Molecular and crystal structures of chitosan/HI type I salt determined by X-ray fiber diffraction.
The three-dimensional structure of chitosan/HI type I salt was determined by the X-ray fiber diffraction technique and linked-atom least-squares refinement method. Two polymer chains and four iodide ions (I(-)) crystallized in a monoclinic unit cell with dimensions a = 9.46(2), b = 9.79(2)], c (fiber axis)=10.33(2)A, beta = 105.2(2) degrees and a space group P2(1). Chitosan chains adopted an extended twofold helical conformation that was stabilized by O-3...O-5 hydrogen bonds, and the O-6 atom adopted nearly gt orientation. Polymer chains zigzag along the b-axis and directly connect to each other by N-2...O-6 hydrogen bonds. Two columns of iodide ions were shown to pack at the bending points of the zigzag sheets, and their locations are closely related to those of water columns in the hydrated chitosan. The iodide ions stabilized the salt structure by forming hydrogen bonds with the N-2 and O-6 atoms of the polymer chains together with an electrostatic interaction between N-2 and the iodide ions.


Understanding childhood adversity in West Sulawesi, Indonesia.
Despite research indicating the long-term impact of adverse childhood experiences (ACE), few studies identify cultural variations in perceptions of ACE in low-resource settings. This study explores culturally-rooted notions of ACE and sources of vulnerability in two culturally distinct districts in West Sulawesi, Indonesia. Data from 50 stakeholders were collected from four focus group discussions and nine semi-structured key informant interviews in Mamasa and Mamuju districts in West Sulawesi. All interviews were conducted in Bahasa Indonesia, recorded, transcribed verbatim, and translated into English. Constant comparative analysis was used to identify key themes. Primary ACE were violence, abandonment due to parents migrating for work, and malnourishment. While individual child characteristics appeared to play a minimal role in vulnerability to ACE, factors at the community and familial levels such as widespread poverty and low levels of parental education led to early transitions to adulthood through child marriage and employment. Cultural norms, particularly adherence to customary law, impacted both vulnerability and responses to violence against children. ACE interventions should expand beyond individual and family-level interventions to address these structural and cultural barriers to resilience.


Mitochondrial abnormalities in choroid plexus of Leigh disease.
Morphological study of the choroid plexuses in three patients with Leigh disease revealed a marked increase in the number of mitochondria in almost all of the choroidal epithelial cells. This finding is considered the morphological expression of a biochemical defect in the mitochondrial metabolism underlying Leigh disease, and the probable explanation for increased CSF levels of lactate and pyruvate in this disease.


Noninvasive Monitoring of the Response of Human Lungs to Low-Dose Lipopolysaccharide Inhalation Challenge Using MRI: A Feasibility Study.
Development of antiinflammatory drugs for lung diseases demands novel methods for noninvasive assessment of inflammatory processes in the lung. To investigate the feasibility of hyperpolarized 129 Xe MRI, 1 H T1 time mapping, and dynamic contrast-enhanced (DCE) perfusion MRI for monitoring the response of human lungs to low-dose inhaled lipopolysaccharide (LPS) challenge compared to inflammatory cell counts from induced-sputum analysis. Prospective feasibility study. Ten healthy volunteers underwent MRI before and 6 hours after inhaled LPS challenge with subsequent induced-sputum collection. 1.5T/hyperpolarized 129 Xe MRI: Interleaved multiecho imaging of dissolved and gas phase, ventilation imaging, dissolved-phase spectroscopy, and chemical shift saturation recovery spectroscopy. 1 H MRI: Inversion recovery fast low-angle shot imaging for T1 mapping, time-resolved angiography with stochastic trajectories for DCE MRI. Dissolved-phase ratios of 129 Xe in red blood cells (RBC), tissue/plasma (TP) and gas phase (GP), ventilation defect percentage, septal wall thickness, surface-to-volume ratio, capillary transit time, lineshape parameters in dissolved-phase spectroscopy, 1 H T1 time, blood volume, flow, and mean transit time were determined and compared to cell counts. Wilcoxon signed-rank test, Pearson correlation. The percentage of neutrophils in sputum was markedly increased after LPS inhalation compared to baseline, P = 0.002. The group median RBC-TP ratio was significantly reduced from 0.40 to 0.31, P = 0.004, and 1 H T1 was significantly elevated from 1157.6 msec to 1187.8 msec after LPS challenge, P = 0.027. DCE MRI exhibited no significant changes in blood volume, P = 0.64, flow, P = 0.17, and mean transit time, P = 0.11. Hyperpolarized 129 Xe dissolved-phase MRI and 1 H T1 mapping may provide biomarkers for noninvasive assessment of the response of human lungs to LPS inhalation. By its specificity to the alveolar region, hyperpolarized 129 Xe MRI together with 1 H T1 mapping adds value to sputum analysis. 1 Technical Efficacy Stage: 2 J. Magn. Reson. Imaging 2020;51:1669-1676.


The thiol-disulfide oxidoreductase system in the cold-adapted bacterium Pseudoalteromonas haloplanktis TAC 125: discovery of a novel disulfide oxidoreductase enzyme.
In prokaryotes, protein disulfide bond oxidation, reduction and isomerization are catalyzed by members of the thioredoxin superfamily, characterized by the conserved C-X-X-C motif in their active site. Thioredoxins and glutaredoxins contribute to the reducing power in the cytoplasm, while the Dsb system catalyzes disulfide bonds formation in the periplasmic space. This paper addresses the question of disulfide bonds formation in a cold-adapted micro-organism, Pseudoalteromonas haloplanktis TAC 125 (PhTAC125) by characterizing the DsbA system. We found distinctive features respect mesophilic counterparts that highlighted for the first time the occurrence of two adjacent chromosomal DsbA genes organised in a functional operon. The sophisticated transcriptional regulation mechanism that controls the expression of these two genes was also defined. The two DsbA proteins, named PhDsbA and PhDsbA2, respectively, were expressed in Escherichia coli and characterized. Results reported in this paper provide some insights into disulfide bonds formation in a micro organism isolated in the Antarctic sea water.


Ultrasound guidance for medical thoracoscopy: a novel approach.
Commonly, a pneumothorax is induced before medical thoracoscopy to facilitate safe entry into the pleural space. Evaluate the use of transthoracic ultrasound to locate a safe entry site for trocar placement during medical thoracoscopy without induction of a preprocedure pneumothorax. The study was designed as a prospective cohort study, performed in the setting of a tertiary care hospital with an active interventional pulmonology program. It included 20 consecutive patients referred for medical thoracoscopy. Ultrasound identified entry sites in all 20 patients. All sites were successfully used, despite the presence of adhesions in 3 patients. There were no complications. Ultrasound could safely and reliably identify entry sites for trocar placement during medical thoracoscopy, even in patients with pleural adhesions. The use of ultrasound may replace the practice of pneumothorax induction before medical thoracoscopy.


Prevalence of intubation rescue by air medical personnel during transfers from rural emergency departments.
Non-emergency-trained providers in rural emergency departments (ED) often lack the skills required for emergency resuscitations and rely on air medical transport teams to provide the initial airway stabilization of these patients. In this study, we determined the prevalence with which endotracheal intubations are required of air medical personnel upon arrival to rural EDs including intubations that were first attempted by the local provider. A retrospective database review was conducted of all air medical transfers from rural hospitals for a 28-month period. Those patients requiring an airway were categorized according to which provider initiated the intubation procedure. The prevalence of intubations performed by air medical and local providers was recorded as the percent of the total number of intubations. There were a total of 217 patients from 11 rural EDs requiring airway support. Air medical personnel were responsible for 85% of the intubations. Alternative airway support was necessary in 5% of the patients after unsuccessful intubation attempts. The failed intubations tended to be slightly older and female. Our study suggests that the vast majority of the intubations for patients requiring a helicopter evacuation from these rural settings are performed by the air medical personnel.


COVID-19 Detection from Chest X-rays Using Trained Output Based Transfer Learning Approach.
The recent Coronavirus disease (COVID-19), which started in 2019, has spread across the globe and become a global pandemic. The efficient and effective COVID-19 detection using chest X-rays helps in early detection and curtailing the spread of the disease. In this paper, we propose a novel Trained Output-based Transfer Learning (TOTL) approach for COVID-19 detection from chest X-rays. We start by preprocessing the Chest X-rays of the patients with techniques like denoising, contrasting, segmentation. These processed images are then fed to several pre-trained transfer learning models like InceptionV3, InceptionResNetV2, Xception, MobileNet, ResNet50, ResNet50V2, VGG16, and VGG19. We fine-tune these models on the processed chest X-rays. Then we further train the outputs of these models using a deep neural network architecture to achieve enhanced performance and aggregate the capabilities of each of them. The proposed model has been tested on four recent COVID-19 chest X-rays datasets by computing several popular evaluation metrics. The performance of our model has also been compared with various deep transfer learning models and several contemporary COVID-19 detection methods. The obtained results demonstrate the efficiency and efficacy of our proposed model.


Comparison of hearing outcomes after treatment for early-stage external auditory canal cancer.
The purpose of this study was to compare the hearing outcomes after surgery with reconstruction of the external auditory canal in combination with tympanoplasty, radiotherapy (RT) alone, and surgery followed by postoperative RT for T1 to 2N0M0 external auditory canal cancer. We performed a retrospective, single-institution review of consecutive patients with early-stage external auditory canal cancer treated with surgery and/or RT between April 1997 and August 2013. Audiological data included the pretreatment hearing thresholds and those obtained at 12 months after the completion of therapy. When we compared the functional gains for air-conduction pure-tone average thresholds and mean air-bone gaps, those in the surgery group (n = 10) were significantly higher than those in the RT (n = 13) and surgery + RT (n = 5) groups. Adequate techniques for reconstruction of the external auditory canal in combination with tympanoplasty after complete surgical resection is useful for hearing preservation in addition to good survival outcomes. © 2015 Wiley Periodicals, Inc. Head Neck 38: E1110-E1116, 2016.


Development of dominant bile duct stenoses in patients with primary sclerosing cholangitis treated with ursodeoxycholic acid: outcome after endoscopic treatment.
Primary sclerosing cholangitis is characterized by progressive fibrotic inflammation and obliteration of intra- and/or extrahepatic bile ducts. In a prospective study of 106 patients treated for up to 13 years with ursodeoxycholic acid, the development of major bile duct stenoses and the efficacy of endoscopic measures have been evaluated. Of 106 patients ten had major duct stenoses at entry, and during a median follow-up period of 5.0 years another 43 developed a dominant stenosis. Fifty-two patients with dominant stenoses were treated endoscopically by repeated balloon dilatations and five patients were temporarily stented. Complications of endoscopic procedures were pancreatitis (5.2%), bacterial cholangitis (3.3%) and bile duct perforation (0.5%). Five years after the first dilatation of a dominant stenosis the Kaplan-Meier survival rates free of liver transplantation were 100% in stage 2, 72% in stage 3 and 50% in stage 4 disease. The actuarial survival free of liver transplantation of the whole group at 3, 5 and 7 years were 0.987, 0.935 and 0.891 and the corresponding survival rates predicted with the Mayo multicenter survival model were 0.860, 0.775 and 0.737 (P<0.001). In advanced disease, occlusion of major bile ducts with time occurs in the majority of patients. Endoscopic opening of dominant stenoses is effective and appears to be a valuable addition to the medical treatment of such patients.


A trans-membrane pore can account for protein movement across zymogen granule membranes.
We have reported that the membrane of zymogen granules, secretion vesicles from the exocrine pancreas, is permeable to its contained proteins by measuring both the loss and accumulation of protein in response to mass action forces [1-3] However, the mechanism of transport has remained unknown. Here we consider evidence that this transport occurs through trans-membrane pores. Using freeze-fracture electron microscopic methods, Cabana et al. [4] have reported the presence of a 15 nm intramembrane particle in zymogen granule membrane which contains a 5 nm (+/- 0.1 nm, S.D.) diameter lucent center. In this article, we propose that this structure is a pore through which proteins can be transported, and test this hypothesis by comparing the predicted phenomenological permeability coefficient for transport by diffusion via this structure, to that calculated from protein flux measurements on granules using an X-ray microscope. The predicted and experimental values were essentially identical and hence support the hypothesis that this structure could be a protein transporting channel.


History of deep vein thrombosis is a discriminator for concomitant atrial fibrillation in pulmonary embolism.
Pulmonary embolism (PE) is the consequence of deep vein thrombosis (DVT) in 70% of all cases. Although, PE and DVT are commonly related to risk factors of Virchow's triad, both entities are linked to cardiovascular risk factors, but risk factors seem differently important in both entities. We aimed to investigate clinical profile and outcome of patients with PE history stratified by concomitant DVT. Data from the observational multi-center thrombEVAL-study were analyzed. The sample (N=2,318) comprised 295 PE patients, of whom 69.2% (N=204) had DVT. Individuals without DVT were older and had higher prevalence of concomitant atrial fibrillation (AF), chronic lung diseases, coronary artery disease, heart failure and hypertension. Multivariable regression revealed an independent association of AF (Odds Ratio (OR) 3.17, 95% CI 1.63-6.18, P<0.001) and coronary artery disease (OR 2.31, 95% CI 1.15-4.66, P=0.019) with PE without DVT. There was higher frequency of permanent AF in individuals without DVT, whereas paroxysmal AF was more prevalent in individuals with DVT. All AF subtypes were independently associated with PE without DVT with increasing ORs from paroxysmal to permanent AF. PE patients with and without DVT did not differ in survival (P=0.32) and cost-relevant clinical outcome (P=0.26) during follow-up. AF in PE patients was associated with cost-relevant clinical outcome (Hazard Ratio (HR) 1.78, 95% CI 1.03-3.09, P=0.040), but no significant difference in survival (HR 0.93, 95% CI 0.35-2.50, P=0.88) was observed. History of DVT is a significant discriminator for clinical profile of PE patients. Individuals without DVT had more often cardiac and pulmonary disease with strongest association with AF. Data advocate a potential link between AF and PE. URL: http://www.clinicaltrials.gov, Unique identifier NCT01809015.


Immunocytochemical assays of amylase and chymotrypsinogen in rat pancreas secretory granules. Efficacy of using immunogold-labeled ultrathin cryosections to estimate relative protein concentrations.
An exploration was conducted as to whether the relative concentration of two intracellular proteins could be evaluated quantitatively from their labeling densities in ultrathin cryosections labeled with the immunogold technique. As a model rat pancreatic cells were used in which the content of amylase (Am) and chymotrypsinogen (Ch) was experimentally altered. Rats were fed either normal laboratory chow or food containing soybean trypsin inhibitor (STI), which affects the Am/Ch ratio in the tissues. The changes in Am and Ch protein levels and enzyme activities were measured biochemically in cell suspension homogenates or in zymogen granule fractions. Within 5 days a maximal change in the Am/Ch was observed as a result of adaptation to the STI diet. The Am/Ch ratio determined biochemically was compared with that from counts of gold particles bound to the respective protein in immunogold-labeled cryosections. The two data sets matched fairly well, indicating that the intensity of the immunoreaction is a reliable reflection of antigen concentration in this system.


Adsorption of hydrogen sulfide by biochars derived from pyrolysis of different agricultural/forestry wastes.
The characteristics and mechanisms of hydrogen sulfide (H₂S) adsorption on three different biochars derived from agricultural/forestry wastes through pyrolysis at various temperatures (100 to 500 ºC) were investigated. In this study, the H₂S breakthrough capacity was measured using a laboratory-characterized using pH and Fourier transform infrared spectroscopy analysis. The results obtained demonstrate that all biochars were effective in H2S sorption. The sorption capacity of the biochar for H₂S removal is related to the pyrolysis temperature and pH of the surface. Certain threshold ranges of the pyrolysis temperature (from 100 to 500 ºC) and pH of the surface are presented. It also concluded that the sorption capacity (for removing H₂S) of rice hull-derived biochar is the largest in three biochars (camphor-derived biochar, rice hull-derived biochar, and bamboo-derived biochar). These observations will be helpful in designing biochar as engineered sorbents for the removal of H₂S. This paper focuses on the adsorption of hydrogen sulfide (H₂S) by biochars derived from wastes. The characteristics and mechanisms of hydrogen sulfide (H₂S) adsorption on three different boichars derived from agricultural/forestry wastes through pyrolysis at various temperatures were investigated. In this study, the H₂S breakthrough capacity was measured using laboratory characterization with pH and Fourier-transform infrared spectroscopy analysis. The results obtained demonstrate that all biochars were effective in H₂S sorption. The sorption capacity of the biochar for H₂S removal is related to the pyrolysis temperature and pH of the surface.


Acute invasive fungal rhinosinusitis: our experience with 18 cases.
Acute invasive fungal rhinosinusitis (AIFRS) is a rapidly progressive life threatening infection that is seen most commonly among immunocompromised patients. We present a case series of 18 patients clinically and histopathologically diagnosed with AIFRS with a mean follow-up of 9.11 ± 2.51 months (range 6-17). Demographic data, apparent symptoms and signs, underlying disorders, and outcomes are discussed. The mean age was 39.56 ± 20.66 years (range 2-75). The most common underlying diseases were diabetes mellitus (50 %) and leukemia (44.44 %). Mucosal biopsy confirmed fungal invasion of the nasal mucosa in all cases. The main fungi were Rhizopus oryzae (55.56 %), Absidia mucor (16.67 %), and Aspergillus fumigatus (27.78 %). Headache and facial pain (77.8 %), facial paresthesia (55.6 %), and ophthalmoplegia (33.3 %) were the most common symptoms and signs. Computed tomography and endoscopic findings showed various stages of sinonasal (100 %), pterygopalatine fossa (55.56 %), orbital (44.45 %), and cerebral (5.56 %) involvement. All patients underwent serial surgical debridement (3.78 ± 1.80 times; range 2-8) simultaneously with systemic antifungal therapy and proper management of the underlying disease. The most extreme case with brain involvement survived and recovered with no evidence of recurrent disease following treatment. All patients were considered cured after two endoscopic negative histopathologic evaluations. Three patients (16.67 %) died, one from uncontrolled leukemia and two due to renal failure. AIFRS is a potentially fatal condition, however, early diagnosis and management of the underlying disease accompanied with systemic antifungal and aggressive serial surgical intervention appears to be effective in reducing mortality in most patients.


Quality of life in older adults receiving hemodialysis: a qualitative study.
Patient priorities for quality of life change with age. We conducted a qualitative study to identify quality of life themes of importance to older adults receiving dialysis and the extent to which these are represented in existing quality of life instruments. We conducted semi-structured interviews with 12 adults aged ≥ 75 years receiving hemodialysis to elicit participant perspectives on what matters most to them in life. We used framework analysis methodology to process interview transcripts (coding, charting, and mapping), identify major themes, and compare these themes by participant frailty status. We examined for representation of our study's subthemes in the Kidney Disease Quality of Life (KDQOL-36) and the World Health Organization Quality of Life for Older Adults (WHOQOL-OLD) instruments. Among the 12 participants, average age was 81 (4.2) years, 7 African-American, 6 women, and 6 met frailty criteria. We identified two major quality of life themes: (1) having physical well-being (subthemes: being able to do things independently, having symptom control, maintaining physical health, and being alive) and (2) having social support (subthemes: having practical social support, emotional social support, and socialization). Perspectives on the subthemes often varied by frailty status. For example, being alive meant surviving from day-to-day for frail participants, but included a desire for new life experiences for non-frail participants. The majority of the subthemes did not correspond with domains in the KDQOL-36 and WHOQOL-OLD instruments. Novel instruments are likely needed to elicit the dominant themes of having physical well-being and having social support identified by older adults receiving dialysis.


[Drug therapy of infectious diarrhea: part 1: acute diarrhea].
Diarrhea is one of the most commonly occurring diseases. This article gives a review of the current state of the treatment of acute infectious diarrhea (part 1) and chronic infectious diarrhea (part 2) as well as of the most important pathogens. Following a presentation of the general principles of the therapy of diarrhea, the targeted antimicrobial therapy of the most important bacterial gastrointestinal infections is described. This includes salmonellosis, shigellosis and Campylobacter infections, infections with pathogenic Escherichia coli strains, yersiniosis and cholera. Due to the increasing incidence and changes in the severity of the disease and important new aspects in the treatment of diarrhea caused by toxigenic Clostridium difficile strains, these disease entities will be described in detail. Symptomatic therapy is still the most important aspect of the treatment of infectious diarrhea. For severely ill patients with a high frequency of stools (> 8/day), immunodeficiency, advanced age or significant comorbidities, empirical antibiotic therapy should be considered. Increasing resistance, in particular against fluoroquinolones must also be taken into consideration. Due to the risk of excessive pathogen proliferation and concomitant intestinal toxin production with protracted or multiple complications during the disease, therapy with motility inhibitors is not recommended. With respect to the treatment of Clostridium difficile infections a promising novel aspect arose in 2012. The macrocyclic antibiotic fidaxomycin can reduce the rate of recurrent disease with the same effectiveness as vancomycin. Furthermore, evidence for the benefits of allogenic stool transplantation is increasing. The treatment of acute diarrhea is still primarily supportive. The benefits of general empirical antibiotic therapy for acute diarrhea are not evidence-based.


A rapid method for pKa determination of drugs using pressure-assisted capillary electrophoresis with photodiode array detection in drug discovery.
We developed a rapid screening method for determination of pK(a) of candidate drugs by pressure-assisted capillary electrophoresis (CE) coupled with a photodiode array detector. Application of pressure during CE analysis allowed completion of one CE run in less than 1 min, and the obtained pH-metric mobility shifts as well as pH-metric UV spectrum were analyzed by a nonlinear regression fitting software to determine pK(a) values. The difference between pK(a) values by this method and by other conventional methods is within 0.25 units for 82 ionic functional groups of 77 drugs. The pK(a) values of 96 compounds in dimethylsulfoxide solution on a 96-well microplate could be measured in 1 day. Our method provides rapid and accurate determination of pK(a) value.


[Transplantation of mouse bone marrow mesenchymal stem cells into the xenogeneic testis].
To study transplantation of mouse bone marrow mesenchymal stem cells (MSCs) into the xenogeneic testis. (1) The tibias and femurs were dissected from 5-6-week-old mice. The marrow in the tibias and femurs was flushed out with medium. MSCs were isolated, cultured and purified in vitro by Percoll density gradient centrifugation combined with adherent method. (2) MSCs of the third generation were adopted and marked with Hoechest33342 for observation, and then made into cell-suspending fluid. (3) The marked MSCs were transplanted into the testis of the xenogeneic mouse by testis net injection. The biopsies of the testis tissues were carried out at different time and made into frost slices at three sites for observation. (1) A lot of purified MSCs were obtained at the third generation. (2) The nucleoli of the marked MSCs showed light-yellow under the fluoroscope. (3) Xenogeneic transplantation of mouse bone marrow MSCs by testis net injection was successful, without immunoreaction. On the 1 st day after transplantation, MSCs only concentratively distributed in the medial slices, the nucleoli being light-yellow; On the 1 st and 3 rd day, MSCs dispersively distributed in the medial slices; On the 6th, 9th and 12th day, MSCs presented in all the slices of the three sites, some ranging tubally; On the 15th and 18th day, the fluorescence of MSCs weakened; On the 21 st day, the fluorescence of MSCs disappeared. Transplantation of mouse bone marrow MSCs into the xenogeneic testis by testis net injection is effective and feasible, without immunoreaction. MSCs can survive after transplantation.


Differential molecular dynamics and transmembrane fluidity gradients in canine myocardial sarcolemma and sarcoplasmic reticulum.
The molecular dynamics of highly purified preparations of canine myocardial sarcolemma (SL) and sarcoplasmic reticulum (SR) were quantified by electron spin resonance spectroscopy (ESR). Canine myocardial SL and SR have substantially different motional regimes in their membrane interiors as demonstrated by alterations in the relative peak height ratios, peak widths and peak splittings in ESR spectra of 16-doxylstearate incorporated into SL and SR. Quantification of the apparent order parameters (S) of 16-doxylstearate in SL and SR by analyses of ESR spectra demonstrated that the interior of the SL membrane was substantially more immobilized than the interior of the SR membrane (e.g. S = 0.168 +/- 0.002 for SL and S = 0.128 +/- 0.003 for SR). In contrast, only modest differences in membrane dynamics near the hydrophobic-hydrophilic interface were present in SL and SR as ascertained by ESR spectra of the probe 5-doxylstearate incorporated into these membranes. Myocardial sarcolemma contained heretofore unsuspected amounts of cholesterol (1.4 +/- 0.1 mumol cholesterol/mg protein) while sarcoplasmic reticulum contained only small amounts of cholesterol (0.17 +/- 0.06 mumol cholesterol/mg protein). Model systems employing binary mixtures of plasmenylcholine/cholesterol and phosphatidylcholine/cholesterol demonstrated that the observed alterations in molecular dynamics were due, in large part, to the differential cholesterol content in these two subcellular membrane compartments. Taken together, these results demonstrate that these two functionally distinct myocardial subcellular membranes have markedly disparate molecular dynamics and transmembrane fluidity gradients which may facilitate their performance of specific functional roles during excitation-contraction coupling in myocardium.


Clinical findings and diagnosis in human granulocytic anaplasmosis: a case series from Massachusetts.
To describe clinical findings and the use of a tick-associated pathogen panel in a series of patients with human granulocytic anaplasmosis (HGA) at a suburban Boston hospital. Medical records were reviewed for inpatients and outpatients at Newton-Wellesley Hospital with a positive polymerase chain reaction (PCR) result for Anaplasma phagocytophilum during the study period March 1 through November 30, 2009. A PCR panel was used to test for tick-borne pathogens. Postal ZIP code data from the patients' areas of residence were used to estimate the area of disease transmission. Thirty-three cases were confirmed during the 2009 transmission season, and 14 of these patients (42%) required hospitalization. Thrombocytopenia and/or leukopenia were observed at the time of presentation in 25 of 30 patients (86%) in whom both white blood cell and platelet counts were determined, and 28 of 33 patients (85%) reported fever. Rash occurred in only 2 of the 33 patients (6%), and 25 (76%) reported one or more respiratory or gastrointestinal symptom. Cases were geographically distributed diffusely throughout the hospital catchment area, with one possible focus of infection identified in Weston, MA. Due to a lack of clinical data reporting to the Massachusetts Department of Public Health, only 20 of 32 HGA cases (63%) fulfilled the case confirmation criteria. Diagnosis of HGA requires a high suspicion for infection even in endemic areas. Use of a tick-associated pathogen panel that includes PCR assays for several organisms could improve detection of underrecognized tick-borne diseases in endemic areas. Lack of epidemiological follow-up to confirm corroborating clinical findings prevents accurate case reporting and assessment of the true HGA burden.


The effect of heparan sulfate on promoting amyloid fibril formation by β-casein and their binding research with multi-spectroscopic approaches.
As a molecular chaperone, β-casein is difficult to form amyloid fibrils under physiological conditions due to its chaperone activity. Heparan sulfate (HS) has drawn attention of technologists all over the word because of its relation to amyloid deposits in some amyloidosis diseases. For better understanding the relationship between the β-casein and HS, the multi-spectroscopic studies were employed. The data of thioflavin T (ThT) binding assay, transmission electron microscopy (TEM) and circular dichroism (CD) demonstrated that HS promoted fibril formation by β-casein in the amount and the growth speed. The results of steady-state UV-vis absorption spectra, fluorescence spectroscopy and fluorescence lifetime revealed that the β-casein-HS complexes were formed and HS quenched the fluorescence of β-casein by a static quenching mechanism. On the basis of fluorescence analysis, the value of binding constant was equal to 1.17 × 107 L mol-1 at 338.15 K and there was about one binding site between them. According to thermodynamic parameters obtained, it was deduced that a spontaneous reaction happened, and protein-ligand complex was stabilized by hydrogen bonds and hydrophobic interaction. Furthermore, using fluorescence resonance energy transfer (FRET) assay, the value of binding distance between HS and Trp143 of β-casein was calculated to be 0.93 nm. Finally, on the basis of synchronous fluorescence experiment, the polarity increasing and hydrophobicity decreasing around Trp143 occurred during the period of fibril formation by β-casein.


Cortical morphology in patients with the deficit and non-deficit syndrome of schizophrenia: a worldwide meta- and mega-analyses.
Converging evidence suggests that schizophrenia (SZ) with primary, enduring negative symptoms (i.e., Deficit SZ (DSZ)) represents a distinct entity within the SZ spectrum while the neurobiological underpinnings remain undetermined. In the largest dataset of DSZ and Non-Deficit (NDSZ), we conducted a meta-analysis of data from 1560 individuals (168 DSZ, 373 NDSZ, 1019 Healthy Controls (HC)) and a mega-analysis of a subsampled data from 944 individuals (115 DSZ, 254 NDSZ, 575 HC) collected across 9 worldwide research centers of the ENIGMA SZ Working Group (8 in the mega-analysis), to clarify whether they differ in terms of cortical morphology. In the meta-analysis, sites computed effect sizes for differences in cortical thickness and surface area between SZ and control groups using a harmonized pipeline. In the mega-analysis, cortical values of individuals with schizophrenia and control participants were analyzed across sites using mixed-model ANCOVAs. The meta-analysis of cortical thickness showed a converging pattern of widespread thinner cortex in fronto-parietal regions of the left hemisphere in both DSZ and NDSZ, when compared to HC. However, DSZ have more pronounced thickness abnormalities than NDSZ, mostly involving the right fronto-parietal cortices. As for surface area, NDSZ showed differences in fronto-parietal-temporo-occipital cortices as compared to HC, and in temporo-occipital cortices as compared to DSZ. Although DSZ and NDSZ show widespread overlapping regions of thinner cortex as compared to HC, cortical thinning seems to better typify DSZ, being more extensive and bilateral, while surface area alterations are more evident in NDSZ. Our findings demonstrate for the first time that DSZ and NDSZ are characterized by different neuroimaging phenotypes, supporting a nosological distinction between DSZ and NDSZ and point toward the separate disease hypothesis.


[Environmental monitoring in the foci of parasitic diseases in the town of Belgorod-Dnestrovsk, Odessa region].
The paper shows a role of environmental objects in the formation of parasitic diseases in Belgorod-Dnestrovsk, developmental changes in intestinal parasitic cenoses in children from closed institutions. To attract parasitologists to the diagnosis of acute intestinal infections of unspecified etiology increases the percent (49.5 to 82.0) of positive results. The findings show what environmental objects should be monitored in the sanitary and parasitological contexts: soil, vegetables, greens in helminthiasis; utensils in preschool children' establishments in contact helminthiasis; drinking water, vegetables, greens in giardiasis.


Block polyelectrolytes and colloidal stability.
We simulate interactions between charged flat surfaces in the presence of block polymers, where the end blocks carry a charge opposite to the surfaces. Using a recently developed simulation technique, we allow full equilibrium, i.e. the chemical potential of the polyelectrolyte is retained as the separation is changed. In general, the block polyions will adsorb strongly enough to overcharge the surfaces. This results in a double layer repulsion at large separation, with a concomitant free energy barrier. At short separations, the surfaces are pulled together by bridging forces. We make some efforts to theoretically design the polymers to be efficient flocculants, i.e. minimize the free energy barrier and still allow for a long-ranged bridging attraction. Here, we also take into account the possibility of nonequilibrium circumstances, which may be relevant in practice. Our results suggest that short chains, with small charged end blocks and a (relatively speaking) long neutral mid block, are likely to promote rapid flocculation.


Impact of an Educational Program to Reduce Healthcare Resources in Community-Acquired Pneumonia: The EDUCAP Randomized Controlled Trial.
Additional healthcare visits and rehospitalizations after discharge are frequent among patients with community-acquired pneumonia (CAP) and have a major impact on healthcare costs. We aimed to determine whether the implementation of an individualized educational program for hospitalized patients with CAP would decrease subsequent healthcare visits and readmissions within 30 days of hospital discharge. A multicenter, randomized trial was conducted from January 1, 2011 to October 31, 2014 at three hospitals in Spain. We randomly allocated immunocompetent adults patients hospitalized for CAP to receive either an individualized educational program or conventional information before discharge. The educational program included recommendations regarding fluid intake, adherence to drug therapy and preventive vaccines, knowledge and management of the disease, progressive adaptive physical activity, and counseling for alcohol and smoking cessation. The primary trial endpoint was a composite of the frequency of additional healthcare visits and rehospitalizations within 30 days of hospital discharge. Intention-to-treat analysis was performed. We assigned 102 patients to receive the individualized educational program and 105 to receive conventional information. The frequency of the composite primary end point was 23.5% following the individualized program and 42.9% following the conventional information (difference, -19.4%; 95% confidence interval, -6.5% to -31.2%; P = 0.003). The implementation of an individualized educational program for hospitalized patients with CAP was effective in reducing subsequent healthcare visits and rehospitalizations within 30 days of discharge. Such a strategy may help optimize available healthcare resources and identify post-acute care needs in patients with CAP. Controlled-Trials.com ISRCTN39531840.


Global regulator engineering enhances bioelectricity generation in Pseudomonas aeruginosa-inoculated MFCs.
The electricigens with high-electroactivity is essential for resolving the low electricity power output (EPT) of microbial fuel cells (MFCs). However, the manipulation by single functional genes shows limitation because electroactivity is a complex phenotype controlled by multiple genes. Herein, global regulator engineering (GRE) was developed to optimize the electroactivity of an isolated strain (Pseudomonas aeruginosa P3-A-11) using an exogenous global regulator IrrE (ionizing radiation resistance E linkage group) as an object. The GRE was implemented through in vitro random mutagenesis by error-prone PCR and in vivo high-through screening comprised of cultures color assay, PYO measurement and MFCs operation. Four mutants with higher electroactivity were obtained, among which, the mutant 11/M2-59 not only displayed the maximal power density, but also exhibited stronger salt tolerance, consequently showing good performance of MFCs in the presence of salt. Apart from the reduced internal resistance, the increase in phenazines amounts primarily contributed to EPT improvement, which was realized by enhancing the core biosynthesis pathway and affecting other pathways (such as central metabolism pathway, quorum sensing system, regulatory network). Notably, IrrE exerted its positive effect on electroactivity even without native regulators (such as PmpR and RpoS). In addition, the significant fluctuations in expression levels of stress-responsive genes mediated by GRE were closely associated with the enhanced salt tolerance. This work demonstrated that GRE was an effective approach for simultaneously optimizing multiple phenotypes (such as electroactivity and stress tolerance), and thus would provide more opportunities to create high-efficiency electricigens and further promoted the practical application of MFCs.


A comparison of covered vs bare expandable stents for the treatment of aortoiliac occlusive disease.
This trial was conducted to determine if covered stents offer a patency advantage over bare-metal stents in the treatment of aortoiliac arterial occlusive disease. The Covered Versus Balloon Expandable Stent Trial (COBEST), a prospective, multicenter, randomized controlled trial, was performed involving 168 iliac arteries in 125 patients with severe aortoiliac occlusive disease who were randomly assigned to receive a covered balloon-expandable stent or bare-metal stent. Patient demographic data, clinical signs and symptoms, TransAtlantic Inter-Society Consensus (TASC) classification, and preprocedure and postprocedure ankle-brachial index measurements were recorded. The primary end points included freedom from binary restenosis and stent occlusion of the treated area, as determined by ultrasound imaging or quantitative visual angiography, or both. Postprocedural follow-up was at 1, 6, 12, and 18 months. Aortoiliac lesions treated with a covered stent were significantly more likely to remain free from binary restenosis than those that were treated with a bare-metal stent (hazard ratio [HR], 0.35; 95% confidence interval (CI), 0.15-0.82; P = .02). Freedom from occlusion was also higher in lesions treated with covered stents than in those treated with a bare-metal stent (HR, 0.28; 95% CI, 0.07-1.09); however, this did not reach statistical significance (P = .07). Subgroup analyses demonstrated a significant difference in freedom from binary restenosis for covered stents in TASC C and D lesions compared with a bare stent (HR, 0.136; 95% CI, 0.042-0.442). This difference was not demonstrated for TASC B lesions (HR, 0.748; 95% CI, 0.235-2.386). COBEST demonstrates covered and bare-metal stents produce similar and acceptable results for TASC B lesions. However, covered stents perform better for TASC C and D lesions than bare stents in longer-term patency and clinical outcome.


Detection of interferon in the sera of patients with psoriasis, and its enhancement by PUVA treatment.
Interferon-like activity could be detected in the sera of nineteen psoriatic patients (interferon titres between 8 and 64 i.u./ml), whereas healthy controls were negative. Following PUVA treatment the interferon levels increased significantly (interferon titres between 32 and 128 i.u./ml). An enhanced interferon production could explain some of the immunological phenomena connected with PUVA treatment.


The rhinal cortex: 'gatekeeper' of the declarative memory system.
Almost all studies probing neural activity underlying the declarative memory system in humans have investigated either memory encoding or retrieval. Here, however, we suggest integrating encoding and retrieval operations into a single operation executed by the rhinal cortex. The more familiar an item is, the less rhinal processing it requires and the less vigorously it is encoded into memory. Given the anatomical position and the functional properties of the rhinal cortex, this operation fulfills an essential task: it optimally allocates limited encoding resources away from familiar information and towards novel information. We propose a rhinal processing stage that optimizes the declarative memory system by fully integrating encoding and retrieval operations into a single 'gatekeeper' operation.


Use of a physiologically based pharmacokinetic-pharmacodynamic model for initial dose prediction and escalation during a paediatric clinical trial.
To build and verify a physiologically based pharmacokinetic (PBPK) model for radiprodil in adults and link this to a pharmacodynamic (PD) receptor occupancy (RO) model derived from in vitro data. Adapt this model to the paediatric population and predict starting and escalating doses in infants based on RO. Use the model to guide individualized dosing in a clinical trial in 2- to 14-month-old children with infantile spasms. A PBPK model for radiprodil was developed to investigate the systemic exposure of the drug after oral administration in fasted and fed adults; this was then linked to RO via a PD model. The model was then expanded to include developmental physiology and ontogeny to predict escalating doses in infants that would result in a specific RO of 20, 40 and 60% based on average unbound concentration following a twice daily (b.i.d.) dosing regimen. Dose progression in the clinical trial was based on observed concentration-time data against PBPK predictions. For paediatric predictions, the elimination of radiprodil, based on experimental evidence, had no ontogeny. Predicted b.i.d. doses ranged from 0.04 mg/kg for 20% RO, 0.1 mg/kg for 40% RO to 0.21 mg/kg for 60% RO. For all infants recruited in the study, observed concentration-time data following the 0.04 mg/kg and subsequent doses were within the PBPK model predicted 5th and 95th percentiles. To our knowledge, this is the first time a PBPK model linked to RO has been used to guide dose selection and escalation in the live phase of a paediatric clinical trial.


Symmetry Breaking of B2N((-, 0, +)): An Aspect of the Electric Potential and Atomic Charges.
In this study, the three forms of B2N((-, 0, +))-radical, anion and cation-have been compared in terms of electric potential and atomic charges, ESP, rather than the well-known cut of the potential energy surface (PES). We have realized that the double minimum of the BNB radical is related to the lack of the correct permutational symmetry of the wave function and charge distribution. The symmetry breaking (SB) for B2N((0, +)) exhibits energy barrier in the region of (5-150) cm(-1). The SB barrier goes through a dynamic change with no centrosymmetric form which depends on the wave function or charge distribution. In spite of A ˜ 2 Σ g + exited state, the B ˜ 2 ∏ g excited configuration contributes to the ground state ( B ˜ 2 ∏ g - X ˜ 2 Σ u + ) for forming radicals. The SB did not occur for the anion form (B2N((-))) in any electrostatic potential and charges distribution. Finally, we have modified the Columbic term of the Schrödinger equation to define the parameters "αα' and ββ'" in order to investigate the SBs subject.


The psychological effects of prenatal diagnostic procedures: maternal anxiety before and after invasive and noninvasive procedures.
(1) To examine the change in anxiety before and after prenatal diagnostic procedures in women undergoing invasive (amniocentesis) and noninvasive (ultrasound) procedures; and (2) to examine predictors of anxiety before the diagnostic procedure. A short-term follow-up study was conducted on a sample of pregnant women in the second trimester. Questionnaires were administered to women scheduled for amniocentesis (n = 37) and ultrasonography (n = 37) before and immediately after the procedure. The following questionnaires were administered: the State-Trait Anxiety Inventory, the Affect Intensity Measure, the COPE inventory, and the Optimism-Pessimism Scale. Prior to the administration of the prenatal diagnostic procedure, measured anxiety levels were the same in both groups of women (p > 0.05). An interaction effect of a two-way ANOVA revealed that anxiety decreased after the procedure in the ultrasound but not the amniocentesis group (F(1, 72) = 5.01, p = 0.028). Although coping styles and affect intensity were found to be related to anxiety (p < 0.05), they were not significant predictors of anxiety before the diagnostic procedure when controlling for trait anxiety and procedure type. Anxiety levels associated with noninvasive but not after invasive, prenatal diagnostics tests decrease immediately following the procedure.


In vitro studies on liposomal amphotericin B obtained by supercritical carbon dioxide-mediated process.
Nanotechnology in drug delivery is a rapidly expanding field. Nanosized liposomal preparations are already in use for efficient drug delivery with better therapeutic indices. Existing methods of liposome preparation are limited by problems of scale-up, difficulty in controlling size, and intercalation efficiency. Here we prepare amphotericin B-intercalated liposomes by a novel process where amphotericin B and purified phosphatidyl choline are solubilized in suitable solvent and precipitated in supercritical fluid carbon dioxide (known as a gas antisolvent technique), to obtain microsized particles that are subsequently introduced into a buffer solution. The morphology of liposomes was characterized through a phase-contrast microscope, and the particle size distribution studied by laser technique showed nanosize with a narrow range of size distribution (between 0.5 and 15 microm) and a higher intercalation efficiency. In vitro studies conducted using Aspergillus fumigatus (MTCC 870) strain proved to be efficient in the retardation of the growth of the organism.


[The portacaval anastomosis in the liver cirrhosis model in the rat--significance of special preoperative preparation for decreasing surgical fatalities].
Following portocaval anastomosis a high lethality (87%) can be observed in a standardized micronodular liver cirrhosis model. By a special preoperative regimen this lethality can be reduced to 10%.


Hematological Malignancies and Arterial Thromboembolism.
Established guidelines exist for prevention and treatment of venous thromboembolism in hematological malignancies, but none for arterial thromboembolism. However, arterial and venous thromboembolism share the same provoking features-including altered procoagulant factors and defective fibrinolytic system. The morbidity for arterial thromboembolism is increasing in hematological malignancies, with the advent of immunomodulatory and targeted therapy. However, survival rate for hematological malignancy is improving. Consequently, as patients with hematological malignancies live longer, comorbidities including diabetes, hypertension and dyslipidemia, may accentuate arterial thrombosis. Thus far, the scientific literature on prophylaxis and treatment for arterial thromboembolism in hematological malignancies is limited. This review highlights the pathogenesis, incidence and clinical features of arterial thromboembolism in hematological malignancies.


Neonatal Nicotine Exposure Leads to Hypothalamic Gliosis in Adult Overweight Rats.
Astrocytes and microglia, the immune competent cells of central nercous system, can be activated in response to metabolic signals such as obesity and hyperleptinaemia. In rats, maternal exposure to nicotine during lactation leads to central obesity, hyperleptinaemia, leptin resistance and alterations in hypothalamic neuropeptides in the offspring during adulthood. In the present study, we studied the activation of astrocytes and microglia, as well as the pattern of inflammatory mediators, in adult offspring of this experimental model. On postnatal day 2 (P2), osmotic minipumps releasing nicotine (NIC) (-6 mg/kg/day) or saline for 14 days were s.c. implanted in dams. Male offspring were killed on P180 and hypothalamic immunohistochemistry, retroperitoneal white adipose tissue (WAT) polymerase chain reaction analysis and multiplex analysis for plasma inflammatory mediators were carried out. At P180, NIC astrocyte cell number was higher in the arcuate nucleus (ARC) (medial: +82%; lateral: +110%), in the paraventricular nucleus (PVN) (+144%) and in the lateral hypothalamus (+121%). NIC glial fibrillary acidic protein fibre density was higher in the lateral ARC (+178%) and in the PVN (+183%). Interleukin-6 was not affected in the hypothalamus. NIC monocyte chemotactic protein 1 was only higher in the periventricular nucleus (+287%). NIC microglia (iba-1-positive) cell number was higher (+68%) only in the PVN, as was the chemokine (C-X3-C motif) receptor 1 density (+93%). NIC interleukin-10 was lower in the WAT (-58%) and plasma (-50%). Thus, offspring of mothers exposed to nicotine during lactation present hypothalamic astrogliosis at adulthood and microgliosis in the PVN.


Heterogeneity in the angiogenic response of a BT474 human breast cancer to a novel vascular endothelial growth factor-receptor tyrosine kinase inhibitor: assessment by voxel analysis of dynamic contrast-enhanced MRI.
To investigate the heterogeneity in the angiogenic response of a human breast cancer xenograft to a novel vascular endothelial growth factor (VEGF)-receptor tyrosine kinase inhibitor, AG-013736, using dynamic contrast-enhanced MR imaging (DCE-MRI). Changes in pharmacokinetic parameters in a seven-day interval were compared between AG-treated and control groups, using Gd-DTPA and albumin-(Gd-DTPA)30. A voxel-by-voxel analysis was performed to produce parametric spatial pharmacokinetic parametric maps and histograms. Histogram segmentation was used to quantify the heterogeneity in tumor response to therapy, and compared with conventional descriptive measures of distribution in terms of their capacity to separate control from AG-treated tumors. The albumin-(Gd-DTPA)30 endothelial transfer constant, Kps, showed changes with AG-013736 treatment and tumor growth. The changes were highly heterogeneous for individual segments of the histogram with different Kps values, and the overall patterns in which the frequency distribution changed differed significantly between the two groups. A change in the number of voxels with Kps ranging from 0.03 to 0.14 mL/min/(100 mL tissue) was the most sensitive variable for separating control from AG-treated tumors (P = 0.0008). Parametric maps of the kinetic parameters also showed spatial heterogeneity in tumor response to treatment. The Kps maps depicted rapid development of central necrosis as a result of AG-013736 treatment. Maps of v(p) demonstrated a marked increase at peripheral regions of necrotic areas. Similar trends were noted in the Gd-DTPA rate constant Ktrans distribution. This study demonstrates the value of histogram analysis of maps of pharmacokinetic parameters for assessing heterogeneity in tumor response to antiangiogenic therapy. Changes in the number of voxels within certain segments of the Kps histogram were the most sensitive variable for separating control from AG-treated tumors.


Well-Being in Anesthesiology Graduate Medical Education: Reconciling the Ideal with Reality.
Addressing burnout through well-being initiatives in anesthesiology residency training has been well described. Our intervention of in-person mindfulness-based stress reduction (MBSR) and cognitive-behavioral therapy (CBT) as a means of addressing burnout among anesthesiology trainees proved unfeasible given attitudinal and logistic variables. We subsequently found success with a "confessions session" model structured as a modification of the Delphi method; this led to organizational changes associated with reduced resident burnout and well-being measured through internal (GME) and external (the Accreditation Council for Graduate Medical Education (ACGME)) annual anonymous surveys.


A rural worker infected with a bovine-prevalent genotype of Campylobacter fetus subsp. fetus supports zoonotic transmission and inconsistency of MLST and whole-genome typing.
Whole-genome characterisation in clinical microbiology enables to detect trends in infection dynamics and disease transmission. Here, we report a case of bacteraemia due to Campylobacter fetus subsp. fetus in a rural worker under cancer treatment that was diagnosed with cellulitis; the patient was treated with antibiotics and recovered. The routine typing methods were not able to identify the microorganism causing the infection, so it was further analysed by molecular methods and whole-genome sequencing. The multi-locus sequence typing (MLST) revealed the presence of the bovine-associated ST-4 genotype. Whole-genome comparisons with other C. fetus strains revealed an inconsistent phylogenetic position based on the core genome, discordant with previous ST-4 strains. To the best of our knowledge, this is the first C. fetus subsp. fetus carrying the ST-4 isolated from humans and represents a probable case of zoonotic transmission from cattle.


Respiratory quotient of stria vascularis of guinea pig in vitro.
The respiratory quotient of the stria vascularis was measured in vitro by means of Cartesian diver microgasometry. A value of about 1.2 was found when the incubation medium was phosphate-buffered serum substitute with glucose as the sole substrate. This value suggests that endogenous lipids and amino acids do not contribute significantly to strial respiratory metabolism and that carbohydrate is the primary fuel in vitro. A high activity of the hexose monophosphate pathway may be responsible for raising the respiroatyr quotient above unity.


In vivo administration of antibodies against type I collagen in rat: the specific accumulation in spleen.
[125I]-labelled rabbit antibodies against rat type I collagen and non-immune IgG were injected into rat circulation. The kinetics of their clearance and the biodistribution in different organs were studied. Both preparations showed very similar clearance rate, the kinetics fitting bi-exponential approximation with characteristic parameters t1,1/2 = 201 +/- 20 min before 320 min and t2,1/2 = 1350 +/- 450 min at times over 320 min for antibodies and 258 +/- 45 min and 890 +/- 140 min for IgG. The specific affinity of the circulating antibodies did not decrease within 24 hours. The antibodies were specifically accumulated in spleen, where their accumulation was 5-fold higher than that of non-immune IgG. Accumulation of antibodies was maximal 3 hours after the injection. The localization ratio (i.e. the ratio of the amount of the antibodies per g of tissue to that per g of blood) reached a maximum 24 hours after the injection and remained stable for 120 hours. Immunofluorescent staining of spleen sections resulted in a bright fluorescence of dense collagenous structures in the trabeculae and in the wall of the central follicular arterium, bright spot fluorescence in the marginal zone of the follicle, and diffuse fluorescence in the red pulp. These findings suggest an unusually high accessibility of collagen type I in spleen to circulating blood plasma components.


The Krüppel-like zinc-finger protein ZNF224 represses aldolase A gene transcription by interacting with the KAP-1 co-repressor protein.
Transcription factors belonging to the Krüppel-like zinc finger family of proteins participate in the regulation of cell differentiation and development. Although many of these proteins have been identified, little is known about their structure and function. We recently purified ZNF224, a new Krüppel-like zinc finger protein, that contains a Krüppel-associated box (KRAB) domain at the NH2 terminus, and 19 Cys2-His2 zinc-finger domains at the COOH terminus. Using chromatin immunoprecipitation and transient transfection assays, we demonstrate that ZNF224 binds in vivo to the distal promoter of the aldolase A gene and represses its transcription. The results of transient co-transfection experiments show that ZNF224-mediated transcription repression requires the 45-amino acid long KRAB A domain. The ability of KRAB-containing ZNF224 protein to repress transcription depends on specific interaction with the KAP-1 co-repressor molecule. Finally, using selective treatment with the HDAC1 inhibitor trichostatin A, we demonstrate that ZNF224-mediated repression requires histone deacetylases.


Habitual exercise is associated with cognitive control and cognitive reappraisal success.
Habitual exercise is associated with enhanced domain-general cognitive control, such as inhibitory control, selective attention, and working memory, all of which rely on the frontal cortex. However, whether regular exercise is associated with more specific aspects of cognitive control, such as the cognitive control of emotion, remains relatively unexplored. The present study employed a correlational design to determine whether level of habitual exercise was related to performance on the Stroop test measuring selective attention and response inhibition, the cognitive reappraisal task measuring cognitive reappraisal success, and associated changes in prefrontal cortex (PFC) oxygenation using functional near-infrared spectroscopy. 74 individuals (24 men, 50 women, age 18-32 years) participated. Higher habitual physical activity was associated with lower Stroop interference (indicating greater inhibitory control) and enhanced cognitive reappraisal success. Higher habitual exercise was also associated with lower oxygenated hemoglobin (O2Hb) in the PFC in response to emotional information. However, NIRS data indicated that exercise was not associated with cognitive control-associated O2Hb in the PFC. Behaviorally, the findings support and extend the previous findings that habitual exercise relates to more successful cognitive control of neutral information and cognitive reappraisal of emotional information. Future research should explore whether habitual exercise exerts causal benefits to cognitive control and PFC oxygenation, as well as isolate specific cognitive control processes sensitive to change through habitual exercise.


Study of O-glycan sialylation in C6 cultured glioma cells: regulation of a beta-galactoside alpha 2,3 sialyltransferase activity by Ca2+/calmodulin antagonists and phosphatase inhibitors.
We have demonstrated that the alpha 2,3 sialyltransferase (alpha 2,3 ST) from C6 cultured glioma cells was inhibited in vivo by W-7 and related Ca2+/Calmodulin (Ca/CaM) antagonists while protein kinase C effectors had no effect. Dephosphorylation of alpha 2,3 ST by the wide specificity alkaline phosphatase led to inactivation indicating that the enzyme is phosphorylated. The serine/threonine protein phosphatase inhibitors okadaic acid and Calyculin A led also to an inhibition of alpha 2,3 ST activity. In addition, Ca/CaM antagonists and phosphatase inhibitors led both to an inhibition of a alpha 2,3 sialoglycoprotein from C6 glioma cells as demonstrated with lectin affinity blotting. A concerted regulatory mechanism with phosphorylation/dephosphorylation of alpha 2,3 ST is then postulated.


The Clinical and Economic Impact of Preoperative Transthoracic Echocardiography in Elderly Patients with Hip Fractures.
The purpose of this study was to evaluate preoperative transthoracic echocardiography (TTE) on cardiac intervention, length of stay (LOS), inpatient mortality, and costs. A retrospective series of 43 preoperative TTE and 161 non-TTE (control) hip fracture patients (> 65 years) was reviewed. The data collected included ASA score, comorbidities, indication for TTE, perioperative cardiac intervention, LOS, inpatient mortality, and cost. One of 43 (2.4%) (TTE) had a cardiac intervention (PTCA/CABG). Zero interventions occurred in the control group. The average time to operation was 1.5 days (TTE) and 0.93 days (control) (p < 0.001). The average LOS was 7.2 days (TTE) and 6.0 days (control), (p = 0.04). Patients (American Society of Anesthesiologists Physical Status Classification 3 and 4 patients), LOS was 7.3 days (TTE) and 6.3 days (control) (p = 0.18). Inpatient mortality was 2.3% (TTE) and 3% (control) (p = 0.493). There was no correlation between TTE and anesthesia. Hospital costs were different between groups (TTE $24,445 and control $18,429, p = 0.02). Preoperative TTE in elderly patients with hip fractures resulted in a low cardiac intervention rate. Patients undergoing preoperative TTE prior to surgery had longer times to operation, LOS, and higher hospital costs. The utility of TTE as a preoperative screening tool is limited in the geriatric hip fracture population and does not appear to effect perioperative mortality rates.


Tissue retention and subcellular distribution of continuously infused melatonin in rats under near physiological conditions.
The fate and disposition of the melatonin released into the circulation is still poorly understood, and almost all current knowledge is derived from measurements made after a single and often a very large dose of labelled melatonin. In continuous infusion experiments in freely moving rats, we have recently demonstrated that considerable amounts of melatonin must be endogenously released in order to achieve and maintain approximately a 10-fold elevation of the low daytime plasma levels of this hormone. We have now applied this infusion paradigm to study the fate and tissue accumulation of [3H]-melatonin continuously infused under near physiological conditions into the jugular vein for a period of 2 hr. The retention of [3H]-melatonin and chloroform-insoluble [3H]-melatonin-metabolites was measured in almost all body tissues and their subcellular compartments immediately at the end of the infusion period and 6 hr later. At the end of the 2 hr infusion period, about 45% of the administered melatonin was recovered as water-soluble metabolites in the urine and about 20% in the small intestine. Some accumulation of [3H]-melatonin-derived water-soluble radioactivity was also noticed in the liver, colon, adrenals, and pituitary, as well as in the feces. The subcellular distribution of this radioactivity differed between tissues. During the period of 6 hr after the termination of infusion, a considerable amount of melatonin-derived radioactivity was found to become increasingly attached to the proteous interlayer of chloroform extracts of tissues and subcellular fractions, from where it could only be liberated by protease treatment.


Augmentation rhinoplasty with autologous grafts.
The authors present their experience with nasal reconstructive surgery using autologous grafts. Currently, grafts are classified into four categories: skin grafts, cartilage grafts, bone grafts, and composite grafts (Table 1). A sample of 132 patients with nasal defects requiring reconstruction was selected. Two cases, presenting nasal valve collapse and deformities after aesthetic nasal surgery, were analyzed in detail. Overall, satisfying aesthetic results (balanced tip projection, dorsum fullness, widening of internal nasal valve angle, minor scar retraction) were achieved for both patient and surgeon in 79% of the cases.


Randomized Controlled Trial Evaluating a Standardized Strategy for Uveitis Etiologic Diagnosis (ULISSE).
To prospectively assess the efficiency of a standardized diagnostic approach, compared to an open strategy, for the etiologic diagnosis of uveitis. Noninferiority, prospective, multicenter, clustered randomized controlled trial. Consecutive patients with uveitis, who visited 1 of the participating departments of ophthalmology, were included. In the standardized group, all patients had a minimal evaluation regardless of the type of uveitis (complete blood count, erythrocyte sedimentation rate, C-reactive protein, tuberculin skin test, syphilis serology, and chest radiograph) followed by more complex investigations according to ophthalmologic findings. In the open group, the ophthalmologist could order any type of investigation. Main outcome was the percentage of etiologic diagnoses at 6 months. Nine hundred and three patients with uveitis were included from January 2010 to May 2013 and the per-protocol population comprised 676 patients (open 373; standardized 303). Mean age at diagnosis was 46 years. Anatomic distribution of uveitis was as follows: anterior (60.8% and 72.3%, P = .0017), intermediate (11.7% and 12.3%, P = .8028), posterior (17.8% and 8.2%, P = .0004), and panuveitis (15.3% and 15.2%, P = .9596). An etiologic diagnosis was established in 54.4% of cases in the open group and 49.5% in the standardized group (P = .2029). The difference between both strategies (standardized minus open) was -4.9% (95% CI [-12.5%; 2.6%]). There were more investigations in the open group than in the standardized group (5371 vs 3759, P < .0001). The standardized strategy appears to be an efficient diagnostic approach for the etiologic diagnosis of uveitis, although its noninferiority cannot be proved.


Safety and tolerability of fluoroquinolones.
Clinical trials in patients with community- and hospital-acquired infections have established that the clinical effectiveness and safety of fluoroquinolones are similar to beta-lactam and macrolide agents. The most common drug-related adverse effects (AEs) with fluoroquinolone therapy involve the gastrointestinal tract and central nervous system and are usually transient and mild to moderate in severity. However, serious toxic reactions have led to the limited and restrictive use of trovafloxacin in the United States and the withdrawal of temafloxacin and grepafloxacin from worldwide markets. In addition, postmarketing spontaneous AE reports have imposed updates in the precautions and warning sections of product package inserts of selected fluoroquinolones. This article reviews the AEs associated with the fluoroquinolones and compares the safety profiles of ciprofloxacin, levofloxacin, gatifloxacin, and moxifloxacin.


Gestational diabetes and impaired glucose tolerance of pregnancy in Riyadh.
A total of 1088 pregnant women was screened for abnormalities in glucose metabolism at the initial antenatal visit; those with specific risk factors were screened again after 28 weeks gestation. In 210 (19.3%) plasma glucose measured 2 h after a 75 g glucose load was greater than or equal to 7.8 mmol/l (140 mg/dl). Follow-up glucose tolerance tests revealed an overall prevalence of abnormal tests of 10.3% (112 of 1088) according to the WHO criteria, of which 21 (1.9%) were diagnosed as gestational diabetes, and 91 (8.4%) as impaired glucose tolerance. This high prevalence was significantly related to age, parity and body mass index. Screening and diagnosis using criteria set by the WHO were found acceptable and are recommended to help resolve the international disagreement on diagnostic criteria of glucose metabolism abnormalities in pregnancy.


Maximising access to timely trauma care across population of Karachi and its districts: a geospatial approach to develop a trauma care network.
To develop and propose a cost-effective trauma care network for Karachi, Pakistan, by calculating maximum timely trauma care (TTC) coverage achieved with the addition of potential designated private and public level 1 and level 2 trauma centres (TCs). A lower middle-income country metropolis, Karachi is Pakistan's largest city with a population of 16 million and a total of 56 hospitals as per government registry data. 41 potential TCs selected using a two-level, contextually-relevant TC designation criteria adapted from various international guidelines. Maximum TTC coverage achievable with the addition of potential TCs. Proposed trauma care network composition to achieve maximum TTC coverage. Coverage with five public level 1 hospitals alone is 74.4%. Marginal benefit with stepwise addition of five potential private level 1 TCs, four public level 2 TCs and two private level 2 TCs is 12.2%, 7.1% and 3.1%, respectively. Maximum possible TTC coverage is 96.7%. Poorest coverage with the proposed 16 hospital network is noted in Malir district while 100% coverage is achieved in the centrally located South, Central and East districts. Addition of private level 1 and private and public level 2 hospitals to the trauma care network is necessary. Implementation of the proposed trauma care network requires strong stewardship from the government and coordinated effort of multiple stakeholders is needed to ensure standard TC designation. The study exhibits an effective method to scientifically plan and develop a cost-effective trauma system which can be applied in other resource-limited geographical areas.


Effects of robotic therapy on upper-extremity function in children with cerebral palsy: A systematic review.
To systematically examine the effects of robotic therapy on upper extremity (UE) function in children with cerebral palsy (CP). A systematic literature search was conducted in Pubmed, CINAHL, Cochrane, PsychInfo, TRIP, and Web of Science up to July 2013. Studies of children with CP, using robotic therapy and measures of UE were included. Nine articles using three different robotic systems were included. Of these, seven were case studies. Overall, robotic therapy showed the potential effects as all studies reported at least one positive outcome: a moderate effect in improving reaching duration, smoothness, or decreased muscle tone, and a small to large effect in standardized clinical assessment (e.g. Fugl-Meyer). This review confirms the potential for robotic therapy to improve UE function in children with CP. However, the paucity of group design studies summons the need for more rigorous research before conclusive recommendations can be made.


Time-resolved study of tryptophan fluorescence in vesicle reconstituted cytochrome oxidase. Effect of redox transition.
Time-resolved study of fluorescence decay of the tryptophan residue in bovine cytochrome c oxidase in phospholipid vesicles is reported for the first time. The effect of the redox state of the protein on its conformation has been investigated using time-resolved decay of tryptophan fluorescence in the oxidised and reduced protein. The fluorescence decay was best fitted using a discrete three exponential model. Amplitude distribution of lifetimes also showed three distinct regions in the analysis of decay profiles by the maximum entropy method (MEM). Results of the time resolved studies showed that the amplitudes as well as the lifetimes of the tryptophan fluorescence remain the same for the oxidised and the reduced states of cytochrome c oxidase, indicating that the environment around tryptophan residues remains more or less unaltered on reduction of the protein. The results suggest that there is no global conformational change in the protein on electron transfer and support the possibility of the existence of local fluctuations in the protein during the redox cycle.


Elevated expression of BAFF receptor, BR3, on monocytes correlates with B cell activation and clinical features of patients with primary Sjögren's syndrome.
We reported that the production of BAFF (B cell-activating factor) and IL-6, both of which are involved in survival and differentiation of B cells, is dysregulated in monocytes of patients with primary Sjögren's syndrome (pSS). In this study, we investigate the relationship between possible aberrations of pSS monocytes and clinical features of pSS patients and the contribution of monocytes to B cell activation, a mechanism involved in the pathogenesis of pSS. Expression of BAFF-receptor (BR3) on peripheral monocytes from patients with pSS (n = 67) and healthy controls (HC: n = 37) was analyzed by FACS. Peripheral monocytes were stimulated with BAFF, and IL-6 production by the cells was measured by ELISA. Peripheral B cells were cultured with BAFF-stimulated monocytes in the presence or absence of anti-IL-6 receptor antibody, and IgG production by the cells was measured by ELISA. Patients' serological data were collected from their clinical records. Patients' disease activity was quantified based on their EULAR Sjögren's syndrome disease activity index (ESSDAI) scores. The proportion of peripheral BR3-positive monocytes (BR3+/CD14+) was significantly increased in pSS patients compared to HC. Moreover, IL-6 production by BAFF-stimulated monocytes was remarkably higher than HC and was significantly correlated with BR3+/CD14+ ratios of patients. In addition, BR3 expression on pSS monocytes was elevated in anti-Ro/SSA and/or anti-La/SSB positive compared to negative patients. Remarkably, BR3 expression on peripheral monocytes was positively and significantly correlated with patients' serum IgG and IgM levels and ESSDAI scores. Moreover, the amount of IgG produced by B cells was markedly higher in pSS patients compared to HC when the cells were co-cultured with BAFF-stimulated autologous monocytes in vitro. Notably, addition of anti-IL-6 receptor antibody into the co-culture system led to inhibition of IgG production by B cells. Our data suggest that elevated BR3 expression in monocytes is associated with clinical features in pSS patients and that enhanced production of IL-6 by BAFF-stimulated monocytes plays a part in the overproduction of IgG by B cells in pSS. These results suggest that BAFF signaling pathways through BR3 in monocytes are possible therapeutic targets for pSS.


The TEL-AML1 real-time quantitative polymerase chain reaction (PCR) might replace the antigen receptor-based genomic PCR in clinical minimal residual disease studies in children with acute lymphoblastic leukaemia.
Prospective studies in children with B-precursor acute lymphoblastic leukaemia (ALL) have shown that polymerase chain reaction (PCR)-based detection of minimal residual disease (MRD) using immunoglobin (Ig) and T-cell receptor (TCR) gene rearrangements as targets can be used to identify patients with a high relapse risk. The disadvantage of this approach is that for each patient preferably two different targets have to be identified. The t(12;21)(p13;q22) with the TEL-AML1 fusion gene is present in approximately 25% of children with B-precursor ALL. In these patients, sensitive reverse transcription (RT)-PCR analysis of the TEL-AML1 fusion transcript might be a more simple and less laborious alternative approach. However, it is unknown how stable the mRNA is and whether the number of transcripts per leukaemic cell remains constant during follow-up. We investigated whether the MRD results obtained using RT-PCR of TEL-AML1 transcripts correlated with the clinically validated genomic PCR for Ig and TCR gene rearrangements. Therefore, we used real-time quantitative (RQ)-PCR analysis for both types of targets and assessed the MRD levels in 36 follow-up bone marrow samples (obtained during the first 1.5 years after diagnosis) from 13 patients with B-precursor ALL. In 34/36 bone marrow samples the Ig/TCR RQ-PCR and TEL-AML1 RQ-PCR revealed equal levels of MRD and these results had a strong correlation (P < 0.0001, R2 = 0.84). Therefore, we conclude that the TEL-AML1 RQ-PCR can, in principle, replace Ig/TCR RQ-PCR in B-precursor ALL with t(12;21).


Sub-clinical neurobehavioral abnormalities associated with low level of mercury exposure through fish consumption.
In order to assess early neurotoxic effects associated with relatively low levels of mercury absorbed through fish eating, two groups of 22 adult male subjects, habitual consumers of tuna fish, and 22 controls were examined using a cross-sectional field study. The assessment included neurobehavioral tests of vigilance and psychomotor function, hand tremor measurements and serum prolactin assessment. Mercury in urine (U-Hg) and serum prolactin (sPRL) were measured in all exposed subjects and controls, whereas measurements of the organic component of mercury in blood (O-Hg) were available for only 10 exposed and six controls. U-Hg was significant higher among exposed subjects (median 6.5 microg/g of creatinine, range 1.8-21.5) than controls (median 1.5 microg/g of creatinine, range 0.5-5.3). The median values of O-Hg were 41.5 microg/l among the tuna fish eaters and 2.6 microg/l in the control group. Both U-Hg and O-Hg were significantly correlated with the quantity of fish consumed per week. Significant differences in sPRL were found between exposed (12.6 ng/ml) and controls (9.1 ng/ml). Individual sPRL were significantly correlated with both U-Hg and O-Hg levels. The neurobehavioral performance of subjects who consumed tuna fish regularly was significantly worse on color word reaction time, digit symbol reaction time and finger tapping speed (FT). After considering the education level and other covariates, the multiple stepwise regression analysis indicated that O-Hg concentration was most significantly associated with individual performance on these tests, accounting for about 65% of the variance in test scores.


Processes in DNA damage response from a whole-cell multi-omics perspective.
Technological advances have made it feasible to collect multi-condition multi-omic time courses of cellular response to perturbation, but the complexity of these datasets impedes discovery due to challenges in data management, analysis, visualization, and interpretation. Here, we report a whole-cell mechanistic analysis of HL-60 cellular response to bendamustine. We integrate both enrichment and network analysis to show the progression of DNA damage and programmed cell death over time in molecular, pathway, and process-level detail using an interactive analysis framework for multi-omics data. Our framework, Mechanism of Action Generator Involving Network analysis (MAGINE), automates network construction and enrichment analysis across multiple samples and platforms, which can be integrated into our annotated gene-set network to combine the strengths of networks and ontology-driven analysis. Taken together, our work demonstrates how multi-omics integration can be used to explore signaling processes at various resolutions and demonstrates multi-pathway involvement beyond the canonical bendamustine mechanism.


Role of endocytosis and cathepsin-mediated activation in Nipah virus entry.
The recent discovery that the Nipah virus (NiV) fusion protein (F) is activated by endosomal cathepsin L raised the question if NiV utilize pH- and protease-dependent mechanisms of entry. We show here that the NiV receptor ephrin B2, virus-like particles and infectious NiV are internalized from the cell surface. However, endocytosis, acidic pH and cathepsin-mediated cleavage are not necessary for the initiation of infection of new host cells. Our data clearly demonstrate that proteolytic activation of the NiV F protein is required before incorporation into budding virions but not after virus entry.


Myocardial infarction in pregnancy.
Three cases of myocardial infarction occurring in the third trimester of pregnancy are presented. The method of delivery in each case was individualised with no maternal or fetal mortality. Each patient had significant risk factors for coronary arterial disease or thrombosis. One patient had normal coronary arteries demonstrated at coronary arteriography.


The dynamics of dissolved inorganic nitrogen species mediated by fresh submarine groundwater discharge and their impact on phytoplankton community structure.
Submarine groundwater discharge (SGD)-driven nutrient inputs have long been speculated to sustain the high frequency of red tide occurrence in Tolo Harbour, Hong Kong, for its larger flux and higher nutrient loadings than river discharge. Based on analysis of high resolution time series biogeochemical and climatological data from 2000 to 2015, fresh SGD-derived dissolved inorganic nitrogen (DIN) is found to be a significant regulator of the annual cycle of phytoplankton community structure in the harbour. In the wet season, fresh SGD supplies nutrients with NH4+:NO3- ratio < 1 to the seawater, meanwhile creates an intensive vertical stratification environment. As a result, diatom which is a NO3- specialist, is prone to be the major group in the harbour. Fresh SGD delivers a same orders of magnitude of DIN as river and precipitation, but it is more important to phytoplankton community structure dynamics because fresh groundwater has smaller NH4+:NO3- ratio that significantly changes the ratio in the harbour. In the dry season, with the decline of fresh SGD and the ease of stratification, vertical mixing uplifts the nutrient (NH4+:NO3- ratio > 1) released from the bottom sediment leading to a NH4+ dominant environment in water column. Dinoflagellate and other groups then become dominant species of phytoplankton in the harbour. Fresh SGD has a major influence on the NH4+:NO3- ratio in the seawater compared to tide-driven SGD, even though the latter contributes a larger proportion SGD. Tide-driven SGD also produces NH4+ and NO3-, but NH4+:NO3- ratio are mainly subject to the beach environment (bare/mangrove beach), which does not change much seasonally, thus dominant DIN species do not change significantly throughout a year. In a conclusion, fresh SGD plays the most important role among all the endmembers in regulating the DIN composition in Tolo Harbour and its fluctuation mediates the phytoplankton community structure.


Acyl-CoA: 1-acyl-glycero-3-phosphoethanolamine O-acyltransferase and liver plasma membrane fluidity.
Investigations have been carried out on the influence of membrane lipid composition and physical state on acyl-CoA: 1-acyl-glycerol-3-phosphoethanolamine O-acyltransferase activity in rat liver plasma membranes. The lipid composition of the membranes was modified either by way of lipid transfer proteins or by partial delipidation with exogenous phospholipases and subsequent enrichment of the membranes with different phospholipids. The results indicated that membrane rigidification by enrichment of the membranes with DPPC or SM reduced the transfer of oleic and palmitic acid to lysophosphatidylethanolamine, whereas all phospholipids inducing membrane fluidization lead to acyltransferase activation. The eventual role of membrane fluidity in the deacylation-reacylation cycle is discussed.


Cervical root resorption following bleaching of endodontically treated teeth.
One year following root canal treatment and internal etching and bleaching of anterior teeth in dogs, the animals were sacrificed and the teeth prepared for stereomicroscopic or light microscopic examination. Evidence of cervical root resorption and ankylosis was noted on several teeth. The bleaching factors associated with the teeth exhibiting resorption were heat with 30% hydrogen peroxide. Resorption was not related to walking bleach or to internal etching alone.


Strategies to Improve Perioperative Palliative Care Integration for Seriously Ill Veterans.
Seriously ill patients are at higher risk for adverse surgical outcomes. Palliative care (PC) interventions for seriously ill surgical patients are associated with improved quality of patient care and patient-centered outcomes, yet, they are underutilized perioperatively. To identify strategies for improving perioperative PC integration for seriously ill Veterans from the perspectives of PC providers and surgeons. We conducted semistructured, in-depth individual and group interviews with Veteran Health Administration PC team members and surgeons between July 2020 and April 2021. Participants were purposively sampled from high- and low-collaboration sites based on the proportion of received perioperative palliative consults. We performed a team-based thematic analysis with dual coding (inter-rater reliability above 0.8). Interviews with 20 interdisciplinary PC providers and 13 surgeons at geographically distributed Veteran Affairs sites converged on four strategies for improving palliative care integration and goals of care conversations in the perioperative period: 1) develop and maintain collaborative, trusting relationships between palliative care providers and surgeons; 2) establish risk assessment processes to identify patients who may benefit from a PC consult; 3) involve both PC providers and surgeons at the appropriate time in the perioperative workflow; 4) provide sufficient resources to allow for an interdisciplinary sharing of care. The study demonstrates that individual, programmatic, and organizational efforts could facilitate interservice collaboration between PC clinicians and surgeons.


Genomic and phenotypic delineation of congenital microcephaly.
Congenital microcephaly (CM) is an important birth defect with long term neurological sequelae. We aimed to perform detailed phenotypic and genomic analysis of patients with Mendelian forms of CM. Clinical phenotyping, targeted or exome sequencing, and autozygome analysis. We describe 150 patients (104 families) with 56 Mendelian forms of CM. Our data show little overlap with the genetic causes of postnatal microcephaly. We also show that a broad definition of primary microcephaly -as an autosomal recessive form of nonsyndromic CM with severe postnatal deceleration of occipitofrontal circumference-is highly sensitive but has a limited specificity. In addition, we expand the overlap between primary microcephaly and microcephalic primordial dwarfism both clinically (short stature in >52% of patients with primary microcephaly) and molecularly (e.g., we report the first instance of CEP135-related microcephalic primordial dwarfism). We expand the allelic and locus heterogeneity of CM by reporting 37 novel likely disease-causing variants in 27 disease genes, confirming the candidacy of ANKLE2, YARS, FRMD4A, and THG1L, and proposing the candidacy of BPTF, MAP1B, CCNH, and PPFIBP1. Our study refines the phenotype of CM, expands its genetics heterogeneity, and informs the workup of children born with this developmental brain defect.


A genetic analysis of glucoamylase activity in the diastatic yeast Saccharomyces cerevisiae NCYC 625.
The wild diastatic yeast Saccharomyces cerevisiae NCYC 625 has been shown to be homozygous for the glucoamylase-specifying gene STA2. spoII-1-mapping has positioned STA2 on chromosome II. Expression of STA2 is suppressed in some but not all diploids capable of sporulation, and is also inhibited by unlinked nuclear suppressor genes (SGL) found in some S. cerevisiae tester strains. EMS-induced glucoamylase-negative mutants often contain STA2-suppressor mutations. Depending on the allelic status of GEP1, a nuclear gene which also appears able to antagonise SGL-mediated suppression, STA2 expression can be blocked in petite mutants.


[Isocitrate dehydrogenases of trematodes parasitizing cattle and the feasibility of inhibiting them using anthelmintic preparations].
Activities and properties of NADF-dependent isocitrate dehydrogenases in cytosol and mitochondrial fractions from trematodes Eurytrema pancreaticum and Calicophoron ijimai were examined. Cytosol and mitochondrial enzymes were activated by ions Mn2+ and Mg2+ and inhibited by ions of heavy metals and p-chloromercuribenzoate. The effect of anthelmintic preparations on activity of enzymes was investigated.


Effects of maximal exercise stress on left ventricular function in patients with coronary artery disease using first pass radionuclide angiocardiography: a rapid, noninvasive technique for determining ejection fraction and segmental wall motion.
Angiographically determined changes in segmental wall motion (SWM) and ejection fraction (EF) are sensitive indices of left ventricular (LV) function. To compare the effects of exercise on LV function, first pass radionuclide angiocardiography was used before and during maximal upright bicycle stress in patients with nonsignificantly stenosed coronary arteries, and in those with greater than 75% stenosis. Gamma camera acquisitions were made in the 30 degree RAO projection using a 20 mCi I.V. bolus of 99mTc-pertechnetate. In the control group (seven normals, one nonsignificant (CAD) the EF significantly increased between rest and exercise (0.65 +/- 0.03 to 0.81 +/- 0.03 (mean +/- SEM), p less than 0.005). In this group SWM measured over the two anterior and two inferoposterior segments uniformly increased. In the 11 patients with a history of angina and significant coronary artery obstruction, the EF did not change in three and significantly decreased in the remaining eight (0.57 +/- 0.04 to 0.45 +/- 0.03, p less than 0.005). In all 11 patients SWM either decreased or did not increase in the areas supplied by the significantly stenosed coronary arteries. Upright maximal stress angiocardiography appears to be well-suited for diagnosing ischemic heart disease and localizing the area of ischemic dysfunction.


Sirt6 protects cardiomyocytes against doxorubicin-induced cardiotoxicity by inhibiting P53/Fas-dependent cell death and augmenting endogenous antioxidant defense mechanisms.
Sirt6, a class III NAD+-dependent deacetylase of the sirtuin family, is a highly specific H3 deacetylase and plays important roles in regulating cellular growth and death. The induction of oxidative stress and death is the critical mechanism involved in cardiomyocyte injury and cardiac dysfunction in doxorubicin-induced cardiotoxicity, but the regulatory role of Sirt6 in the fate of DOX-impaired cardiomyocytes is poorly understood. In the present study, we exposed Sirt6 heterozygous (Sirt6+/-) mice and their littermates as well as cultured neonatal rat cardiomyocytes to DOX, then investigated the role of Sirt6 in mitigating oxidative stress and cardiac injury in the DOX-treated myocardium. Sirt6 partial knockout or silencing worsened cardiac damage, remodeling, and oxidative stress injury in mice or cultured cardiomyocytes with DOX challenge. Cardiomyocytes infected with adenoviral constructs encoding Sirt6 showed reversal of this DOX-induced damage. Intriguingly, Sirt6 reduced oxidative stress injury by upregulating endogenous antioxidant levels, interacted with oxidative stress-stirred p53, and acted as a co-repressor of p53 in nuclei. Sirt6 was recruited by p53 to the promoter regions of the target genes Fas and FasL and further suppressed p53 transcription activity by reducing histone acetylation. Sirt6 inhibited Fas/FasL signaling and attenuated both Fas-FADD-caspase-8 apoptotic and Fas-RIP3 necrotic pathways. These results indicate that Sirt6 protects the heart against DOX-induced cardiotoxicity by upregulating endogenous antioxidants, as well as suppressing oxidative stress and cell death signaling pathways dependent on ROS-stirred p53 transcriptional activation, thus reducing Fas-FasL-mediated apoptosis and necrosis. •Sirt6 is significantly decreased in DOX-insulted mouse hearts and cardiomyocytes. •Sirt6 attenuates DOX-induced cardiac atrophy, dysfunction and oxidative stress. • Sirt6 reduces oxidative stress injury by upregulating endogenous antioxidants. • Sirt6 interacts with p53 as a co-repressor to suppress p53 transcriptional regulation and inhibits Fas-FasL-mediated apoptosis and necrosis downstream of p53.


In vivo interaction between Trypanosoma gambiense and leucocytes in mice.
Naturally occurring phagocytosis of Trypanosoma gambiense by mouse eosinophils and neutrophils was reported. In vivo and in vitro experiments using monoclonal antibodies confirmed that the phagocytosis is triggered by G1 class antibodies against variable surface antigen. Ultrastructural observation revealed the mode of entry and the intracellular fate of T. gambiense: initial attachment, pseudopodia formation and complete invagination. This phagocytosis resulted in the killing of T. gambiense by mouse eosinophils and neutrophils, suggesting that eosinophils and neutrophils give at least partial protection against infection with T. gambiense in combination with the specific antibodies.


Enchytraeus crypticus as model species in soil ecotoxicology.
Enchytraeids are ecologically relevant soil organisms, due to their activity in decomposition and bioturbation in many soil types worldwide. The enchytraeid reproduction test (ERT) guidelines ISO 16387 and OECD 220 are exclusive to the genus Enchytraeus and recommend using the species E. albidus with a 6-week test period. The suggested alternative, E. crypticus has a shorter generation time which may enable the ERT to be twice as fast. To confirm the suitability of a 3-week test period for E. crypticus, the toxicity of five chemicals, with distinct properties and modes of action, was assessed in LUFA 2.2 soil. In all controls the validity criteria were met, as survival of E. crypticus was above 92% and more than 772 juveniles were produced. The good performance supports its appropriateness as model species. Reproduction was more sensitive than survival, with only cadmium and 3,5-dichloroaniline causing significant lethal effects in the tested concentration ranges. The effect concentration causing 50% reduction in the number of juveniles (EC50) was 35 mg kg(-1) for cadmium, <1.0 mg kg(-1) for carbendazim, 145 mg kg(-1) for phenanthrene, 275 mg kg(-1) for pentachloroaniline and 102 mg kg(-1) for 3,5-dichloroaniline. To evaluate the sensitivity of E. crypticus, the present results were compared to literature data for E. albidus. In conclusion, E. crypticus is a suitable model species in soil ecotoxicology, with advantages such as good control performance and speed, leading to a reliable and faster ERT.


Cyclic AMP stimulates platelet-derived growth factor B chain mRNA expression in murine macrophage cell lines.
Prostaglandin E(2) plays a role in cytokine production presumably by altering intracellular levels of cAMP. In this paper, we report on the differential expression of cytokine genes in murine macrophages in response to stimulation with activators of cAMP. Macrophages were cultured with or without cAMP activators in the presence or absence of LPS. Prior to treatment, macrophages do not express interleukin-1beta, but do express low levels of tumour necrosis factor alpha and platelet-derived growth factor B chain mRNAs. After culture with cAMP-inducers, including PGE(2), dibutyryl cAMP and forskolin, PDGF B chain mRNA is induced. Forskolin, for example, induced expression PDGF B chain mRNA to a level ranging from 25% to 200% of the level induced by LPS in 6 h. In contrast, cAMP-inducers enhance the expression of IL-1beta and TNF-alpha mRNAs, but only in the presence of LPS. The combination of forskolin and LPS does not appear to act synergistically on PDGF B chain mRNA levels, suggesting that LPS-stimulated effects are not mediated through a cAMP-dependent pathway. Furthermore, macrophages differentially express cytokine genes in response to treatment with inducers of intracellular cAMP.