Non-β-Lactam Allosteric Inhibitors Target Methicillin-Resistant Staphylococcus aureus: An In Silico Drug Discovery Study.
Penicillin-binding proteins (PBPs) catalyze the final stages for peptidoglycan cell-wall bio-synthesis. Mutations in the PBP2a subunit can attenuate β-lactam antibiotic activity, resulting in unimpeded cell-wall formation and methicillin-resistant Staphylococcus aureus (MRSA). A double mutation in PBP2a (i.e., N146K and E150K) is resistant to β-lactam inhibitors; however, (E)-3-(2-(4-cyanostyryl)-4-oxoquinazolin-3(4H)-yl) benzoic acid (QNZ), a heterocyclic antibiotic devoid of a β-lactam ring, interacts non-covalently with PBP2a allosteric site and inhibits PBP enzymatic activity. In the search for novel inhibitors that target this PBP2a allosteric site in acidic medium, an in silico screening was performed. Chemical databases including eMolecules, ChEMBL, and ChEBI were virtually screened for candidate inhibitors with a physicochemical similarity to QNZ. PBP2a binding affinities from the screening were calculated based on molecular docking with co-crystallized ligand QNZ serving as a reference. Molecular minimization calculations were performed for inhibitors with docking scores lower than QNZ (calc. -8.3 kcal/mol) followed by combined MD simulations and MM-GBSA binding energy calculations. Compounds eMol26313223 and eMol26314565 exhibited promising inhibitor activities based on binding affinities (ΔGbinding) that were twice that of QNZ (-38.5, -34.5, and -15.4 kcal/mol, respectively). Structural and energetic analyses over a 50 ns MD simulation revealed high stability for the inhibitors when complexed with the double mutated PBP2a. The pharmacokinetic properties of the two inhibitors were predicted using an in silico ADMET analysis. Calculated binding affinities hold promise for eMol26313223 and eMol26314565 as allosteric inhibitors of PBP2a in acidic medium and establish that further in vitro and in vivo inhibition experimentation is warranted.


Role of APOBEC3 proteins in proteasome inhibitor-mediated reactivation of latent HIV-1 viruses.
Proteasome inhibitors (PIs) have been identified as an emerging class of HIV-1 latency-reversing agents (LRAs). These inhibitors can reactivate latent HIV-1 to produce non-infectious viruses. The mechanism underlying reduced infectivity of reactivated viruses is unknown. In this study, we analysed PI-reactivated viruses using biochemical and virological assays and demonstrated that these PIs stabilized the cellular expression of HIV-1 restriction factor, APOBEC3G, facilitating its packaging in the released viruses. Using infectivity assay and immunoblotting, we observed that the reduction in viral infectivity was due to enhanced levels of functionally active APOBEC3 proteins packaged in the virions. Sequencing of the proviral genome in the target cells revealed the presence of APOBEC3 signature hypermutations. Our study strengthens the role of PIs as bifunctional LRAs and demonstrates that the loss of infectivity of reactivated HIV-1 virions may be due to the increased packaging of APOBEC3 proteins in the virus.


Unplanned Complex Suicide-A Consideration of Multiple Methods.
Detailed death investigations are mandatory to find out the exact cause and manner in non-natural deaths. In this reference, use of multiple methods in suicide poses a challenge for the investigators especially when the choice of methods to cause death is unplanned. There is an increased likelihood that doubts of homicide are raised in cases of unplanned complex suicides. A case of complex suicide is reported where the victim resorted to multiple methods to end his life, and what appeared to be an unplanned variant based on the death scene investigations. A meticulous crime scene examination, interviews of the victim's relatives and other witnesses, and a thorough autopsy are warranted to conclude on the cause and manner of death in all such cases.


Direct binding of GTP cyclohydrolase and tyrosine hydroxylase: regulatory interactions between key enzymes in dopamine biosynthesis.
The signaling functions of dopamine require a finely tuned regulatory network for rapid induction and suppression of output. A key target of regulation is the enzyme tyrosine hydroxylase, the rate-limiting enzyme in dopamine synthesis, which is activated by phosphorylation and modulated by the availability of its cofactor, tetrahydrobiopterin. The first enzyme in the cofactor synthesis pathway, GTP cyclohydrolase I, is activated by phosphorylation and inhibited by tetrahydrobiopterin. We previously reported that deficits in GTP cyclohydrolase activity in Drosophila heterozygous for mutant alleles of the gene encoding this enzyme led to tightly corresponding diminution of in vivo tyrosine hydroxylase activity that could not be rescued by exogenous cofactor. We also found that the two enzymes could be coimmunoprecipitated from tissue extracts and proposed functional interactions between the enzymes that extended beyond provision of cofactor by one pathway for another. Here, we confirm the physical association of these enzymes, identifying interacting regions in both, and we demonstrate that their association can be regulated by phosphorylation. The functional consequences of the interaction include an increase in GTP cyclohydrolase activity, with concomitant protection from end-product feedback inhibition. In vivo, this effect would in turn provide sufficient cofactor when demand for catecholamine synthesis is greatest. The activity of tyrosine hydroxylase is also increased by this interaction, in excess of the stimulation resulting from phosphorylation alone. Vmax is elevated, with no change in Km. These results demonstrate that these enzymes engage in mutual positive regulation.


Biomimetic nanopatterns as enabling tools for analysis and control of live cells.
It is becoming increasingly evident that cell biology research can be considerably advanced through the use of bioengineered tools enabled by nanoscale technologies. Recent advances in nanopatterning techniques pave the way for engineering biomaterial surfaces that control cellular interactions from the nano- to the microscale, allowing more precise quantitative experimentation capturing multi-scale aspects of complex tissue physiology in vitro. The spatially and temporally controlled display of extracellular signaling cues on nanopatterned surfaces (e. g., cues in the form of chemical ligands, controlled stiffness, texture, etc.) that can now be achieved on biologically relevant length scales is particularly attractive enabling experimental platform for investigating fundamental mechanisms of adhesion-mediated cell signaling. Here, we present an overview of bio-nanopatterning methods, with the particular focus on the recent advances on the use of nanofabrication techniques as enabling tools for studying the effects of cell adhesion and signaling on cell function. We also highlight the impact of nanoscale engineering in controlling cell-material interfaces, which can have profound implications for future development of tissue engineering and regenerative medicine.


The menisci of the lumbar zygapophysial joints.
In a study of 82 lumbar zygapophysial joints three types of intra-articular structures were identified. They were adipose tissue pads and fibro-adipose meniscoids, both located at the superior and inferior poles of the joint, and connective tissue rims, located along the dorsal and ventral margins. Every lumbar zygapophysial joint contained at least one of these structures and 47 contained more than one type. Connective tissue rims are short, central projections of the joint capsule and do not enter between the articular surfaces. Adipose tissue pads are covered by synovium and fill the subcapsular space at the superoventral and inferodorsal poles of the joint. Fibro-adipose meniscoids, also covered by synovium, project from the joint capsule at the superior and inferior poles and enter between the articular surfaces. Adipose tissue pads and fibro-adipose meniscoids are probably derived from a common primitive mesenchymal meniscus which primarily differentiates into a fatty structure. The fibrous component of fibro-adipose meniscoids then secondarily develops as a result of compression of the tip of the fatty structure between the articular surfaces. The function of these intra-articular structures is not evident but may be related to protection of the articular processes as they subluxate during flexion and extension. Meniscus entrapment in the lumbar zygapophysial joints has been proposed as a cause of acute locked back, but the present morphological data are inconsistent with this view.


Long-term preservation and storage of mycobacteria.
Under contract with the National Institute of Allergy and Infectious Diseases, the Trudeau Mycobacterial Culture Collection has been greatly expanded to provide for the scientific community a collection of representative strains of mycobacteria of biomedical importance. Problems concerned with the preparation, bottling, and distribution of such organisms have been dealt with and are detailed in this paper. Examination of collected data revealed that the temperature of storage and not the suspending menstruum was more important for prolonged survival of mycobacteria stored at subzero temperatures. For optimum results, mycobacteria may be suspended either in Dubos Tween-albumin broth or in Middlebrook 7H-9 liquid medium supplemented with ADC enrichment (commercial sources used) and stored at -70 C. Either of these suspending fluids supplied a growth-supporting medium for cultures which must be shipped long distances, often without refrigeration. To avoid sublimation of suspending medium during prolonged storage at -70 C (a problem inherent in many screw-capped containers), we have chosen to use vaccine-stoppered serum bottles sealed with aluminum crimp caps. The methods described have provided suspensions with (i) excellent viability over prolonged periods of storage, (ii) stable metabolic activities, and (iii) highly reproducible inocula for animal experiments.


Analysis and optimization of the robot setup for robotic-ultrasound-guided radiation therapy.
Robotic ultrasound promises continuous, volumetric, and non-ionizing tracking of organ motion during radiation therapy. However, placement of the robot is critical because it is radio-opaque and might severely influence the achievable dose distribution. We propose two heuristic optimization strategies for automatic placement of an ultrasound robot around a patient. Considering a kinematically redundant robot arm, we compare a generic approach based on stochastic search and a more problem-specific segmentwise construction approach. The former allows for multiple elbow configurations while the latter is deterministic. Additionally, we study different objective functions guiding the search. Our evaluation is based on data for ten actual prostate cancer cases and we compare the resulting plan quality for both methods to manually chosen robot configurations previously proposed. The mean improvements in the treatment planning objective value with respect to the best manually selected robot position and a single elbow configuration range from 8.2 to 32.8% and 8.5 to 15.5% for segmentwise construction and stochastic search, respectively. Considering three different elbow configurations, the stochastic search results in better objective values in 80% of the cases, with 30% being significantly better. The optimization strategies are robust with respect to beam sampling and transducer orientation and using previous optimization results as starting point for stochastic search typically results in better solutions compared to random starting points. We propose a robust and generic optimization scheme, which can be used to optimize the robot placement for robotic ultrasound guidance in radiation therapy. The automatic optimization further mitigates the impact of robotic ultrasound on the treatment plan quality.


T Cell-Macrophage Fusion Triggers Multinucleated Giant Cell Formation for HIV-1 Spreading.
HIV-1-infected macrophages participate in virus dissemination and establishment of virus reservoirs in host tissues, but the mechanisms for virus cell-to-cell transfer to macrophages remain unknown. Here, we reveal the mechanisms for cell-to-cell transfer from infected T cells to macrophages and virus spreading between macrophages. We show that contacts between infected T lymphocytes and macrophages lead to cell fusion for the fast and massive transfer of CCR5-tropic viruses to macrophages. Through the merge of viral material between T cells and macrophages, these newly formed lymphocyte-macrophage fused cells acquire the ability to fuse with neighboring noninfected macrophages. Together, these two-step envelope-dependent cell fusion processes lead to the formation of highly virus-productive multinucleated giant cells reminiscent of the infected multinucleated giant macrophages detected in HIV-1-infected patients and simian immunodeficiency virus-infected macaques. These mechanisms represent an original mode of virus transmission for viral spreading and a new model for the formation of macrophage virus reservoirs during infection.IMPORTANCE We reveal a very efficient mechanism involved in cell-to-cell transfer from infected T cells to macrophages and subsequent virus spreading between macrophages by a two-step cell fusion process. Infected T cells first establish contacts and fuse with macrophage targets. The newly formed lymphocyte-macrophage fused cells then acquire the ability to fuse with surrounding uninfected macrophages, leading to the formation of infected multinucleated giant cells that can survive for a long time, as evidenced in vivo in lymphoid organs and the central nervous system. This route of infection may be a major determinant for virus dissemination and the formation of macrophage virus reservoirs in host tissues during HIV-1 infection.


Recurrent Tuberculosis in a Young Child.
A young child, 19 months of age, presented with a second episode of tuberculosis after full recovery from initial tuberculosis disease 6 months earlier. Mycobacterium tuberculosis strains isolated from both episodes were genotyped and differed from one another. We present the first case of proven tuberculosis reinfection in a likely immunocompetent child, living in a high-risk environment favorable for exposition to M. tuberculosis but in a low-incidence country.


Effects of interruption of the enterohepatic circulation of bile acids on the transport of very low density-lipoprotein triglycerides.
An increase in plasma very low density lipoprotein-triglycerides (VLDL-TG) is seen frequently during treatment with bile acid-binding resins. The purpose of this study was to determine whether this increment in VLDL-TG is due mainly to an increase in synthesis of VLDL, or to an enhanced catabolism. Three types of patients were studied: (1) 7 normotriglyceridemic subjects. (2) 4 obese patients, and (3) 9 hypertriglyceridemic patients. Before treatment they underwent a study of VLDL-TG kinetics that employed multicompartmental analysis of specific activity curves following injection of 3H-glycerol. The patients were then treated with a bile acid-binding resin, either cholestyramine or colestipol, for several weeks to several months. At the end of the treatment period, they were readmitted to the hospital for a second study of VLDL-TG kinetics. The patients showed a variable response to resin therapy. Many had an increase in concentrations of VLDL-TG, but others had no change or even a slight decrease. However, analysis of the data showed a high correlation between change in production rates of VLDL-TG and change in concentration. Also, when the data for the 20 patients were combined, there was a statistically significant increase in both synthetic rates and concentrations of VLDL-TG; in contrast, the fractional catabolic rate (FCR) was unchanged by therapy. Therefore, our data show that when treatment with bile acid sequestrants causes an increase in plasma VLDL-TG, the increase is due to an increment in production and not to a decrease in catabolism.


Longitudinal Trajectories of Alcohol Consumption with All-Cause Mortality, Hypertension, and Blood Pressure Change: Results from CHNS Cohort, 1993-2015.
Previous studies have demonstrated a J-shaped association of alcohol consumption with all-cause mortality and hypertension, but the majority of these studies focus on a single measurement of alcohol intake and were conducted in a Western population. Whether long-term trajectories of alcohol consumption are associated with all-cause mortality, hypertension, and a change in blood pressure remains to be elucidated. In the large, population-based China Health and Nutrition Survey cohort from between 1993 and 2015, group-based trajectory modeling was conducted to identify distinct alcohol-consumption trajectory classes. We investigated their association with all-cause mortality and hypertension using Cox regression and binary logistics regression models. A restricted cubic spline was performed to determine the nonlinear relationships of mean alcohol intake with mortality and hypertension. Multivariate-adjusted generalized linear mixed-effects models were conducted to assess the change in blood pressure among alcohol-consumption trajectory classes. Among the 5298 participants, 48.4% were women and the mean age was 62.6 years. After 22 years of follow-up, 568 (10.7%) of the participants died and 1284 (24.2%) developed hypertension. Long-term light and moderate drinkers had a lower risk of death than the non-drinkers, and a restricted cubic spline showed a J-shaped relationship between mean alcohol intake and mortality. Although blood pressure increased slower in light and moderate drinkers, a reduced risk of hypertension was only observed in the former. The long-term heavy drinkers had the highest blood pressure and death rate. Light alcohol intake might be protective even in the long run, while heavy drinking reversed the beneficial effect. The causality of such a connection needs to be further investigated.


The effectiveness of weekly iron supplementation regimen in improving the iron status of Chinese children and pregnant women.
It is evident that intermittent iron supplementation is better than daily supplementation in two aspects: iron absorption is more efficient and has insignificant side effects in contrast to the daily dose. The significantly higher daily iron loss observed in the daily iron supplemented groups rats also suggests alterations in total body iron metabolism. Based on serum ferritin distribution patterns, intermittent iron supplementation avoids temporary iron overload with daily iron supplemented. We conclude that weekly iron supplementation scheme is safer and easier to administer. This feasible strategy for the control of iron deficient anemia in pregnant women and children would be an effective iron-supplementation program (Baily et al., 1993).


The role of preclinical models in radiopharmaceutical therapy.
Radiopharmaceutical therapy (RPT) is a treatment modality that involves the use of radioactively labeled targeting agents to deliver a cytotoxic dose of radiation to tumor while sparing normal tissue. The biologic function of the target and the biologic action of the targeting agent is largely irrelevant as long as the targeting agent delivers cytotoxic radiation to the tumor. Preclinical RPT studies use imaging and ex vivo evaluation of radioactivity concentration in target and normal tissues to obtain biodistribution and pharmacokinetic data that can be used to evaluate radiation absorbed doses. Since the efficacy and toxicity of RPT depend on radiation absorbed dose, this quantity can be used to translate results from preclinical studies to human studies. The absorbed dose can also be used to customize therapy to account for pharmacokinetic and other differences among patients so as to deliver a prespecified absorbed dose to the tumor or to dose-limiting tissue. The combination of RPT with other agents can be investigated and optimized by identifying the effect of other agents on tumor or normal tissue radiosensitivity and also on how other agents change the absorbed dose to these tissues. RPT is a distinct therapeutic modality whose mechanism of action is well understood. Measurements can be made in preclinical models to help guide clinical implementation of RPT and optimize combination therapy using RPT.


Genome wide gene-expression analysis of facultative reproductive diapause in the two-spotted spider mite Tetranychus urticae.
Diapause or developmental arrest, is one of the major adaptations that allows mites and insects to survive unfavorable conditions. Diapause evokes a number of physiological, morphological and molecular modifications. In general, diapause is characterized by a suppression of the metabolism, change in behavior, increased stress tolerance and often by the synthesis of cryoprotectants. At the molecular level, diapause is less studied but characterized by a complex and regulated change in gene-expression. The spider mite Tetranychus urticae is a serious polyphagous pest that exhibits a reproductive facultative diapause, which allows it to survive winter conditions. Diapausing mites turn deeply orange in color, stop feeding and do not lay eggs. We investigated essential physiological processes in diapausing mites by studying genome-wide expression changes, using a custom built microarray. Analysis of this dataset showed that a remarkable number, 11% of the total number of predicted T. urticae genes, were differentially expressed. Gene Ontology analysis revealed that many metabolic pathways were affected in diapausing females. Genes related to digestion and detoxification, cryoprotection, carotenoid synthesis and the organization of the cytoskeleton were profoundly influenced by the state of diapause. Furthermore, we identified and analyzed an unique class of putative antifreeze proteins that were highly upregulated in diapausing females. We also further confirmed the involvement of horizontally transferred carotenoid synthesis genes in diapause and different color morphs of T. urticae. This study offers the first in-depth analysis of genome-wide gene-expression patterns related to diapause in a member of the Chelicerata, and further adds to our understanding of the overall strategies of diapause in arthropods.


Multi-phase defense by the big-headed ant, Pheidole obtusospinosa, against raiding army ants.
Army ants are well known for their destructive raids of other ant colonies. Some known defensive strategies include nest evacuation, modification of nest architecture, blockade of nest entrances using rocks or debris, and direct combat outside the nest. Since army ants highly prefer Pheidole ants as prey in desert habitats, there may be strong selective pressure on Pheidole to evolve defensive strategies to better survive raids. In the case of P. obtusospinosa Pergande (Hymenoptera: Formicidae), the worker caste system includes super majors in addition to smaller majors and minor workers. Interestingly, P. obtusospinosa and the six other New World Pheidole species described to have polymorphic major workers are all found in the desert southwest and adjacent regions of Mexico, all co-occurring with various species of Neivamyrmex army ants. Pheidole obtusospinosa used a multi-phase defensive strategy against army ant raids that involved their largest major workers. During army ant attacks, these super majors were involved in blocking the nest entrance with their enlarged heads. This is the first description of defensive head-blocking by an ant species that lacks highly modified head morphology, such as a truncated or disc-shaped head. P. obtusospinosa super majors switched effectively between passive headblocking at the nest entrance and aggressive combat outside the nest. If this multi-phase strategy is found to be used by other Pheidole species with polymorphic majors in future studies, it is possible that selective pressure by army ant raids may have been partially responsible for the convergent evolution of this extra worker caste.


Hereditary early-onset Parkinson's disease caused by mutations in PINK1.
Parkinson's disease (PD) is a neurodegenerative disorder characterized by degeneration of dopaminergic neurons in the substantia nigra. We previously mapped a locus for a rare familial form of PD to chromosome 1p36 (PARK6). Here we show that mutations in PINK1 (PTEN-induced kinase 1) are associated with PARK6. We have identified two homozygous mutations affecting the PINK1 kinase domain in three consanguineous PARK6 families: a truncating nonsense mutation and a missense mutation at a highly conserved amino acid. Cell culture studies suggest that PINK1 is mitochondrially located and may exert a protective effect on the cell that is abrogated by the mutations, resulting in increased susceptibility to cellular stress. These data provide a direct molecular link between mitochondria and the pathogenesis of PD.


SLE and Tuberculosis: A Case Series and Review of Literature.
Systemic Lupus Erythematosus (SLE) and Tuberculosis (TB) are intricately related with an increase in the risk of TB in SLE. Primary mechanisms pertaining to the increased susceptibility for TB are the inherent immunodeficient state of SLE and use of immunosuppressant agents in the treatment of SLE. We report a case series of five female patients of SLE with TB who presented between January 2015 and December 2015 in a tertiary care teaching hospital in North Eastern India. All the patients were young to middle aged females having SLE with or without lupus nephritis who were on immunosuppressive therapy with corticosteroids, mycophenolate mofetil or cyclophosphamide. Two of the cases had sputum positive pulmonary tuberculosis while rest had Extra-Pulmonary TB (EPTB). The response to anti-tubercular therapy led to clinical improvement in all the cases except one who had an adverse outcome. Our series further substantiates the increased risk of TB in SLE thus, prompting further research towards better management of these two disease entities in conjunction.


Prospective trial of the detection of urolithiasis on ultralow dose (sub mSv) noncontrast computerized tomography: direct comparison against routine low dose reference standard.
In this prospective trial we compared ultralow dose computerized tomography reconstruction algorithms and routine low dose computerized tomography for detecting urolithiasis. A total of 48 consenting adults prospectively underwent routine low dose noncontrast computerized tomography immediately followed by an ultralow dose series targeted at a 70% to 90% reduction from the routine low dose technique (sub mSv range). Ultralow dose series were reconstructed with filtered back projection, and adaptive statistical and model based iterative reconstruction techniques. Transverse (axial) and coronal images were sequentially reviewed by 3 relatively inexperienced trainees, including a radiology resident, a urology fellow and an abdominal imaging fellow. Three experienced abdominal radiologists independently reviewed the routine low dose filtered back projection images, which served as the reference standard. The mean effective dose for the ultralow dose scans was 0.91 mSv (median 0.82), representing a mean ± SD 78% ± 5% decrease compared to the routine low dose. Overall sensitivity and positive predictive value per stone for ultralow dose computerized tomography at a 4 mm threshold was 0.91 and 0.98, respectively. Sensitivity, specificity, positive and negative predictive values, and accuracy per patient were 0.87, 1.00, 1.00, 0.94 and 0.96, respectively. At a 4 mm threshold the sensitivity and positive predictive value per stone of the ultralow dose series for filtered back projection, and adaptive statistical and model based iterative reconstruction was 0.89 and 0.96, 0.91 and 0.98, and 0.93 and 1.00, respectively. Sensitivity, specificity, positive and negative predictive values, and accuracy per patient at the 4 mm threshold were 0.82, 1.00, 1.00, 0.91 and 0.94 for filtered back projection, 0.85, 1.00, 1.00, 0.93 and 0.95 for adaptive statistical iterative reconstruction, and 0.94, 1.00, 1.00, 0.97 and 0.98 for model based iterative reconstruction, respectively. Sequential review of coronal images changed the final stone reading in 13% of cases and improved diagnostic confidence in 49%. At a 4 mm renal calculus size threshold ultralow dose computerized tomography is accurate for detection when referenced against routine low dose series with dose reduction to below the level of a typical 2-view plain x-ray of the kidneys, ureters and bladder. Slight differences were seen among the reconstruction algorithms. There was mild improvement with model based iterative reconstruction over filtered back projection and adaptive statistical iterative reconstruction. Coronal images improved detection and diagnostic confidence over axial images alone.


Relevance of multidrug-resistant Pseudomonas aeruginosa infections in cystic fibrosis.
Multidrug-resistant (MDR) Pseudomonas aeruginosa is an important issue for physicians who take care of patients with cystic fibrosis (CF). Here, we review the latest research on how P. aeruginosa infection causes lung function to decline and how several factors contribute to the emergence of antibiotic resistance in P. aeruginosa strains and influence the course of the infection course. However, many aspects of the practical management of patients with CF infected with MDR P. aeruginosa are still to be established. Less is known about the exact role of susceptibility testing in clinical strategies for dealing with resistant infections, and there is an urgent need to find a tool to assist in choosing the best therapeutic strategy for MDR P. aeruginosa infection. One current perception is that the selection of antibiotic therapy according to antibiogram results is an important component of the decision-making process, but other patient factors, such as previous infection history and antibiotic courses, also need to be evaluated. On the basis of the known issues and the best current data on respiratory infections caused by MDR P. aeruginosa, this review provides practical suggestions to optimize the diagnostic and therapeutic management of patients with CF who are infected with these pathogens.


Oral non-squamous malignant tumors; diagnosis and treatment.
Some 90% of oral cancers consist of squamous cell carcinomas that arise from the oral mucosa. The remaining 10% of malignancies consist of malignant melanomas, carcinomas of the intraoral salivary glands, sarcomas of the soft tissues and the bones, malignant odontogenic tumors, non-Hodgkin's lymphomas and metastases from primary tumors located elsewhere in the body. These malignancies will be briefly reviewed and discussed. The emphasis is on diagnosis and management.


Preventing dehydration-related hospitalizations: a mixed-methods study of parents, inpatient attendings, and primary care physicians.
The goal of this study was to identify the proportion of dehydration-related ambulatory care-sensitive condition hospitalizations, the reasons why these hospitalizations were preventable, and factors associated with preventability. A cross-sectional survey of primary care providers (PCPs), inpatient attending physicians, and parents was conducted in a consecutive series of children with ambulatory care-sensitive conditions admitted to an urban hospital over 14 months. Eighty-five children were diagnosed with dehydration. Their mean age was 1.6 years; most had public (74%) or no (17%) insurance, and were nonwhite (91%). The proportion of hospitalizations assessed as preventable varied from 12% for agreement among all 3 sources to 45% for any source. Parents identified inadequate prevention (50%), poor self-education (34%), and poor quality of care (38%) as key factors. PCPs identified parents providing insufficient home rehydration (33%), not visiting the clinic (25%), and not calling earlier (16%) as reasons. Inpatient attending physicians cited home rehydration (40%), delays in seeking care (40%), and lacking a PCP (20%) as contributors. Physicians (PCPs and inpatient attending physicians) were more likely than parents to describe the admission as inappropriate (75% vs 67% vs 0%; P < .01). Parental dissatisfaction with their child's PCP and a history of avoiding primary care due to costs or insurance problems were associated with significantly higher odds of preventable hospitalization. Up to 45% of dehydration-related hospitalizations may be preventable. Inadequate parental education by physicians, insufficient home rehydration, deferring clinic visits, insurance and cost barriers, inappropriate admissions, poor quality of care, and parental dissatisfaction with PCPs are the reasons that these hospitalizations might have been prevented.


Molecularly-mediated assemblies of plasmonic nanoparticles for Surface-Enhanced Raman Spectroscopy applications.
In recent years, Surface-Enhanced Raman Spectroscopy (SERS) has experienced a tremendous increase of attention in the scientific community, expanding to a continuously wider range of diverse applications in nanoscience, which can mostly be attributed to significant improvements in nanofabrication techniques that paved the way for the controlled design of reliable and effective SERS nanostructures. In particular, the plasmon coupling properties of interacting nanoparticles are extremely intriguing due to the concentration of enormous electromagnetic enhancements at the interparticle gaps. Recently, great efforts have been devoted to develop new nanoparticle assembly strategies in suspension with improved control over hot-spot architecture and cluster structure, laying the foundation for the full exploitation of their exceptional potential as SERS materials in a wealth of chemical and biological sensing. In this review we summarize in an exhaustive and systematic way the state-of-art of plasmonic nanoparticle assembly in suspension specifically developed for SERS applications in the last 5 years, focusing in particular on those strategies which exploited molecular linkers to engineer interparticle gaps in a controlled manner. Importantly, the novel advances in this rather new field of nanoscience are organized into a coherent overview aimed to rationally describe the different strategies and improvements in the exploitation of colloidal nanoparticle assembly for SERS application to real problems.


Rapid and sensitive detection of antibiotic resistance on a programmable digital microfluidic platform.
The widespread dissemination of CTX-M extended spectrum β-lactamases among Escherichia coli bacteria, both in nosocomial and community environments, is a challenge for diagnostic bacteriology laboratories. We describe a rapid and sensitive detection system for analysis of DNA containing the blaCTX-M-15 gene using isothermal DNA amplification by recombinase polymerase amplification (RPA) on a digital microfluidic platform; active matrix electrowetting-on-dielectric (AM-EWOD). The devices have 16,800 electrodes that can be independently controlled to perform multiple and simultaneous droplet operations. The device includes an in-built impedance sensor for real time droplet position and size detection, an on-chip thermistor for temperature sensing and an integrated heater for regulating the droplet temperature. Automatic dispensing of droplets (45 nL) from reservoir electrodes is demonstrated with a coefficient of variation (CV) in volume of approximately 2%. The RPA reaction is monitored in real-time using exonuclease fluorescent probes. Continuous mixing of droplets during DNA amplification significantly improves target DNA detection by at least 100 times compared to a benchtop assay, enabling the detection of target DNA over four-order-of-magnitude with a limit of detection of a single copy within ~15 minutes.


Rapid radioimmunoassay for parathyroid hormone: its use in hypercalcemic crisis.
Patients with severe hypercalcemia often require intensive medical therapy to avoid a fatal outcome. When therapy with saline and loop diuretics cannot control the hypercalcemia, use of the toxic drug mithramycin is most appropriate for malignancy-associated hypercalcemia, but parathyroidectomy is indicated for parathyroid crisis. Although differentiation of these two syndromes is possible in many cases on clinical grounds, the determination of immunoreactive parathyroid hormone values provides a direct assessment of parathyroid function. Radioimmunoassay for parathyroid hormone has traditionally required four to seven days, but more rapid results would be desirable in this medical emergency. We report a modification of a previously published midregion-specific PTH assay that provides accurate results within 24 hours and describe its use in six patients with life-threatening hypercalcemia.


Comparison between hospitalized and screening controls in studies assessing breast cancer risk.
In case-control studies, selection of an appropriate group of controls is a critical step which may affect the outcome of the analysis. We studied the differences in reproductive, lifestyle, and anthropometric variables between controls from a hospital-based study and controls from a screening clinic. Odds ratios for breast cancer were calculated using the two types of controls, and the impact on the resulting odds ratio was studied. Some interesting differences in odds ratios obtained with the two sets of controls were found. Among premenopausal screening subjects, the odds ratio for breast cancer did not change across quartiles of body mass,while among hospital subjects, the risk of breast cancer significantly decreased with increasing body mass. For liquor and beer consumption, a three-fold increase in breast cancer risk was observed among premenopausal hospital subjects, whereas no association was found among screening subjects. Among postmenopausal women, a significant decrease in breast cancer risk with ovariectomy was only observed in the hospital-based study. Our results suggest that in examining the association between body mass, alcohol consumption, or ovariectomy and breast cancer risk, the choice of control group used should be carefully considered.


Impact of mammography screening interval on breast cancer diagnosis by menopausal status and BMI.
Controversy remains regarding the frequency of screening mammography. Women with different risks for developing breast cancer because of body mass index (BMI) may benefit from tailored recommendations. To determine the impact of mammography screening interval for women who are normal weight (BMI < 25), overweight (BMI 25-29.9), or obese (BMI ≥ 30), stratified by menopausal status. Two cohorts selected from the Breast Cancer Surveillance Consortium. Patient and mammography data were linked to pathology databases and tumor registries. The cohort included 4,432 women aged 40-74 with breast cancer; the false-positive analysis included a cohort of 553,343 women aged 40-74 without breast cancer. Stage, tumor size and lymph node status by BMI and screening interval (biennial vs. annual). Cumulative probability of false-positive recall or biopsy by BMI and screening interval. Analyses were stratified by menopausal status. Premenopausal obese women undergoing biennial screening had a non-significantly increased odds of a tumor size > 20 mm relative to annual screeners (odds ratio [OR] = 2.07; 95 % confidence interval [CI] 0.997 to 4.30). Across all BMI categories from normal to obese, postmenopausal women with breast cancer did not present with higher stage, larger tumor size or node positive tumors if they received biennial rather than annual screening. False-positive recall and biopsy recommendations were more common among annually screened women. The only negative outcome identified for biennial vs. annual screening was a larger tumor size (> 20 mm) among obese premenopausal women. Since annual mammography does not improve stage at diagnosis compared to biennial screening and false-positive recall/biopsy rates are higher with annual screening, women and their primary care providers should weigh the harms and benefits when deciding on annual versus biennial screening.


A vagally mediated histaminergic component of food-related drinking in the rat.
Histamine elicited drinking in a dose-related manner typically within 5 min after subcutaneous injection in male albino rats. Threshold for increased drinking was 1.25 mg/kg, and 2.5 mg/kg elicited half of the maximal drinking response that followed 20 mg/kg. Histamine was not differentially potent for drinking in the day or night phase of the diurnal cycle. Bilateral subdiaphragmatic vagotomy, with the hepatic branch left intact, severely attenuated drinking in response to systemic histamine: Vagotomized rats drank later and less than did normal rats after doses of histamine between 1.25 and 40 mg/kg. This attenuation was attributed to the destruction of vagal afferent fibers because histamine-elicited drinking was not affected by blockade of vagal efferents with the peripheral anticholinergic atropine methyl nitrate. Drugs antagonistic to peripheral H2 histamine receptors specifically inhibited drinking in response to histamine: Intraperitoneal cimetidine or metiamide delayed and decreased drinking after sc histamine and temporarily decreased drinking after hypovolemia produced by sc polyethylene glycol, but these H2 antagonists failed to inhibit drinking after water deprivation, cellular dehydration, or isoproterenol. Finally, cimetidine or metiamide inhibited drinking in temporal association with a meal of liquid or solid food without decreasing food intake. These results constitute the first evidence for a peripheral histaminergic determinant of food-related drinking in the rat.


[Aggregation of fragmented chromatin associated with the synthesis of products of its treatment with nuclease].
Isolated cell nuclei were incubated with nucleases followed by extraction of chromatin with a low salt buffer. With an increase of nuclear chromatin degradation with DNAse I or micrococcal nuclease, solubilization of deoxyribonucleoprotein (DNP) by a low salt buffer increases, reaching a maximum upon hydrolysis with 2-4% nuclear DNA and then decreases appreciably after extensive treatment with nucleases. Soluble fragmented chromatin aggregates in the course of treatment with DNAase. I. Addition to gel chromatin preparations of exogenous products of nuclease treatment of isolated nuclei leads to its aggregation. Pretreatment of nuclear chromatin with RNAase prevents solubilization of DNP by low ionic strength solutions. Some experimental data obtained with the use of severe nuclease treatment are discussed; for a correct interpretation of these data the aggregation of fragmented chromatin by products of its nuclease degradation should be taken into consideration.


Proposed reclassification of melanoma: a meeting held at the XVIth International Pigment Cell Congress, 3rd November 1996.
The current classification of cutaneous melanoma was developed in 1972 and revised in 1982. Since that time, new concepts and terminology have evolved that require consideration of a further revision. Regional meetings of interested parties have been held to review the Classification and there will be an open meeting on the topic at the 1997, 4th World Conference on Melanoma in Sydney, Australia. This paper reports on a meeting to discuss some of the concepts that will form part of that process, held on November 3, 1996 as part of the XVIth International Pigment Cell Conference. A questionnaire is included that will allow the interested reader to provide comments on the topic.


The effects of self-movement, observation, and imagination on mu rhythms and readiness potentials (RP's): toward a brain-computer interface (BCI).
Current movement-based brain-computer interfaces (BCI's) utilize spontaneous electroencephalogram (EEG) rhythms associated with movement, such as the mu rhythm, or responses time-locked to movements that are averaged across multiple trials, such as the readiness potential (RP), as control signals. In one study, we report that the mu rhythm is not only modulated by the expression of self-generated movement but also by the observation and imagination of movement. In another study, we show that simultaneous self-generated multiple limb movements exhibit properties distinct from those of single limb movements. Identification and classification of these signals with pattern recognition techniques provides the basis for the development of a practical BCI.


Verification of an improved hip joint center prediction method.
In motion analysis, the hip joint center (HJC) is used to define the proximal location of the thigh segment and is also the point about which hip moments are calculated. The HJC cannot be palpated; its location must be calculated. Functional methods have been proposed but are difficult to perform by some clinical populations. Therefore, regression methods are utilized, but yield large errors in estimating the HJC location. These prediction methods typically utilize the anterior and posterior superior iliac spines, where excessive adipose tissue makes correctly locating difficult. A new regression method (Hara) utilizes leg length and has been shown to improve HJC location in cadavers and less error than previous pelvic based regression methods, such as those proposed by Harrington et al. This study compared the accuracy of the HJC location calculated with both of the Harrington methods and the Hara method. The coronal knee angle was calculated for each method using a static motion analysis trial, and compared to the tibiofemoral angle measured on a gold standard digital full-leg coronal radiograph. This study demonstrated that the Hara method was more accurate than either of the Harrington methods. The mean error between the gold standard x-ray measurement and the motion analysis calculation for the Harrington (stepwise and LOOCV), the Harrington (linear regression), and Hara regression methods, respectively were 6.0°, 4.0°, and 1.8°. Accurately modeling the HJC is critical for data interpretation and patient care. This study confirmed that the Hara HJC regression method is valid in an in-vivo setting.


Goat herpesviruses: biological and physicochemical properties.
Two herpesvirus isolates from goats are known which cause afflictions of the digestive tract in kids and, in some cases, abortion. An antigenic relationship of these goat herpesviruses with infectious bovine rhinotracheitis/infectious pustular vulvo-vaginitis virus (bovid herpesvirus 1, BHV-1) was reported and because of the species-specific pathogenicity, the goat isolates were named caprine herpesvirus 1. In this report the two isolates are further characterized and compared with BHV-1. Although the caprine herpesviruses share many biological and physicochemical properties with BHV-1, they can be differentiated from the bovine viruses with respect to growth cycle, one-way cross-neutralization and, most importantly, the restriction endonuclease fragments of their DNAs. The molecular weight of the caprine herpesvirus DNA, based on electron microscopic length measurement is 90 X 10(6), similar to that of BHV-1 (95 X 10(6]. On the basis of these genomic differences, we propose that DNA restriction endonuclease patterns of the caprine herpesviruses should be designated as prototypic of bovid herpesvirus 6 (BHV-6).


The genome sequence of the anaerobic, sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough.
Desulfovibrio vulgaris Hildenborough is a model organism for studying the energy metabolism of sulfate-reducing bacteria (SRB) and for understanding the economic impacts of SRB, including biocorrosion of metal infrastructure and bioremediation of toxic metal ions. The 3,570,858 base pair (bp) genome sequence reveals a network of novel c-type cytochromes, connecting multiple periplasmic hydrogenases and formate dehydrogenases, as a key feature of its energy metabolism. The relative arrangement of genes encoding enzymes for energy transduction, together with inferred cellular location of the enzymes, provides a basis for proposing an expansion to the 'hydrogen-cycling' model for increasing energy efficiency in this bacterium. Plasmid-encoded functions include modification of cell surface components, nitrogen fixation and a type-III protein secretion system. This genome sequence represents a substantial step toward the elucidation of pathways for reduction (and bioremediation) of pollutants such as uranium and chromium and offers a new starting point for defining this organism's complex anaerobic respiration.


Microtubule inhibitor D-24851 induces p53-independent apoptotic cell death in malignant glioma cells through Bcl-2 phosphorylation and Bax translocation.
D-24851 is a recently developed microtubule inhibitor that induces G2/M cell-cycle arrest and has an antitumor effect in many cancer cell types. It is expected to be a promising chemotherapeutic agent against a broad range of tumors. However, the precise mechanisms underlying its antitumor effect remain to be determined. Here, we investigated the in vitro effect of D-24851 on tumor growth and the apoptosis mechanism in human malignant glioma cells. Because both p53-dependent and -independent pathways of apoptosis have been reported, we used cell lines with wild-type p53 (U87-MG and D54) and cell lines with mutant p53 (U373-MG and T98G) and compared their responses to D-24851. D-24851 substantially inhibited the proliferation of the four glioma cell lines tested in a dose- and time-dependent manner. The inhibitory effect of D-24851 on tumor growth was associated with cell-cycle arrest in G2/M, subsequently inducing apoptosis. D-24851 treatment induced phosphorylated Bcl-2 and translocated Bax from the cytoplasm to the mitochondria, resulting in apoptotic cell death. These events took place regardless of the p53 status of tumor cells. Our results indicated that D-24851 effectively induces apoptosis through Bcl-2 phosphorylation and Bax translocation in human malignant glioma cells in a p53-independent manner. The results of this study make D-24851 even more promising as a therapeutic agent, especially because many malignant gliomas have a heterogeneous p53 status.


[Medical treatment of heart failure at a tertiary hospital of São Paulo].
To study how patients with heart failure (HF) are treated in a tertiary hospital in São Paulo. One hundred patients with HF during ambulatory care were analyzed. Seventy-six were men, and the average population age was 56.8 years old. All patients were submitted to echocardiogram, which identified ventricular diameters ranging between 48 and 89 mm (average 65.9) and ejection fraction (EF) between 0.22 and 0.59 (average 0.43). The cause of HF was ischemic in 42 cases, dilated cardiomyopathy in 28, valvular heart disease in 12, Chagas' disease in 10 and systemic hypertension in 8 patients. The prescribed treatment was analyzed, with attention to the prescription and dosage of angiotensin converting enzyme (ACE) inhibitors. We also analyzed whether the cause and/or the degree of HF influenced the treatment chosen. Eighty-seven patients received ACE inhibitors, 31 received doses below those recommended in the large trials. Digoxin was prescribed in 69 cases, diuretics in 85, and aspirin in 33. When dividing the patients according to EF, the group with EF below 0.45 was prescribed more often ACE inhibitors (91.5% vs 80.4%) and had more often usage of adequate doses (61% vs 48.7%). In this sample the majority of the patients were treated according to modern recommendations and tolerated well ACE inhibitors, however 1/3 did not receive ACE inhibitors in the recommended doses. Treatment based on betablockers or angiotensin II inhibitors were not routinely employed.


Schizophrenia and smoking: evidence for a common neurobiological basis?
Several previous investigations have suggested that the gene for the alpha 7-nicotinic receptor may play a role in the pathogenesis of schizophrenia and may be responsible for the heavy smoking among schizophrenic patients. In a study of 129 healthy controls and 127 schizophrenic, schizoaffective, and bipolar patients we have aimed 1) to confirm the potential association between schizophrenia and the alpha 7-nicotinic receptor, 2) to test the diagnostic specificity of alpha 7-receptor subunits with respect to psychiatric diagnoses, and 3) to investigate potential receptor differences between smokers and nonsmokers in the general population. Our analysis included the two dinucleotide polymorphisms D15S1360 and L76630 that are localized in a genomic fragment containing the alpha 7-nicotinic receptor gene CHRNA7. Highly significant differences (P < 0.0001) between the allele distributions of patients and controls were detected for these two markers with all three diagnostic subgroups contributing to the discrimination. An independently ascertained replication sample of 24 patients confirmed this finding. Our results suggested an unspecific vulnerability that depended on the severity of overall psychopathology in terms of the co-occurrence of psychopathology with no clear-cut boundary between the diagnostic entities. In comparison with healthy controls, this vulnerability was lowest among schizophrenics, intermediate among bipolars, and highest among schizoaffectives. As to the question of alpha 7-receptor differences between smokers and nonsmokers among the healthy control subjects, our analysis revealed no significant differences, thus indicating that the differences between patients and controls are more than just a smoker/nonsmoker distinction. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:173-177, 2000.


Noninvasive brain-computer interface enables communication after brainstem stroke.
Brain-computer interfaces (BCIs) provide communication that is independent of muscle control, and can be especially important for individuals with severe neuromuscular disease who cannot use standard communication pathways or other assistive technology. It has previously been shown that people with amyotrophic lateral sclerosis (ALS) can successfully use BCI after all other means of independent communication have failed. The BCI literature has asserted that brainstem stroke survivors can also benefit from BCI use. This study used a P300-based event-related potential spelling system. This case study demonstrates that an individual locked-in owing to brainstem stroke was able to use a noninvasive BCI to communicate volitional messages. Over a period of 13 months, the participant was able to successfully operate the system during 40 of 62 recording sessions. He was able to accurately spell words provided by the experimenter and to initiate dialogues with his family. The results broadly suggest that, regardless of the precipitating event, BCI use may be of benefit to those with locked-in syndrome.


Evaluation of the DNA damage in lymphocytes, sperm and buccal cells of workers under environmental and occupational boron exposure conditions.
Industrial production and use of boron compounds have increased during the last decades, especially for the manufacture of borosilicate glass, fiberglass, metal alloys and flame retardants. This study was conducted in two districts of Balıkesir; Bandırma and Bigadic, which geographically belong to the Marmara Region of Turkey. Bandırma is the production and exportation zone for the produced boric acid and some borates and Bigadic has the largest B deposits in Turkey. 102 male workers who were occupationally exposed to boron from Bandırma and 110 workers who were occupationally and environmentally exposed to boron from Bigadic participated to our study. In this study the DNA damage in the sperm, blood and buccal cells of 212 males was evaluated by comet and micronucleus assays. No significant increase in the DNA damage in blood, sperm and buccal cells was observed in the residents exposed to boron both occupationally and environmentally (p = 0.861) for Comet test in the sperm samples, p = 0.116 for Comet test in the lymphocyte samples, p = 0.042 for micronucleus (MN) test, p = 0.955 for binucleated cells (BN), p = 1.486 for condensed chromatin (CC), p = 0.455 for karyorrhectic cells (KHC), p = 0.541 for karyolitic cells (KLY), p = 1.057 for pyknotic cells (PHC), p = 0.331 for nuclear bud (NBUD)). No correlations were seen between blood boron levels and tail intensity values of the sperm samples, lymphocyte samples, frequencies of MN, BN, KHC, KYL, PHC and NBUD. The results of this study came to the same conclusions of the previous studies that boron does not induce DNA damage even under extreme exposure conditions.


Age Related Differences in Monocyte Subsets and Cytokine Pattern during Acute COVID-19-A Prospective Observational Longitudinal Study.
The COVID-19 pandemic drastically highlighted the vulnerability of the elderly population towards viral and other infectious threats, illustrating that aging is accompanied by dysregulated immune responses currently summarized in terms like inflammaging and immunoparalysis. To gain a better understanding on the underlying mechanisms of the age-associated risk of adverse outcome in individuals experiencing a SARS-CoV-2 infection, we analyzed the impact of age on circulating monocyte phenotypes, activation markers and inflammatory cytokines including interleukin 6 (IL-6), IL-8 and tumor necrosis factor (TNF) in the context of COVID-19 disease progression and outcome in 110 patients. Our data indicate no age-associated differences in peripheral monocyte counts or subset composition. However, age and outcome are associated with differences in monocyte activation status. Moreover, a distinct cytokine pattern of IL-6, IL-8 and TNF in elderly survivors versus non-survivors, which consolidates over the time of hospitalization, suggests that older patients with adverse outcomes experience an inappropriate immune response, reminiscent of an inflammaging driven immunoparalysis. Our study underscores the value, necessity and importance of longitudinal monitoring in elderly COVID-19 patients, as dynamic changes after symptom onset can be observed, which allow for a differentiated insight into confounding factors that impact the complex pathogenesis following an infection with SARS-CoV-2.


Ex Vivo OCT-Based Multimodal Imaging of Human Donor Eyes for Research into Age-Related Macular Degeneration.
A progression sequence for age-related macular degeneration (AMD) learned from optical coherence tomography (OCT)-based multimodal (MMI) clinical imaging could add prognostic value to laboratory findings. In this work, ex vivo OCT and MMI were applied to human donor eyes prior to retinal tissue sectioning. The eyes were recovered from non-diabetic white donors aged ≥80 years old, with a death-to-preservation time (DtoP) of ≤6 h. The globes were recovered on-site, scored with an 18 mm trephine to facilitate cornea removal, and immersed in buffered 4% paraformaldehyde. Color fundus images were acquired after anterior segment removal with a dissecting scope and an SLR camera using trans-, epi-, and flash illumination at three magnifications. The globes were placed in a buffer within a custom-designed chamber with a 60 diopter lens. They were imaged with spectral domain OCT (30° macula cube, 30 µm spacing, averaging = 25), near-infrared reflectance, 488 nm autofluorescence, and 787 nm autofluorescence. The AMD eyes showed a change in the retinal pigment epithelium (RPE), with drusen or subretinal drusenoid deposits (SDDs), with or without neovascularization, and without evidence of other causes. Between June 2016 and September 2017, 94 right eyes and 90 left eyes were recovered (DtoP: 3.9 ± 1.0 h). Of the 184 eyes, 40.2% had AMD, including early intermediate (22.8%), atrophic (7.6%), and neovascular (9.8%) AMD, and 39.7% had unremarkable maculas. Drusen, SDDs, hyper-reflective foci, atrophy, and fibrovascular scars were identified using OCT. Artifacts included tissue opacification, detachments (bacillary, retinal, RPE, choroidal), foveal cystic change, an undulating RPE, and mechanical damage. To guide the cryo-sectioning, OCT volumes were used to find the fovea and optic nerve head landmarks and specific pathologies. The ex vivo volumes were registered with the in vivo volumes by selecting the reference function for eye tracking. The ex vivo visibility of the pathology seen in vivo depends on the preservation quality. Within 16 months, 75 rapid DtoP donor eyes at all stages of AMD were recovered and staged using clinical MMI methods.


Sensory and motor interdependence in postural adjustments.
The sensory reafference from a movement depends upon the movement, and the movement chosen depends upon the available senses, as demonstrated by vestibular patients who abandon certain movements. Often, one variable is assumed to be dependent whereas the other is independent; however, sensory and motor dynamics in posture are interdependent as conditions upon each other. This paper applies conditional dynamics to characterize the global structure of interdependence between sensory states and motor strategies in fast postural adjustments. The mathematical formalism incorporates rich but disparate experimental, clinical, and theoretical results about sensory and motor control of posture. The control structures presented include relatively stable anatomical, physiological, and functional structures, both continuous and discrete, leading to a composite functional logic for the coordination of these structures in sensorimotor control. Results include sensorimotor control structures for postural adjustments for healthy subjects and certain types of vestibular patients. The sensorimotor control structures for patients with absent vestibular function suggest implications for management of the deficit.


Preperitoneal laparoscopic lateral repair in pelvic organ prolapse - a novel approach.
The aim of this study is to present a novel approach for a paravaginal defect treatment. This extraperitoneal approach can be performed in patients with comorbidities and on obese patients. The main advantages are: not requiring the pneumoperitoneum and the Trendelenburg position and the avoidance of peritoneal adhesions. This study presents the results in 27 patients with cystocele caused by a lateral defect pelvic organ prolapse quantification (POP Q) stage II or higher. The procedure was performed with a modified Richardson and Burch technique using a preperitoneal approach. Three follow-up examinations were conducted two, six weeks, and six months after the operation. A quality of life assessment was conducted before and after surgery using the short form of the PFIQ-7. All patients had a POP Q II cystocele and 59% had concomitant stress urinary incontinence. In all patients cystocele was reduce to asymptomatic POP Q stage I or 0. Mean operation time was approximately 80 minutes. In six months post-operation follow up, one case of recurrence was noted. The patients' quality of life revealed a statistical improvement from an average of 6.8 points before, to an average of 0.7 points after the operation (p < 0.05) in the PFIQ-7. Preperitoneal laparoscopic lateral repair is a relatively fast procedure and it is also feasible for obese women and for patients with a cardiopulmonary risk. Neither the Trendelenburg position nor the pneumoperitoneum are required. Postoperatively, the patients witnessed a reduction of the cystocele and complaints connected with their previous condition.


Strategic promiscuity helps avoid inbreeding at multiple levels in a cooperative breeder where both sexes are philopatric.
In cooperative breeders, the tension between the opposing forces of kin selection and kin competition is at its most severe. Although philopatry facilitates kin selection, it also increases the risk of inbreeding. When dispersal is limited, extra-pair paternity might be an important mechanism to avoid inbreeding, but evidence for this is equivocal. The red-winged fairy-wren is part of a genus of cooperative breeders with extreme levels of promiscuity and male philopatry, but is unique in that females are also strongly philopatric. Here, we test the hypothesis that promiscuity is an important inbreeding avoidance mechanism when both sexes are philopatric. Levels of extra-pair paternity were substantial (70% of broods), but did not arise through females mating with their helpers, but via extra-group mating. Offspring were more likely to be sired by extra-pair males when the social pair was closely related, and these extra-pair males were genetically less similar to the female than the social male and thus, inbreeding is avoided through extra-pair mating. Females were consistent in their choice of the extra-pair sire over time and preferred early moulting males. Despite neighbouring males often being close kin, they sired 37% of extra-pair offspring. However, females that gained paternity from neighbours were typically less related to them than females that gained paternity further away. Our study is the first to suggest that mating with both closely related social partners and neighbours is avoided. Such sophistication in inbreeding avoidance strategies is remarkable, as the extreme levels of promiscuity imply that social context may provide little cue to relatedness.


Quantitative relationship of sick building syndrome symptoms with ventilation rates.
Data from published studies were combined and analyzed to develop best-fit equations and curves quantifying the change in sick building syndrome (SBS) symptom prevalence in office workers with ventilation rate. For each study, slopes were calculated, representing the fractional change in SBS symptom prevalence per unit change in ventilation rate per person. Values of ventilation rate, associated with each value of slope, were also calculated. Linear regression equations were fitted to the resulting data points, after weighting by study size. Integration of the slope-ventilation rate equations yielded curves of relative SBS symptom prevalence vs. ventilation rate. Based on these analyses, as the ventilation rate drops from 10 to 5 l/s-person, relative SBS symptom prevalence increases approximately 23% (12% to 32%), and as ventilation rate increases from 10 to 25 l/s-person, relative prevalence decreases approximately 29% (15% to 42%). Variations in SBS symptom types, building features, and outdoor air quality may cause the relationship of SBS symptom prevalence with ventilation rate in specific situations to differ from the average relationship predicted in this paper. On average, providing more outdoor air ventilation will reduce prevalence rates of sick building syndrome (SBS) symptoms. However, given the costs of energy use, including increased risks of climate change, it is important to balance the benefits and risks of increased ventilation. This paper provides initial estimates of how the incremental health benefits per unit of increased ventilation diminish at higher levels of ventilation.


The association of ACTN3 R577X polymorphism with sports specificity in Japanese elite athletes.
The α-actinin-3 proteins regulate muscle function and are located in the Z-line of the fast skeletal muscle. A common null polymorphism of R577X in α-actinin-3 gene (ACTN3) results in its complete absence in fast-twitch muscles. The ACTN3 R577X polymorphism is associated with sprint/power performance in athletes. However, little is known about how this polymorphism impacts sports other than sprint/power-oriented sports in Japanese elite athletes. The aim of our study was to examine the association between ACTN3 R577X polymorphism and elite athlete status in various sports categorized as power/sprint, endurance, artistic, martial arts, and ball game sports. The subjects included 906 Japanese elite athletes and 649 Japanese controls. We analysed the genotype frequency of the ACTN3 R577X polymorphism in sprint/power (n = 120), endurance (n = 150), artistic (n = 45), martial arts (n = 94), and ball game (n = 497) sports athletes. A higher number of sprint/power athletes were R allele carriers compared to the controls, and the endurance and artistic athletes (OR = 1.69, 1.83, and 2.36, 95% CI: 1.02-2.79, 1.02-3.31, and 1.08-5.13, respectively). The frequency of RR genotype was higher in sprint/power, martial arts, and ball game sports athletes (OR = 1.61, 1.84, and 1.39, 95% CI: 1.04-2.50, 1.11-2.95, and 1.05-1.83, respectively) compared to control. Furthermore, there is a significant linear trend with increasing R allele according to athletic status (P for trend < 0.05). The ACTN3 R allele is positively associated with sports performance requiring explosive power such as sprint/power, martial arts, and ball game sports categories.


Associations between physical activity and health parameters in adolescent pupils in Egypt.
Physical activity (PA) could be protective against hypertension, atherosclerosis, coronary heart disease and cardiovascular disease. This quantitative study assessed the association between a PA intervention and three anthropometric parameters (weight, body mass index, body fat) and four physiological parameters (cholesterol level, systolic blood pressure, diastolic blood pressure, heart rate) among secondary school pupils (N = 160) in Egypt through the school term. The pupils were randomised to an intervention group (80 pupils) and controls (80 pupils). Measurements were obtained for all pupils twice: at baseline; and then again after three months. The PA intervention programme comprised an 'afterschool' one hour of moderate exercise three times a week for three months. Both the controls and the intervention pupils attended the 'normal' exercise schedule provided by the school; in addition, the intervention group attended afterschool PA programme from about 2-3 o'clock in the afternoon. At baseline, employing pupil's BMI, 27.5% and 28.8% of the intervention and control pupils respectively were classified as overweight. After three months, the percentage of overweight decreased to 12.5% in the intervention pupils, while it increased to 37.3% in the controls. At the end of the three months period, there were significant improvements across most anthropometric and physiological parameters of the intervention pupils when compared with the control children. The correlation coefficient of the improvements for the boys and the girls was 0.97, indicating clearly that the intervention was having nearly the same beneficial effect for boys and girls. A moderate PA programme for a modest period of 3 months could be effective in maintaining or enhancing pupil's anthropometric and physiological parameters in comparison to the controls where there was deterioration in both parameters. Policy makers and secondary schools in Egypt might need to pay more attention to PA programmes conducted on school days, in order to motivate pupils to attend such programmes. There is also an urgent need to look at current PA systems within schools in Egypt in order to assess PA outside school times.


Glycosylation of phytepsin and expression of dad1, dad2 and ost1 during onset of cell death in germinating barley scutella.
Dysfunction and downregulation of dad (defending against death) has been linked to programmed cell death (PCD) in animals and plants. As DAD is an essential subunit of the oligosaccharyltransferase that is located in the ER membrane, the results have raised the possibility that downregulation of N-linked glycosylation could be involved in the regulation of PCD. Here we show that the 16 kDa subunit of phytepsin, a vacuolar proteinase, is normally processed and glycosylated at the onset of DNA fragmentation in germinating barley scutella. Two cDNA clones encoding dad (dad1, dad2), and one cDNA encoding another subunit of the same oligosaccharyltransferase complex (ost1) were isolated from barley. Northern analysis of germinating scutella show that the expression of only dad1 is declining before onset of DNA fragmentation. In contrast to this, the expression of both dad2 and ost1 increase before onset of DNA fragmentation.


A new scanning heterodyne interferometer scheme for mapping both surface structure and effective local reflection coefficient.
A new scanning microscope scheme which can map both phase and amplitude change of the probe beam is introduced. We will show that the true surface structure can be imaged by using the results of phase measurements while the amplitude image represents the map of the magnitude of the effective local reflection coefficient (ELRC). Relation between the surface structure and the ELRC is discussed. Spatial resolution is 0.67 microm which is limited by diffraction and the precision for measuring point-to-point variation of the average height of the surface structure is a few nanometers. Potential of this microscopy on surface diagnostics is discussed.


Recent advances on CD4+ T cells in atherosclerosis and its implications for therapy.
Atherosclerosis is an arterial inflammatory disease and the primary cause of cardiovascular disease. T helper (Th) cells are an important part in atherosclerotic plaque as they can be either disease promoting or protective. A body of evidence points to a pro-atherosclerotic role of Th1 cells, whereas the role of Th2, Th17 and iNKT cells seems more complex and dependent on surrounding factors, including the developmental stage of the disease. Opposed to Th1 cells, there is convincing support for an anti-atherogenic role of Tregs. Recent data identify the plasticity of Th cells as an important challenge in understanding the functional role of different Th cell subsets in atherosclerosis. Much of the knowledge of Th cell function in atherosclerosis is based on findings from experimental models and translating this into human disease is challenging. Targeting Th cells and/or their specific cytokines represents an attractive option for future therapy against atherosclerosis, although the benefits and the risk of modulation of Th cells with these novel drug targets must first be carefully assessed.


Lithium carbonate addition in tricyclic antidepressant-resistant unipolar depression. Correlations with the neurobiologic actions of tricyclic antidepressant drugs and lithium ion on the serotonin system.
Preliminary reports suggested that the addition of lithium carbonate to the regimen of patients treated with, but not responding to, a tricyclic antidepressant (TCA) drug can induce a rapid alleviation of depression. We examined the effect of lithium carbonate addition in 39 patients with unipolar depression whose conditions were not improved by at least three weeks' TCA drug administration. In 30 of 42 observations, lithium carbonate brought about a greater than 50% improvement within 48 hours. In a second study, the effects of lithium carbonate addition were compared in five amitriptyline hydrochloride-pretreated and five placebo-pretreated patients who showed no improvement after a three-week treatment. All five patients receiving amitriptyline showed a greater than 50% improvement 48 hours after lithium carbonate addition, whereas only one patient in the placebo group showed a marked response. In a third study the effect of lithium carbonate withdrawal was studied in nine TCA-resistant patients who had shown a marked improvement 48 hours after lithium addition. Only five of these patients had a relapse five days after lithium discontinuation. Since animal studies have shown that TCA drugs sensitize forebrain neurons to serotonin and that lithium enhances the activity of serotonin-containing neurons, we propose that the antidepressant effect of lithium addition in TCA-resistant patients might be mediated by enhancing serotonin neurotransmission.


Prothrombin and partial thromboplastin times as preoperative screening tests.
Preoperative tests of coagulation function have been suggested to detect patients who are likely to have abnormal bleeding during and after surgery. A study was designed to determine the yield of prothrombin time (PT) and partial thromboplastin time (PTT), both in discovering patients who are at risk for abnormal bleeding and in inducing changes in patient care or outcome. Of 750 patients on three surgical services, 611 (81%) patients had no indication of a bleeding disorder on history or physical examination. Of the 139 patients who had clinical indications, 25 (18.0%) patients had an abnormal PT or PTT. Of the 611 patients without clinical indications, 480 patients had the PT or PTT determined, and 13 (2.7%) patients had abnormal results. One (0.2%) of the 480 patients might have benefited from the test result (this patient required a second operation to control arterial bleeding). The prolonged PT or PTT was of no apparent clinical importance in the remaining 12 patients without indications of bleeding disorders preoperatively. The low yield of the PT and PTT in detecting unsuspected bleeding disorders preoperatively was further obscured by the larger number of apparently false-positive results.


Stromal Microenvironment Shapes the Intratumoral Architecture of Pancreatic Cancer.
Single-cell technologies have described heterogeneity across tissues, but the spatial distribution and forces that drive single-cell phenotypes have not been well defined. Combining single-cell RNA and protein analytics in studying the role of stromal cancer-associated fibroblasts (CAFs) in modulating heterogeneity in pancreatic cancer (pancreatic ductal adenocarcinoma [PDAC]) model systems, we have identified significant single-cell population shifts toward invasive epithelial-to-mesenchymal transition (EMT) and proliferative (PRO) phenotypes linked with mitogen-activated protein kinase (MAPK) and signal transducer and activator of transcription 3 (STAT3) signaling. Using high-content digital imaging of RNA in situ hybridization in 195 PDAC tumors, we quantified these EMT and PRO subpopulations in 319,626 individual cancer cells that can be classified within the context of distinct tumor gland "units." Tumor gland typing provided an additional layer of intratumoral heterogeneity that was associated with differences in stromal abundance and clinical outcomes. This demonstrates the impact of the stroma in shaping tumor architecture by altering inherent patterns of tumor glands in human PDAC.


Symptomatic gallstones in patients with spinal cord injury.
Patients with spinal cord injury (SCI) have an increased prevalence of cholelithiasis. The goal of this study was to clarify the presentation and management of symptomatic gallstone disease in patients with SCI. We performed a retrospective study of presentation of gallstone complications in patients with SCI who underwent cholecystectomy for complications of gallstone disease. The West Roxbury Veterans Administration Medical Center SCI registry (605 patients) was searched for patients who had undergone cholecystectomy more than 1 year after SCI (35 patients). Gallbladder disease profiles for the 35 patients undergoing cholecystectomy for complications of gallstone disease were prepared, including demographics, clinical presentation, diagnostic studies, operative and pathologic findings, and postoperative complications. All patients were white. Thirty-four were male and the mean age was 50 years (range 35 to 65 years). The majority of patients (66%) complained of right upper quadrant abdominal pain, even those patients with SCI at high (i.e., cervical) levels. Of the 35 patients in our study group, 22 (63%) had biliary colic and chronic cholecystitis, nine (26%) had acute cholecystitis (gangrenous cholecystitis in two), two (6%) had choledocholithiasis symptoms or cholangitis, and two (6%) had gallstone pancreatitis. Major perioperative morbidity occurred in two (6%) of the 35 patients (pulmonary embolus; intraoperative hemorrhage), and there were no deaths. In the great majority of patients with SCI, cholelithiasis presents with chronic pain and not with life-threatening complications. Our findings suggest that presentation is no more acute in patients with SCI than in the general population. Characteristic symptoms and signs are not necessarily obscured by SCI injury, regardless of the level.


Attitudes toward Pursuing Genetic Testing among Parents of Children with Autism Spectrum Disorder in Taiwan: A Qualitative Investigation.
The diagnosis of autism spectrum disorder (ASD) cases is increasing in Taiwan. Genetic testing for children with ASD offers several potential benefits and is available with out-of-pocket expenses. Parents play a pivotal role in having their children with ASD tested; therefore, understanding their perceptions of, and perceived barriers to genetic testing is vital. Semi-structured interviews were conducted with 39 parents of children with ASD in Taiwan. Interviews were recorded and transcribed verbatim. NVivo 12 software (QSR International, Doncaster, Australia) was used to facilitate an inductive coding methodology. The majority of participants (74.4%) supported ASD genetic testing for their children with ASD, citing reasons such as clarifying ASD etiology, well-informed family planning, contributing to ASD research, and early ASD detection and intervention. Others indicated that they were either against such testing (17.9%), or unsure (7.7%) about whether to take their children with ASD for genetic testing. Those who were opposed reported that their main concerns related to perceptions of no value of genetic testing, potential for family conflict, and financial difficulties. Most of the parents of children with ASD that we interviewed expressed favorable views of ASD genetic testing. There exists a need to increase parental access to education and counseling, and to include testing coverage in Taiwanese national health insurance.


The rumble sound generated by a tidal bore event in the Baie du Mont Saint Michel.
A tidal bore is a sharp rise in free-surface elevation propagating upstream in an estuarine system at the leading edge of the flood tide. It generates a powerful noise that was sometimes compared to the sounds of a horse cavalcade. Herein the sounds generated by a tidal bore event in the Baie du Mont Saint Michel were carefully recorded. The data showed three distinct periods. These were the incoming tidal bore when the sound amplitude increased with the approaching bore front, the passage of the tidal bore in front of the microphone where the impacts of the bore on the bank, rocks, or jetty generated powerful noises, and the upstream propagation of the bore when the flood flow motion caused additional loud noises. During the arrival of the tidal bore, the sound levels were less energetic and a lower-pitch sound was noted than during the subsequent record. For the breaking bore process, the analysis of the sound record indicated a dominant frequency around 76-77 Hz. The low-pitch rumble had a frequency comparable to the collective bubble oscillations, suggesting that air entrapment in the bore roller might play a major role in the acoustic signature.


Molecular mapping of QTLs conferring stay-green in grain sorghum (Sorghum bicolor L. Moench).
Drought resistance is of enormous importance in crop production. The identification of genetic factors involved in plant response to drought stress provides a strong foundation for improving drought tolerance. Stay-green is a drought resistance trait in sorghum (Sorghum bicolor L. Moench) that gives plants resistance to premature senescence under severe soil moisture stress during the post-flowering stage. The objective of this study was to map quantitative trait loci (QTLs) that control the stay-green and chlorophyll content in sorghum. By using a restriction fragment length polymorphism (RFLP) map, developed from a recombinant inbred line (RIL) population, we identified four stay-green QTLs, located on three linkage groups. The QTLs (Stg1 and Stg2) are on linkage group A, with the other two, Stg3 and Stg4, on linkage groups D and J, respectively. Two stay-green QTLs, Stg1 and Stg2, explaining 13-20% and 20-30% of the phenotypic variability, respectively, were consistently identified in all trials at different locations in two years. Three QTLs for chlorophyll content (Chl1, Chl2, and Chl3), explaining 25-30% of the phenotypic variability were also identified under post-flowering drought stress. All coincided with the three stay-green QTL regions (Stg1, Stg2, and Stg3) accounting for 46% of the phenotypic variation. The Stg1 and Stg2 regions also contain the genes for key photosynthetic enzymes, heat shock proteins, and an abscisic acid (ABA) responsive gene. Such spatial arrangement shows that linkage group A is important for drought- and heat-stress tolerance and yield production in sorghum. High-resolution mapping and cloning of the consistent stay-green QTLs may help to develop drought-resistant hybrids and to understand the mechanism of drought-induced senescence in plants.


Determination of worries and expectations of family practice patients.
One of the cornerstones of family practice is the development of the physician-patient relationship within the context of the consultation. Each consultation is modified both by the prior expectations of the participants and by illness-related worries held by the patient. This paper describes the development of an instrument to record the worries and expectations of patients visiting their family physicians and changes occurring as a result of the consultation. The instrument comprises a card sort composed of 26 cards, on each of which is typed a statement relating to a worry or expectation that a patient might hold. One hundred patients were asked to sort these cards before and after the consultation. The major worries of the group were about discomfort, the effects of illness on the family, the prospect of a physical examination, and about explaining the problem to the physician. Prominent expectations were for an explanation of diagnosis and treatment and a friendly and understanding manner from the physician.


Modeling weight loss of cheese during ripening and the influence of dairy system, parity, stage of lactation, and composition of processed milk.
The yield, flavor, and texture of ripened cheese result from numerous interrelated microbiological, biochemical, and physical reactions that take place during ripening. The aims of the present study were to propose a 2-compartment first-order kinetic model of cheese weight loss over the ripening period; to test the variation in new informative phenotypes describing this process; and to assess the effects on these traits of dairy farming system, individual farms within dairy system, animal factors, and milk composition. A total of 1,211 model cheeses were produced in the laboratory using individual 1.5-L milk samples from Brown Swiss cows reared on 83 farms located in Trento Province. During ripening (60 d; temperature 15°C, relative humidity 85%), the weight of all model cheeses was measured, and cheese yield (cheese weight/processed milk weight, %CY) was calculated at 7 intervals from cheese-making (0, 1, 7, 14, 28, 42, and 60 d). Using these measures, a 2-compartment first-order kinetic model (3-parameter equation) was developed for modeling %CY during the ripening period, as follows: [Formula: see text] , where %CYt is the %CY at ripening time t; %CYi and %CYf are the modeled %CY traits at time 0 d (%CYi = initial %CY) and at the end of a ripening period sufficient to reach a constant wheel weight (%CYf = final %CY after 60 d ripening in the case of small model cheeses); kCY is the instant rate constant for cheese weight loss (%/d). Cheese weight and protein and fat losses were calculated as the % difference between the model cheeses at 0 and after 60 d of ripening. The variation in cheese pH was calculated as the % difference between pH at 0 and after 60 d. Dairy system, individual herd within dairy system, and the cow's parity and lactation stage (tested with a linear mixed model) strongly affected almost all the traits collected during model cheese ripening. Milk fat, protein, lactose, pH, and somatic cell score also greatly affected almost all the traits, although kCY was affected only by milk protein. After including milk composition in the linear mixed model, the importance of all the herd and animal sources of variation was greatly reduced for all traits. The proposed model and novel traits could be tested, first, with the aim of establishing new monitoring procedures enabling the dairy industry to improve milk quality-based payment systems at the herd level and, second, with a view to exploring possible genetic improvements to dairy cow populations.


Pediatric renal transplantation under tacrolimus-based immunosuppression.
Tacrolimus has been used as a primary immunosuppressive agent in adult and pediatric renal transplant recipients, with reasonable outcomes. Methods. Between December 14, 1989 and December 31, 1996, 82 pediatric renal transplantations alone were performed under tacrolimus-based immunosuppression without induction anti-lymphocyte antibody therapy. Patients undergoing concomitant or prior liver and/or intestinal transplantation were not included in the analysis. The mean recipient age was 10.6+/-5.2 years (range: 0.7-17.9). Eighteen (22%) cases were repeat transplantations, and 6 (7%) were in patients with panel-reactive antibody levels over 40%. Thirty-four (41%) cases were with living donors, and 48 (59%) were with cadaveric donors. The mean donor age was 27.3+/-14.6 years (range: 0.7-50), and the mean cold ischemia time in the cadaveric cases was 26.5+/-8.8 hr. The mean number of HLA matches and mismatches was 2.8+/-1.2 and 2.9+/-1.3; there were five (6%) O-Ag mismatches. The mean follow-up was 4.0+/-0.2 years. The 1- and 4-year actuarial patient survival was 99% and 94%. The 1- and 4-year actuarial graft survival was 98% and 84%. The mean serum creatinine was 1.1+/-0.5 mg/dl, and the corresponding calculated creatinine clearance was 88+/-25 ml/min/1.73 m2. A total of 66% of successfully transplanted patients were withdrawn from prednisone. In children who were withdrawn from steroids, the mean standard deviation height scores (Z-score) at the time of transplantation and at 1 and 4 years were -2.3+/-2.0, -1.7+/-1.0, and +0.36+/-1.5. Eighty-six percent of successfully transplanted patients were not taking anti-hypertensive medications. The incidence of acute rejection was 44%; between December 1989 and December 1993, it was 63%, and between January 1994 and December 1996, it was 23% (P=0.0003). The incidence of steroid-resistant rejection was 5%. The incidence of delayed graft function was 5%, and 2% of patients required dialysis within 1 week of transplantation. The incidence of cytomegalovirus was 13%; between December 1989 and December 1992, it was 17%, and between January 1993 and December 1996, it was 12%. The incidence of early Epstein-Barr virus-related posttransplant lymphoproliferative disorder (PTLD) was 9%; between December 1989 and December 1992, it was 17%, and between January 1993 and December 1996, it was 4%. All of the early PTLD cases were treated successfully with temporary cessation of immunosuppression and institution of antiviral therapy, without patient or graft loss. These data demonstrate the short- and medium-term efficacy of tacrolimus-based immunosuppression in pediatric renal transplant recipients, with reasonable patient and graft survival, routine achievement of steroid and anti-hypertensive medication withdrawal, gratifying increases in growth, and, with further experience, a decreasing incidence of both rejection and PTLD.


Accounting for animal movement in estimation of resource selection functions: sampling and data analysis.
Patterns of resource selection by animal populations emerge as a result of the behavior of many individuals. Statistical models that describe these population-level patterns of habitat use can miss important interactions between individual animals and characteristics of their local environment; however, identifying these interactions is difficult. One approach to this problem is to incorporate models of individual movement into resource selection models. To do this, we propose a model for step selection functions (SSF) that is composed of a resource-independent movement kernel and a resource selection function (RSF). We show that standard case-control logistic regression may be used to fit the SSF; however, the sampling scheme used to generate control points (i.e., the definition of availability) must be accommodated. We used three sampling schemes to analyze simulated movement data and found that ignoring sampling and the resource-independent movement kernel yielded biased estimates of selection. The level of bias depended on the method used to generate control locations, the strength of selection, and the spatial scale of the resource map. Using empirical or parametric methods to sample control locations produced biased estimates under stronger selection; however, we show that the addition of a distance function to the analysis substantially reduced that bias. Assuming a uniform availability within a fixed buffer yielded strongly biased selection estimates that could be corrected by including the distance function but remained inefficient relative to the empirical and parametric sampling methods. As a case study, we used location data collected from elk in Yellowstone National Park, USA, to show that selection and bias may be temporally variable. Because under constant selection the amount of bias depends on the scale at which a resource is distributed in the landscape, we suggest that distance always be included as a covariate in SSF analyses. This approach to modeling resource selection is easily implemented using common statistical tools and promises to provide deeper insight into the movement ecology of animals.


The effect of caffeine on normal and denervated gastrocnemius muscles of Uromastix hardwickii.
The effect of time and 5 mM caffeine was studied on the maximum twitch tension of the normal and 11-13 days denervated gastrocnemius muscles of Uromastix hardwickii. The maximum twitch tension was found to decrease with time and it remained about 68% of initial after 60 minutes at 20 degrees C. Five mM caffeine only produced potentiation in normal and denervated muscles while there was no contracture in either muscle. Although caffeine increased the overall contraction and relaxation times in control muscles, it was more effective on the contraction time of the denervated muscles in which the contraction time increased with time. Denervated muscles were seen to be less sensitive to caffeine as the contraction times increased with time (at about 30-40 minutes) in these muscles while in normal control muscles the contraction time increased immediately. The action of caffeine in relation to these effects is discussed.


Postischemic endothelium-dependent vascular reactivity is preserved in adhesion molecule-deficient mice.
Previous studies utilizing monoclonal antibodies directed against specific leukocyte-endothelial cell adhesion proteins have suggested that neutrophils mediate endothelial cell injury in a number of vascular beds after ischemia-reperfusion (I/R). In the present study, we investigated superior mesenteric artery (SMA) vascular reactivity to acetylcholine (ACh) and sodium nitroprusside (SNP) after occlusion and reperfusion in wild-type (C57BL/6) mice and in gene-targeted mice that are deficient in either CD11/CD18, intercellular adhesion molecule 1 (ICAM-1), or P-selectin. All mice were 4 wk of age, and the SMA was occluded for 45 min and then reperfused for 45 min. Segments of SMA were isolated and suspended in organ chambers and contracted with phenylephrine (10(-5) M), and the maximal vasorelaxation to ACh (10(-6) M) and SNP (10(-6) M) was measured. SMA from sham-operated C57BL/6 mice relaxed 83.5 +/- 3.3% to ACh and 91.7 +/- 3.4% to SNP. In contrast, segments of SMA from C57BL/6 mice subjected to I/R demonstrated a marked impairment in vasorelaxation to ACh (51.3 +/- 4.7%, P < 0.01 vs. sham) without any impairment in the vasoreactivity to SNP (86.1 +/- 5.5%). In CD11/CD18-deficient mice, SMA reactivity to ACh (84.7 +/- 2.3%) and SNP (91.2 +/- 2.8%) was unaffected by I/R. Similarly, SMA rings from ICAM-1-deficient mice exhibited normal vasorelaxation to ACh and SNP with maximal vasorelaxation of 83.1 +/- 2.9 and 87.4 +/- 3.0%, respectively. We also observed profound preservation of endothelium-dependent vasorelaxation after I/R in P-selectin-deficient mice. These findings indicate that leukocyte-endothelial cell adhesion molecule deficiency is associated with the preservation of endothelium-dependent vascular responses after I/R.


Early morbidity encountered in the dietary-related mouse model of Barrett's esophagus: a question of zinc?
Recently, a mouse model for Barrett's esophagus based on a zinc-deficient diet supplemented with deoxycholic bile acids has been published. The aim of this study was to attempt to reproduce these data and extend them by employing genetically modified mice and intraperitoneal iron supplementation. The study design encompassed six experimental groups (wild type, Apc-mutant and Smad4-mutant mice, with or without iron injections), with all animals fed with the zinc-deficient diet supplemented with deoxycholic bile acids. All treatments were started at 3-5 weeks of age (the majority [78%] at 5 weeks). Animals were scheduled for euthanasia at two distinct time points, namely at 3 and 6 months of age. All mice showed signs of considerable distress already 4 weeks after the start of the modified diets, and had to be euthanized before the first evaluation time point (mean age 9.3 weeks, range 5-15 weeks). No differences were observed between wild type and genetically modified mice, or between animals with or without iron supplementation. On histological examination, we could not detect any lesions (Barrett's esophagus-like or tumors) other than esophagitis. In the currently presented experimental settings, we were not able to reproduce the mouse model according to which Barrett's-like lesions could be detected in animals fed with the zinc-deficient diet supplemented with deoxycholic bile acids.