Intrinsic curvature of DNA influences LacR-mediated looping.
Protein-mediated DNA looping is a common mechanism for regulating gene expression. Loops occur when a protein binds to two operators on the same DNA molecule. The probability of looping is controlled, in part, by the basepair sequence of inter-operator DNA, which influences its structural properties. One structural property is the intrinsic or stress-free curvature. In this article, we explore the influence of sequence-dependent intrinsic curvature by exercising a computational rod model for the inter-operator DNA as applied to looping of the LacR-DNA complex. Starting with known sequences for the inter-operator DNA, we first compute the intrinsic curvature of the helical axis as input to the rod model. The crystal structure of the LacR (with bound operators) then defines the requisite boundary conditions needed for the dynamic rod model that predicts the energetics and topology of the intervening DNA loop. A major contribution of this model is its ability to predict a broad range of published experimental data for highly bent (designed) sequences. The model successfully predicts the loop topologies known from fluorescence resonance energy transfer measurements, the linking number distribution known from cyclization assays with the LacR-DNA complex, the relative loop stability known from competition assays, and the relative loop size known from gel mobility assays. In addition, the computations reveal that highly curved sequences tend to lower the energetic cost of loop formation, widen the energy distribution among stable and meta-stable looped states, and substantially alter loop topology. The inclusion of sequence-dependent intrinsic curvature also leads to nonuniform twist and necessitates consideration of eight distinct binding topologies from the known crystal structure of the LacR-DNA complex.


Pharmacokinetic Studies around the Mono- and Difunctionalization of a Bioavailable Cyclic Decapeptide Scaffold.
We previously reported the design of several cyclic decapeptides based on a generic scaffold that achieved favorable oral bioavailability and exposure. With the goal to further investigate the potential of this approach, we describe herein the effect of mono- and difunctionalization of this scaffold. A series of cyclic decapeptides were therefore subjected to a range of in vitro assays and pharmacokinetic (PK) studies to investigate whether the introduction of polar or charged groups could be tolerated by the "engineered" scaffold while maintaining good PK profiles. Whereas the introduction of charged amino acids proved-besides maintaining low clearance-to conceal the inherent PK properties of the scaffold, the introduction of polar amino acids (i.e., threonine and pyridyl alanine) led to several cyclic decapeptides exhibiting excellent PK profiles together with a solubility that was significantly improved relative to that of previously reported cyclic decapeptides.


[Clinical evaluation of hypopharyngeal cancer].
Clinical results of cases of hypopharyngeal carcinoma treated from 1966 to 1990 at Kyushu University Hospital were investigated. In 1966-1972, all cases were treated with only radical operation. We used FAR therapy since 1972, and preoperative chemotherapy (CDDP and Peplomycin) in addition to FAR therapy since 1984. The crude five-year survival rate of the operative therapy-group (25 cases in 1966-1972) was 24%, and that of the FAR therapy-group with or without operation (47 cases in 1972-1980) was 38%. Although it was cumulative survival rate by Kaplan-Meier's method, the five-year survival rate of the group (16 cases in 1984-1990) treated with a combination of FAR therapy, preoperative-chemotherapy, and operation was 76%. The improvement in the survival rate and the role of preoperative treatment (FAR therapy and chemotherapy) are discussed.


Cytometric analysis and clinical features in a Moroccan cohort with severe combined immunodeficiency.
Severe combined immunodeficiency (SCID) is a form of primary immunodeficiency disease (PID). It is characterized by a serious abnormality of the cellular and sometimes humoral system due to a deficiency in development of T cells, B cells and/or NK cells. The early diagnosis of SCID improves the prognosis. Typically, the initial consideration of SCID is made based on low lymphocyte counts. Notwithstanding, the heterogeneity of lymphocyte count presentation makes the diagnosis of SCID a significant challenge. The objective of this cross-sectional retrospective study was to analyze the lymphocyte subpopulation counts along with clinical manifestations within a Moroccan cohort diagnosed as SCID compared to children diagnosed with non-PID diseases. Thirty-five SCID confirmed patients were selected in the period between 2008 and 2018 and compared with non-PID patients. Results of peripheral blood T, B, and NK lymphocyte subpopulation counts were measured by flow cytometry for each SCID subtype. As expected, T cell count was less than 300 cells/μL in most patients with SCID (85.5%). Unexpectedly, significantly higher T cell counts were detected in some patients with a confirmed clinical diagnosis and family history of SCID. 5.7% of our SCID Moroccan cohort had T cell numbers in the range between 300 and 500 cells/μL. 8.7% of our SCID Moroccan cohort had T cell numbers higher than 500 cells/μL. Of the SCID subtypes, the proportion of SCID with B cell deficiencies was highly represented in our cohort. 71.4% of Moroccan SCID patients (25 out of 35 patients) were of T-B-subtype. Furthermore, 40% of the patients (14 out of 35 patients) had a T-B-NK+ profile and 31.4% had a T-B-NK- profile (11 out of 35 patients). The most common clinical manifestations observed in our SCID cohort were pneumonia, failure to thrive, candidiasis, diarrhea, bronchitis and urinary tract infections. Our results not only highlight the relatively frequent presence of atypical SCID in the Moroccan population with unexpectedly high T cell numbers, but also describes the incidence pattern of common SCID subtypes in Morocco. Physicians in Morocco may find this local region-specific difference in SCID important for making improved early diagnosis of this disease.


Operational Theories in Phase Space: Toy Model for the Harmonic Oscillator.
We show how to construct general probabilistic theories that contain an energy observable dependent on position and momentum. The construction is in accordance with classical and quantum theory and allows for physical predictions, such as the probability distribution for position, momentum, and energy. We demonstrate the construction by formulating a toy model for the harmonic oscillator that is neither classical nor quantum. The model features a discrete energy spectrum, a ground state with sharp position and momentum, an eigenstate with a nonpositive Wigner function as well as a state that has tunneling properties. The toy model demonstrates that operational theories can be a viable alternative approach for formulating physical theories.


Acoustically induced streaming flows near a model cod otolith and their potential implications for fish hearing.
The ears of fishes are remarkable sensors for the small acoustic disturbances associated with underwater sound. For example, each ear of the Atlantic cod (Gadus morhua) has three dense bony bodies (otoliths) surrounded by fluid and tissue, and detects sounds at frequencies from 30 to 500 Hz. Atlantic cod have also been shown to localize sounds. However, how their ears perform these functions is not fully understood. Steady streaming, or time-independent, flows near a 350% scale model Atlantic cod otolith immersed in a viscous fluid were studied to determine if these fluid flows contain acoustically relevant information that could be detected by the ear's sensory hair cells. The otolith was oscillated sinusoidally at various orientations at frequencies of 8-24 Hz, corresponding to an actual frequency range of 280-830 Hz. Phase-locked particle pathline visualizations of the resulting flows give velocity, vorticity, and rate of strain fields over a single plane of this mainly two-dimensional flow. Although the streaming flows contain acoustically relevant information, the displacements due to these flows are likely too small to explain Atlantic cod hearing abilities near threshold. The results, however, may suggest a possible mechanism for detection of ultrasound in some fish species.


A history of dermatologic surgery in the United States.
Dermatologic surgery has a long and distinguished history in the United States. To examine the specific contributions of American dermatologic surgeons. The medical literature on cutaneous reconstructive and cosmetic surgery for the last century and a half was researched. Numerous American dermatologic surgeons have had a major impact on scientific and technological discoveries in cutaneous surgery. Dermatologic surgeons have been significantly involved in cutaneous surgery since the second half of the 19th century. Dermatologic surgeons have contributed many important advances to the fields of chemical peeling, cryosurgery, dermabrasion, electrosurgery, hair transplantation, soft tissue augmentation, tumescent liposuction, laser surgery, phlebology, Mohs chemosurgery, cutaneous reconstruction, wound healing, botulium toxin, blepharoplasty, and rhytidectomy. Dermatologic surgeons in the United States have contributed significantly to the history of reconstructive and cosmetic surgery. Dermatologic surgeons have been leaders in advancing this field and are poised to continue in the future.


Lung Cancer Screening in Asbestos-Exposed Populations.
Asbestos exposure is the most important cause of occupational lung cancer mortality. Two large randomized clinical trials in the U.S. and Europe conclusively demonstrate that annual low-dose chest CT (LDCT) scan screening reduces lung cancer mortality. Age and smoking are the chief risk factors tested in LDCT studies, but numerous risk prediction models that incorporate additional lung cancer risk factors have shown excellent performance. The studies of LDCT in asbestos-exposed populations shows favorable results but are variable in design and limited in size and generalizability. Outstanding questions include how to: (1) identify workers appropriate for screening, (2) organize screening programs, (3) inform and motivate people to screen, and (4) incorporate asbestos exposure into LDCT decision-making in clinical practice. Conclusion: Screening workers aged ≥50 years with a history of ≥5 years asbestos exposure (or fewer years given intense exposure) in combination with either (a) a history of smoking at least 10 pack-years with no limit on time since quitting, or (b) a history of asbestos-related fibrosis, chronic lung disease, family history of lung cancer, personal history of cancer, or exposure to multiple workplace lung carcinogens is a reasonable approach to LDCT eligibility, given current knowledge. The promotion of LDCT-based screening among asbestos-exposed workers is an urgent priority.


Community control in a Gesellschaft world: a comment.
This comment discusses Nassi's analysis in terms of culture conflict as discussed by Earl Johnson (1977). Johnson's analysis, which examines gemeinschaft to gesellschaft societal changes resulting from industrialization provides a wholistic, evolutionary view of changing modes of community control. It is concluded that Johnson's analysis offers a more radical perspective than Nassi's because it deals not merely with power and economic relations, but also with fundamental values which transcend generations and singular societies.


Relating causal and probabilistic approaches to contextuality.
A primary goal in recent research on contextuality has been to extend this concept to cases of inconsistent connectedness, where observables have different distributions in different contexts. This article proposes a solution within the framework of probabi- listic causal models, which extend hidden-variables theories, and then demonstrates an equivalence to the contextuality-by-default (CbD) framework. CbD distinguishes contextuality from direct influences of context on observables, defining the latter purely in terms of probability distributions. Here, we take a causal view of direct influences, defining direct influence within any causal model as the probability of all latent states of the system in which a change of context changes the outcome of a measurement. Model-based contextuality (M-contextuality) is then defined as the necessity of stronger direct influences to model a full system than when considered individually. For consistently connected systems, M-contextuality agrees with standard contextuality. For general systems, it is proved that M-contextuality is equivalent to the property that any model of a system must contain 'hidden influences', meaning direct influences that go in opposite directions for different latent states, or equivalently signalling between observers that carries no information. This criterion can be taken as formalizing the 'no-conspiracy' principle that has been proposed in connection with CbD. M-contextuality is then proved to be equivalent to CbD-contextuality, thus providing a new interpretation of CbD-contextuality as the non-existence of a model for a system without hidden direct influences. This article is part of the theme issue 'Contextuality and probability in quantum mechanics and beyond'.


Investigation and reporting practices for drownings: implications for injury prevention research in new york state.
The New York State Department of Health conducted a study of drownings among New York State residents. A total of 883 drownings between 1988 and 1994 met study criteria. Medical examiner, coroner, police, and hospital records were obtained to supplement death certificate data. Although police reports were an important source of information, acquiring police records was labor intensive and time consuming and would be unnecessary if ME/Cs used standard death investigation guidelines and a uniform reporting form. The authors compared International Classification of Diseases-9 E codes listed on death certificates with information from supplementary data sources. Appropriate International Classification of Diseases-9 E codes were often not used in cases for which a specific code was available. In some cases, a code was assigned that failed to correctly reflect information provided on the death certificate. More frequently, the description provided by the medical examiner or coroner did not permit the use of a more specific code. The role of alcohol could not be assessed for 38% of drowning victims because toxicology testing was not performed, results were not made available by the medical examiner or coroner, or samples were collected 24 hours or more after the time of the drowning. Investigation and reporting practices that would benefit injury prevention research are suggested.


Comparative Analysis of the Impact of Urolithins on the Composition of the Gut Microbiota in Normal-Diet Fed Rats.
The gut microbiota consists of a community of microorganisms that inhabit the large intestine. These microbes play important roles in maintaining gut barrier integrity, inflammation, lipid and carbohydrate metabolism, immunity, and protection against pathogens. However, recent studies have shown that dysfunction in the gut microbiota composition can lead to the development of several diseases. Urolithin A has recently been approved as a functional food ingredient. In this study, we examined the potentials of urolithin A (Uro-A) and B (Uro-B) in improving metabolic functions and their impact on gut microbiota composition under a metabolically unchallenged state in normal rats. Male Wistar rats (n = 18) were randomly segregated into three groups, with Group 1 serving as the control group. Groups 2 and 3 were administered with 2.5 mg/kg Uro-A and Uro-B, respectively, for four weeks. Our results showed that both Uro-A and B improved liver and kidney functions without affecting body weight. Metagenomic analysis revealed that both Uro-A and B induced the growth of Akkermansia. However, Uro-A decreased species diversity and microbial richness and negatively impacted the composition of pathogenic microbes in normal rats. Taken together, this study showed the differential impacts of Uro-A and B on the gut microbiota composition in normal rats and would thus serve as a guide in the choice of these metabolites as a functional food ingredient or prebiotic.


Comorbidity in adults with epilepsy--United States, 2010.
Epilepsy, a spectrum disorder characterized by recurring seizures, affects approximately 2.3 million U.S. adults. Epilepsy poses challenges because of uncontrolled seizures, treatment complexity, social disadvantages (e.g., unemployment), and stigma. Persons with epilepsy are at increased risk for early mortality and for comorbidities that can complicate epilepsy management, increase health-care costs, and shorten the lifespan. Numerous studies have described higher rates of psychiatric comorbidity (e.g., depression and anxiety) in persons with epilepsy. However, fewer studies have examined nonpsychiatric comorbidity in a nationally representative U.S. sample of adults with epilepsy. To assess the prevalence of nonpsychiatric comorbidities, CDC analyzed data from the 2010 National Health Interview Survey (NHIS). Adults with epilepsy had a higher prevalence of cardiovascular, respiratory, some inflammatory, and other disorders (e.g., headache, migraine, and various other types of pain) than adults without epilepsy. Public health agencies can work with health-care providers, the Epilepsy Foundation, and other partners to ensure that adults with epilepsy have access to health promotion resources and chronic disease self-management programs.


Is fake news contributing to increased Covid-19 BAME deaths?
We write this letter as doctors and proud members of the Black, Asian and Minority Ethnic (BAME) community from a South Asian background. Recent Office for National Statistics (ONS) data suggest that the BAME population is disproportionately affected by Covid-19. Observations and experiences from within our family and wider community led us to explore how cultural aspects may account for these figures. Both intrinsic and extrinsic factors are likely to contribute to this unfortunate statistic.


Genetically engineered Salmonella typhimurium as an imageable therapeutic probe for cancer.
Tumor-targeting bacteria have been studied in terms of their ability to visualize the infection pathway (through imaging probes) or to carry therapeutic molecules to tumors. To integrate these monitoring and therapeutic functions, we engineered attenuated Salmonella typhimurium defective in guanosine 5'-diphosphate-3'-diphosphate synthesis to carry cytotoxic proteins (cytolysin A) and express reporter genes. We successfully visualized the therapeutic process with these engineered bacteria in mice and found that they often mediated complete tumor (CT-26) eradication on cytotoxic gene induction. Furthermore, treatment with the engineered bacteria markedly suppressed metastatic tumor growth.


The relationship between on-the-job accidents and patient-hazard incidents for a group of respiratory therapists during a 5-year period.
Safety has not traditionally been stressed in the health care industry to the same extent that it has been in other industries. This is ironic, because accidents in hospitals may endanger not only employees but patients as well. The safety records of 26 respiratory therapists who had had either on- or off-the-job accidents or who had been involved in patient-hazard incidents over a 5-year period were analyzed. There was a positive relationship between employees' involvement in on-the-job accidents and involvement in patient-hazard incidents. This indicates that, for this group, involvement in an on-the-job accident increased the possibility that an employee would be involved in a patient-hazard incident. Although this does not indicate a cause-and-effect relationship between the two factors, it does show that there was a definite relationship between them. Employees should be counseled for safety violations, not just in general terms but in terms specifically related to therapeutic procedures.


Leishmania major telomerase TERT protein has a nuclear/mitochondrial eclipsed distribution that is affected by oxidative stress.
In its canonical role the reverse transcriptase telomerase recovers the telomeric repeats that are lost during DNA replication. Other locations and activities have been recently described for the telomerase protein subunit TERT in mammalian cells. In the present work, using biochemistry, molecular biology, and electron microscopy techniques, we found that in the human parasite Leishmania major, TERT (and telomerase activity) shared locations between the nuclear, mitochondrial, and cytoplasmic compartments. Also, some telomerase activity and TERT protein could be found in ∼ 100-nm nanovesicles. In the mitochondrial compartment, TERT appears to be mainly associated with the kinetoplast DNA. When Leishmania cells were exposed to H2O2, TERT changed its relative abundance and activity between the nuclear and mitochondrial compartments, with the majority of activity residing in the mitochondrion. Finally, overexpression of TERT in Leishmania transfected cells not only increased the parasitic cell growth rate but also increased their resistance to oxidative stress.


Extrauterine abdominal pregnancy: report of a case.
A healthy, 34-year-old, gravida 3, para 1,011, patient presented for cesarean delivery in her 35th week of gestation with a diagnosis of complete placenta previa. During her 26th week of gestation, the patient was admitted to a high-risk obstetric unit with the diagnosis of premature rupture of membranes. Numerous ultrasonographic studies were conducted throughout her 10-week hospital stay, confirming the admitting diagnosis. A routine cesarean section was planned, and preparations were made for a potential increase in blood loss related to the placenta previa. The procedure began under spinal anesthesia and, upon incision of the abdomen, an extrauterine pregnancy was identified. The patient was immediately anesthetized and intubated at the request of the surgeon. During the 3-hour surgical procedure, the patient sustained massive blood loss, transfusions, central line placement, and aggressive pharmacological therapy. The patient was extubated the day after surgery, and was discharged approximately 1 week later. The only major complication was compartment syndrome of the left upper extremity related to the infiltration of vasopressors requiring fasciotomy and closure 2 days later. The incidence, morbidity/mortality, and anesthetic implications of abdominal pregnancy are reviewed.


Factors influencing orthodontic treatment time for non-surgical Class III malocclusion.
To identify variables and their effect size on orthodontic treatment time of Class III malocclusion. Forty-five Class III malocclusion cases were selected from 2008 patients' records. Clinical charts, cephalometric radiographs, and pre and posttreatment dental casts were evaluated. Age, sex, PAR index at T1 and T2, overjet, missing teeth, extractions, number of treatment phases, missed appointments, appliance breakages, and cephalometric variables SNA, SNB, ANB, Wits, SnGoGn, CoA, CoGn, IMPA, 1.PP were investigated by multiple linear regression analysis and stepwise method at p<0.05. The sample was also divided into two groups: Group 0-2 (patients who had missed two clinical appointments or less) and Group >2 (patients who missed more than 2 appointments), to detect the influence of this data on treatment time and the quality of the treatment (PAR T2). Average treatment time was 30.27 months. Multiple regression analysis showed that missed appointment (R2=0.4345) and appliance breakages (R2=0.0596) are the only variables able to significantly predict treatment duration. Treatment time for patients who missed more than 2 appointments was nearly one year longer. However, no significant influence on PAR T2 was observed for those patients. Orthodontic treatment duration in Class III patients is mainly influenced by factors related to patient compliance. Patients who missed more appointments did not show worse orthodontic finishing, but longer treatment. No occlusal, cephalometric, or demographic variable obtained before treatment was able to give some significant prediction about treatment time in Class III patients.


Risk factors for renal scar formation in infants with first episode of acute pyelonephritis: a prospective clinical study.
We prospectively determined the risk factors for renal scar formation after the first episode of acute pyelonephritis as confirmed on dimercapto-succinic acid scintigraphy in children younger than 1 year. A total of 213 infants with acute pyelonephritis were enrolled in the study. Infants with urological abnormalities other than vesicoureteral reflux were excluded from analysis. Followup scanning was performed 6 months after acute pyelonephritis and voiding cystourethrography was performed after the acute phase of infection. Possible risk factors were evaluated including gender, peak fever, duration of fever before and after treatment with antibiotics, white blood cell count, C-reactive protein concentration, presence of vesicoureteral reflux and reflux grade. Six months after acute pyelonephritis 37 of 213 (17.4%) infants and 41 of 248 (16.5%) renal units with acute photon defects on initial dimercapto-succinic acid scintigraphy had renal scars. The rates of scar formation were significantly higher in infants with vesicoureteral reflux than in those without (39.4% vs 7.5%, p <0.001, OR 9.433) and in renal units with vesicoureteral reflux than in those without (39.4% vs 8.2%, p <0.001, OR 7.237). Renal scar formation was related to reflux grade (none-8.2%, grade I-20%, grade II-22.7%, grade III-40%, grade IV-70%, grade V-55.6%, p <0.001) but not to any other clinical or laboratory variables. The presence of vesicoureteral reflux was the only independent risk factor for renal scar formation after acute pyelonephritis in infants. The prevalence of renal scarring was significantly correlated with reflux grade. Voiding cystourethrography is necessary in infants after the first acute pyelonephritis episode is confirmed on dimercapto-succinic acid renal scintigraphy.


High-resolution magnetic resonance imaging and 18FDG-PET findings of the cervical spinal cord before and after decompressive surgery in patients with compressive myelopathy.
Evaluation of cervical spinal cord (CSC) of patients with compressive myelopathy by magnetic resonance imaging (MRI) and high-resolution (18F)fluoro-deoxyglucose (18FDG) positron emission tomography (PET). To determine changes in morphology, intramedullary signal intensity, and glucose metabolic rate in CSC after decompression, and to assess the utility of 18FDG-PET in evaluation of patients with cervical myelopathy. The significance of CSC enlargement after decompression and signal intensity changes within the cord remain elusive. No data are available on metabolic activity of the compressed CSC. Only a few studies have examined correlation between high-resolution MRI and 18FDG-PET neuroimaging in cervical myelopathy. We studied 24 patients who underwent cervical decompressive surgery in terms of postoperative neurologic improvement and changes in MRI and 18FDG-PET. Neurologic status was assessed by the Japanese Orthopedic Association scoring system (17-point scale). Signal intensity change in the cord was qualitatively assessed on both T1- and T2-weighted images. The transverse area of the CSC on MRIs and glucose metabolic rate (standardized uptake value [SUV]) from 18FDG-PET were measured digitally. Neurologic improvement correlated with preoperative CSC transverse area at maximal compression (P < 0.01) and at follow-up (P < 0.001) and with mean SUV before surgery (P < 0.01) and at follow-up (P < 0.05). Preoperative signal intensity change on MRIs (low intramedullary signal intensity abnormality on T1-weighted image and high intramedullary on T2-weighted image) correlated negatively with neurologic improvement rate (P < 0.05). The transverse area of the CSC was significantly smaller after surgery in patients with preoperative MRI signal intensity changes (P < 0.05). The SUV at follow-up tended to normalize in association with neurologic improvement. Our results showed that postoperative neurologic improvement in patients with cervical compressive myelopathy correlated with increased transverse area of the spinal cord, signal intensity change on both T1- and T2-weighted image, and the mean SUV.


Physiology of the Yellow-Green 6 Gene in Tomato: A Possible Interrelationship between the Phenotypic Expressions of the Yellow-Green 6 Gene Mutation and the Gibberellins.
The yellow-green 6 (yg(6)) mutation in tomato (Lycopersicon esculentum Mill.) is controlled by a single recessive gene with pleiotropic effects. The syndrome of characters associated with the mutation are enhanced stem elongation, reduced chlorophyll content and absence of detectable anthocyanins. We now have shown that the mutant also has fewer lateral roots than the wild type and higher l-phenylalanine ammonia-lyase (E.C. 184.108.40.206) activity than the normal tomato. These traits of the mutant closely resemble those induced in many plants by the application of gibberellic acid which suggests that the phenotypic expressions of the mutation might in some manner be related to the endogenous level or activity of the gibberellins. In support of this premise, data are presented which show that the characters of the mutant can be induced in the wild type tomato by application of gibberellic acid. Conversely, several traits of the wild type can be induced in the mutant by an inhibitor of gibberellin hiosynthesis, Phosfon. In addition, an embryoless barley half-seed bioassay for the gibberellins and gas-liquid chromatography indicated that the mutant contained at least three times as much total gibberellin as the wild type plant.


Routine use of commercial antibiotic-loaded bone cement in primary total joint arthroplasty: a critical analysis of the current evidence.
Antibiotic-loaded cement (ABLC) has been widely utilized as an adjuvant treatment for patients with periprosthetic joint infection (PJI) but has also evolved to play a prophylactic role against infection in primary total joint arthroplasties (TJA). Nevertheless, there is currently a paucity of studies that systematically investigated this concept. This review aimed at answering the following questions: (I) Can routine use of ABLC help reduce the current infection rates in primary TJA? (II) What are the risks associated with this approach? And (III) can routine use be justified in primary TJA from an economic standpoint? Multiple databases were queried including PubMed, EMBASE, EBSCO Host, and SCOPUS. Studies published between January 1, 1990 and March 31, 2018 were reviewed. Inclusion criteria were studies reporting: (I) clinical outcomes of routine use of ABLC in primary hip and knee arthroplasty with 2-year minimum follow-up, (II) complications related to the use of ABLC, (III) cost of using ABLC. The final analysis included 24 studies. Data from multiple studies demonstrate contradictory results for infection rates when ABLC is used in all primary procedures with a majority of studies showing similar infection rates between ABLC and plain cement. The main concerns associated with routine use of ABLC are negative effects on the mechanical stability of cement, possible systemic and local toxicity of the absorbed antibiotic, and development of resistant bacterial strains. However, current literature has not clinically validated these concerns. Lastly, with an estimated increase in 117 million dollars with the routine use of ABLC in only 50% of TJAs performed each year, it is difficult to justify the use of ABLC without clear superiority in reducing infection. The use of ABLC has undeniably changed the way orthopaedic surgeons deal with PJI today. However, the large-scale, prophylactic use of ABLC in primary TJAs requires further research and justification.


Management of Myositis-Related Interstitial Lung Disease.
Interstitial lung disease (ILD) is a frequent pulmonary manifestation and an important cause of morbidity and mortality in patients with idiopathic inflammatory myopathy. Myositis-related ILD presents a therapeutic challenge for clinicians, as there are no available guidelines to help with management decisions. This review covers the existing evidence on the pharmacologic and nonpharmacologic management of myositis-related ILD, highlighting the lack of randomized controlled data to guide treatment. Given the absence of existing guidelines to inform treatment decisions, we provide a comprehensive summary, including dosing, side effects, and suggested monitoring of the commonly used immunosuppressive agents and a proposed treatment algorithm based on the existing literature.


New autosomal recessive cerebellar ataxia disorder in a large inbred Lebanese family.
A large inbred Lebanese pedigree with congenital spastic ataxia, microcephaly, optic atrophy, short stature, speech defect, abnormal osmiophilic pattern of skin vessels, cerebellar atrophy, and severe mental retardation transmitted as an autosomal recessive trait has been studied. None of the children had any evidence of a metabolic disease, and the analysis of respiratory chain complex abnormalities was unremarkable. Only one child had a history of perinatal difficulties. Differential diagnosis and the possibility that this disorder is a hitherto unreported one are discussed.


Risk stratification with exercise N(13)-ammonia PET in adults with anomalous right coronary arteries.
In adults with an interarterial and intramural course of an anomalous right coronary artery from the left sinus (AAORCA), surgical unroofing is recommended in the setting of myocardial ischaemia. However, data regarding functional testing are limited, and the management of adults without ischaemia is unclear. To evaluate these patients, we employed an exercise N(13)-ammonia positron emission tomography (PET) protocol. We hypothesised that patients with typical angina and exertional dyspnoea would be more likely to have ischaemia and that patients without ischaemia could be managed conservatively. Between July 2008 and December 2014, we retrospectively identified 27 consecutive patients >18 years old with an interarterial and intramural course of an AAORCA who had exercise N(13)-ammonia PET. The majority of patients had anatomic delineation with cardiac CT (25, 93%), and most patients had chest pain (24, 89%). Myocardial ischaemia with PET was common (13, 48%), and ischaemia was more likely in patients with typical angina and exertional dyspnoea (p<0.05). Surgery was performed in 12 patients including 11 patients with ischaemia. At a median follow-up of 245 days, there were no deaths in patients with surgery or in patients managed conservatively. In patients with an interarterial and intramural course of an AAORCA, typical angina and exertional dyspnoea are associated with ischaemia on exercise N(13)-ammonia PET. Referral for surgical unroofing in symptomatic patients with ischaemia on exercise N(13)-ammonia PET and initial conservative management in patients without ischaemia seems appropriate, though larger studies with long-term follow-up are needed.


Congenital myopathies and related disorders.
Considerable progress has been made in molecular genetic research and in identifying the underlying pathogenesis of congenital myopathies, with implications for genetic counseling. Therefore an overview of such advances in the last two years is most timely and relevant for a more precise delineation of these disorders. New mutations have been described on the ryanodine receptor gene, including the carboxyl-terminus region, and experimental models developed to explain their role in central core disease. Phenotype-genotype correlations for nemaline myopathy have improved our understanding of those related to gene mutations. In multi-minicore disease, collaborative studies support genetic heterogeneity and autosomal-recessive inheritance. Research on X-linked myotubular myopathies has revealed a high percentage of mothers of sporadic cases as carriers. Although not initially included within the congenital myopathies, desmin-related or myofibrillar myopathies are described here because they are closely related to other congenital myopathies with intracytoplasmic inclusions. Western blot for myotubularin and desmin has been proposed as a useful diagnostic test for both X-linked myotubular myopathy and desmin-related myopathy, and in-vitro and mouse models for the latter have provided insights into its pathogenesis. Several entities still await genetic characterization. Here we focus on clinical features, inheritance, and molecular genetics. Advances in immunohistochemistry and molecular genetics in congenital muscular dystrophies have enriched our knowledge of this heterogeneous group of disorders, leading to more accurate classification and differentiation between the various congenital myopathies.


Neonatal screening for congenital hypothyroidism and phenylketonuria at Siriraj Hospital, Mahidol University, Bangkok, Thailand--a pilot study.
A newborn screening program for congenital hypothyroidism (CH) and phenylketonuria (PKU), a pilot study, was initiated at the Medical Genetics Unit, Department of Pediatrics, Siriraj Hospital Faculty of Medicine, Mahidol University in Bangkok, Thailand from January 1994 to December 1998, using dried blood spots (DBS). A total of 18,739 infants (out of 85,150 livebirths) were screened (22 % coverage). Three cases of congenital hypothyroidism (CH) were identified (incidence of 1: 6,246, livebirths), by enzyme linked immunosorbent (ELISA) and fluoroimmunoassays using a cut-off level of TSH >20 microlU/ml: the recall rate of 0.24%. The screening for PKU was done by fluorometric (Guthrie) and enzyme linked immunosorbent (ELISA) methods; using cut-off levels of phenylalanine > 4 mg/dl and > 3.6 mg/dl respectively, with a recall rate of 0.13%. There was no PKU found. Our study, a voluntary program, emphasizes the importance of parental education and consent; specimen collection and handling; appropriate follow-up and referral to specialists for treatment and counseling. Routine newborn screening for CH and PKU is being established to ascertain the maximum coverage, using recommendations and guidelines from this pilot study.


Organizational culture and ethics work satisfaction.
Nurse executives have opportunities for creating constructive organizational cultures and for supporting ethical healthcare practice. Because culture and ethics are so central to effective nursing practice, this study explored relationships between them, measuring ethics as work satisfaction pertaining to ethics. This article provides initial assessment of one dimension of ethics, ethics work satisfaction, and examines its relationship to organizational culture.


Effects of elevated selenium and salinity concentrations in root zone on selenium and salt secretion in saltgrass (Distichlis spicata L.).
The effects of elevated selenium (Se) and salinity concentrations in the root zone on Se and salt secretion and accumulation were studied for an inland (Kesterson) and a coastal (Bodega Bay) saltgrass in sand culture and under greenhouse conditions. The results of this study indicate that the secretory mechanism of the saltgrass exhibits high ion specificity. Plants of both ecotypes were more efficient at Cl- and Na+ secretion than at SO4(2-) and Se secretion, suggesting that the saltgrass secretion mechanism is adapted primarily to saline environments high in NaCl. The saltgrass plants of the Kesterson ecotype secreted more Se than the Bodega Bay plants when treated with Se alone. However, the Bodega Bay plants secreted more Se than the Kesterson plants when the plants were treated with Se+NaCl. These differences reflect a ecotypical difference in which the Kesterson plants are more adaptive to the seleniferous soil, and the Bodega Bay plants are more adaptive to a coastal saline soil high in NaCl. Sulfate availability inhibited both Se accumulation and Se secretion in the plants of both ecotypes by approximately 98%. Ion secretion molar ratios of Na:Cl were calculated and the results suggest that Na+ secretion is dependent on the availability of Cl-. Selenium was taken up by plants with little discrimination, and thus it may be regarded as a master of chemical mimicry sharing similar physical and chemical properties with sulfur (S). Selenium and salt accumulation indices and secretion efficiency indices were calculated and found that the accumulation indices were higher for Se than for S, suggesting that Se uptake may be more passive and less regulated by active transport than S. Secretion efficiency was much higher for Na+ and Cl- than for Se and SO4(2-), but the efficiency indices between ecotypes were comparable, suggesting that the secretion mechanism in this species is designed mainly for adaptation of high NaCl concentrations. About 85% of the secreted Se was found in selenate form. This finding suggests that the mechanism of Se translocation and Se secretion in the saltgrass resembles the movement of selenate in xylem found in nonhalophytic plants. Less than 15% of organic Se was detected in the secretions and only trace amount of selenite was detected. Overall, the Se secretion mechanism in saltgrass does not seem to contribute substantial Se speciation in the plant soil system.


Additive and synergistic effects of a low-molecular-weight, heparin-like molecule and low doses of cyclosporin in preventing arterial graft rejection in rats.
Arteriosclerotic intimal proliferation is one of the main long-term complications of organ transplantation. Low-molecular-weight, heparin-like molecules prevent myointimal proliferation in arterial wall injury and limit rejection in skin allografts. Cyclosporin limits rejection but has no major effect on intimal proliferation. Therefore, an experimental protocol was designed to test whether heparin-like molecules interacted with low doses of cyclosporin to prevent arterial wall immune system injury and response in a model of arterial graft rejection in normotensive and hypertensive rats. Aortic allografts were performed in spontaneously hypertensive rats (SHRs) and Wistar-Kyoto (WKY) normotensive control rats. Four groups of 10 allografted (SHR and WKY) rats were used: one group was treated with placebo, one with low doses of cyclosporin (2 mg/kg body wt per day), one with low-molecular-weight, heparin-like molecule (1 mg/kg body wt per hour), and one with low doses of cyclosporin plus low-molecular-weight, heparin-like molecule. Ten SHRs and 10 WKYs were isografted and served as the control groups. All rats were killed 8 weeks after aortic grafting. Structural parameters of the grafted segment were measured by morphometric analysis on formalin-fixed sections with specific stains. The classical signs of immune system injury and response were present in the untreated allografts in SHRs and WKYs: inflammatory infiltration of the adventitia, medial injury, and intimal proliferative response. Low doses of cyclosporin had a significant beneficial effect on immune medial injury by increasing medial thickness and the number of remaining smooth muscle cells and decreasing the extracellular matrix injury. Cyclosporin had no protective effect on intimal proliferation.(ABSTRACT TRUNCATED AT 250 WORDS)


Developmental disorders in children born to women with sickle cell disease: A report from the Boston Birth Cohort.
Children exposed to maternal sickle cell disease (SCD) have many theoretical risks for developmental disorders, but little is known about long-term outcomes for these children. We used the Boston Birth Cohort to compare developmental outcomes between children exposed to maternal SCD and matched, unexposed controls. Children with exposure to maternal SCD had increased risk of attention deficit hyperactivity disorder (OR 5.12, 95% CI 1.36-19.19, p = 0.02) and obesity (OR 2.74, 95% CI 1.10-6.87, p = 0.03). In utero and/or environmental exposures may help explain these findings. Further studies of outcomes of children born to women with SCD are needed.


Properties of the narrative scoring scheme using narrative retells in young school-age children.
To evaluate the clinical utility of the narrative scoring scheme (NSS) as an index of narrative macrostructure for young school-age children. Oral retells of a wordless picture book were elicited from 129 typically developing children, ages 5-7. A series of correlations and hierarchical regression equations were completed using microstructural measures of vocabulary and grammar to predict NSS scores. While the NSS was significantly correlated with age and each of the microstructural measures, the hierarchical regression analyses revealed a unique relationship between vocabulary and narrative macrostructure. The NSS is an efficient and informative tool for documenting children's development of narrative macrostructure. The relationship between the NSS and microstructural measures demonstrates that it is a robust measure of children's overall oral narrative competence and a powerful tool for clinicians and researchers. The unique relationship between lexical diversity and the NSS confirmed that a special relationship exists between vocabulary and narrative organization skills in young school-age children.


Molecular and cellular phenotypic profiles of gastric noninvasive neoplasia.
According to the Padova international classification, 52 gastric noninvasive neoplasias (NIN) were classified as follows: 20 low-grade NIN (L-NIN); 9 high-grade NIN including suspicion for carcinoma without invasion (H-NIN); and 23 high-grade NIN including carcinoma without invasion (Ca-NIN). The molecular and cellular phenotypic profiles were investigated and compared. The APC gene was mutated in seven (35%) L-NIN, two (22%) H-NIN, and two (9%) Ca-NIN tumors; APC mutations were significantly more frequent in L-NIN compared with Ca-NIN tumors (p < 0.05). Mutations of the p53 gene were found in five (22%) Ca-NIN tumors but were not observed in L-NIN or H-NIN tumors (p < 0.05). Loss of heterozygosity involving at least one chromosomal locus was detected in 14 (61%) Ca-NIN tumors but was not detected in L-NIN or H-NIN tumors. High-frequency microsatellite instability (MSI-H) was detected in one (5%) L-NIN tumor and in six (26%) Ca-NIN tumors. The frequencies of loss of heterozygosity and MSI-H were significantly higher in Ca-NIN than in L-NIN or H-NIN tumors (p < 0.05). Nuclear accumulation of p53 protein was detected in no L-NIN tumors, 1 (11%) H-NIN tumor, and 10 (44%) Ca-NIN tumors (p < 0.01). All tumors with loss of hMLH1 expression exhibited MSI-H (p < 0.01). Cellular phenotypic analysis revealed that seven (35%) L-NIN tumors and one (4%) Ca-NIN tumor had complete-type intestinal metaplastic phenotype and that one (5%) L-NIN tumor and one (4%) Ca-NIN tumor had a gastric foveolar epithelial phenotype, whereas the remaining tumors exhibited an ordinary phenotype. Thus, the complete-type intestinal metaplastic phenotype was more characteristic of L-NIN tumors than of H-NIN or Ca-NIN tumors (p < 0.01). In summary, the Padova international classification correlated with both the molecular and cellular phenotypic profiles. In practice, p53 and hMLH1 immunohistochemistry discriminated Ca-NIN from L-NIN and H-NIN tumors.


Recycling the versatile Pipecolic linker.
The Pipecolic linker is a new highly versatile handle which immobilizes on solid support through a carboxylic acid function a wide range of amines, alcohols, and hydrazines. The anchoring step on pipecolic resin is very easy and efficient, and compounds are released with high purities upon acidic treatment. During this treatment, an oxazolonium intermediate is hydrolyzed, yielding the cleavage of ester or amide bond and the release of free carboxylic acid of the starting linker. In this study, we report the possibility of recycling the pipecolic resin after the use of several trifluoroacetic acid (TFA) cleavage cocktails. We demonstrate that it can be reused up to five times without significant loading decrease.


Differential diagnosis of monoclonal gammopathies.
Several disorders are associated with a monoclonal immunoglobulin detected by serum or urine electrophoresis, the most common being a monoclonal gammopathy of undetermined significance, multiple myeloma, Waldenstrom's macroglobulinemia, and amyloidosis. The clinical features of these conditions, as well as other similar entities, are described in this review. The objective is to demonstrate the importance of electrophoretic studies in the differential diagnosis of plasma cell dyscrasias and in guiding the decision for rational therapies.


Disparities in COVID-19-related stressful life events in the United States: Understanding who is most impacted.
Impacts of the COVID-19 pandemic in the United States have been exacerbated by pre-existing inequities in resources and opportunities, leaving the most vulnerable to face a multitude of hardships. The goal of the current study was to characterise COVID-19-related stressful life events in specific life domains and to identify the sociodemographic characteristics of individuals who are more likely to experience such events. Participants (n = 372, 57% female) in a follow-up study of the NICHD Study of Early Child Care and Youth Development completed the Epidemic-Pandemic Impacts Inventory (June-August 2020) to assess COVID-19-related stressors. Sociodemographic factors (gender, race/ethnicity, socioeconomic status and wealth) were examined simultaneously as predictors of the number of stressful life events in separate categories of work/finances, home life, social activity, health and healthcare, adjusted for covariates (household size, community COVID-19 transmission risk). In negative binomial regression analyses, being female (vs. male) predicted a 31%, 64%, 13% and 94% increase in the number of stressful life events in domains of work/finances, home life, social activity and healthcare, respectively, whereas each one standard deviation increase in wealth predicted a 17%, 16% and 21% reduction in the number of stressful life events in domains of work/finances, COVID-19 infection and healthcare, respectively. Findings highlight the pronounced and far-reaching impacts of the COVID-19 pandemic on women as well as the unique role wealth may play in lessening such impacts. This new knowledge may be leveraged to develop intervention and policy-related strategies to remediate impacts of COVID-19-related stressors on those most vulnerable.


Solution structure and dynamics of epidermal growth factor and transforming growth factor alpha.
Circular dichroism (CD) and Fourier transform infrared spectroscopic studies have shown that the secondary structure of transforming growth factor alpha (TGF-alpha) is very similar to that of epidermal growth factor (EGF). The infrared spectra revealed a minor difference between the two proteins, in particular in the beta-sheet structure. A large difference was observed with CD between the two proteins in the apparent conformation each adopts when the disulfide bonds are reduced. Reduced TGF-alpha showed a distinct alpha-helical conformation only at a high trifluoroethanol concentration, whereas reduced EGF assumed an alpha-helical conformation in the absence of trifluoroethanol. This indicates that these two proteins adopt different secondary structures in the absence of disulfide bonds, although they assume similar folding structures in their presence. These data suggest that the disulfide bonds to a large degree dictate the conformation of these two proteins. Additionally, differences in the dynamic behavior between EGF and TGF-alpha were also observed. Infrared experiments showed that the hydrogen-deuterium exchange rate is much higher for TGF-alpha than for EGF, indicating that TGF-alpha is a more flexible molecule. The rate of reduction of the disulfide bonds by dithiothreitol was also faster for TGF-alpha. Therefore, it can be concluded that although EGF and TGF-alpha have a similar overall conformation, TGF-alpha is a more flexible molecule than EGF.


Circulating microparticles, lupus anticoagulant and recurrent miscarriages.
Pregnancy is a pro-inflammatory/hypercoagulable state. Impairment of trophoblastic invasion and placental microthrombi are involved mechanisms in the pathogenesis of recurrent miscarriages (RM). Although in RM related to antiphospholipid antibodies (aPL) non-thrombotic mechanism seems to play an important role as well, we focused this review on the thrombotic mechanisms of RM related to aPL. Thus, in cases of RM related to aPL, placental injury produces inflammatory changes in endothelial cells. Endothelial dysfunction has been shown by increased plasma levels of ICAM-1/VCAM-1 and E-selectin. In RM associated with aPL, the thrombogenic mechanisms included different pathways: protein C inhibition, annexin-5 displacement, blocking of beta(2)GP1 anticoagulant activity and tissue factor upregulation. A new marker/causative agent of RM by itself or in relation to lupus anticoagulant (LA) has risen: circulating microparticles. Microparticles are a heterogeneous group of small, membrane-coated vesicles with a diameter of 0.1-1 microm. Microparticles are released from the cellular membrane during cell activation/apoptosis. Exposition of phospholipids in the outer membrane leaflet could explain the role of microparticles in the thrombotic events. Microparticles have been associated with RM. Microparticles are increased in women with RM when compared with healthy pregnant women. A relationship between aPL and activated endothelial cells (EC) occurs, as well as between EC and circulating microparticles. Interestingly, microparticles induced coagulation in vitro via tissue factor, especially in plasmas with LA. Finally, the relationship between EC, microparticles, LA and RM is revised.


Electronic structure of 3d[M(H2O)6](3+) ions from Sc(III) to Fe(III): a quantum mechanical study based on DFT computations and natural bond orbital analyses.
The metal-donor atom bonding along the series of 3d[M(H2O)6](3+) ions from Sc(3+) to Fe(3+) has been investigated by density-functional calculations combined with natural localized bond orbital analyses. The M-OH(2) bonds were considered as donor-acceptor bonds, and the contributions coming from the metal ion's 3d sigma-, 3d pi-, and 4s sigma-interactions were treated individually. In this way, the total amount of charge transferred from the water oxygen-donor atoms toward the appropriate metal orbitals could be analyzed in a straightforward manner. One result obtained along these lines is that the overall extent of ligand-to-metal charge transfer shows a strong correlation to the hydration enthalpies of the aqua metal ions. If the contributions to the total ligand-to-metal ion charge transfer are divided into sigma- and pi-contributions, it turns out that Cr(3+) is the best sigma-acceptor, but its pi-accepting abilities are the weakest along the series. Fe(3+) is found to be the best pi-acceptor among the 3d hexaaqua ions studied. Its aptitude to accept sigma-electron density is the second weakest along the series and only slightly higher than that of Sc(3+) (the least sigma-acceptor of all ions) because of the larger involvement of the Fe(3+) 4s orbital in sigma-bonding. The strengths of the three types of bonding interactions have been correlated with the electron affinities of the different metal orbitals. Deviations from the regular trends of electron affinities along the series were found for those [M(H2O)6](3+) ions that are subject to Jahn-Teller distortions. In these cases (d(1) = [Ti(H2O)6](3+), d(2) = [V(H2O)6](3+), and d(4) = [Mn(H2O)6](3+)), ligand-to-metal charge transfer is prevented to go into those metal orbitals that contain unpaired d electrons. A lowering of the complex symmetry is observed and coupled with the following variations: The Ti(3+)- and V(3+)-hexaaqua ions switch from T(h)() to C(i)() symmetry while the Mn(3+)-hexaaqua ion moves to D(2)(h)() symmetry. The loss of orbital overlap leading to a diminished ligand-to-metal charge transfer toward the single occupied metal orbitals is compensated by amplified bonding interactions of the ligand orbitals with the unoccupied metal orbitals to some extent.


Using VIPT-jump to distinguish between different folding mechanisms: application to BBL and a Trpzip.
Protein folding involves a large number of sequential molecular steps or conformational substates. Thus, experimental characterization of the underlying folding energy landscape for any given protein is difficult. Herein, we present a new method that can be used to determine the major characteristics of the folding energy landscape in question, e.g., to distinguish between activated and barrierless downhill folding scenarios. This method is based on the idea that the conformational relaxation kinetics of different folding mechanisms at a given final condition will show different dependences on the initial condition. We show, using both simulation and experiment, that it is possible to differentiate between disparate kinetic folding models by comparing temperature jump (T-jump) relaxation traces obtained with a fixed final temperature and varied initial temperatures, which effectively varies the initial potential (VIP) of the system of interest. We apply this method (hereafter refer to as VIPT-jump) to two model systems, tryptophan zipper (Trpzip)-2c and BBL, and our results show that BBL exhibits characteristics of barrierless downhill folding, whereas Trpzip-2c folding encounters a free energy barrier. In addition, using the T-jump data of BBL we are able to provide, via Langevin dynamics simulations, a realistic estimate of its conformational diffusion coefficient.


Comparisons and combinations in anti-anginal therapy.
The availability of three distinct classes of anti-anginal agents makes it possible, in theory, to choose a single drug to counteract the specific mechanism responsible for the syndrome in an individual patient, or to combine therapy where appropriate. In practice, other factors, such as concomitant disease, cost and side-effects, prove to have an over-riding influence on the choice of treatment which, in any case, often has to be decided upon on the basis of trial and error.


[Pharmaco-ethological analysis of agonistic behavior between resident and intruder mice: effects of adrenergic beta-blockers].
The present study was conducted to investigate the effects of adrenergic beta-blockers on agonistic behavior using quantitative ethological methods. In order to generate the agonistic behavior we employed the resident-intruder paradigm: An intruder male mouse is introduced into the home cage of a resident male mouse that has been cohabiting with a female for 5 weeks. The following drugs were administered orally to either resident or intruder mice: dl-Propranolol, oxprenolol, and carteolol. The injection-test interval was 60 min. Each test was recorded using a video TV monitor system, and at a later time several behavioral elements shown by both resident and intruder mice were measured. dl-Propranolol (5, 10, 20 mg/kg), oxprenolol (30, 50, 75 mg/kg), and carteolol (30, 50, 75 mg/kg) significantly suppressed the resident's aggressive episodes (offensive sideways posture, tail rattle, and attack bite) when resident mice were treated. By contrast, when intruder mice were treated with beta-blockers, aggressive episodes by untreated residents were not affected. The results suggest that dl-propranolol, oxprenolol, and carteolol have specific effects on the hostility of resident mice.


Introducing a combined Leishman-Giemsa stain as a new staining technique for avian blood smears.
The use of conventional stains including Giemsa, Wright and Leishman have become an essential tool for differential diagnosis of cells in peripheral blood. The aim of the study was to develop a new combination of Leishman-Giemsa (L&G) stain for avian blood smears and to compare its efficacy with conventional staining methods. Three sets of peripheral blood smears, one smear for L&G stain and two other smears for Leishman and Giemsa stains, created from 50 broiler chickens blood samples. All the three sets of slides were blind screened by two expert clinical pathologists and scored based on the staining characteristics (4 parameters) such as nuclear features of red blood cell (RBC) and white blood cell (WBC), cytoplasmic features and cytoplasmic granularity of WBC. The average grading score assigned by two experts for each staining method were compared. The average grading score of two conventional Leishman and Giemsa staining methods were significantly lower (P<0.001) than new L&G staining method in avian nuclear features of the RBC and WBC. The L&G stain gave a better clarity of nuclear features of avian RBC and WBC. The new L&G staining technique created significant differences (P<0.001) in cytoplasmic features of avian WBC compared to the other two methods. For the first time, the results of the present study showed that the avian blood cells are more desirable stained with a new combination of L&G stain. In addition, it gives a better nuclear and cytoplasmic differential staining than the conventional Giemsa and Leishman stains when used alone.


T Cell Membrane Heterogeneity Aids Antigen Recognition and T Cell Activation.
T cells are critical for co-ordinating the immune response. T cells are activated when their surface T cell receptors (TCRs) engage cognate antigens in the form of peptide-major histocompatibility complexes (pMHC) presented on the surface of antigen presenting cells (APCs). Large changes in the contact interface between T cells and APCs occur over the course of tens of minutes from the initial contact to the formation of a large-scale junction between the two cells. The mature junction between a T cell and APC is known as the immunological synapse, and this specialized plasma membrane structure is the major platform for TCR signaling. It has long been known that the complex organization of signaling molecules at the synapse is critical for appropriate activation of T cells, but within the last decade advances in microscopy have opened up investigation into the dynamics of T cell surface topology in the immune synapse. From mechanisms mediating the initial contact between T cells and APCs to roles in the organization of molecules in the mature synapse, these studies have made it increasingly clear that local membrane topology has a large impact on signaling processes. This review focuses on the functional consequences of the T cells' highly dynamic and heterogeneous membrane, in particular, how membrane topology leads to the reorganization of membrane proteins on the T cell surface.


The psychometric properties of a caregiving-related strain scale in oncology.
Caregivers are rarely assessed for caregiving-related strain. This study explored the psychometric properties of the caregiving-related physical, emotional, and financial strain questions in the National Alliance for Caregiving's (NAC) survey and a new total score (NAC-3). Comparisons were made to existing caregiver quality of life, distress, and burden instruments through an online, cross-sectional survey of U.S. adult cancer caregivers. Findings from 299 caregivers showed that the NAC-3 was moderately to strongly associated with each of the comparison instruments. Similar concepts were also shown to be more strongly correlated (except physical health construct) and dissimilar concepts items had weaker correlations. Findings suggest that the total score and individual items are psychometrically sound for use as compared to established caregiving-related instruments. The total score and items provide a concise option, potentially beneficial for clinical use, and can be compared to nationally representative samples of caregivers through NAC's surveys.


Rapid determination of pathogens in mastitic milk of dairy cows using Gram staining.
This study aimed to determine whether causative pathogens in mastitic milk can be determined by Gram staining after the centrifugation of milk. Gram staining was performed using unconcentrated and concentrated milk cells. Using this method, we found that the background of microscopic image of unconcentrated milk cells was complex and bacteria were difficult to detect. In contrast, the background of the smears in the concentrated milk cells was translucent, and bacterial and somatic cells were clearly visible. The sensitivity and specificity of the Gram staining of concentrated milk cells were 84.4% and 86.0% and 50.0% and 94.5% for the detection of gram-positive and gram-negative bacteria, respectively. The presented method provides a simple and inexpensive means of determining mastitis-causing pathogens.


The role of conserved charged residues in the bidirectional rotation of the bacterial flagellar motor.
Many bacteria rotate their flagella both counterclockwise (CCW) and clockwise (CW) to achieve swimming toward attractants or away from repellents. Highly conserved charged residues are important for that motility, which suggests that electrostatic interactions are crucial for the rotor-stator function. It remains unclear if those residues contribute equally to rotation in the CCW and CW directions. To address this uncertainty, in this study, we expressed chimeric rotors and stators from Vibrio alginolyticus and Escherichia coli in E. coli, and measured the rotational speed of each motor in both directions using a tethered-cell assay. In wild-type cells, the rotational speeds in both directions were equal, as demonstrated previously. Some charge-neutralizing residue replacements in the stator decreased the rotational speed in both directions to the same extent. However, mutations in two charged residues in the rotor decreased the rotational speed only in the CCW direction. Subsequent analysis and previous results suggest that these amino acid residues are involved in supporting the conformation of the rotor, which is important for proper torque generation in the CCW direction.


Tacit knowledge of caring and embodied selfhood.
The tacit knowledge paradigm is gaining recognition as an important source of knowledge that informs clinical decision-making. It is, however, limited by an exclusive focus on knowledge acquired through clinical practice, and a consequent neglect of the primordial and socio-cultural significance of embodied selfhood, precisely what provides the foundational structure of tacit knowledge of caring and facilitates its manifestation. Drawing on findings from a qualitative study of 43 dementia care practitioners in Ontario, Canada that utilised research-based drama and focus group methodology, we argue that embodied selfhood is fundamental to tacit knowledge of caring. Results are analysed drawing upon the theoretical precepts of embodied selfhood that are rooted in Merleau-Ponty's (1962) reconceptualisation of perception and Bourdieu's (1977, 1990) notion of habitus. We conclude with a call for further exploration of the body as a site of the production of tacit knowledge.


Clinical utility of serum macrophage migration inhibitory factor in men with prostate cancer as a novel biomarker of detection and disease progression.
Macrophage migration inhibitory factor (MIF) has been shown to play an important role in the growth and metastasis of prostate cancer. The objective of this study was to determine whether the serum level of MIF could be used as a diagnostic biomarker for prostate cancer as well as a predictor of disease progression. A total of 369 men who underwent systematic prostate biopsy from January 2000 to April 2004 and 30 healthy controls were included in this study. The serum MIF level was measured using an enzyme linked immunosorbent assay. Associations between the serum MIF level and several clinicopathological factors were analyzed. Among 359 patients, 137 were diagnosed as having prostate cancer. The mean values of serum MIF in the control, benign and cancer-groups were 2.1, 3.5 and 10.8 ng/ml, respectively. The MIF levels of patients with prostate cancer were significantly higher than those of the other two groups. A comparison of MIF and PSA values in all patients showed a positive correlation (correlation coefficient r(2)=0.56, p<0.0001). The MIF value in patients with prostate cancer was significantly associated with clinical T stage, Gleason score and percentage of positive biopsy core (PPBC). MIF levels in patients with metastasis were significantly elevated compared with those in patients without metastasis. Among patients undergoing radical prostatectomy, the level of MIF in those with pathologically confirmed extraprostatic disease was significantly higher than that in patients with organ-confined disease. Moreover, multivariate analysis showed that MIF values, PSA values and PPBC were independent predictors for extraprostatic disease. These findings suggest that an increased serum MIF level is closely associated with the progression of prostate cancer, and thus the serum MIF level could be useful as a novel biomarker for the detection of prostate cancer as well as a predictor of disease progression.


The Vocational Rehabilitation Index: a guide to accident victims' requirements for return-to-work assistance.
The Vocational Rehabilitation Index (VRI) is an easily administered, seven-item, ordinally scaled assessment, developed from analysis of variables associated with early return to work in a representative sample of 194 persons who received compensation for injuries at work or in road traffic accidents. The VRI discriminates between persons who return to work and those who do not. It can be used to identify, amongst non-returners, those whose return to work might be assisted by referral to rehabilitation. It may also indicate the kind of assistance that would be most helpful.


Microphase Separation Engineering toward 3D Porous Carbon Assembled from Nanosheets for Flexible All-Solid-State Supercapacitors.
Although hierarchitectures could energize carbon materials to address the challenges encountered in emerging flexible energy storage, how to make the trade-offs among specific surface area, pore configuration, and conductivity is still a lingering issue. Herein, 3D porous carbon assembled by nanosheets (HCAs) with tunable hierarchical porous structure is acquired from amphiphilic coal tar pitch and chitosan by means of a facile microphase separation strategy without any templates. The polar molecular chains of chitosan and the surrounding pitch molecules with strong π-π* bonds self-assemble respectively to form hierarchical pores and a network of nanosheets in a stepped pyrolysis process. Due to the combined effects of the meso-dominant porous structure, high specific surface area, and nitrogen-rich nature, the as-assembled symmetric all-solid-state supercapacitor with a wide voltage range of 0-1.8 V delivers a specific capacitance of 296 F g-1 at 0.2 A g-1 and an energy density of 27 Wh kg-1 at a power density of 450 W kg-1. The strategy of microphase separation is proposed originally to design and to fabricate carbon materials with multilevel nanoarchitectural trade-offs for high-performance supercapacitors.


Expression and functional significance of nicotinamide N-methyl transferase in skeletal muscles of patients with chronic obstructive pulmonary disease.
Nicotinamide N-methyl transferase (NNMT) is highly expressed in quadriceps muscles of patients with chronic obstructive pulmonary disease (COPD). However, its expression in the diaphragm of these patients has not been assessed. The functional significance of NNMT induction in skeletal muscles of patients with COPD is also unknown. (1) To compare NNMT expressions in the diaphragm and quadriceps muscles of patients with COPD. (2) To identify the influence of proinflammatory cytokines on NNMT expression. (3) To assess the influence of NNMT on indices of myogenesis (satellite cell migration and proliferation) and the defense against oxidative stress. Costal diaphragm muscle biopsies were acquired from 13 patients with moderate and severe COPD and 8 control subjects. Quadriceps muscle biopsies were obtained from 12 patients with COPD and 14 control subjects. NNMT expressions were significantly elevated in the diaphragm and quadriceps muscles of patients with COPD; however, the relative induction of NNMT expression was greater in the quadriceps muscle (10-fold) than it was in the diaphragm (2-fold). NNMT expressions correlated negatively with the severity of COPD and limb muscle wasting. In skeletal myoblasts, NNMT expression was significantly induced by IL-6, transforming growth factor beta, and tumor necrosis factor-alpha. Overexpression of NNMT in myoblasts triggered a significant increase in proliferation and migration, but had no influence on cell death. Carbonyl formation, induced by exposing myoblasts to H(2)O(2), was significantly attenuated when NNMT was overexpressed. Up-regulation of NNMT expression in the skeletal muscles of patients with COPD may represent an adaptive response designed to improve myogenesis and defend against oxidative stress.


Maternal exposure to benzo[a]pyrene alters development of T lymphocytes in offspring.
Childhood cancer has been increasing significantly over the past two decades in the United States, suggesting that environmental exposures may be playing a causative role. One such cause may be maternal smoking during pregnancy. Suspected carcinogens in cigarette smoke and environmental pollution include N-nitrosamines and polycyclic aromatic hydrocarbons, which may be several micrograms per exposure. Previously, we have shown that mouse progeny of mothers exposed to benzo[a]pyrene (B[a]P) during midpregnancy had abnormalities in their humoral and cell-mediated immune response. Immunodeficiency was detectable during gestation, at one week after birth and persisted for 18 months. Tumor incidences in progeny were eight to 10-fold higher than in controls. The present study compared frequencies of CD4+, CD8+, V gamma 2+, and V beta 8+ T cells in progeny following in utero exposure to B[a]P. The significant reduction in newborn CD4+CD8+, CD4+CD8+V beta 8+ thymocytes and CD4+ splenocytes from 1-week-old progeny, suggests that B[a]P induces abnormal changes in developing T cells. These early alterations may lead to postnatal T cell suppression, thus providing a more suitable environment for the growth of tumors later in life. These results suggest that developmental immunosuppression mediated by B[a]P may play a critical role in the relationship between maternal exposures and childhood carcinogenesis.


Effect of thidiazuron, a cytokinin-active urea derivative, in cytokinin-dependent ethylene production systems.
Cytokinins are known to stimulate ethylene production in mungbean hypocotyls synergistically with indoleacetic acid (IAA), in mungbean hypocotyls synergistically with Ca(2+), and in wilted wheat leaves. Thidiazuron, a substituted urea compound, mimicked the effect of benzyladenine (BA) in all three systems. In the Ca(2+) + cytokinin system and the IAA + cytokinin systems of mungbean hypocotyls, thiadiazuron was slightly more active than BA at equimolar concentration. In mungbean hypocotyls exogenously applied IAA was rapidly conjugated into IAA asparate, and this conjugation process was effectively inhibited by thidiazuron, as by cytokinins. In the wilted wheat leaves system, 10 micromolar thidiazuron exerted stress ethylene production equal to that exerted by 1 millimolar BA, indicating that thidiazuron is more active than BA by two orders. The structure-activity relationship of thidiazuron and its thiadiazolylurea analogs in stimulating Ca(2+)-dependent ethylene production in mungbean hypocotyls was found to agree well with the structure-activity relationship of these derivatives in promoting the growth of callus tissues. These results indicate that thidiazuron and its derivatives are highly active to mimic the adenine-type cytokinin responses in promoting ethylene production and that the structure-activity relationship in promoting the growth of callus and in promoting ethylene production is similar.


Outcome of Ahmed valve implantation when preoperative IOP less than 21 mm Hg.
To examine the results of Ahmed glaucoma drainage device (GDD) implantation in patients with a preoperative intraocular pressure (IOP) of 20 mm Hg or less. Noncomparative retrospective case series. Sixty-six surgeries in 63 patients were analyzed. Mean follow-up was 51 months. The mean drop in IOP was 3.8 mm Hg (23.3%, P<0.0001) at 12 months and 3.9 mm Hg (24.0%, P<0.0001) at final follow-up. The number of glaucoma medications used postoperatively was significantly less than preoperatively. Although the mean visual acuity was poorer postoperatively this did not reach statistical significance. Surgical success was defined as IOP > or =5 mm Hg and 20% lower than preoperatively with or without hypotensive therapy, visual acuity perception of light or better and no further glaucoma surgery. About 57.6% and 53.0% of patients were considered a success at 12 months and final follow-up, respectively. Fourteen patients (21.2%) underwent additional glaucoma surgery, 8 of which had a second Ahmed GDD inserted whereas 6 had cyclodestruction. All failures were due to inadequate IOP control or further glaucoma surgery, with no cases considered failures due to vision loss or hypotony. Perioperative complications developed in 6 cases (9.1%) whereas long-term problems related to surgery occurred in 8 cases (12.1%). In patients with IOPs of 20 mm Hg or less who require IOPs in the low teens and below Ahmed GDD surgery does seem to be an effective option. The success rates of surgery, however, must be balanced against the risk of complications. In addition, the relative merits of Ahmed GDD implantation versus mitomycin trabeculectomy and/or nonvalved GDDs requires further investigation.


Estimating Unreported COVID-19 Cases with a Time-Varying SIR Regression Model.
Potential unreported infection might impair and mislead policymaking for COVID-19, and the contemporary spread of COVID-19 varies in different counties of the United States. It is necessary to estimate the cases that might be underestimated based on county-level data, to take better countermeasures against COVID-19. We suggested taking time-varying Susceptible-Infected-Recovered (SIR) models with unreported infection rates (UIR) to estimate factual COVID-19 cases in the United States. Both the SIR model integrated with unreported infection rates (SIRu) of fixed-time effect and SIRu with time-varying parameters (tvSIRu) were applied to estimate and compare the values of transmission rate (TR), UIR, and infection fatality rate (IFR) based on US county-level COVID-19 data. Based on the US county-level COVID-19 data from 22 January (T1) to 20 August (T212) in 2020, SIRu was first tested and verified by Ordinary Least Squares (OLS) regression. Further regression of SIRu at the county-level showed that the average values of TR, UIR, and IFR were 0.034%, 19.5%, and 0.51% respectively. The ranges of TR, UIR, and IFR for all states ranged from 0.007-0.157 (mean = 0.048), 7.31-185.6 (mean = 38.89), and 0.04-2.22% (mean = 0.22%). Among the time-varying TR equations, the power function showed better fitness, which indicated a decline in TR decreasing from 227.58 (T1) to 0.022 (T212). The general equation of tvSIRu showed that both the UIR and IFR were gradually increasing, wherein, the estimated value of UIR was 9.1 (95%CI 5.7-14.0) and IFR was 0.70% (95%CI 0.52-0.95%) at T212. Despite the declining trend in TR and IFR, the UIR of COVID-19 in the United States is still on the rise, which, it was assumed would decrease with sufficient tests or improved countersues. The US medical system might be largely affected by severe cases amidst a rapid spread of COVID-19.


Two insulin-responsive glucose transporter isoforms and the insulin receptor are developmentally expressed in rabbit preimplantation embryos.
Glucose is the most important energy substrate for mammalian blastocysts. Its uptake is mediated by glucose transporters (GLUT). In muscle and adipocyte cells insulin stimulates glucose uptake by activation of the insulin receptor (IR) pathway and translocation of GLUT4. GLUT4 is expressed in bovine preimplantation embryos. A new insulin-responsive isoform, GLUT8, was recently described in mouse blastocysts. Thus, potentially, two insulin-responsive isoforms are expressed in early embryos. The mechanism of insulin action on embryonic cells, however, is still not clear. In the present study expression of IR, GLUT1, 2, 3, 4, 5 and 8 was studied in rabbit preimplantation embryos using RT-PCR, Western blotting and immunohistochemistry. The rabbit mRNA sequences for the complete coding region of IR, GLUT4 and a partial GLUT8 sequence were determined by RACE-PCR and sequencing. GLUT4 was expressed in 3-day-old morulae and in 4- and 6-day-old blastocysts. IR and GLUT8 transcripts were detectable only in blastocysts. Blastocysts also expressed GLUT1 and 3, but not GLUT2 and 5. Transcript numbers of GLUT4 and 8 were higher in trophoblast than in embryoblast cells. Translation of IR, GLUT4 and 8 proteins in blastocysts was confirmed by Western blotting. GLUT4 was localized mainly in the membrane and in the perinuclear region in trophoblast cells while in embryoblast cells its localization was predominantly in the perinuclear cytoplasm. The possible function(s) of two insulin-responsive isoforms, GLUT4 and GLUT8, in rabbit preimplantation embryos needs further investigation. It may not necessarily be linked to insulin-stimulated glucose transport.


Fauna and Larval Habitat Characteristics of Mosquitoes (Diptera: Culicidae) in Kashan County, Central Iran, 2019.
Mosquitoes are responsible for spreading devastating parasites and pathogens causing some important infectious diseases. The present study was done to better understand and update the fauna of Culicidae and to find out the distribution and the type of their larval habitats in Kashan County. This study was done in four districts of Kashan County (Central, Qamasr, Niasar and Barzok). Mosquito larvae were collected from 23 active larval habitats using a standard 350ml capacity mosquito dipper from April to late December 2019. The collected larvae were transferred to containers containing lactophenol, and after two weeks individually mounted in Berlese's fluid on a microscope slide and identified to species by morphological characters and valid keys. In this study, a total of 9789 larvae were collected from urban and rural areas in Kashan County. The identified genera were Anopheles, Culiseta and Culex. In this study larvae of An. turkhudi, Cx. perexiguus, Cx. mimeticus, Cx. deserticola and Cs. subochrea were collected for the first time from Kashan County. The results of this study indicate the presence and activity of different mosquito species in Kashan County that some of them are vectors of arbovirus and other vector-borne diseases.


Flavanone Glycoside Biosynthesis in Citrus: Chalcone Synthase, UDP-Glucose:Flavanone-7-O-Glucosyl-Transferase and -Rhamnosyl-Transferase Activities in Cell-Free Extracts.
Previous indirect evidence suggested that the biosynthesis of flavonoids in Citrus may not proceed via the usual chalcone synthase reaction and that glycosylation occurs during chalcone formation and not afterward, as has been reported in other species. We detected chalcone-synthase and UDP-glucose:flavanone-7-O-glucosyl-transferase activities in cell-free extracts of Citrus. The glucosylated flavanone was further rhamnosylated when exogenous UDP-glucose and NADPH were added to the extract. Chalcone-synthase activity was detected in cell-free extracts derived from young leaves and fruits. Young fruits (2 millimeter diameter) had the highest chalcone synthase activity. UDP-glucose:flavanone-7-O-glucosyl-transferase activity was measured in cell-free extracts derived from young leaves and fruits of Citrus mitis and Citrus maxima. The highest UDP-glucose:flavanone-7-O-glucosyl-transferase activity was found in young C. maxima leaves. These data indicate that Citrus contains a flavonoid pathway similar to that studied in other species.


α-Acetolactate decarboxylase immobilized in chitosan: A highly stable biocatalyst to prevent off-flavor in beer.
In this work, we studied the covalent immobilization of α-acetolactate decarboxylase (ALDC) on glutaraldehyde activated chitosan beads for application in beers to prevent off-flavor. The immobilized preparation was characterized regarding yield, efficiency, activity recovery, thermal and pH stabilities, and operational stability in beer. The concentration of activating agent was evaluated and at the best condition an immobilization yield of 84% and activity recovery of 9.1% were verified. Optimal pH was 6.0 for free and immobilized enzymes, however, the immobilized enzyme showed higher relative activity at the tested pH, with emphasis on the pH range from 5.0 to 6.5. Besides, immobilized enzyme showed stabilization factor of almost sevenfold at 60°C compared to free form. The immobilized biocatalyst was tested in repeated batches in beer for α-acetolactate conversion to acetoin, and after 12 cycles the enzyme presented 80% of its initial activity. To date, this is the most stable immobilized preparation of ALDC reported.


Decision aids to facilitate decision making around behavior change in the field of health promotion: A scoping review.
To broadly synthesize literature regarding decision aids (DAs) supporting decision making about diet, physical activity, sleeping and substance use a scoping review was performed. Multiple sources were used: (1) Scientific literature searches, (2) excluded references from a Cochrane review regarding DAs for treatments and screenings, and (3) results from additional searches. Interventions had to (1) support informed decision making and (2) provide information and help to choose between at least two options. Two researchers screened titles and abstracts. Relevant information was extracted descriptively. Thirty-five scientific articles and four DAs (grey literature) were included. Results were heterogeneous. Twenty-nine (94%) studies described substance use DAs. All DAs offered information and value and/or preference clarification. Many other elements were included (e.g., goal-setting). DA's effects were mixed. Few studies used standardized measures, e.g., decisional conflict (n = 4, 13%). Some positive behavioral effects were reported: e.g., smoking abstinence (n = 1). This research shows only some positive behavioral effects of DAs. However, studies reported heterogeneous results/outcomes, impeding knowledge synthesis. Areas of improvement were identified, e.g., establishing which intervention elements are effective regarding health behavior decision making. DAs can potentially be beneficial in supporting people to change health behaviors - especially regarding smoking.


Serum levels of TNF-alpha, sIL-2R, IL-6, and IL-8 are increased and associated with elevated lipid peroxidation in patients with Behçet's disease.
Behçet's disease (BD) is asystemic immunoinflammatory disorder and the aetiopathogenesis is to be specified. Cytokines play a role in immune response and in many inflammatory diseases. The aim of this case-control study is to investigate serum pro-inflammatory cytokine tumour necrosis factor (TNF)-alpha, interleukin-1beta (IL-1beta), soluble IL-2 receptor (sIL-2R), IL-6, and chemokine IL-8 levels in patients with BD. We also determined the end product of lipid peroxidation (malondialdehyde (MDA)) in BD patients as an index for oxidative stress. A total of 37 patients (19 men, 18 women) with BD (active, n = 17; inactive, n = 20) and 20 age-matched and sex-matched healthy control subjects (11 men, nine women) included in this cross-sectional, blinded study. Serum TNF-alpha, IL-1beta, sIL-2R, IL-6 and IL-8 levels were determined by a spectrophotometer technique using the immulite chemiluminescent immunometric assay. Lipid peroxidation was evaluated by Wasowicz et aL The levels of cytokines and lipid peroxidation in the active period were compared with the inactive period of the disease. Results are expressed as mean +/- standard error. IL-1beta levels were below the detection limits of the assay (< 5 pg/ml) in all samples. Mean levels of MDA (8.1+/-0.7 micromol/l), sIL-2R (800+/-38 U/ml), IL-6 (12.6+/-1.1 pg/ml), IL-8 (7.2+/-0.4 pg/ml), and TNF-alpha (7.9+/-0.5 pg/ml) in active BD patients were significantly higher than those in inactive patients (4.3+/-0.5 micromol/l, p < 0.01; 447+/-16 U/ml, p < 0.001; 8.3+/-0.6 pg/ml, p = 0.006; 5.3+/-0.1 pg/ml, p < 0.001; and 5.1 0.2 pg/ml, p < 0.001; respectively) or control subjects (2.1+/-0.2 micromol/l, p < 0.001; 446+/-20 U/ml, p < 0.001; 6.4+/-0.2 pg/ml, p < 0.001; 5.4+/-0.1 pg/ml, p < 0.001; and 4.7+/-0.1 pg/ml, p < 0.001, respectively). On the contrary, only the mean IL-6 level was significantly different between inactive BD and control subjects (p = 0.02). All acute phase reactants were significantly higher in active BD than in inactive period (for each, p < 0.01). High levels of sIL-2R, IL-6, IL-8 and TNF-alpha indicate the activation of immune system in BD. Serum sIL-2R, IL-6, IL-8 and TNF-alpha seem to be related to disease activity. Increased lipid peroxidation suggests oxidative stress in BD and therefore tissue damage in such patients. Amelioration of clinical manifestations would be envisaged by targeting these cytokines, chemokines and lipid peroxidation with pharmacological agents.


Genotype combination contributes to psoriasis: An exhaustive algorithm perspective.
Researchers have learned that nearly all conditions and diseases have a genetic component. With the benefit of technological advances, many single-nucleotide polymorphisms (SNPs) have been found to be associated with the risk of complex disorders by using genome wide association studies (GWASs). Disease-associated SNPs are sometimes shared by healthy controls and cannot clearly distinguish affected individuals from unaffected ones. The combined effects of multiple independent SNPs contribute to the disease process, but revealing the relationship between genotype and phenotype based on the combinations remains a great challenge. In this study, by considering the disease prevalence rate, we conducted an exhaustive process to identify whether a genotype combination pattern would have a decisive effect on complex disorders. Based on genotype data for 68 reported SNPs in 8,372 psoriasis patients and 8,510 healthy controls, we found that putative causal genotype combination patterns (CGCPs) were only present in psoriasis patients, not in healthy subjects. These results suggested that psoriasis might be contributed by combined genotypes, complementing the traditional modest susceptibility of a single variant in a single gene for a complex disease. This work is the first systematic study to analyze genotype combinations based on the reported susceptibility genes, considering each individual among the cases and controls from the Chinese population, and could potentially advance disease-gene mapping and precision medicine due to the causality relationship between the candidate CGCPs and complex diseases.


Roles of N-terminal region residues Lys11, Arg13, and Arg24 of antithrombin in heparin recognition and in promotion and stabilization of the heparin-induced conformational change.
The N-terminal region residues, Lys11, Arg13, and Arg24, of the plasma coagulation inhibitor, antithrombin, have been implicated in binding of the anticoagulant polysaccharide, heparin, from the identification of natural mutants with impaired heparin binding or by the X-ray structure of a complex of the inhibitor with a high-affinity heparin pentasaccharide. Mutations of Lys11 or Arg24 to Ala in this work each reduced the affinity for the pentasaccharide approximately 40-fold, whereas mutation of Arg13 to Ala led to a decrease of only approximately 7-fold. All three substitutions resulted in the loss of one ionic interaction with the pentasaccharide and those of Lys11 or Arg24 also in 3-5-fold losses in affinity of nonionic interactions. Only the mutation of Lys11 affected the initial, weak interaction step of pentasaccharide binding, decreasing the affinity of this step approximately 2-fold. The mutations of Lys11 and Arg13 moderately, 2-7-fold, altered both rate constants of the second, conformational change step, whereas the substitution of Arg24 appreciably, approximately 25-fold, reduced the reverse rate constant of this step. The N-terminal region of antithrombin is thus critical for high-affinity heparin binding, Lys11 and Arg24 being responsible for maintaining appreciable and comparable binding energy, whereas Arg13 is less important. Lys11 is the only one of the three residues that is involved in the initial recognition step, whereas all three residues participate in the conformational change step. Lys11 and Arg13 presumably bind directly to the heparin pentasaccharide by ionic, and in the case of Lys11, also nonionic interactions. However, the role of Arg24 most likely is indirect, to stabilize the heparin-induced P-helix by interacting intramolecularly with Glu113 and Asp117, thereby positioning the crucial Lys114 residue for optimal ionic and nonionic interactions with the pentasaccharide. Together, these findings show that N-terminal residues of antithrombin make markedly different contributions to the energetics and dynamics of binding of the pentasaccharide ligand to the native and activated conformational states of the inhibitor that could not have been predicted from the X-ray structure.


Discovery and current research status of feline morbillivirus.
Feline morbillivirus (FeMV) is an emerging virus that was first discovered in Hong Kong in 2012. FeMV is epidemiologically associated with kidney and other lower urinary tract diseases in cats. Phylogenetic analysis of its genome sequence indicates that FeMV is the most closely related to the members of genus morbillivirus, although FeMV is relatively distant in the phylogenetic analysis, and its target tissues and pathogenicity are different from the other members of the genus. The origin and routes of dissemination of FeMV are not clear since genetic types are not always correlated to the geographical distribution of the isolates. Since the discovery of the virus, several reports showed the epidemiological association of FeMV infection with kidney and lower urinary tract diseases in cats. However, the pathogenicity of FeMV is not clear yet due to paucity of the isolated virus strains and chronic nature of the subjected diseases. Diagnosis of FeMV infection has been performed using both nucleic acid and serological methods. However, there are no standard diagnostic methods to detect antibodies against FeMV, which will be useful to study epidemiology and pathogenicity of FeMV. Besides FeMV is an interesting subject as an additional member to the morbilliviruses possessing unusual characteristics comparing to the other morbilliviruses, further studies of FeMV is important in the veterinary field since it may lead to new therapies or prevention of chronic kidney diseases of cats.


Epac-mediated Ca(2+) mobilization and exocytosis in inner medullary collecting duct.
PKA has traditionally been thought as the binding protein of cAMP for mediating arginine vasopressin (AVP)-regulated osmotic water permeability in kidney collecting duct. It is now known that cAMP also exerts its effects via Epac (exchange protein directly activated by cAMP) and that intracellular Ca(2+) mobilization is necessary for AVP-induced apical exocytosis in inner medullary collecting duct (IMCD). The role of Epac as an effector of cAMP action in addition to PKA was investigated using confocal fluorescence microscopy in perfused IMCD. PKA inhibitors (1 microM H-89 or 10 microM KT-5720) at concentrations known to inhibit aquaporin-2 (AQP2) phosphorylation did not prevent AVP-induced Ca(2+) mobilization and oscillations. Epac-selective cAMP agonist (8-pCPT-2'-O-Me-cAMP) mimicked AVP in triggering Ca(2+) mobilization and oscillations, which was blocked by ryanodine but not by Rp-cAMP (a competitive antagonist of cAMP binding to PKA). 8-pCPT-2'-O-Me-cAMP also triggered apical exocytosis in the presence of a PKA inhibitor. Immunolocalization of AQP2 in perfused IMCD demonstrated that 8-pCPT-2'-O-Me-cAMP induces apical targeting of AQP2 and that AQP2 is abundant in junctional regions of basolateral membrane. Immunofluorescence study also confirmed the presence of Epac (isoform I) in IMCD. These results indicate that activation of Epac by an exogenous cAMP analog triggers intracellular Ca(2+) mobilization and apical exocytotic insertion of AQP2 in IMCD.


The effect of L-ornithine hydrochloride ingestion on performance during incremental exhaustive ergometer bicycle exercise and ammonia metabolism during and after exercise.
L-Ornithine has an important role in ammonia metabolism via the urea cycle. This study aimed to examine the effect of L-ornithine hydrochloride ingestion on performance during incremental exhaustive ergometer bicycle exercise and ammonia metabolism during and after exercise. In all, 14 healthy young adults (age: 22.2±1.0 years, height: 173.5±4.6 cm, body mass: 72.5±12.5 kg) who trained regularly conducted incremental exhaustive ergometer bicycle exercises after -ornithine hydrochloride supplementation (0.1 g/kg, body mass) and placebo conditions with a cross-over design. The exercise time (sec) of the incremental ergometer exercise, exercise intensity at exhaustion (watt), maximal oxygen uptake (ml per kg per min), maximal heart rate (beats per min) and the following serum parameters were measured before ingestion, 1 h after ingestion, just after exhaustion and 15 min after exhaustion: ornithine, ammonia, urea, lactic acid and glutamate. All indices on maximal aerobic capacity showed insignificant differences between both the conditions. Plasma ammonia concentrations just after exhaustion and at 15 min after exhaustion were significantly more with ornithine ingestion than with placebo. Plasma glutamate concentrations were significantly higher after exhaustion with ornithine ingestion than with placebo. It was suggested that, although the ingestion of L-ornithine hydrochloride before the exercise cannot be expected to improve performance, it does increase the ability to buffer ammonia, both during and after exercise.