Electrodeposition of carrier-free 57Co on rhodium as an approach to the preparation of Mössbauer sources.
Electrodeposition of carrier-free (57)Co on a rhodium matrix as the first step of preparing Mössbauer sources was studied. To optimize the plating parameters, the influences of current density, volume and pH of the electrolyte solution, shape, thickness, and surface area of the rhodium cathode, mode of cathode pretreatment, concentration of (57)Co and duration of electrolysis were investigated.


Electron microscope detection of an endogenous infection of retrovirus-like particles in L20B cells.
L20B cells are a cell line commonly used for the isolation of poliovirus. The current study indicates that L20B cells are chronically infected with a retrovirus-like particle that replicates in the cytoplasm and buds through the plasma membrane. The findings indicate that care is needed in the use of L20B cells for certain virus isolation studies and emphasize the importance of electron microscope studies as an adjunct to the development of diagnostic virology protocols.


Spatial Analysis of Infections by Toxoplasma gondii and Neospora caninum (Protozoa: Apicomplexa) in Small Ruminants in Northern Italy.
The objectives of this study were: (i) To investigate possible geographical or environmental factors influencing the infections by Toxoplasma gondii and Neospora caninum in sheep and goats in northern Italy; (ii) to identify areas at risk of infection to set up preventive measures. Forty-three sheep and goat farms were included. Their locations were plotted and associated with T. gondii and N. caninum seroprevalence, then the distribution of farms' prevalence was evaluated by spatial analysis. Significant clusters for both low and high prevalence were obtained, and a generalized linear model with ordinal logistic regression was implemented to verify if spatial clustering could be due to climate factors (temperature, rainfall, and their interaction). Clusters of high (80.0%) and low prevalence (28.12%) resulted for T. gondii seroprevalence in sheep farms. No significant clusters resulted for goat farms. Clusters of high (38.68%) and low prevalence (21.23%) resulted for N. caninum seroprevalence in sheep farms. One high-prevalence cluster (15.62%) resulted for goat farms. For goats, spatial analysis and analysis on climatic data showed the absence of environmental significant risk factors associated with T. gondii or N. caninum infection. On the contrary, for sheep, annual temperature, rainfall, and their association affected the risk of T. gondii and N. caninum infection. Particularly, high temperatures and abundant rainfalls were related to T. gondii seroprevalence, while low temperatures and scarce rainfalls were related to N. caninum seroprevalence.


Current density imaging sequences with separation of mobile-ion current from immobile-ion current.
Three sequences for electric current density imaging (CDI) are studied by testing their ability to selectively detect the electric current of different types of charge carriers. Two of the methods are standard DC-CDI and AC-CDI sequences, and a new AC-DC-CDI sequence is also introduced. The methods rely on the application of square voltage pulses in combination with refocusing RF pulses that can, depending on their position with respect to the voltage pulses, eliminate resistive, capacitive or neither of the current components from the detection pathway. It is demonstrated by experiments on a model system that the DC-CDI sequence detects only the resistive current of mobile ions, the AC-DC-CDI sequence detects only the capacitive current of fixed ions, while the AC-CDI sequence detects both current components. The three sequences introduce a new MRI contrast mechanism that depends on the ion mobility of the sample. As such, the methods may play an important role in the study of electric conductivity phenomena in biological systems.


p53 protein expression levels as bioindicator of individual exposure to ionizing radiation by flow cytometry.
Ionizing radiation (IR) can cause various lesions in DNA, which induce the increase of p53 expression levels in order to repair radiation induced damage. Thus, the correlation between the increase of p53 expression and an irradiation may constitute a fast and powerful method of individual monitoring in cases of accidental or suspected exposures to IR. In this context, the aim of this research was to evaluate changes in lymphocyte p53 expression levels, based on flow cytometry, after in vitro irradiation of peripheral blood samples. For the measurement of such expression levels of p53 protein, an investigation was carried out in order to establish a methodology of analysis based on flow cytometry. Hence, relationships among levels of expression of p53 protein with the absorbed dose have been verified. The results presented in this report emphasized flow cytometry as an important tool for the fast evaluation of p53 protein expression levels as bioindicator of individual exposure to acute ionizing radiation.


An acute pleuropneumonia in a pig caused by Chromobacterium violaceum.
A 2.5-month-old, 30 kg Duroc pig died 10 days after showing clinical signs of dyspnoea and diarrhoea. Acute necrotizing and fibrinous pleuropneumonia with locally extensive lesions was found. Chromobacterium violaceum was isolated from pneumonic lung tissues and intratracheal inoculation of a pure culture into two SPF pigs reproduced lesions similar to those found in the natural infection.


Novel Radiobiological Gamma Index for Evaluation of 3-Dimensional Predicted Dose Distribution.
To propose a gamma index-based dose evaluation index that integrates the radiobiological parameters of tumor control (TCP) and normal tissue complication probabilities (NTCP). Fifteen prostate and head and neck (H&N) cancer patients received intensity modulated radiation therapy. Before treatment, patient-specific quality assurance was conducted via beam-by-beam analysis, and beam-specific dose error distributions were generated. The predicted 3-dimensional (3D) dose distribution was calculated by back-projection of relative dose error distribution per beam. A 3D gamma analysis of different organs (prostate: clinical [CTV] and planned target volumes [PTV], rectum, bladder, femoral heads; H&N: gross tumor volume [GTV], CTV, spinal cord, brain stem, both parotids) was performed using predicted and planned dose distributions under 2%/2 mm tolerance and physical gamma passing rate was calculated. TCP and NTCP values were calculated for voxels with physical gamma indices (PGI) >1. We propose a new radiobiological gamma index (RGI) to quantify the radiobiological effects of TCP and NTCP and calculate radiobiological gamma passing rates. The mean RGI gamma passing rates for prostate cases were significantly different compared with those of PGI (P<.03-.001). The mean RGI gamma passing rates for H&N cases (except for GTV) were significantly different compared with those of PGI (P<.001). Differences in gamma passing rates between PGI and RGI were due to dose differences between the planned and predicted dose distributions. Radiobiological gamma distribution was visualized to identify areas where the dose was radiobiologically important. RGI was proposed to integrate radiobiological effects into PGI. This index would assist physicians and medical physicists not only in physical evaluations of treatment delivery accuracy, but also in clinical evaluations of predicted dose distribution.


Surgical Correction of Distal Junctional Kyphosis.
Distal junctional kyphosis (DJK) is defined as the development of a kyphotic angulation over 10 degrees below a fusion construct and has been described as a complication of the treatment of adolescent idiopathic scoliosis, Scheuermann kyphosis, adult spinal deformity, and cervical deformity. There are some inherent risk factors to DJK: multilevel fusions, damage to the midline soft tissues including interspinous/supraspinous ligaments, T5-T12 thoracic kyphosis, T11-L2 thoracolumbar kyphosis, and increased mismatch between cervical lordosis and T1 slope. A 53-year-old male presented with cervicalgia, inability to sustain horizontal gaze, and kyphosis-enabled forward head posture. He underwent C3-T1 posterior decompression and fusion as treatment for cervical myelopathy 18 months prior. Neurologic examination was normal, with appreciable protrusion of the T1 vertebral spinous process. Surgery was initiated through subperiosteal exposure of C2-T6, followed by removal of previously set instrumentation, placement of new screws, and posterior column osteotomies of selected segments. Final steps involved compression across excised portions, locking pedicle screws, and a multirod insertion after closure of the posterior column osteotomies by compression maneuvers. Correction for DJK encompasses sagittal alignment restoration, a stable construct, and a good biological environment for healing. Failure of DJK realignment can occur if the patient's ligaments deteriorate distal to the construct or fractures develop in vertebral bodies at the lowest instrumented vertebra or lowest instrumented vertebra +1. One year after surgery, the patient's condition improved, evidenced from both patient self-report and a standing posture radiograph.


Pseudotumor complicating a well-fixed ceramic-on-polyethylene total hip arthroplasty.
A suspicious area surrounding hardware components, usually following arthroplasty, should raise awareness to a possible particle related disease. Even in the presence of confusing or less typical symptoms, clinicians and surgeons must join efforts to diagnose and treat the patient effectively and in the appropriate time.


Generation of induced pluripotent stem cells (iPSCs) stably expressing CRISPR-based synergistic activation mediator (SAM).
Human fibroblasts were engineered to express the CRISPR-based synergistic activation mediator (SAM) complex: dCas9-VP64 and MS2-P65-HSF1. Two induced pluripotent stem cells (iPSCs) clones expressing SAM were established by transducing these fibroblasts with lentivirus expressing OCT4, SOX2, KLF4 and C-MYC. We have validated that the reprogramming cassette is silenced in the SAM iPSC clones. Expression of pluripotency genes (OCT4, SOX2, LIN28A, NANOG, GDF3, SSEA4, and TRA-1-60), differentiation potential to all three germ layers, and normal karyotypes are validated. These SAM-iPSCs provide a novel, useful tool to investigate genetic regulation of stem cell proliferation and differentiation through CRISPR-mediated activation of endogenous genes.


Neurofilament Light Chain Predicts Disease Severity and Progression in Multiple System Atrophy.
Neurofilament light chain (NFL), a potential biomarker of multiple system atrophy (MSA), has been reported in several studies. The objective of this study was to investigate whether plasma NFL levels are correlated with the progression of motor and cognition function in MSA. Patients with MSA were part of a prospective cohort study with assessments at baseline and after 1 year. Plasma NFL was quantified using ultrasensitive Simoa technology. A total of 91 patients with MSA and 60 healthy controls (HCs) were enrolled. NFL levels increased from baseline to 1-year follow-up (P = 0.010). Baseline plasma NFL levels were significantly associated with motor severity and progression in patients with MSA (P < 0.05) but not with cognitive progression (P > 0.05). Plasma NFL is a reliable biomarker for the disease severity of MSA and monitoring the progression of MSA, but not the progression of cognition. © 2021 International Parkinson and Movement Disorder Society.


Large surface area activated charcoal and the inhibition of aspirin absorption.
Activated charcoal's adsorptive capacity, and therefore potential efficacy, is generally related to its surface area. In our study, the efficacy of two activated charcoal preparations, Actidose-Aqua 1,500 m2/g and Super Char, 3,000 m2/g, were compared on the basis of their ability to inhibit aspirin absorption. Twelve healthy male subjects fasted for eight hours before and four hours after a 20 mg/kg oral dose of aspirin. One hour after aspirin dosing, each subject received either no charcoal, 25 g Actidose-Aqua, or 25 g Super Char in a randomized crossover design. Each aspirin dose was separated from the previous dose by at least seven days. Total urine volumes were collected over 12-hour intervals, beginning 12 hours before the aspirin dose and continuing for 72 hours after dosing. Urine salicylate concentration was measured with a colorimetric assay. The fraction of aspirin dose recovered in the urine was 0.96 +/- 0.13, 0.78 +/- 0.18, and 0.50 +/- 0.20 for the control, Actidose-Aqua, and Super Char treatment phases, respectively. In vitro, Super Char was found to bind more salicylic acid than Actidose-Aqua at pH 8.1. We conclude that both activated charcoal preparations significantly reduced the gastrointestinal absorption of aspirin (P less than .05) and that Super Char was significantly more effective than Actidose-Aqua in reducing the gastrointestinal absorption of aspirin (P less than .01).


Effect of leaf extract of Capparis zeylanica Linn. on spatial learning and memory in rats.
This study was designed to investigate the nootropic activity of Capparis zeylanica Linn. leaves in rats. The raw material of Capparis zeylanica leaves was successively extracted with petroleum ether and methanol using a Soxhlet apparatus and macerated to form an aqueous extract. The methanolic and aqueous extracts were evaluated for their effect on spatial learning and memory in rats using the Morris water maze task. Three doses (50, 100 and 150 mg/kg; p.o.) of methanolic and aqueous extracts of Capparis zeylanica were administered for 7 successive days to separate groups of animals. Results showed that both the extracts significantly enhanced memory, as shown by decrease in escape latency time. Furthermore, methanolic and aqueous extracts in all doses tested significantly increased the time spent in the target quadrant during the probe trial, indicating retention of spatial memory of the location of a previously placed platform in the target quadrant. These findings indicate that methanolic and aqueous extracts of Capparis zeylanica Linn. leaves have potent nootropic activity. The anti-oxidant property of Capparis zeylanica may contribute favorably to the memory enhancement effect. However, further studies are needed to identify the exact mechanism of action.


Detection of colorectal carcinomas by intraoperative RIS in addition to preoperative RIS: surgical and immunohistochemical findings.
The immunoscintigraphic detection of tumour foci less than 1 cm in diameter fails even with single photon emission tomography (SPET) owing to low contrast against background activity. In an attempt to improve detection of macroscopically invisible tumour spread, intraoperative scintimetry (IOSM) with a hand-held gamma-probe was performed in addition to SPET 24-30 and 41-48 h after injection of the technetium-99m carcinoembryonic antigen (CEA MoA) on 12 patients with colorectal carcinoma and 3 patients with different neoplastic and inflammatory diseases. Tumour specimens were measured in vitro in a gamma well counter. For comparison, the presence and amount of CEA in the tumour cells were evaluated immunohistochemically. After modification, the gamma-probe originally designed for iodine-131 was 20 times more sensitive; activities of 99mTc located close to the collimator hole were measured with absolute sensitivity of 100 cps = 2.5 kBq 99mTc. The unfavourably high background activity affected both the in vitro and in vivo analysis: SPET results had been considered positive in 8 of 15 cases. In vitro tumour/non-tumour (t/nt) ratios greater than 2.0 were found in 4 cases. In vivo IOSM resulted in t/nt ratios greater than 2.0 in only 3 patients. In most cases, there was no coincidence of elevated t/nt ratios from the different methods. A correlation was derived between positive immunoscintimetric in vitro findings and immunohistochemically proven interstitial localization of CEA in tumor cells. In conclusion, the measurement technique of IOSM seems adequate, but clinical success will depend on a more specific enrichment of MoA in tumour tissue.(ABSTRACT TRUNCATED AT 250 WORDS)


A Computational Model of Protein Induced Membrane Morphology with Geodesic Curvature Driven Protein-Membrane Interface.
Continuum or hybrid modeling of bilayer membrane morphological dynamics induced by embedded proteins necessitates the identification of protein-membrane interfaces and coupling of deformations of two surfaces. In this article we developed (i) a minimal total geodesic curvature model to describe these interfaces, and (ii) a numerical one-one mapping between two surface through a conformal mapping of each surface to the common middle annulus. Our work provides the first computational tractable approach for determining the interfaces between bilayer and embedded proteins. The one-one mapping allows a convenient coupling of the morphology of two surfaces. We integrated these two new developments into the energetic model of protein-membrane interactions, and developed the full set of numerical methods for the coupled system. Numerical examples are presented to demonstrate (1) the efficiency and robustness of our methods in locating the curves with minimal total geodesic curvature on highly complicated protein surfaces, (2) the usefulness of these interfaces as interior boundaries for membrane deformation, and (3) the rich morphology of bilayer surfaces for different protein-membrane interfaces.


Potentiation of the apoptotic signaling pathway in both the striatum and hippocampus and neurobehavioral impairment in rats exposed chronically to a low-dose of cadmium.
Cadmium (Cd) is a highly toxic heavy metal. It accumulates in biological tissues, especially in fish which constitutes a first rank food for humans, particularly in the coastal areas. This study investigates the effect of long-term exposure to low Cd concentration (17 μg/kg/day) in rat striatum and hippocampus. In this study, the neurobehavioral ability changes were assessed by applying cognitive standard testing at the end of the rats' exposure period. In addition, the examination of mitochondrial swelling was performed at the same time of evaluation of its redox status in the brain regions studied through stress parameters (GSH, MDA, GST, and CAT). This study examined also whether this long-term exposure can modify the apoptotic signaling pathway via assessment of apoptotic markers (caspase-8 and 9, Bax, Bcl-2, and Cyt-c) in cell lysates. The results of this study showed changes in neurobehavioral abilities of animals and a stronger mitochondrial swelling associated with a significant decrease in antioxidant systems (GSH, GST, and CAT) and conversely an increase in the lipoperoxidation end product (MDA) in both the striatal and hippocampal mitochondria. In addition, the results revealed a significant increase in pro-apoptotic intracellular components such as caspase-9, Cyt-c, and Bax, and showed also an evident decrease in Bcl-2 levels. In conclusion, our results reported that chronic exposure to Cd produces behavioral and cognitive perturbations, enhances oxidative stress associated with mitochondrial edema and Cyt-c leakage, and, ultimately, potentiates apoptosis signaling pathway in both brain regions in rats.


A case of very late-onset systemic lupus erythematosus and updated pooled analysis of late-onset cases in the literature.
Systemic lupus erythematosus predominantly affects young women during childbearing age. It is rare after the age of 90, and there is no report of very late onset in persons over 90 in China. Here, we reported a case of a female patient, the onset age of whom was 90, analyzing her clinical features and treatments. And when conducting a pooled analysis of late-onset cases in the literature, we found that the organ damages and severity of lupus disease of old-onset SLE were not benign, and considering their ages, comorbidities and high rate of mortalities, appropriate interventions and close follow-up for this age group are needed.


Chlorine and Bromine Isotope Analysis of Polychlorinated Biphenyls and Polybrominated Diphenyl Ethers Using Gas Chromatography-Quadrupole Mass Spectrometry.
A compound-specific chlorine/bromine isotope analysis (Cl-/Br-CSIA) method was developed using gas chromatography-quadrupole mass spectrometry for polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs), which are toxic to human health and are frequently detected in various abiotic and biotic media. For PCB congeners, the molecular ion method for a concentration of 0.5-10.0 ppm, a dwell time of 20-100 ms, a relative EM voltage of 200 V, an electric current of 34 μA, and an ionization energy of 70 eV was determined as the most suitable scheme, which obtained standard deviations (SDs) of chlorine isotope ratios ranging from 0.00008 to 0.00068. As for the PBDE congeners, the lowest SDs, ranging from 0.00050 to 0.00172, were determined using the top four ion method with a concentration of 5-10 ppm and a dwell time of 20-50 ms. Both the chlorine and bromine isotope ratios showed strong concentration dependencies. Therefore, external standardization or detecting chlorine and bromine isotope ratios at a uniform concentration level is necessary to eliminate the concentration effect. In addition, 13C-correction is critical to remove interference from carbon isotopes. This newly developed Cl-/Br-CSIA method successfully determined the chlorine/bromine isotope ratios of PCBs/PBDEs in technical mixtures and traced the chlorine/bromine isotope ratio variations of PCBs/PBDEs in photodegradation experiments, thereby suggesting that it is a promising tool for assessing the sources and transformation processes of PCBs and PDBEs in the environment.


3D harmonic phase tracking with anatomical regularization.
This paper presents a novel algorithm that extends HARP to handle 3D tagged MRI images. HARP results were regularized by an original regularization framework defined in an anatomical space of coordinates. In the meantime, myocardium incompressibility was integrated in order to correct the radial strain which is reported to be more challenging to recover. Both the tracking and regularization of LV displacements were done on a volumetric mesh to be computationally efficient. Also, a window-weighted regression method was extended to cardiac motion tracking which helps maintain a low complexity even at finer scales. On healthy volunteers, the tracking accuracy was found to be as accurate as the best candidates of a recent benchmark. Strain accuracy was evaluated on synthetic data, showing low bias and strain errors under 5% (excluding outliers) for longitudinal and circumferential strains, while the second and third quartiles of the radial strain errors are in the (-5%,5%) range. In clinical data, strain dispersion was shown to correlate with the extent of transmural fibrosis. Also, reduced deformation values were found inside infarcted segments.


[Isolation of several species of the genus Aspergillus from soil of intrahospital ornamental plants].
The earth of ornamental plants is one of the main reservoirs of Aspergillus type of fungi in hospital areas. We studied 174 ornamental interior plants from a hospital at Santiago. Samples were obtained from the soil surface and sowed in Sabouraud-glucose agar, adding streptomycin and G-penicillin. After 72 h of culture, at least one strain of Aspergillus was isolated from 140 samples (80.5%). The most frequently isolated strain was A fumigatus (129 samples), followed by A niger (75 samples). A fumigatus and A niger were the only isolated strains in 65 and 11 samples respectively. These findings confirm that ornamental plants can be important reservoirs of Aspergillus strains, a potential infectious agent for immunocompromised patients, in hospital areas.


Outbreak studies of a GII-3 and a GII-4 norovirus revealed an association between HBGA phenotypes and viral infection.
Noroviruses are the major viral pathogen of epidemic acute gastroenteritis. Two outbreaks of norovirus gastroenteritis that occurred in China in 2003 and 2006, caused by a GII-4 and a GII-3 strain, respectively, were studied to investigate potential association between viral infection and histo-blood types of hosts. The histo-blood group antigen (HBGA) phenotypes of 146 subjects (16 from the GII-3 and 130 from the GII-4 outbreaks) were determined in a saliva-based EIA. Our results showed that the secretor status of individuals was strongly associated with infection in the two outbreaks (P=0.0007, OR=0.044, 95% CI, 0.003-0.765); none of the nonsecretor in either outbreak developed symptomatic infection. The infection rate of individuals with the ABH and Lewis blood types varied between the two outbreaks. In the GII-4 outbreak, association of ABH blood types with noroviral infection (P=0.001, Chi-square=16.13) has been observed, in which the type A individuals had an increased risk of infection [61% in the symptomatic group (n=41) vs. 30% in the asymptomatic group (n=89), P=0.0001], while the type O individuals showed a decreased infection rate (17% vs. 48% in the two groups, P=0.0048). In the GII-3 outbreak, however, individuals with the H antigen only appeared to have a higher rate of infection (33% vs. 14%, P=0.059). Our study provided further evidence in the association between noroviral infection and the HBGA types of hosts. While the nonsecretor phenotype appears naturally resistant to these two strains, additional determinants on the HBGAs also may play roles in host range of the two strains.


In acute ischemic stroke, EVT alone vs. IV alteplase plus EVT did not differ for functional outcome at 90 d.
LeCouffe NE, Kappelhof M, Treurniet KM, et al. A randomized trial of intravenous alteplase before endovascular treatment for stroke. N Engl J Med. 2021;385:1833-44. 34758251.


Approach to the diagnosis of interstitial lung disease.
Interstitial lung diseases (ILDs) encompass a wide range of diffuse pulmonary disorders, characterized by a variable degree of inflammatory and fibrotic changes of the alveolar wall and eventually the distal bronchiolar airspaces. ILDs may occur in isolation or in association with systemic diseases. The clinical evaluation of a patient with ILD includes a thorough medical history and detailed physical examination; obligatory diagnostic testing includes laboratory testing, chest radiography, and high-resolution computed tomography and comprehensive pulmonary function testing and blood gas analysis. To optimize the diagnostic yield, a dynamic interaction between the pulmonologist, radiologist, and pathologist is mandatory.


Preparation, mass diffusion, and biocompatibility analysis of porous-channel controlled calcium-alginate-gelatin hybrid microbeads for in vitro culture of NSCs.
The Ca-alginate/gelatin (CAG) microbeads were prepared and evaluated through assays for their mechanical strength, permeability, and the feasibility as a cell carrier for in vitro culture of neural stem cells. The effects of different concentrations of sodium alginate, gelatin, and calcium chloride on the mechanical strength of CAG microbeads were determined using a self-made puncture force tester. Following this, the microbeads were immersed in DMEM media for a specified period to test its decay resistance. A diffusion model including a calculation formula of diffusion coefficient was built to investigate the diffusion of glucose and bovine serum albumin (BSA) through the wall of the microbeads. Furthermore, the feasibility of the microbeads for in vitro culture was identified using neural stem cells from Kunming mouse. Through a systematic approach and comprehensive analysis, the optimal gelatin conditions for microbead preparation were determined; the final combination of parameters of 1.5 % (wt%) sodium alginate (SA), 0.5 % (wt%) gelatin, and 4 % (wt%) CaCl2 were the best conditions for NSC cultures. This experiment demonstrated that CAG microbeads had good cytocompatibility that made it suitable for the culture and successfully maintained stemness of neural stem cells.


Trends in the characteristics of human functional genomic data on the gene expression omnibus, 2001-2017.
The gene expression omnibus (GEO) is the world's largest public repository of functional genomic data. Despite its broad use in secondary genomic analyses, the temporal trends in the characteristics of genomic data on GEO, including experimental procedures, geographic origin, funder(s), and related disease, have not been examined. We identified 75,376 Series deposited to the GEO during 2001-2017 and built a database of all human genomic data (39,076 Series, 51.8% of all Series). Using the associated publications, we obtained funding information and identified the related disease area. Of the Series with classified disease areas, the two most common were cancer (n = 12,688, 32.5%) and immunologic diseases (n = 2,393, 6.1%), while the percentages of all other disease areas were below 5%, including neurological diseases (n = 1733, 4.4%), infectious diseases (n = 1225, 3.1%), diabetes (n = 828, 2.1%), and cardiovascular diseases (n = 299, 0.8%). In recent years, there has been a significant increase in the use of high-throughput sequencing (HTS), protein array and multiple-platform technologies, as well as in the proportion of North American deposits. Compared to those from other regions, North American deposits appeared to lead the shift from array-based to HTS technologies (odds ratio [OR], 95% confidence intervals [CI] = 3.39, 3.23-3.55, P = 9.40E-323), and were less likely to focus on a major disease area (OR = 0.64, 95% CI: 0.61-0.67, P = 5.02E-107), suggesting a greater emphasis on basic science in North America. Furthermore, the Series utilizing HTS were less likely to be disease-classified compared to other technologies (OR = 0.39, 95% CI: 0.37-0.41, P = 1.00E-322), suggesting a preferential use or adoption of HTS in basic science settings. Finally, funding from the NHGRI, NCI, NIEHS, and NCCR resulted in a higher number of GEO Series per grant than other NIH institutes, demonstrating different preferences on genomic studies among awardees of NIH institutes. Our findings demonstrate geographic, technological, and funding disparities in the trends of GEO deposit characteristics.


Inflammation and atherosclerosis: disease modulating therapies.
Advances in the mechanistic understanding of atheroma initiation, repair, progression, and rupture have solidified the pivotal role played by the immune system in the pathophysiology of atherosclerotic vascular disease. These mechanistic findings have been extended into humans, with a strong evidence basis for the independent association between elevated blood markers of inflammation and future cardiovascular (CV) events. Investigations with statins as well as more conventional anti-inflammatory medications provide indirect evidence to support the concept that modifying immune responses can improve CV outcomes; however, robust evidence to support the use of anti-inflammatory treatment strategies to manage atherosclerotic vascular disease is still lacking. Such evidence may emerge from a new wave of clinical trials directly exploring the effects of targeted immune modulation on CV risk. These trials will provide key additional insights into atherosclerosis and will help determine the fate of immune modulation as a new treatment strategy in atherosclerotic vascular disease.


Determining the impact of roasting degree, coffee to water ratio and brewing method on the sensory characteristics of cold brew Ugandan coffee.
In today's market, there is a growing demand for high-quality coffee with distinctive sensory characteristics, such a cold brew (CB) coffee. The objectives of this study were to determine the impact of a) degree of roasting, b) coffee to water ratio (C2WR), and c) brewing method on the sensory characteristics of CB coffee from d) various Ugandan coffee beans, both Robusta and Arabica. Four distinct coffee samples, sourced from lowland and mountainous regions in Uganda, were roasted and tested using a factorial design. A highly trained sensory panel evaluated the samples using 42 attributes. Results showed that all factors studied had an impact on most of the attributes. Not surprisingly, Robusta coffees generally had more bitter taste than Arabica coffees and the dark roast samples generally were more bitter than the medium roast coffees. Also, coffee samples that were brewed using a higher C2WR generally were more bitter than the coffees using a lower C2WR. However, although most of the main effects had a significant impact, their effects were mitigated by their interaction with other factors. For example, Medium roast Robusta that was slow-dripped with a high C2WR had a more bitter taste than the corresponding Arabica samples. However, when the Medium roast Robusta was steeped with a high C2WR it had a similar bitter intensity with the corresponding Arabica samples. Thus, although major impacts are critical, individual sample combinations must be considered when evaluating coffee samples for their impact on the sensory characteristics.


Anti-CD28 antibody-induced kidney allograft tolerance related to tryptophan degradation and TCR class II B7 regulatory cells.
B7/CTLA-4 interactions negatively regulate T-cell responses and are necessary for transplant tolerance induction. Tolerance induction may therefore be facilitated by selectively inhibiting the B7/CD28 pathway without blocking that of B7/CTLA-4. In this study, we selectively inhibited CD28/B7 interactions using a monoclonal antibody modulating CD28 in a rat model of acute kidney graft rejection. A short-term treatment abrogated both acute and chronic rejection. Tolerant recipients presented few alloantibodies against donor MHC class II molecules, whereas untreated rejecting controls developed anti-MHC class I and II alloantibodies. PBMC from tolerant animals were unable to proliferate against donor cells but could proliferate against third-party cells. The depletion of B7+, non-T cells fully restored this reactivity whereas purified T cells were fully reactive. Also, NK cells depletion restored PBMC reactivity in 60% of tolerant recipients. Conversely, NK cells from tolerant recipients dose-dependently inhibited alloreactivity. PBMC anti-donor reactivity could be partially restored in vitro by blocking indoleamine-2,3-dioxygenase (IDO) and iNOS. In vivo, pharmacologic inhibition of these enzymes led to the rejection of the otherwise tolerated transplants. This study demonstrates that an initial selective blockade of CD28 generates B7+ non-T regulatory cells and a kidney transplant tolerance sustained by the activity of IDO and iNOS.


The analgesic effect of continuous subacromial bupivacaine infusion after arthroscopic shoulder surgery: a randomized controlled trial.
The benefits of subacromial local anesthetic infusion are controversial. The aim of this study was to evaluate the efficacy of subacromial bupivacaine infusion after arthroscopic acromionplasty and rotator cuff procedures. Ninety patients with subacromial impingement disease scheduled for elective shoulder arthroscopy were enrolled in this randomized, prospective and placebo-controlled study. The patients received subacromially either 5.0 mg/ml bupivacaine or 9.0 mg/ml saline at the rate of 2 ml/h post-operatively for 48 h. The primary outcome measure was the use of rescue oxycodone. The consumption of opioids, ibuprofen, paracetamol and codeine, and the intensity of pain were recorded. Patients receiving bupivacaine infusion used significantly less oxycodone compared with patients receiving saline [15 vs. 20 mg (median) oxycodone intravenously on the day of surgery, 0 vs. 10 mg (median) perorally on the first post-operative day]. There was no significant difference in the use of ibuprofen, paracetamol and codeine, except that paracetamol was used more in patients receiving saline during the day of surgery (P = 0.009). The pain scores of patients receiving bupivacaine were lower at 18 h (P = 0.008). Average pain scores for the worst pain experienced did not differ between the groups. There were three (4%) premature discontinuations because of technical problems. Considering the low need of opioids, expenses and technical problems in these patients, subacromial 5.0 mg/ml bupivacaine infusion 2 ml/h confers only moderate and probably clinically insignificant efficacy compared with placebo. This efficacy might be overweighed by the disadvantages.


The role of calcium, magnesium, and zinc in carcinogenesis.
The effects of calcium, magnesium, and zinc supplementation and of magnesium depletion on carcinogenesis are comprehensively reviewed, including epidemiologic and experimental investigations. Some data on the effects of neoplasia on the homeostasis of these metals are also presented. Despite many conflicting results, this review reveals that calcium supplementation is more likely to enhance than inhibit chemical carcinogenesis; magnesium or zinc supplementation tends to inhibit carcinogenesis; magnesium deficiency increases the incidence of neoplasia in humans and animals; parenteral administration of magnesium along with a carcinogen produces local anticarcinogenic effects, while zinc's activity tends to be systemic; and there is a simple correlation between the inhibition of carcinogenesis by the magnesium and zinc supplementation and the reduction of carcinogen binding to cells and DNA. The mechanisms of these effects are not clear. They may involve molecular interactions between metal and carcinogen at different enzymatic and regulatory sites of target cells undergoing neoplastic transformation, as well as stimulation of the host immune system.


Determination of trace elements in human liver biopsy samples by ICP-MS and TXRF: hepatic steatosis and nickel accumulation.
Human liver biopsy samples, collected from 52 individuals, were analysed by inductively coupled plasma-mass spectrometry (ICP-MS) and total reflection X-ray fluorescence (TXRF) spectrometry in a retrospective study (i.e. patient selection and liver biopsy were not for the purpose of element analysis). The freeze-dried samples (typically 0.5-2 mg dry weight) were digested in a laboratory microwave digestion system and solutions with a final volume of 1 mL were prepared. The concentrations of Cr, Mn, Fe, Ni, Cu, Zn, Rb, and Pb were determined by use of a Thermo Elemental X7 ICP-MS spectrometer. TXRF measurements were performed with an Atomika Extra IIA spectrometer. Yttrium was employed as an internal standard, prepared by dissolution of 5N-purity yttria (Y(2)O(3)) in our laboratory. The accuracy was tested by analysis of NIST 1577a Bovine Liver certified reference material. The concentrations of Fe, Cu, Zn, and Rb determined in human liver biopsy samples were in good agreement with data published by other authors. The distribution of nickel in the samples was surprisingly uneven-nickel concentrations ranged from 0.7 to 12 microg g(-1) (dry weight) in 38 samples and in several samples were extremely high, 36-693 microg g(-1). Analysis of replicate procedural blanks and control measurements were performed to prevent misinterpretation of the data. For patients with steatosis (n=14) Ni concentrations were consistently high except for two who had levels close to those measured for the normal group. As far as we are aware no previous literature data are available on the association of steatosis with high concentration of nickel in human liver biopsies taken from living patients.


Low-field one-dimensional and direction-dependent relaxation imaging of bovine articular cartilage.
The structure of articular cartilage is separated into three layers of differently oriented collagen fibers, which is accompanied by a gradient of increasing glycosaminoglycan (GAG) and decreasing water concentration from the top layer towards the bone interface. The combined effect of these structural variations results in a change of the longitudinal and transverse relaxation times as a function of the distance from the cartilage surface. In this paper, this dependence is investigated at a magnetic field strength of 0.27 T with a one-dimensional depth resolution of 50 μm on bovine hip and stifle joint articular cartilage. By employing this method, advantage is taken of the increasing contrast of the longitudinal relaxation rate found at lower magnetic field strengths. Furthermore, evidence for an orientational dependence of relaxation times with respect to an axis normal to the surface plane is given, an observation that has recently been reported using high-field MRI and that was explained by preferential orientations of collagen bundles in each of the three cartilage zones. In order to quantify the extent of a further contrast mechanism and to estimate spatially dependent glycosaminoglycan concentrations, the data are supplemented by proton relaxation times that were acquired in bovine articular cartilage that was soaked in a 0.8 mM aqueous Gd++ solution.


A triple blind, placebo controlled, randomised controlled trial of betahistine dihydrochloride in the treatment of primary tinnitus.
To evaluate the effectiveness of betahistine in the treatment of primary tinnitus. To evaluate the effectiveness of betahistine in the treatment of primary tinnitus. Universidade estadual Paulista Julio de Mesquita Filho, Botucatu Medical School, São paulo, Brazil. Adult patients with primary tinnitus who had not undergone treatment for tinnitus in the last 6 months were included. Patients with profound sensorineural deafness, hearing aid users and patients with metabolic, neurological, psychiatric or decompensated cardiovascular diseases were excluded. in the betahistine group, patients received betahistine 24 mg every 12 h for 90 days; in the control group, patients received placebo tablets every 12 h for 90 days. Primary outcome measure: Tinnitus Handicap Inventory (THI). Clinical Global Impression Improvement (CGI-I) and a question of 'Yes' or 'No' to participants about their perception of improvement in symptoms. Of 284 participants initially identified, 62 were randomised (betahistine group n = 31; control group n = 31). Median age (IQR) 54 (48-60) years, with a balanced number of men and women. There was no difference in THI outcome between the study groups (median difference, -2 points; 95% CI, -8 to 6 points); the THI after the intervention was a median (IQR) 4 (-4 to 14) lower points in the betahistine group, and a median (IQR) 2 (-6 to 10) in the control group. There was no statistical difference in secondary outcome measures. Adverse events were mild and there was no statistical difference between groups. Betahistine dihydrochloride was ineffective in the treatment of primary tinnitus in adults.


Ecological processes underlying community assembly of aquatic bacteria and macroinvertebrates under contrasting climates on the Tibetan Plateau.
Understanding the role of climatic variation on biodiversity is of chief importance due to the ongoing biodiversity loss and climate change. Freshwaters, one of the most threatened ecosystems in the world, offer a valuable context to study biodiversity patterns of distinct organism groups in relation to climatic variation. In the Tibetan Plateau biodiversity hotspot - Hengduan Mountain region, we studied the effects of climate and local physico-chemical factors on stream microorganisms (i.e. bacteria) and macroorganisms (i.e. macroinvertebrates) in two parallel catchments with contrasting precipitation and temperature, that is, the Nujiang and Lancang Rivers. Diversities and community structures were better explained by climatic and local environmental variables in the drier and colder catchment and at higher elevations, than in the warmer and wetter conditions and at lower elevations. This suggests that communities may be more strongly assembled by deterministic processes in the former, comparatively harsher conditions, compared to the latter, more benign conditions. Macroinvertebrates were more strongly affected by climatic and local environmental factors compared to bacteria, but the diversities and community structures of the two groups showed spatially similar responses to overall abiotic variation, being especially evident with their community structures' responses to climate. Furthermore, bacterial and macroinvertebrate diversities were positively correlated in the drier and colder catchment, implying that these biologically and ecologically distinct organism groups are likely to be driven by similar processes in areas with such climatic conditions. We conclude that changes in climatic and local environmental conditions may affect the diversity of macroorganisms more strongly than that of microorganisms, at least in subtropical mountainous stream ecosystems studied here, but simultaneous responses of both groups to environmental changes can also be expected.


Selective hippocampal neurodegeneration in transgenic mice expressing small amounts of truncated Aβ is induced by pyroglutamate-Aβ formation.
Posttranslational amyloid-β (Aβ) modification is considered to play an important role in Alzheimer's disease (AD) etiology. An N-terminally modified Aβ species, pyroglutamate-amyloid-β (pE3-Aβ), has been described as a major constituent of Aβ deposits specific to human AD but absent in normal aging. Formed via cyclization of truncated Aβ species by glutaminyl cyclase (QC; QPCT) and/or its isoenzyme (isoQC; QPCTL), pE3-Aβ aggregates rapidly and is known to seed additional Aβ aggregation. To directly investigate pE3-Aβ toxicity in vivo, we generated and characterized transgenic TBA2.1 and TBA2.2 mice, which express truncated mutant human Aβ. Along with a rapidly developing behavioral phenotype, these mice showed progressively accumulating Aβ and pE3-Aβ deposits in brain regions of neuronal loss, impaired long-term potentiation, microglial activation, and astrocytosis. Illustrating a threshold for pE3-Aβ neurotoxicity, this phenotype was not found in heterozygous animals but in homozygous TBA2.1 or double-heterozygous TBA2.1/2.2 animals only. A significant amount of pE3-Aβ formation was shown to be QC-dependent, because crossbreeding of TBA2.1 with QC knock-out, but not isoQC knock-out, mice significantly reduced pE3-Aβ levels. Hence, lowering the rate of QC-dependent posttranslational pE3-Aβ formation can, in turn, lower the amount of neurotoxic Aβ species in AD.


[His-bundle stimulation and alternative RV stimulation sites].
The increasing evidence that right ventricular (RV) apical stimulation is detrimental for cardiac mechanics and increases the risk of new onset heart failure and mortality promotes strategies to avoid ventricular pacing and - if not applicable due to atrioventricular block - stipulates the search for alternative techniques of pacing the heart from the right ventricle. Despite more than 15 years of clinical research it remains unclear whether pacing from the right ventricular outflow tract or septum, dual site RV stimulation or selective pacing of the His bundle results in hemodynamic and prognostic benefit over the traditional method of apical RV pacing. The article reviews pertinent literature and tries to demonstrate pathophysiological mechanisms.


Objective evaluation of upper limb claudication: use of isokinetic dynamometry.
The objective of this work is to present an objective, practical, and reproducible method for evaluating the functional limitation caused by occlusive arterial disease in upper limbs: a stress test using an isokinetic dynamometer. Twenty-three patients with unilateral subclavian artery occlusive disease were included in the study, forming group 1. Seven patients of similar age, with atherosclerotic or Takayasu's disease in the aorto-iliac segment, without subclavian artery occlusive disease, were included as a control group (group 2). For all tests, we utilized the CYBEX 6000 isokinetic dynamometer. The elbow was tested using a series of 30 repetitions of extension and flexion of the arm, performed up to a maximum of 270 repetitions (9 series) or until the limit of the tested limb was reached. We initially compared all the limbs without arterial disease of both groups to analyze whether they presented similar functional performance. No significant difference was found for any of the parameters studied. Considering that upper limbs without arterial disease have a similar response to exercise in these analyzed parameters, we compared the upper limbs in group 1 (with and without subclavian artery occlusive disease). For all parameters, the limbs with subclavian artery occlusive disease presented significantly lower values than the control limbs (P < 0.05), which can be objectively attributed to the ischemia (there were different responses in different individuals, which allows the quantification of the limitation caused by subclavian artery occlusive disease). In this study, we present a new stress test for impairment in patients with subclavian artery occlusive disease that might facilitate the classification of patients according to their functional impairment, and thus result in a better choice of therapeutic approach for each case as well as reporting of objective parameters that allow comparisons of the results of different treatments, including for long-term follow-up.


Distribution of sequence differences in influenza N9 neuraminidase of tern and whale viruses and crystallization of the whale neuraminidase complexed with antibodies.
Neuraminidase genes from A/tern/Australia/G70C/75 (H11N9) and A/whale/Maine/1/84 (H13N9) influenza viruses have been sequenced. Seventy-two nucleotide changes were found, 17 of which result in changes in the amino acid sequence of the neuraminidase; 3 in the stalk region and 14 in the heads. To our surprise, all of the sequence changes in the head region are located on the base of the neuraminidase tetramer, resulting in conservation of antigenic sites on top of the neuraminidase which vary extensively in human influenza virus neuraminidase. Whale N9 neuraminidase, like tern N9 neuraminidase, possesses high levels of hemagglutinating activity but, unlike the tern neuraminidase, failed to form large well-ordered crystals. However, when the neuraminidase was complexed with Fab fragments of monoclonal antibodies, which were made against the tern N9 neuraminidase, large crystals of the complexes were obtained which diffract X-rays to beyond 3 A.


Efficacy and Safety of HSK3486 for Anesthesia/Sedation in Patients Undergoing Fiberoptic Bronchoscopy: A Multicenter, Double-Blind, Propofol-Controlled, Randomized, Phase 3 Study.
Fiberoptic bronchoscopy is a complex procedure with the need for sufficient patient anesthesia/sedation while maintaining safety. This trial aimed to evaluate the efficacy, safety, and pharmacokinetics of HSK3486 during fiberoptic bronchoscopy. This multicenter, double-blind, randomized, non-inferiority, parallel-group phase 3 trial was conducted in patients who underwent fiberoptic bronchoscopy. Patients randomly received HSK3486 0.4 mg/kg (N = 134) or propofol 2.0 mg/kg (N = 133). The primary efficacy endpoint was the successful rate of fiberoptic bronchoscopy, and secondary efficacy endpoints included successful induction of anesthesia/sedation, duration, time to being fully alert, and time to patient discharge. Safety assessments and drug concentrations were also measured. A total of 267 patients completed fiberoptic bronchoscopy, with a success rate of 100% and a 95% confidence interval of - 2.8 to 2.8% for the difference between the groups, which met the predesigned criteria of > - 8%, confirming the non-inferiority of anesthesia/sedation produced by HSK3486 compared to propofol. Among the secondary efficacy endpoints, only time to full alertness (median 8.50 vs. 6.00 min, P = 0.012) and time to discharge (median 13.00 vs. 9.87 min, P = 0.002) were slightly longer in the HSK3486 group. The incidence of adverse events was significant lower in the HSK3486 group (52.6 vs. 76.5%, P < 0.001) mainly because of less pain on injection (4.4 vs. 39.4%, P < 0.001) compared to the propofol group. HSK3486 had a similar terminal elimination half-life as propofol. HSK3486 exhibited non-inferiority anesthesia/sedation compared to propofol in patients undergoing fiberoptic bronchoscopy, and had a good safety profile with a lower incidence of pain on injection. Clinicaltrials.gov, NCT04111159, registered on 1 October 2019.


Nucleolar targeting of proteins by the tandem array of basic amino acid stretches identified in the RNA polymerase I-associated factor PAF49.
There is accumulating evidence to indicate that the regulation of subnuclear compartmentalization plays important roles in cellular processes. The RNA polymerase I-associated factor PAF49 has been shown to accumulate in the nucleolus in growing cells, but disperse into the nucleoplasm in growth-arrested cells. Serial deletion analysis revealed that amino acids 199-338 were necessary for the nucleolar localization of PAF49. Combinatorial point mutation analysis indicated that the individual basic amino acid stretches (BS) within the central (BS1-4) and the C-terminal (BS5 and 6) regions may cooperatively confer the nucleolar localization of PAF49. Addition of the basic stretches in tandem to a heterologous protein, such as the interferon regulatory factor-3, translocated the tagged protein into the nucleolus, even in the presence of an intrinsic nuclear export sequence. Thus, tandem array of the basic amino acid stretches identified here functions as a dominant nucleolar targeting sequence.


Nonunion of extended trochanteric osteotomies in impaction grafting femoral revisions.
Extended trochanteric osteotomies have been recommended to facilitate femoral component removal, femoral cement removal, and acetabular exposure in cases of difficult revision hip arthroplasty. Complications due to the osteotomy have been rare and no nonunions have been reported when this osteotomy has been used in conjunction with extensively porous-coated implants. It has been suggested that the osteotomy should also work well with impaction grafting revisions. This is a report of two cases of nonunion of extended trochanteric osteotomies in which the impaction grafting technique was used.


The effect of Chlamydia pneumoniae on the expression of peroxisome proliferator-activated receptor-gamma in vascular smooth muscle cells.
This study was designed to investigate the change of peroxisome proliferator-activated receptor gamma (PPARgamma) after the infection of the human coronary artery smooth muscle cells (HCSMCs) with Chlamydia pneumoniae (C. pneumoniae) and the effect of PPARgamma agonist on the expression of PPARgamma of C. pneumoniae-infected HCSMCs. To determine the effect of PPARgamma agonist on the proliferation of C. pneumoniae-infected HCSMCs, rosiglitazone at various concentrations was applied 1 hour before inoculation of HCSMCs. The expression of PPARgamma mRNA in HCSMCs increased from 3 hours after C. pneumoniae infection and reached that of noninfected HCSMCs at 24 hours (p<0.05). The expression of PPARgamma protein in HCSMCs also increased from 3 hours after C. pneumoniae and persisted until 24 hours as compared with that of noninfected HCSMCs (p<0.05). The pretreatment of HCSMCs with rosiglitazone followed by the infection with C. pneumoniae augmented the expression of PPARgamma mRNA and protein (p<0.05) and decreased cell proliferation. Our results showed that the expression of PPARgamma increases in response to C. pneumoniae infection and rosiglitazone further augmented the expression of PPARgamma. It is suggested that rosiglitazone could ameliorate the chronic inflammation in the vessel wall induced by C. pneumoniae by augmenting PPARgamma expression.


[MiR-140-3p Downregulation in Association with PDL-1 Overexpression in Many Cancers: A Review from the Literature Using Predictive Bioinformatics Tools].
Programmed death-ligand 1 (PD-L1) has been speculated to play a critical role in suppression of the immune system and it can be upregulated in cancer cells, which may allow cancers to evade the host immune system. MicroRNAs (miRNAs) are small non-coding RNA molecules (containing about 22 nucleotides), that function in RNA silencing and post-transcriptional regulation of gene expression. MiRNAs were found deregulated (upregulated or downregulated) and implicated in cancer development with various roles which depend on their gene target. Using targetscan web server prediction algorithm, we concluded that miR-140-3p is a targeting mirRNA with conserved consequential pairing of target region for PD-L1. Moreover, by reviewing all the available cancer studies in Pub/Medline about miR-140-3p, was found permanently down regulated. Furthermore, in recent immunotherapy related clinical trials in most cancers, evaluated PD-L1, it is found overexpressed. In the near future, in vitro or in vivo studies need to validate whether there is direct correlation between PD-L1 overexpression and miR-140-3p downregulation as targetscan performed algorithm predicted.


Smith-Magenis syndrome.
Smith-Magenis syndrome (SMS) is a mental retardation syndrome with distinctive behavioral characteristics, dysmorphic features, and congenital anomalies ascribed to an interstitial deletion of chromosome 17p11.2. Severe sleep disturbances and maladaptive daytime behavior have been linked to an abnormal circadian rhythm of melatonin with a diurnal instead of nocturnal secretion of this hormone. SMS provides a demonstration of a biological basis for sleep disorder in a genetic disease. Considering that clock genes mediate the generation of circadian rhythm, haploinsufficiency for a circadian system gene, mapping to chromosome 17p11.2, may cause the inversion of the melatonin circadian rhythm in SMS. The disorder of circadian timing in SMS may also affect entrainment pathway (retinohypothalamic tract), pacemaker functions (suprachiasmatic nuclei), or synthesis and release of melatonin by the pineal gland. Elucidating pathophysiological mechanisms of behavioral phenotypes in genetic disease can provide an original therapeutic approach in SMS: blockade of endogenous melatonin production during the day combined with exogenous melatonin administration in the evening.


Performance of calcaneus quantitative ultrasound and dual-energy X-ray absorptiometry in the discrimination of prevalent asymptomatic osteoporotic fractures in postmenopausal women.
Due to its low cost, portability, and nonionizing radiation, quantitative ultrasound (QUS) of the heel is an alternative to the measurement with dual X-ray absorptiometry (DXA) in the evaluation of bone status. The objective of the study is to compare in asymptomatic postmenopausal women the ability of QUS and DXA to discriminate between those with and without prevalent vertebral fractures (VFs). The study cohort consists of a population of 295 postmenopausal women aged between 60 and 84 (mean age, weight and BMI of 66.3 years, 72.0 kg and 29.4 kg/m(2), respectively). Lateral VFA images and scans of the lumbar spine and proximal femur were obtained by two technologists using a GE Healthcare Lunar Prodigy densitometer. VFs were defined using a combination of Genant semiquantitative (SQ) approach and morphometry. All women had a calcaneous QUS examination. The mean age of the women in our sample was 66.3 (+/-5.3) years, ranging from 60 to 84 years. Eighty-seven (29.3%) women had VFs Genant grade 2 and 3. Patients with VFs had an age and a number of years of menopause higher to those without VFs, but showed lower height, weight, and BMI. All densitometric and ultrasonometric measurements were significantly reduced in women with VFs. The intercorrelations of BMD at different sites were high, and the correlations of BUA with BMD were lower. BUA correlated weakly with total hip BMD (r = 0.36), lumbar spine BMD (r = 0.32), and much less with femur BMD (r = 0.30); all correlations were significant (P < 0.01). Analysis of the AUC for the ROC curves showed lumbar spine T-score below -2.5 to provide consistently the highest AUC (0.64). Age-adjusted ORs after correction for confounding variables (years of menopause, weight, height, and BMI) for QUS and BMD measurements showed that only lumbar spine T-score below -2.5 could predict significantly the presence of VFs (OR, 1.94; 95%CI, 1.02-3.41). Lumbar spine BMD (and not QUS) was able to discriminate asymptomatic postmenopausal women with prevalent VFs from women without VFs and independently contributed to determining the association with fracture. The combination of QUS and BMD did not improve the diagnostic ability of either individual technique.


Hexameric structure of the flagellar master regulator FlhDC from Cupriavidus necator and its interaction with flagellar promoter DNA.
Bacterial flagella are assembled with ∼30 different proteins in a defined order via diverse regulatory systems. In gram-negative bacteria from the Gammaproteobacteria and Betaproteobacteria classes, the transcription of flagellar genes is strictly controlled by the master regulator FlhDC. In Gammaproteobacteria species, the FlhDC complex has been shown to activate flagellar expression by directly interacting with the promoter region in flagellar genes. To obtain the DNA-binding mechanism of FlhDC and determine the conserved and distinct structural features of Betaproteobacteria and Gammaproteobacteria FlhDCs that are necessary for their functions, we determined the crystal structure of Betaproteobacteria Cupriavidus necator FlhDC (cnFlhDC) and biochemically analyzed its DNA-binding capacity. cnFlhDC specifically recognized the promoter DNA of the class II flagellar genes flgB and flhB. cnFlhDC adopts a ring-like heterohexameric structure (cnFlhD4C2) and harbors two Zn-Cys clusters, as observed for Gammaproteobacteria Escherichia coli FlhDC (ecFlhDC). The cnFlhDC structure exhibits positively charged surfaces across two FlhDC subunits as a putative DNA-binding site. Noticeably, the positive patch of cnFlhDC is continuous, in contrast to the separated patches of ecFlhDC. Moreover, the ternary intersection of cnFlhD4C2 behind the Zn-Cys cluster forms a unique protruding neutral structure, which is replaced with a charged cavity in the ecFlhDC structure.


Practical Analysis of Hi-C Data: Generating A/B Compartment Profiles.
Recent advances in next-generation sequencing (NGS) and chromosome conformation capture (3C) analysis have led to the development of Hi-C, a genome-wide version of the 3C method. Hi-C has identified new levels of chromosome organization such as A/B compartments, topologically associating domains (TADs) as well as large megadomains on the inactive X chromosome, while allowing the identification of chromatin loops at the genome scale. Despite its powerfulness, Hi-C data analysis is much more involved compared to conventional NGS applications such as RNA-seq or ChIP-seq and requires many more steps. This presents a significant hurdle for those who wish to implement Hi-C technology into their laboratory. On the other hand, genomics data repository sites sometimes contain processed Hi-C data sets, allowing researchers to perform further analysis without the need for high-spec workstations and servers. In this chapter, we provide a detailed description on how to calculate A/B compartment profiles from processed Hi-C data on the autosomes and the active/inactive X chromosomes.


Photochemistry of phthalimidoadamantane dipeptides: effect of amino acid side chain on photocyclization.
A series of dipeptides 1 was synthesized that at the N-site contained 3-(N-phthalimidoadamantane-1-carboxylic acid and at the C-site different aliphatic or aromatic L- or D-amino acids. The photochemical reaction of dipeptides 1 under acetone-sensitized conditions gave simple decarboxylation products 6, and decarboxylation-induced cyclization products 7, as well as some secondary products 8 and 9 formed by elimination of H2O or ring enlargement, respectively. Molecules 9 undergo secondary photoinduced H-abstractions by the phthalimide chromophore, delivering more complex polycycles 11. The photodecarboxylation-induced cyclization to 7 was observed only with phenylalanine (Phe), proline (Pro), leucine (Leu) and isoleucine (Ile). Contrary to dipeptides with Phe, the cyclization takes place with almost complete racemization at the amino acid chiral center, but diastereoselectively giving only one pair of enantiomers. The conducted investigation is important as it provides the breath and the scope of dipeptide cyclizations activated by phthalimides.


Do age, gender, BMI and disease duration influence the clinical outcomes in patients of knee osteoarthritis treated with serial injections of autologous platelet rich plasma?
To study whether age, gender, body mass index(BMI) and disease duration influence the clinical outcomes in kellgren-Lawrence(K-L) grade II,III knee osteoarthritis(KOA) patients treated with serial injections of platelet rich plasma(PRP). 65 patients were given three monthly intra-articular injections of PRP in this prospective interventional study. The patients were divided into subgroups depending on the factor studied: by age(in years) into young <45(n = 7), middle age 45-60(n = 35), and elderly >60(n = 23): by BMI(in kg/m2) into; normal <25(n = 25), overweight 25-30(n = 27) and obese >30(n = 13) and disease duration; less(n = 32) or more than 1 year(n = 33) symptom duration. Visual analogue scale (VAS) and Western Ontario and McMaster Universities Arthritis Index (WOMAC) were used as outcome measures and assessed before each injection and then at 6 and 9 months post injection. Groups were homogenous with respect to baseline characteristics. Mean VAS and WOMAC scores showed a statistically significant improvement (P < 0.0001) across all groups and subgroups (age,gender,BMI,disease duration) at follow up. On intra-subgroup comparison, we found no significant differences(P > 0.05) among age, BMI or gender subgroups, however the scores were significantly better in patients with disease duration of less than 1 year than those with more than 1 year duration at both 6 and 9 months[P < 0.001(RC = 9.630,95% CI = 4.037-15.222,P = 0.001)]. PRP injections if given serially can improve the short term subjective scores of VAS and WOMAC scores in patients with K-L grade II and III KOA irrespective of age, gender, BMI or disease duration, however, clinical benefits can be maximized if given early in the disease course.


Ethyl 2-diethyl-amino-4-oxo-3,5-diphenyl-4,5-dihydro-3H-pyrrolo-[3,2-d]pyrimidine-7-carboxyl-ate.
In the title compound, C(25)H(26)N(4)O(3), the two fused pyrrolo-[3,2-d]pyrimidine rings form a dihedral angle of 3.7 (2)°. The two substituent phenyl rings are twisted with respect to the pyrrole and pyrimidine rings, making dihedral angles of 57.2 (2) and 69.0 (2)°, respectively. The ethyl and eth-oxy groups are disordered over two positions; the site occupancies are 0.53 (1) and 0.47 (1) for ethyl, and 0.63 (1) and 0.37 (1) for eth-oxy. The crystal packing features C-H⋯O hydrogen bonds.


Repeated transcranial direct current stimulation in prolonged disorders of consciousness: A double-blind cross-over study.
To evaluate effects of 5 sessions of transcranial direct current stimulation (tDCS) over the left dorsolateral prefrontal cortex in patients with prolonged disorders of consciousness (DOC). Seven patients in vegetative state (VS) and 6 in minimally conscious state (MCS), at ≥3months after brain injury, were randomized into two groups: group 1 received one week of active tDCS and 1week of sham stimulation, separated by 1 resting week; group 2 received active and sham stimulation in reverse order. We performed clinical and EEG evaluations before and after the first stimulation session, two hours after the last weekly stimulation, twice during the resting week, and during a 3-month follow-up. We observed small changes of patients' conditions after the first tDCS session and immediately after the 5 active stimulations. Substantial clinical and EEG changes were observed in 5/13 patients (3 in MCS and 2 in VS) starting after entire (active and sham) stimulation protocol and further progressing during the next months. No baseline features distinguished patients who improved from patients who did not improve. Repeated tDCS did not exert remarkable short-term clinical and EEG effects in patients with prolonged DOC. Further studies should ascertain whether tDCS might promote clinical recovery in the long-term period.


Latitudinal Distributions and Controls of Bacterial Community Composition during the Summer of 2017 in Western Arctic Surface Waters (from the Bering Strait to the Chukchi Borderland).
The western Arctic Ocean is experiencing some of the most rapid environmental changes in the Arctic. However, little is known about the microbial community response to these changes. Employing observations from the summer of 2017, this study investigated latitudinal variations in bacterial community composition in surface waters between the Bering Strait and Chukchi Borderland and the factors driving the changes. Results indicate three distinctive communities. Southern Chukchi bacterial communities are associated with nutrient rich conditions, including genera such as Sulfitobacter, whereas the northern Chukchi bacterial community is dominated by SAR clades, Flavobacterium, Paraglaciecola, and Polaribacter genera associated with low nutrients and sea ice conditions. The frontal region, located on the boundary between the southern and northern Chukchi, is a transition zone with intermediate physical and biogeochemical properties; however, bacterial communities differed markedly from those found to the north and south. In the transition zone, Sphingomonas, with as yet undetermined ecological characteristics, are relatively abundant. Latitudinal distributions in bacterial community composition are mainly attributed to physical and biogeochemical characteristics, suggesting that these communities are susceptible to Arctic environmental changes. These findings provide a foundation to improve understanding of bacterial community variations in response to a rapidly changing Arctic Ocean.


On the nomenclature of congenital disorders of glycosylation (CDG).
A new nomenclature of CDG is proposed because the current one is too complex for clinicians and provides no added value.


Healthcare utilization and the quality of life of children and adolescents with sickle cell disease.
Youth diagnosed with sickle cell disease (SCD) are at increased risk of poor health-related quality of life (HRQOL) due to the complexities associated with this disease. The literature notes that predictors such as pain and poor mental health are associated with increased healthcare access; however, the connection between healthcare use and their overall well-being has been understudied. This study investigates whether healthcare utilization predicts the HRQOL in youth with SCD. Patients completed the Pediatric Quality of Life (PedsQL) 3.0 SCD module, whereas the researcher conducted a retrospective chart review to gather patient characteristics such as emergency room (ER) and hospitalization occurrences over the past 12 months. The study consisted of 150 pediatric patients with SCD, ages 8-17 years old, and their parents. Patients with ≥ 4 ER visits and hospitalizations reported worse HRQOL scores than their respective counterparts. Additionally, a higher frequency of ER visits (P = 0.05) and hospitalizations (P = 0.005) predicted lower HRQOL scores. Age (P = 0.04) also emerged as a significant predictor for both regression models, as increased healthcare access among older patients with SCD was associated with poorer HRQOL. This study found that as youth with SCD require ER treatment and/or hospital admission, they are at increased risk for lower HRQOL, specifically as they get older. Findings suggest that attention should be paid to patients who require more frequent healthcare intervention. Improvement in outpatient care of pediatric patients with SCD may help to mitigate ER and inpatient use.


Kinetic study on the equilibrium between membrane-bound and free photoreceptor G-protein.
Formation of the complex between photoreceptor G-protein (G) and photoactivated rhodopsin (RM) leads to a change in the light scattering of the disk membranes (binding signal or signal P). The signal measured on isolated disks (so-called PD signal) is exactly stoichiometric in its final level to bound G-protein but its kinetics are much slower than the RMG binding reaction. In this study on isolated disks, recombined with G-protein, we analyzed the PD-signal level and kinetics as a function of flash intensity and compared it to the RMG-complex formation monitored spectroscopically (by extra metarhodopsin II). The basic observation is that the initial slopes of the PD signals decrease with flash intensity when the signals are normalized to the same final level. This finding prevents an explanation of the scattering signal by a slow postponed reaction of the RMG complex. We propose to interpret the scattering change as a redistribution of G-protein between a membrane-bound and a solved state. The process is driven by the complexation of membrane-bound G to flash-activated rhodopsin (RM). The experimental evidence for this two-state model is the following: The intensity dependence of the initial rate of the PD signal is explained by the model. Under the assumption of a bimolecular reaction of free G with sites at the membrane, equal to rhodopsin in their concentration, the measured rates yield a KD of 10(-5) M. Evaluation of the extra MII kinetics yields a biphasic rise at saturating flashes. The measured rates fit to the supply of free and membrane-bound G-protein for the reaction with RM. Quantitative estimation of the expected scattering intensity changes gives a comprehensive description of binding signal and dissociation signal by the gain and loss of G-protein scattering mass. The temperature dependence of the PD-signal rate leads to an activation energy of the membrane-association process of E alpha = 44 kJ/mol.


Efficacy of fosfomycin and its combination with linezolid, vancomycin and imipenem in an experimental peritonitis model caused by a Staphylococcus aureus strain with reduced susceptibility to vancomycin.
The objective of this study was to evaluate the in vitro and in vivo efficacies of therapies including fosfomycin against clinical Staphylococcus aureus isolates with reduced susceptibility to vancomycin (hGISA). Time-kill curves were performed over 24 h. Peritonitis in C57BL/6 mice was induced by intraperitoneal inoculation of 10(8) CFU/ml. Four hours later (0 h), therapy was started and the treatment groups were: control (not treated), fosfomycin (100 mg/kg/5 h), vancomycin (60 mg/kg/5 h), imipenem (30 mg/kg/5 h), fosfomycin plus linezolid, fosfomycin plus vancomycin and fosfomycin plus imipenem, receiving subcutaneous therapy over 25 h. Bacterial counts in peritoneal fluid, bacteraemia and mortality rates were determined. In vitro, fosfomycin showed a synergistic effect when combined with the other antimicrobials tested. In the animal model, fosfomycin combinations were effective and significantly reduced the bacteraemia rates achieved in the control, imipenem and vancomycin groups (p < 0.05). The best combination in vivo was fosfomycin plus imipenem. Also, fosfomycin plus linezolid was significantly better than vancomycin alone, reducing the bacterial concentration in the peritoneal fluid. In conclusion, in vitro and in vivo, fosfomycin in combination with linezolid, vancomycin or imipenem exerted a good activity. Fosfomycin plus imipenem was the most active combination, decreasing 3 log CFU/ml, and appears to be a promising combination for clinical practice.


Analyzing Perceived Psychological and Social Stress of University Students: A Machine Learning Approach.
The COVID-19 pandemic has worsened the psychological and social stress levels of university students due to physical illness, enhanced dependence on mobile devices and internet, a lack of social activities, and home confinement. Therefore, early stress detection is crucial for their successful academic performance and mental well-being. The advent of machine learning (ML)-based prediction models can have a crucial impact in predicting stress at its early stages and taking necessary steps for the well-being of individuals. This study aims to develop a reliable machine learning-based prediction model for perceived stress prediction and validate the model using real-world data collected through an online survey among 444 university students from different ethnicity. The machine learning models were built using supervised machine learning algorithms. Principal Component Analysis (PCA) and the chi-squared test were employed as feature reduction techniques. Moreover, Grid Search Cross-Validation (GSCV) and Genetic Algorithm (GA) were employed for hyperparameter optimization (HPO). According to the findings, around 11.26% of individuals were identified with high levels of social stress. In comparison, approximately 24.10% of people were found to be suffering from extremely high psychological stress, which is quite alarming for students' mental health. Furthermore, the prediction results of the ML models demonstrated the most remarkable accuracy (80.5%), precision (1.000), F1 score (0.890), and recall value (0.826). The Multilayer Perceptron model was shown to have the maximum accuracy when combined with PCA as a feature reduction approach and GSCV for HPO. The convenience sampling technique used in this study only considers self-reported data, which may have biased results and lack generalizability. Future research should consider a large sample of data and focus on tracking long-term impacts with coping strategies and interventions. The results of this study can be used to develop strategies to mitigate adverse effects of the overuse of mobile devices and promote student well-being during pandemics and other stressful situations.


YAP/TAZ affects the development of pulmonary fibrosis by regulating multiple signaling pathways.
YAP and TAZ are important co-activators of various biological processes in human body. YAP/TAZ plays a vital role in the development of pulmonary fibrosis. Dysregulation of the YAP/TAZ signaling pathway is one of the most important causes of pulmonary fibrosis. Therefore, considering its crucial role, summary of the signal mechanism of YAP/TAZ is of certain guiding significance for the research of YAP/TAZ as a therapeutic target. The present review provided a detailed introduction to various YAP/TAZ-related signaling pathways and clarified the specific role of YAP/TAZ in these pathways. In the meantime, we summarized and evaluated possible applications of YAP/TAZ in the treatment of pulmonary fibrosis. Overall, our study is of guiding significance for future research on the functional mechanism of YAP/TAZ underlying lung diseases as well as for identification of novel therapeutic targets specific to pulmonary fibrosis.


Algorithms for the segmental motor innervation of the extremities.
The pattern of the segmental motor innervation to the extremities is reflected in the kinematics of their joint linkages. More proximally located joints have more complex movements and are controlled by more spinal cord levels than more distal joints with simpler movements. More proximally located joints are also controlled by more cranially located groups of spinal cord levels while more distal ones are innervated by more caudally located groups. Algorithms are presented that summarize these principles in the form of two simple, concise charts and a stick figure. The cord levels given by the algorithms for the innervation of any particular muscle are in good general agreement with recognized authorities. In addition, these algorithms are easily learned and readily applied to clinical problems by placing the patient in the position assumed when playing the piano or operating an automobile. In this posture, the most cranial two spinal cord levels of a group controlling a joint are responsible for "upward" motion of that joint and the most caudal two or three control "downward" motion thereof.


Flapping motion as a fluorescent probe for assembly process involving highly viscous liquid-like cluster intermediates during evaporative crystallization.
Fluorescence probes are widely used to assess the molecular environment based on their photo-physical properties. Specifically, flexible and aromatic photo-functional system (FLAP) is unique viscosity probe owing to the excited-state planarization of anthracene wings. We have previously applied fluorescence spectroscopy to monitor the evaporative crystallization of solvents. The fluorescence color and spectral changes, which depend on the aggregation form, enable direct fluorescence visualization during evaporative crystallization. The fluorescence visualization of the liquid-like cluster intermediate proposed in the two-step nucleation model for the nucleation process has been achieved. However, the physical properties of these clusters, especially the viscosity, molecular motion, and intermolecular interactions, are still unclear. In this study, FLAPs are used as probes for local-viscosity changes and space limitations of the liquid-like cluster state during evaporative crystallization by observing the fluorescence-spectral changes and using hyperspectral-camera (HSC) imaging. Green emission originates from the monomer in the solution owing to the free-flapping motion. The fluorescence color turns blue with increasing viscosity under crowding conditions. If the survival time of the liquid-like cluster state is sufficient, crystalline phase (R-phase) formation proceeds via a 2-fold π-stacked array of the V-shaped molecules. It is difficult to form the V-shaped stacked columnar structures in the liquid-like cluster state region, resulting in the deposition of head-to-tail dimer structures, such as the yellow-emissive phase (Y-phase). In the case of the FLAP, the stacking intermediate does not form during solvent evaporation in the liquid-like cluster; rather, it is deposited in an amorphous form that exhibits blue emission (B-phase). These findings suggest that it is important to the maintenance of the survival time of the liquid-like cluster states to organize and rearrange the stacking forms. We have achieved the fluorescence probing of viscosity changes at local molecular motion with solvent depletion during solvent evaporation for the first time.


Biosynthetic Mesh Reconstruction after Abdominoperineal Resection for Low Rectal Cancer: Cross Relation of Surgical Healing and Oncological Outcomes: A Multicentric Observational Study.
Local wound complications are among the most relevant sequelae after an abdominoperineal resection (APR) for low rectal cancer. One of the proposed techniques to improve the postoperative recovery and to accelerate the initiation of adjuvant chemotherapy is the mesh reinforcement of the perineal wound. The aim of the current study is to compare the surgical and oncological outcomes after APR performed with a biosynthetic mesh reconstruction versus the conventional procedure. From 2015 to 2020, in two tertiary centres, the surgical outcomes, the wound events (i.e., surgical site infections, wound dehiscence and the complete healing time) and the oncological outcomes (i.e., time length to start adjuvant chemo-radiotherapy, an over 8-week delay in chemotherapy and the recurrence rate) were retrospectively analysed in patients undergoing APR reinforced with biosynthetic mesh (Group A) and conventional APR (Group B). Results Sixty-one patients were treated with APR (25 in Group A and 36 in Group B). Patients in Group A presented lower time for: healing (16 versus 24 days, p = 0.015), inferior perineal wound dehiscence rates (one versus nine cases, p = 0.033), an earlier adjuvant therapy start (26 versus 70 days, p = 0.003) and a lower recurrence rate (16.6% vs. 33.3%, p = 0.152). In our series, the use of a biosynthetic mesh for the neo-perineum reconstruction after a Miles' procedure has resulted in safe, reproducible results affected by limited complications, guarantying a rapid start of the adjuvant therapy with clear benefits in oncological outcomes. Further randomized clinical trials with long-term follow-up are needed to validate these results.


Sunlight: a major factor associated with the development of melanocytic nevi in Australian schoolchildren.
Case-control studies have identified melanocytic nevi (MN) as the most important phenotypic risk factor for melanoma. A knowledge of any environmental factors that cause MN may facilitate prevention of melanoma. This study was undertaken to explore the possible role of ambient solar irradiation in the development of MN in children. With a standard protocol developed after international consultation, the same medical observers examined children in three Australian cities (Melbourne, Sydney, and Townsville) that span a wide range of latitude. A total of 1123 Australian schoolchildren 6, 9, 12, and 15 years of age were surveyed. Larger numbers of MN were found (mean 65.4 MN, standard deviation 52.9) than in previous studies of children. Prevalence increased with diminishing latitude (51.1 in Melbourne, 66.5 in Sydney and 77.2 in Townsville), particularly in children 6 and 9 years of age. Although nevus numbers were higher in children with light skin and hair, blue eyes, and freckling, the latitude gradient remained after adjustment for these and other factors in multivariate analysis. Latitude of residence, and by implication ambient UV radiation, is strongly related to nevus prevalence in young Australian children. However, these differences diminish with age and may disappear by 15 years of age.


Purified collagen I oriented membrane for tendon repair: an ex vivo morphological study.
Injured tendons have limited repair ability after full-thickness lesions. Tendon regeneration properties and adverse reactions were assessed ex vivo in an experimental animal model using a new collagen I membrane. The multilamellar membrane obtained from purified equine Achilles tendon is characterized by oriented collagen I fibers and has been shown to sustain cell growth and orientation in vitro. The central third of the patellar tendon (PT) of 10 New Zealand White rabbits was sectioned and grafted with the collagen membrane; the contralateral PT was cut longitudinally (sham-operated controls). Animals were euthanized 1 or 6 months after surgery, and tendons were subjected to histological and Synchrotron Radiation-based Computed Microtomography (SRµCT) examination and 3D structure analysis. Histological and SRµCT findings showed satisfactory graft integration with native tendon. Histological examination also showed ongoing angiogenesis. Adverse side-effects (inflammation, rejection, calcification) were not observed. The multilamellar collagen I membrane can be considered as an effective tool for tendon defect repair and tendon augmentation.


Amiodarone pulmonary toxicity: cytopathology, ultrastructure, and immunocytochemistry.
One hundred ninety cardiac patients were prospectively enrolled in an amiodarone protocol. Over a 10-year period, 16 patients developed new or progressive respiratory symptoms while taking amiodarone. These symptoms included dyspnea associated with abnormal chest radiographs or new or worsening abnormalities on pulmonary function testing. Specimens for microscopic examination were obtained by fiberoptic bronchoscopy with transbronchial lung biopsy (TBB), bronchoalveolar lavage (BAL), open lung biopsy (OLB), or autopsy. Large foamy macrophages with characteristic lamellated cytoplasmic inclusions were noted in all specimens, regardless of other evidence of pulmonary toxicity, suggesting that foamy macrophages represent a routine drug effect. Foamy macrophages were not present in BAL specimens from 53 normal controls and were rarely seen in specimens from 27 patients who had other interstitial lung diseases. When present, the foamy macrophages were less prominent than those seen in specimens from patients receiving amiodarone. Fibrosis was noted in 11 of 16 histological specimens, whereas type II-cell-hyperplasia was observed in 7 of the 16 specimens. Four of the 16 patients with respiratory symptoms died, and their autopsy revealed a combination of foamy macrophages with fibrosis and type II cell hyperplasia reflective of amiodarone pulmonary toxicity. Hyperplastic type II cells were not found in the absence of fibrosis. Immunocytochemistry allowed differentiation between foamy macrophages and type II cells and represents a useful tool for future investigations of the pathogenesis of amiodarone-induced pulmonary disease.


On the origins of the absorption spectroscopy of pterin and Re(CO)3(pterin)(H2O) aqueous solutions. A combined theoretical and experimental study.
The origins of the absorption spectroscopy of pterin and Re(CO)3(pterin)(H2O) complex as a function of pH is studied using the hybrid functional B3LYP and PBE0 in combination with 6-311++G(d,p) and LanL2TZ(f) basis sets. A natural bond analysis was performed to the principal molecular orbitals involved in the electronic transitions of the studied compounds. The low energy band of pterin, which is described as a H→L electronic transition, can be interpreted as an admixture of π→π(*), n→π(*) and n→n electronic transitions involving π(C2-N1, C6-N5, C9-C10) and n(C2) orbitals of the HOMO and π(*)(C6-N5, C7-N8) and n(C4) orbitals of the LUMO. The low energy band of Re(CO)3(pterin)(H2O) can be described as a combination of different MLLCT transitions where electron density residing on different π orbitals of carbonyl-Re bonds and lone pairs of Re is transferred to pterin moiety. Besides MLLCT transitions, IL, LLCT and LLMCT transitions contribute the absorptions of the Re(I) complex in the wavelength region corresponding to the high energy bands. The calculated electronic spectra of the acid and base forms of pterin and Re(CO)3(pterin)(H2O) were simulated from the theoretical results and compared to the experimental absorption spectra with great accuracy both in position and relative intensities of the absorption bands.


[Effect of Na+ and Ca++ ion concentrations in the outer medium surrounding the erythrocyte membrane on the viscosity of blood suspensions].
A small concentration of Na+ ions (0.1 mM) causes an enlargement of the viscosity in suspensions of erythrocytes which is partial reversible by addition of Na+ ions. Ca++ ions have a conserving function. The relative viscosity is changed in the relation of concentrations of ions of Ca++:Na+ = 1:2. Changes in the viscosities are refered to change of structure of the components of membrane, especially for the part of lipids.


Testicular and kidney masses in a HIV-infected man.
A 37-year-old Caucasian male presented with what appeared clinically and diagnostically as testicular and kindey tomour's. However, syphilis serology and histology confirm that the lesions were gumma's. Without routine syphilis serology, the correct diagnosis may have been missed.


A Delicate Connection: c-di-AMP Affects Cell Integrity by Controlling Osmolyte Transport.
Bacteria use second-messenger molecules to adapt to their environment. Several second messengers, among them cyclic di-AMP (c-di-AMP), have been discovered and intensively studied. Interestingly, c-di-AMP is essential for growth of Gram-positive bacteria such as Bacillus subtilis, Listeria monocytogenes, and Staphylococcus aureus. Many studies demonstrated that perturbation of c-di-AMP metabolism affects the integrity of the bacterial cell envelope. Therefore, it has been assumed that the nucleotide is essential for proper cell envelope synthesis. In this Opinion paper, we propose that the cell envelope phenotypes caused by perturbations of c-di-AMP metabolism can be interpreted differently: c-di-AMP might indirectly control cell envelope integrity by modulating the turgor, a physical variable that needs to be tightly adjusted. We also discuss open questions related to c-di-AMP metabolism that need to be urgently addressed by future studies.


Calcitonin gene-related peptide expression levels predict motor neuron vulnerability in the superoxide dismutase 1-G93A mouse model of amyotrophic lateral sclerosis.
In amyotrophic lateral sclerosis (ALS) some motor neurons degenerate while others survive. The molecular mechanisms underlying this selective vulnerability and resistance, respectively, are poorly understood. Since the neuropeptide, calcitonin gene-related peptide (CGRP), is expressed by many but not all motor neurons, we asked if motor neuron CGRP levels predict their vulnerability in the SOD1-G93A mouse model of ALS. In wild type mice three types of somatic motor neurons were distinguished based on their CGRP expression pattern, i.e. highCGRP, lowCGRP, and nonCGRP. Since motor nuclei III, IV and VI contained mostly nonCGRP motor neurons, they defined the oculomotor group. In comparison, the facial group (nuclei V, VII and XII) contained equal numbers of all three types, while the spinomedullary group (ambiguus nucleus and lumbar spinal cord) contained mainly highCGRP motor neurons. Analysis on the transcript level, and of mice lacking the αCGRP isoform, revealed that these group differences in CGRP expression were predominantly based on αCGRP. At disease end-stage in SOD1-G93A mice, group-specific extent of motor neuron loss correlated with CGRP expression as neurons with highCGRP were reduced by 80%, those with lowCGRP by 50%, and nonCGRP motor neurons were not significantly affected in all three groups. Finally, highCGRP motor neuron degeneration preceded lowCGRP motor neuron degeneration during disease progression. Our analysis revealed that the relative abundance of CGRP mRNA and immunoreactivity in motor neurons predicts their vulnerability. CGRP may be an autocrine or paracrine factor promoting motor neuron degeneration in this ALS model.


Confluent hepatic fibrosis in monozygotic twins.
The MRI and CT features of confluent hepatic fibrosis (CHF) are reported in two 6-year-old twins. This is the first case report in children of this little known entity, which may mimic mass lesions, and the first to describe an association of CHF with anti-tuberculous drugs.


Lumbar Multifidus and Erector Spinae Muscle Synergies in Patients with Nonspecific Low Back Pain During Prone Hip Extension: A Cross-sectional Study.
Prone hip extension is used to clinically assess movement control in patients with nonspecific low back pain (LBP). Excessive lumbopelvic movements and altered muscle activation patterns are common in patients with nonspecific LBP. However, no evidence exists regarding lumbar multifidus and lumbar erector spinae muscle synergy patterns in patients with nonspecific LBP during this clinical test. To determine the difference in lumbopelvic motion and change in muscle synergy between patients with nonspecific LBP and healthy individuals. A cross-sectional study design. University physical therapy clinic and laboratory. Seven patients with nonspecific LBP (age [mean ± SD] 29 ± 5 years, 43% female, body mass index [BMI] of 25 ± 2 kg/m2 , Numeric Pain Rating Scale 6 ± 2; Oswestry Disability Index 20% ± 8%) and seven age-, sex-, and BMI-matched healthy individuals (mean age 28 ± 5 years, 43% female, BMI of 22 ± 2 kg/m2 ) were recruited. Each participant performed six repetitions of prone hip extension on each side; kinematic and electromyographic data were collected simultaneously. Kinematic data were used to represent lumbopelvic motion, whereas electromyography (EMG) data were used to represent muscle activity. A paired t-test was used to determine the difference in lumbopelvic motion. Principal component analysis and two-way repeated-measures analysis of variance were used to extract muscle synergies and identify differences in muscle synergy patterns between and within groups. Results demonstrated no significant group difference (P > .05) in amount of lumbopelvic motion. However, healthy individuals synergistically activate lumbar multifidus and lumbar erector spinae with 81.0% variance accounted for (VAF). Patients with nonspecific LBP had an altered synergy with independent activation of lumbar multifidus on the painful side with 32.0% VAF and the lumbar multifidus on the nonpainful side, and both lumbar erector spinae with 52.2% VAF synergistically activated. These findings suggest that clinicians should focus on muscle activation patterns rather than the amount of lumbopelvic motion during clinical observation of prone hip extension. Level III.


Stabilizing peptide fusion for solving the stability and solubility problems of therapeutic proteins.
Protein aggregation is a major stability problem of therapeutic proteins. We investigated whether a novel stabilizing peptide [acidic tail of synuclein (ATS) peptide] could be generally used to make a more stable and soluble form of therapeutic proteins, particularly those having solubility or aggregation problems. We produced ATS fusion proteins by fusing the stabilizing peptide to three representative therapeutic proteins, and then compared the stress-induced aggregation profiles, thermostability, and solubility of them. We also compared the in vivo stability of these ATS fusion proteins by studying their pharmacokinetics in rats. The human growth hormone-ATS (hGH-ATS) and granulocyte colony-stimulating factor-ATS (G-CSF-ATS) fusion proteins were fully functional as determined by cell proliferation assay, and the ATS fusion proteins seemed to be very resistant to agitation, freeze/thaw, and heat stresses. The introduction of the ATS peptide significantly increased the storage and thermal stabilities of hGH and G-CSF. The human leptin-ATS fusion protein also seemed to be very resistant to aggregation induced by agitation, freeze/thaw, and heat stresses. Furthermore, the ATS peptide greatly increased the solubility of the fusion proteins. Finally, pharmacokinetic studies in rats revealed that the ATS fusion proteins are also more stable in vivo. Our data demonstrate that a more stable and soluble form of therapeutic proteins can be produced by fusing the ATS peptide.


Pressure ulcer grading.
This article discusses the different aspects of classifying pressure ulcers. It aims to increase understanding of the grading methods used and how their reliability and validity may be assessed. In turn, this aims to increase the potential for nurses to enhance their assessment and pressure ulcer prevention skills.