional WMH was significantly greater among PD-MCI in the frontal, parietal and occipital regions. Logistic regression analyses demonstrated WMH to be associated with PD-MCI independent of age, education, and vascular risk factors. Increasing WMH volume was associated with lower performance on executive function, memory and language. WMH is an important risk factor for PD-MCI independent of vascular risk factors. PD patients with WMH should be regularly screened for MCI.


Dose-reduced fludarabine, cyclophosphamide and rituximab is well tolerated in older patients with chronic lymphocytic leukemia and has preserved therapeutic efficacy.
Despite its efficacy in prospective trials, full dose fludarabine, cyclophosphamide and rituximab (FCR) may be too toxic for elderly patients with chronic lymphocytic leukemia (CLL) in clinical practice. We retrospectively reviewed the impact of dose reductions in FCR therapy on the outcomes of 42 consecutive patients aged 65-87 (median 72) years. Despite a median cumulative fludarabine dose reduction of 50% from full dose, the objective response and complete response rates were 86% and 38% respectively (frontline 94%/59%; previously treated 80%/24%). Dose reductions of 25-75% were not significantly associated with inferior progression free survival compared to minimal reductions (≤25%) (p=0.49), and did not preclude deep responses, including six cases (14%) of minimal residual disease negativity. Although hematological and infectious toxicities were common, treatment limiting adverse effects were infrequent. Dose attenuated FCR appears to have preserved efficacy and may be a viable therapeutic option for elderly patients with CLL.


Confirmation of C4 gene copy number variation and the association with systemic lupus erythematosus in Chinese Han population.
The distribution of complement component 4 (C4) gene copy number (GCN) has been validated in European populations. Meanwhile, C4 gene has been identified as a susceptibility gene for systemic lupus erythematosus (SLE). However, the association and the possible phenotype significance remain to be determined intensely in the Chinese population. This study was designed to validate the distribution of C4 GCNs in Chinese Han and the correlation between C4 GCNs and SLE using quantitative real-time polymerase chain reaction in 924 SLE patients and 1,007 controls. The results presented distribution of C4 GCNs in healthy populations and also showed that lower C4 GCN was a risk factor for SLE and higher C4 GCN was a protective factor against the disease susceptibility, which was similar to the report in the Caucasian population. Furthermore, we found the association between C4A GCN and disease subphenotypes of arthritis with SLE. We conclude that the association of C4 GCN with SLE was replicated in Chinese Han population, which highlighted the importance of C4 in SLE pathogenesis of diverse populations.


Targeting V(D)J recombinase: putting a PHD to work.
In this issue of Immunity, Liu et al. (2007) show that V(D)J recombinase binds chromatin marked by H3K4 trimethylation. Because this mark associates with active promoters, the finding forges a new link between transcription, epigenetics, and recombinase targeting during lymphocyte development.


PE-only/PE_PGRS proteins of Mycobacterium tuberculosis contain a conserved tetra-peptide sequence DEVS/DXXS that is a potential caspase-3 cleavage motif.
Mycobacterium tuberculosis H37Rv is an intracellular pathogen responsible for causing tuberculosis in humans. The M. tuberculosis genome has been shown to contain a very large and unique family of PE proteins made of two sub-families: PE-only and PE_PGRS proteins. These two subtypes of proteins play a crucial role in the pathogenesis of the microbe. However, despite numerous investigations, the role of these proteins in disease development remains obscure. In this study, sequence analysis with a search for short conserved motifs revealed a conserved tetra-peptide motif DEVS/DXXS at the PE domain of almost every PE-only and PE_PGRS protein. The motif was found at a distance of 43-46 amino acids from the N-terminal of PE_PGRS proteins, and at a distance of between 35 and 82 amino acids of the PE-only proteins. As phosphorylation of the serine residue of this tetra-peptide could yield a motif similar to the caspase-3 binding recognition sequence DEVD/E, the region from a representative PE_PGRS protein (PE_PGRS45) was docked to human caspase-3. Strong interactions of only the protein containing the phosphorylated motif (DEVpS/DXXpS) to caspase-3 were observed. This suggested that the conserved DEVS/DXXS motif could have evolved for phosphorylation and subsequent recognition by caspase-3. These findings have important implications in unravelling the role of these PE proteins in mycobacterial infection.


GM3 synthase deficiency increases brain glucose metabolism in mice.
GM3 synthase (GM3S) deficiency is a rare neurodevelopmental disorder caused by an inability to synthesize gangliosides, for which there is currently no treatment. Gangliosides are brain-enriched, plasma membrane glycosphingolipids with poorly understood biological functions related to cell adhesion, growth, and receptor-mediated signal transduction. Here, we investigated the effects of GM3S deficiency on metabolism and mitochondrial function in a mouse model. By indirect calorimetry, GM3S knockout mice exhibited increased whole-body respiration and an increased reliance upon carbohydrate as an energy source. 18F-FDG PET confirmed higher brain glucose uptake in knockout mice, and GM3S deficient N41 neuronal cells showed higher glucose utilization in vitro. Brain mitochondria from knockout mice respired at a higher rate on Complex I substrates including pyruvate. This appeared to be due to higher expression of pyruvate dehydrogenase (PDH) and lower phosphorylation of PDH, which would favor pyruvate entry into the mitochondrial TCA cycle. Finally, it was observed that blocking glucose metabolism with the glycolysis inhibitor 2-deoxyglucose reduced seizure intensity in GM3S knockout mice following administration of kainate. In conclusion, GM3S deficiency may be associated with a hypermetabolic phenotype that could promote seizure activity.


Complete elimination ('cure') of progressively growing intraocular tumors by local injection of tumor-specific CD8+ T lymphocytes.
The purpose of this study was to examine if anterior chamber tumors could be eliminated by adoptive immunotherapy. Precursor T cells were recovered from lymph nodes draining subconjunctival space or anterior chamber tumors and restimulated in vitro to obtain fully functional tumor-specific CD8+ cytolytic T cells. Subconjunctival space tumor-derived cytolytic T cells eliminated tumors completely when injected simultaneously into the eye with the tumor cells, or when the anterior chamber injection was delayed for up to 3 days. In vitro restimulated cytolytic T cells obtained from lymph nodes draining progressively growing anterior chamber tumors were as effective as those obtained from lymph nodes draining subconjunctival space tumors, indicating that mice with anterior chamber tumors possess precursor cytotoxic T cells with the potential to differentiate into cytolytic T cells capable of eliminating intraocular tumors. By determining the frequency of tumor-specific cytolytic T cells in the effector cell inocula, it was estimated that cures of intraocular tumors are possible when the ratio of cytolytic T cells to tumor cells is less than 1:50, and the total tumor burden at the time of adoptive immunotherapy is less than 80,000 viable tumor cells. It was concluded that terminally differentiated, antigen-specific, cytotoxic T cells injected directly into an intraocular tumor site are capable of eliminating the tumor, and may represent a mode of ocular tumor immunotherapy in the future.


Yielding and thixotropic cellulose microgel-based network in high-content surfactant for stably suspending of functional beads.
Functional particles, such as microcapsules of perfumes, enzymes, or anti-mite agents, are desired to stably suspend in the high-content surfactant solution, providing additional functionalities for household products. Due to the disassociation of high-content surfactant, most linear or branched polymers would fail to modify the rheological properties of the high-content surfactant solution, especially for the suspending ability. In this research, the 2,2,6,6-tetramethylpiperidine-1-oxyl oxidized bacterial cellulose microgel (T-microgel) and hydroxypropyl methylcellulose (HPMC) were employed as "island" and "chain," respectively, which could self-assemble together to fabricate a yield and thixotropic continuous network in a high-content surfactant solution. The suspending ability of microgel in high-content surfactant is better than cellulose nanofiber and carboxymethylcellulose. This is the first time to report a cellulose microgel-based rheological modifier. T-microgel/HPMC synthetic system mixed with high content surfactant presented a typical Carreau-Yasuka fluid. Meanwhile, the effects of the HPMC and surfactant on the rheological properties of the combined system were investigated, and an optimal ratio for the 'island'/'chain' synthetic system was found to modify its yield and thixotropy behavior successfully. The potential application of this combined system was explored and found to work with all kinds of surfactants at high concentrations, which is more advantageous than most commercial suspending agents.


Entropic comparison of Landau-Zener and Demkov interactions in the phase space of a quadrupole billiard.
We investigate two types of avoided crossings in a chaotic billiard within the framework of information theory. The Shannon entropy in the phase space for the Landau-Zener interaction increases as the center of the avoided crossing is approached, whereas for the Demkov interaction, the Shannon entropy decreases as the center of avoided crossing is passed by with an increase in the deformation parameter. This feature can provide a new indicator for scar formation. In addition, it is found that the Fisher information of the Landau-Zener interaction is significantly larger than that of the Demkov interaction.


Combined effects of adrenaline and insulin on active electrogenic Na+-K+ transport in rat soleus muscle.
Both beta 2-adrenoreceptor stimulants (such as adrenaline and salbutamol) and insulin can increase active Na+-K+ transport and hyperpolarise skeletal cells. Thus, adrenaline and insulin, which are otherwise antagonistic regulators of several metabolic processes, have one action in common, namely, stimulation of active ion translocation. This is especially interesting as cyclic AMP stimulates Na+-K+ transport, whereas a lowering of the cytoplasmic concentration of cyclic AMP has been proposed as an early signal in the action of insulin. Here we report the results of experiments in which the active Na+-K+ transport and membrane potential (EM) of rat soleus muscles were studied during the action of supramaximal doses of insulin and beta 2-adrenoreceptor stimulants, alone and in combination. We conclude that the stimulant action of insulin on active electrogenic Na+-K+ transport is unlikely to be evoked by a lowering of the intracellular concentration of cyclic AMP.


An epidemiological study of hand eczema. III. Characterization of hairdressers with and without hand eczema, regarding demographic factors and medical histories.
The aim of this study was to compare, by stratification, subgroups of hairdressers. Three groups were considered: Group I (102 persons) consisted of hairdressers reporting skin affection at the time of completing an earlier mailed questionnaire (point prevalence). Group II reported exanthema prior to completing, while group III reported not having had exanthema at all (51 and 50 persons, respectively). No significant difference was found between these groups with regard to age and duration in work. The mean number of months since start of disease was significantly higher for group I (64 (range: 0.5-552)) than group II (7 (range: 0.6-38)). The frequency of atopic dermatitis was 12.7% (C.I: 6.3-19.2) in group I, compared with 3.9% (C.I: 0.0-9.2) in group II and none in group III. This difference between groups was significant. A significantly higher extent of sick leave was found in group I, compared to group II.


Methylxanthines and drug dependence: a focus on interactions with substances of abuse.
This chapter examines the psychostimulant actions of methylxanthines, with a focus on the consequences of their excessive use. Consumption of methylxanthines is pervasive and their use is often associated with that of substances known to produce dependence and to have abuse potential. Therefore, the consequences of this combined use are taken into consideration in order to evaluate whether, and to what extent, methylxanthines could influence dependence on or abuse of other centrally active substances, leading to either amplification or attenuation of their effects. Since the methylxanthine that mostly influences mental processes and readily induces psychostimulation is caffeine, this review mainly focuses on caffeine as a prototype of methylxanthine-produced dependence, examining, at the same time, the risks related to caffeine use.


Protective effect of erythropoietin against radiocontrast-induced renal tubular epithelial cell injury.
Recombinant human erythropoietin (rhEpo) has been shown to reduce tissue injury following ischemia-reperfusion. We examined whether rhEpo protects in vitro renal tubular epithelial cells against radiocontrast media-induced injury. LLC-PK1 renal tubular epithelial cells were exposed to non-ionic radiocontrast agent iohexol (low-osmolar) or iodixanol (iso-osmolar), with or without rhEpo (200 U/ml). Following a 6-hour exposure, cells were incubated for 24 h in radiocontrast-free culture medium. Cell viability was then assessed by the MTT assay. We also assessed cell apoptosis by the TUNEL assay, and activities of caspase-3, caspase-8, and caspase-9 were determined by a luminescence assay. rhEpo improved viability of iohexol-treated LLC-PK1 cells by 27 +/- 6% (88.1 +/- 1.5 vs. 70.8 +/- 3.3%, p = 0.008). Similarly, rhEpo improved the viability of iodixanol-treated LLC-PK1 cells by 26 +/- 4% (82.5 +/- 2.1vs. 65.7 +/- 1.7%, p = 0.028). rhEpo also decreased apoptosis rates of iohexol-treated LLC-PK1 cells (6.4 +/- 0.9/1,000 cells vs. 14.8 +/- 2.4/1,000 cells, p = 0.028), and iodixanol-treated LLC-PK1 cells (8.0 +/- 1.2/1,000 cells vs. 13.5 +/- 1.9/1,000 cells, p = 0.028). In iohexol-treated LLC-PK1 cells, rhEpo attenuated activation of caspase-3 (p = 0.003), caspase-8 (p = 0.033) and caspase-9 (p = 0.055). rhEpo attenuates in vitro renal tubular epithelial cell injury induced by low- and iso-osmolar radiocontrast media, possibly by reduction of caspases activation and apoptosis rates.


Vision: in the blink of an eye.
Although we blink every 4 to 6 seconds, we notice neither the act of blinking nor the mini-blackouts they cause. A new study using imaging techniques identifies the neural structures in humans involved in suppressing vision processing and visual awareness during blinking.


Antifungal potential of bacterial rhizosphere isolates associated with three ethno-medicinal plants (poppy, chamomile, and nettle).
The objective of the present study was to isolate Actinobacteria, preferably Streptomyces spp. from the rhizosphere soils of three ethno-medicinal plants collected in Serbia (Papaver rhoeas, Matricaria chamomilla, and Urtica dioica) and to screen their antifungal activity against Candida spp. Overall, 103 sporulating isolates were collected from rhizosphere soil samples and determined as Streptomyces spp. Two different media and two extraction procedures were used to facilitate identification of antifungals. Overall, 412 crude cell extracts were tested against Candida albicans using disk diffusion assays, with 42% (43/103) of the strains showing the ability to produce antifungal agents. Also, extracts inhibited growth of important human pathogens: Candida krusei, Candida parapsilosis, and Candida glabrata. Based on the established degree and range of antifungal activity, nine isolates, confirmed as streptomycetes by 16S rRNA sequencing, were selected for further testing. Their ability to inhibit Candida growth in liquid culture, to inhibit biofilm formation, and to disperse pre-formed biofilms was assessed with active concentrations from 8 to 250 μg/mL. High-performance liquid chromatographic profiles of extracts derived from selected strains were recorded, revealing moderate metabolic diversity. Our results proved that rhizosphere soil of ethno-medicinal plants is a prolific source of streptomycetes, producers of potentially new antifungal compounds.


Autologous grafts for treatment of vocal sulcus and atrophy.
To obtain complete glottal closure, and evaluate improvement of vocal fold vibration amplitude and vocal quality in patients with sulcus vocalis. Autogenous fat and/or fascia augmentation was used in 34 patients with sulcus vocalis. Mean follow-up time was 1 year. The perceptual acoustic and phonatory functions and videolaryngostroboscopic data were evaluated before and after the procedure. Most of the individuals reported an improvement of vocal quality, a complete glottal closure, an enhancement of mucosal wave excursion, and significant results of acoustic perceptual and phonatory functions after the surgery. Fat and fascia injections are effective autogenous implants and should be considered options in the treatment of patients with sulcus vocalis and vocal fold atrophy.


Comprehensive assessment of molecular function, tissue characterization, and hemodynamic performance by non-invasive hybrid imaging: Potential role of cardiac PETMR.
Noninvasive cardiovascular imaging plays a key role in diagnosis and patient management including monitoring treatment efficacy. The usefulness of noninvasive cardiovascular imaging has been extensively studied and shown to have high diagnostic reliability and prognostic significance, while the nondiagnostic results frequently encountered with single imaging modality require complementary or alternative imaging techniques. Hybrid cardiac imaging was initially introduced to integrate anatomical and functional information to enhance the diagnostic performance, and lately employed as a strategy for comprehensive assessment of the underlying pathophysiology of diseases. More recently, the utility of computed tomography has grown in diversity, and emerged from being an exploratory technique allowing functional measurement such as stress dynamic perfusion. Cardiac magnetic resonance imaging (CMR) is widely accepted as a robust tool for evaluation of cardiac function, fibrosis, and edema, yielding high spatial resolution and soft-tissue contrast. However, the use of intravenous contrast materials is typically required for accurate diagnosis with these imaging modalities, despite the associated risk of renal toxicity. Nuclear cardiology, established as a molecular imaging technique, has advantages in visualization of the disease-specific biological process at cellular level using numerous probes without requiring contrast materials. Various imaging modalities should be appropriately used sequentially to assess concomitant disease and the progression over time. Therefore, simultaneous evaluation combining high spatial resolution and disease-specific imaging probe is a useful approach to identify the regional activity and the stage of the disease. Given the recent advance and potential of multiparametric CMR and novel nuclide tracers, hybrid positron emission tomography MR is becoming an ideal tool for disease-specific imaging.


QbD guided development of immediate release FDM-3D printed tablets with customizable API doses.
The objective of this work was to develop a fused deposition modeling (FDM) 3D printed immediate release (IR) tablet with flexibility in adjusting the dose of the active pharmaceutical ingredient (API) by scaling the size of the dosage form and appropriate drug release profile steadiness to the variation of dimensions or thickness of the deposited layers throughout the printing process. Polyvinyl alcohol-based filaments with elevated API content (50% w/w) were prepared by hot melt extrusion (HME), through systematic screening of polymeric formulations with different drug loadings, and their printability was evaluated by means of mechanical characterization. For the tablet fabrication step by 3D printing (3DP), the Quality by Design (QbD) approach was implemented by employing risk management strategies and Design of Experiments (DoE). The effects of the tablet design, tablet size and the layer height settings on the drug release and the API content were investigated. Between the two proposed original tablet architectures, the honeycomb configuration was found to be a suitable candidate for the preparation of IR dosage forms with readily customizable API doses. Also, a predictive model was obtained, which assists the optimization of variables involved in the printing phase and thereby facilitates the tailoring process.


Role of acetylcholine, histamine and gastrin in the acid response to intracisternal injection of TRH analog, RX 77368, in the rat.
The role of gastrin, acetylcholine and histamine in the acid response to central vagal activation induced by intracisternal injection of the stable analog, RX 77368, was further investigated in urethane-anesthetized rats with gastric fistula. The gastrin monoclonal antibody 28-2 injected intravenously, at a dose previously shown to prevent gastrin-induced stimulation of acid secretion, did not alter the peak acid response to intracisternal injection of RX 77368 (15 ng). The TRH analog (30 ng) injected into the cisterna magna increased levels of histamine measured in the hepatic portal blood. Cimetidine administered at a dose which completely blocked the stimulation of gastric acid secretion produced by intravenous infusion of histamine, inhibited by 62% the stimulatory effect of intracisternal RX 77368 (30 ng). The M1 muscarinic antagonist, pirenzepine, completely prevented the acid secretion induced by intracisternal RX 77368 (30 ng). These results indicate that the acid response to central vagal activation by the TRH analog in rats involved M1 muscarinic receptors along with histamine release acting on H2 histaminergic receptors whereas gastrin does not appear to play an important role.


Protein engineering the surface of enzymes.
The protein surface is the interface through which a protein senses the external world. Its composition of charged, polar and hydrophobic residues is crucial for the stability and activity of the protein. The charge state of seven of the twenty naturally occurring amino acids is pH dependent. A total of 95% of all titratable residues are located on the surface of soluble proteins. In evolutionary related families of proteins such residues are particularly prone to substitutions, insertions and deletions. We present here an analysis of the residue composition of 4038 proteins, selected from 125 protein families with < 25% identity between core members of each family. Whereas only 16.8% of the residues were truly buried, 40.7% were > 30% exposed on the surface and the remainder were < 30% exposed. The individual residue types show distinct differences. The data presented provides an important new approach to protein engineering of protein surfaces. Guidelines for the optimization of solvent exposure for a given residue are given. The cutinase family of enzymes has been investigated. The stability of native cutinase has been studied as a function of pH, and has been compared with the cutinase activity towards tributyrin. Whereas the onset of enzymatic activity is linked with the deprotonation of the active site HIS188, destabilization of the 3D structure as determined by differential scanning calorimetry is coupled with the loss of activity at very basic pH values. A modeling investigation of the pH dependence of the electrostatic potentials reveals that the activity range is accompanied by the development of a highly significant negative potential in the active site cleft. The 3D structures of three mutants of the Fusarium solani pisi cutinase have been solved to high resolution using X-ray diffraction analysis. Preliminary X-ray data are presented.