Is minor surgery safe during the COVID-19 pandemic? A multi-disciplinary study.
To assess the risk of postoperative SARS-CoV-2 infection during the COVID-19 pandemic. The CONCEPTION study was a cohort, multidisciplinary study conducted at Conception University Hospital, in France, from March 17th to May 11th, 2020. Our study included all adult patients who underwent minor surgery in one of the seven surgical departments of our hospital: urology, digestive, plastic, gynecological, otolaryngology, gynecology or maxillofacial surgery. Preoperative self-isolation, clinical assessment using a standardized questionnaire, physical examination, nasopharyngeal RT-PCR and chest CT scan performed the day before surgery were part of our active prevention strategy. The main outcome was the occurrence of a SARS-CoV-2 infection within 21 days following surgery. The COVID-19 status of patients after discharge was updated during the postoperative consultation and to ensure the accuracy of data, all patients were contacted again by telephone. A total of 551 patients from six different specialized surgical Departments in our tertiary care center were enrolled in our study. More than 99% (546/551) of included patients underwent a complete preoperative Covid-19 screening including RT-PCR testing and chest CT scan upon admission to the Hospital. All RT-PCR tests were negative and in 12 cases (2.2%), preoperative chest CT scans detected pulmonary lesions consistent with the diagnosis criteria for COVID-19. No scheduled surgery was postponed. One patient (0.2%) developed a SARS-CoV-2 infection 20 days after a renal transplantation. No readmission or COVID-19 -related death within 30 days from surgery was recorded. Minor surgery remained safe in the COVID-19 Era, as long as all appropriate protective measures were implemented. These data could be useful to public Health Authorities in order to improve surgical patient flow during a pandemic.


How Commonly Is the Diagnosis of Gastric Low Grade Dysplasia Upgraded following Endoscopic Resection? A Meta-Analysis.
Gastric dysplasia is a well-known precancerous lesion. Though the diagnosis of gastric low grade dysplasia (LGD) is generally made from endoscopic forceps biopsy (EFB), the accuracy is doubtful after numerous EFB-proven gastric LGD were upgraded to gastric high grade dysplasia (HGD) or even carcinoma (CA) by further diagnostic test with the procedure of endoscopic resection (ER). We aimed to evaluate the upgraded diagnosis rate (UDR) and the risk factors by ER in EFB-proven gastric LGD lesions. Two investigators independently searched studies reporting the UDR by ER in EFB-proven gastric LGD lesions from databases and analyzed the overall UDR, HGD-UDR and CA-UDR. The pooled UDR by ER in EFB-proven gastric LGD lesions was 25.0% (95% CI, 20.2%-29.8%), made up of HGD-UDR and CA-UDR by rates of 16.7% (95% CI, 12.8%-20.6%) and 6.9% (95% CI, 4.2%-9.6%) respectively. Lesion size larger than 2 cm, surface with depression and nodularity under endoscopic examinations were the major risk factors associated with UDR. In conclusion, one quarter of EFB-proven gastric LGD lesions will be diagnosed as advanced lesions, including gastric HGD (16.7%) and gastric CA (6.9%) by ER. The diagnosis of those LGD lesions with an endoscopic diameter larger than 2cm, and depressed or nodular surface are more likely to be upgraded after ER.


Characterization of a rat lung microsomal fraction obtained by sepharose 2B ultrafiltration.
A new procedure for obtaining rat lung microsomes essentially free of interfering hemoproteins has been developed. The method includes Sepharose 2B column chromatography of the 12,000 X g supernatant of lung homogenates, followed by ultracentrifugation of the material eluted in the void volume. Microsomes isolated in this manner contain specific levels of cytochromes b5 and P-450 and of NADPH-cytochrome c reductase that are among the highest ever reported for a rat lung microsomal fraction. After treatment of rats with 3-methylcholanthrene, the specific content of cytochrome P-450 in lung microsomes is doubled and that of cytochrome b5 increases 1.5 times. Several spectral differences between hepatic and lung microsomal cytochrome P-450 are apparent. In lung microsomes, the maximum of the reduced CO-bound cytochrome complex in a difference spectrum is at 453 nm for the noninduced hemoprotein and shifts to 451 nm after 3-methylcholanthrene induction. In contrast, no significant change in the ethylisocyanide difference spectra of reduced microsomes is obtained after induction; moreover, the spectra obtained with induced and noninduced cytochrome P-450 are similar to the one shown by hepatic microsomes from polycyclic hydrocarbon-treated rats. Furthermore, spectrophotometric studies on n-octylamine binding to control and induced lung cytochrome P-450 yielded results different from those previously obtained with rabbit liver microsomes. It is concluded that the cytochrome P-450 present in rat lung microsomes before and after 3-methylcholanthrene treatment of the animals is distinctly different from the liver hemoprotein.


Geometrical deformation of vocal fold tissues induced by formalin fixation.
Many existing studies of vocal fold geometry are based on anatomical measurements made on histologically fixed laryngeal tissues using formalin. However, the validity of these geometric data is questionable because of the potentially significant tissue deformation associated with formalin fixation, particularly tissue shrinkage. Previous experiments have shown that valid geometric data may be obtained on vocal fold tissue samples quickly frozen with liquid nitrogen. Based on this finding, the present study attempted to quantify the geometric deformation of formalin-fixed vocal fold tissues with respect to quick-frozen tissues. Six freshly harvested canine larynges were quickly frozen with liquid nitrogen and sectioned at the mid-membranous coronal plane. Each larynx was thawed and divided into halves along the midsagittal plane, one of which was not fixed; the other was fixed with formalin, from which histological tissue sections were also prepared. Measurements of vocal fold geometry were made on digital images of mid-membranous coronal sections of the tissue samples, based on linear dimensions of vocal fold depth and thickness defined functionally for biomechanical applications. Significant shrinkage of the vocal fold (particularly the vocal fold body) and considerable distortion of the vocal fold contour (particularly at the free edge) were observed for the formalin-fixed samples and the histological sections in comparison with the unfixed samples. Results of the present study suggested that significant geometric artifacts are induced by conventional histological fixation of laryngeal tissues using formalin. These artifacts should be carefully considered for interpreting any vocal fold geometric data obtained through formalin fixation.


Inhibition of mitogen-activated proliferation of human peripheral lymphocytes in vitro by propionic acid.
Recurrent infections are common features in patients affected by propionic acidaemia (McKusick 232000) and methylmalonic acidaemia (McKusick 251000). Since these disorders are biochemically characterized by tissue accumulation of propionic acid and methylmalonic acid respectively, it is possible that these compounds may act as immunosuppressants. We therefore investigated the effect of propionate and methylmalonate on cellular growth of human peripheral lymphocytes stimulated in vitro by phytohaemagglutinin, concanavalin A and pokeweed mitogen, a recognized test of cellular immunocompetence. Lymphocytes were cultured in flat-bottomed 96-well microplates at 37 degrees C for 96 h (phytohaemagglutinin and concanavalin A) or 144 h (pokeweed mitogen) in the presence of one mitogen at different concentrations and of one acid added at doses of 1.0, 2.5 or 5.0 mM. Cell blastogenesis was measured by the incorporation of tritiated thymidine into cellular DNA and compared with that of identical cultures with no acid added (controls). A consistent and progressive inhibitory effect of propionic acid with increasing concentrations in culture was identified with all mitogens and was more pronounced with pokeweed mitogen. Lymphocyte blastogenesis was not altered in the presence of methylmalonic acid. The effect of propionate was observed only when the drug was added at the beginning (phytohaemagglutinin-activated) or until 24 h (concanavalin A- and pokeweed mitogen-activated) of culture. The viability of lymphocytes after treatment with the drug, as assessed by the Trypan Blue exclusion test, revealed no change when compared with the same untreated lymphocytes, indicating no lymphocytotoxic activity. In conclusion, propionic acid, which accumulates in tissues of patients with propionic acidaemia, causes 'in vitro' immunosuppression, which may be related to the recurrent infections characteristic of these patients.


Cell aggregation activates small GTPase Rac1 and induces CD44 cleavage by maintaining lipid raft integrity.
Lipid rafts are highly ordered membrane domains that are enriched in cholesterol and glycosphingolipids and serve as major platforms for signal transduction. Cell detachment from the extracellular matrix (ECM) triggers lipid raft disruption and anoikis, which is a barrier for cancer cells to metastasize. Compared to single circulating tumor cells (CTCs), our recent studies have demonstrated that CD44-mediatd cell aggregation enhances the stemness, survival and metastatic ability of aggregated cells. Here, we investigated whether and how lipid rafts are involved in CD44-mediated cell aggregation. We found that cell detachment, which mimics the condition when tumor cells detach from the ECM to metastasize, induced lipid raft disruption in single cells, but lipid raft integrity was maintained in aggregated cells. We further found that lipid raft integrity in aggregated cells was required for Rac1 activation to prevent anoikis. In addition, CD44 and γ-secretase coexisted at lipid rafts in aggregated cells, which promoted CD44 cleavage and generated CD44 intracellular domain (CD44 ICD) to enhance stemness of aggregated cells. Consequently, lipid raft disruption inhibited Rac1 activation, CD44 ICD generation, and metastasis. Our findings reveal two new pathways regulated by CD44-mediated cell aggregation via maintaining lipid raft integrity. These findings also suggest that targeting cell aggregation-mediated pathways could be a novel therapeutic strategy to prevent CTC cluster-initiated metastasis.


Mathematical birth of Early Afterdepolarizations in a cardiomyocyte model.
Early Afterdepolarizations (EADs) are abnormal behaviors that can lead to cardiac failure and even cardiac death. In this paper we investigate the occurrence and development of these phenomena in a reduced Luo-Rudy cardiac model. Through a comprehensive dynamical analysis, we map out the distinct patterns observed in the parametric plane, differentiating between normal beats without EADs and pathological beats with EADs. By examining the bifurcation structure of the model, we elucidate the dynamical elements associated with these patterns and their transitions. Using a fast-slow analysis, we explore the emergence and evolution of EADs in the model. Notably, our approach combines the two commonly used fast-slow approaches (1-slow-2-fast and 2-slow-1-fast), and we show how both approaches together provide a more complete understanding of this phenomenon.


Circus movements and blebbing locomotion in dissociated embryonic cells of an amphibian, Xenopus laevis.
Circus movements, which involve the circumferential rotation of a hyaline cytoplasmic protrusion, occur in cells obtained by EDTA dissociation of gastrula-stage Xenopus laevis embryos. Only a few dissociated blastula-stage cells show circus movements, more early gastrula-stage cells show them, and nearly all late gastrula-stage cells show them. Circus movements cease in cells prior to mitosis and begin again in daughter cells after mitosis is completed. In early gastrulae, only 17% of prospective endodermal cells show circus movements while 79% of prospective mesodern, archenteric roof, and posterior neural ectoderm do so. Isolated cells as well as groups of cells in vitro are often propelled by circus movements. There is an obvious antagonism between cell contact and circus movements. The morphogenetic significance of circus movements and blebbing locomotion is discussed.


Label-free measurements of the diffusivity of molecules in lipid membranes.
An important and characteristic property of a cell membrane is the lateral mobility of protein molecules in the lipid bilayer. This has conventionally been measured by labeling the molecules with fluorescent markers and monitoring their mobility by different fluorescence-based techniques. However, adding the label to the studied molecule may affect the system, so it is an assumption in almost all experiments that the measured mobility of the biomolecule with its label is the same as that of the unlabeled molecule. However, this assumption is rarely tested due to a lack of suitable methods. In this work, a new technique to perform label-free diffusivity measurements is developed and used to measure the effect of the label for two common protein-lipid systems: 1) streptavidin (SA) coupled to a supported lipid bilayer (SLB) through biotinylated lipids and 2) the extracellular part of the T-cell adhesion protein CD2, coupled to an SLB through histidine tags to nickel-chelating lipids. A measurable (≈12%) decrease in diffusivity is found for both labeled proteins, even though the molecular mass of the label is almost 100 times smaller than those of the proteins (≈50 kDa). The results illustrate the importance of being able to study different biophysical properties of cell membranes and their mimics without relying on fluorescent labels, especially if fluorescent labeling is difficult or is expected to affect the nature of the intermolecular interactions being studied.


A neuronally based model of contrast gain adaptation in fly motion vision.
Motion-sensitive neurons in the visual systems of many species, including humans, exhibit a depression of motion responses immediately after being exposed to rapidly moving images. This motion adaptation has been extensively studied in flies, but a neuronal mechanism that explains the most prominent component of adaptation, which occurs regardless of the direction of motion of the visual stimulus, has yet to be proposed. We identify a neuronal mechanism, namely frequency-dependent synaptic depression, which explains a number of the features of adaptation in mammalian motion-sensitive neurons and use it to model fly motion adaptation. While synaptic depression has been studied mainly in spiking cells, we use the same principles to develop a simple model for depression in a graded synapse. By incorporating this synaptic model into a neuronally based model for elementary motion detection, along with the implementation of a center-surround spatial band-pass filtering stage that mimics the interactions among a subset of visual neurons, we show that we can predict with remarkable success most of the qualitative features of adaptation observed in electrophysiological experiments. Our results support the idea that diverse species share common computational principles for processing visual motion and suggest that such principles could be neuronally implemented in very similar ways.


The immune response to mitochondrial autoantigens.
Similar to other autoimmune diseases, there are intense humoral and cellular responses to intracytoplasmic antigens in primary biliary cirrhosis (PBC). The autoantigens against antimitochondrial antibodies (AMAs) in PBC are located on the inner mitochondrial membrane and are identified as members of the 2-oxo-acid dehydrogenase complexes (2-OADCs), of which the E2 subunit of pyruvate dehydrogenase complex (PDC-E2) is the major autoantigen; AMAs are is present in approximately 90 to 95% of PBC sera. An orchestrated immune response against the intrahepatic biliary epithelial cell (BEC) through 2-OADC-specific CD4 (+) helper T cells and CD8 (+) CTL are thought to be the major players in the immunological destruction of BECs in PBC. We believe that a prior/primary event of specific intrahepatic BEC malfunction (possibly caused by environmental insults such as xenobiotics and microorganisms) is responsible for the breaking of self-tolerance to 2-OADC and leads to BEC destruction. The generation of 2-OADC-specific T cells further accelerates the damage of BEC. In addition, immunoglobulin A AMAs may participate in the destruction of BECs by damaging mitochondria.


Novel Sternal Protection Device for Cardiac Surgery Via Median Sternotomy Incision.
Sternal bleeding during cardiac surgery is currently controlled using bone wax or other chemical substances that may result in adverse effects and affect wound healing and recovery. The purpose of this study was to identify a safe, cost-effective, and easy-to-use technique to reduce sternal bleeding and sternal trauma during cardiac surgery. After sternotomy, a sternal protection device was placed over each hemisternal section before insertion of the retractor and remained in situ until the end of surgery. Sternal bleeding and ease of use were assessed and recorded during surgery. Sternal trauma was assessed and recorded within 5 minutes of removal of the device, and overall satisfaction (Global Impression) and any intraoperative adverse events or device malfunction were reported at surgery completion. Patients were followed up 24 hours and 4 weeks after surgery. Twelve patients completed the study. Adverse events reported were not considered related to the device. No sternal trauma was identified in any patient. In 9 of 11 patients, sternal bleeding was reduced after insertion of the device. The device was generally considered easy to use, although some difficulty was encountered when used with the Internal Mammary Artery retractor. Our data suggest that the device is safe and able to reduce sternal bleeding during surgery using sternal retractors. We recommend further studies in a larger population of patients with a control group to evaluate the device's ability to reduce the morbidity associated with sternal bleeding and sternal trauma.


[Prognosis in stomach carcinoma].
Between 1974 and 1981, laparotomies were performed on 233 patients with histologically proven gastric carcinoma. Lymph-node metastases were found in 72.6%, distant metastases in 14.2%. The condition was found to be inoperable in 18.9%, a palliative procedure was performed in 17.6%, a questionably curative operation in 33.5% and a curative one in 30.0%. Postoperative mortality was 15.9%, highest in case of inoperability and palliative operations. During the follow-up period of 5-13 years 137 patients (58.8%) died of metastases or recurrence, two (0.9%) of another malignancy and 30 (12.9%) of other illnesses. The five-year survival rate was 13.3%. There was no difference in prognosis between carcinoma in a residual stomach (after previous partial resection) and a previously unoperated one. Carcinoma of the cardia gave the worst results. No patient with an early cancer died of it.


[Antitoxic properties of monoclonal antibodies against diphtheria toxin].
Antitoxin properties of monoclonal antibodies to diphtheria toxin were studied in chick fibroblast culture and during the dermonecrotic tests in guinea pigs. Antitoxic properties were shown for antibodies specific to B subunit of the toxin but not for the ones specific to a subunit. The mixture of monoclonal B subunit specific antibodies reveals a higher neutralizing activity as compared with the activity produced by any single monoclonal antibody tested. The protective activity demonstrated by antibodies is connected with the preventing of toxin molecules fixation on the cellular receptors.


Regional anaesthesia for carotid endarterectomy.
Summary Regional anaesthesia is a popular choice for patients undergoing carotid endarterectomy (CEA). Neurological function is easily assessed during carotid cross-clamping; haemodynamic control is predictable; and hospital stay is consistently shorter compared with general anaesthesia (GA). Despite these purported benefits, mortality and stroke rates associated with CEA remain around 5% for both regional anaesthesia and GA. Regional anaesthetic techniques for CEA have improved with improved methods of location of peripheral nerves including nerve stimulators and ultrasound together with a modification in the classification of cervical plexus blocks. There have also been improvements in local anaesthetic, sedative, and arterial pressure-controlling drugs in patients undergoing CEA, together with advances in the management of patients who develop neurological deficit after carotid cross-clamping. In the UK, published national guidelines now require the time between the patient's presenting neurological event and definitive treatment to 1 week or less. This has implications for the ability of vascular centres to provide specialized vascular anaesthetists familiar with regional anaesthetic techniques for CEA. Providing effective regional anaesthesia for CEA is an important component in the armamentarium of techniques for the vascular anaesthetist in 2014.


Impaired Limb Proprioception in Adults With Spasmodic Dysphonia.
Focal dystonia of the head and neck are associated with a loss of kinesthetic acuity at muscles distant from the dystonic sites. That is, while the motor deficits in focal dystonia are confined, the associated somatosensory deficits are generalized. This is the first systematic study to examine, if patients diagnosed with spasmodic dystonia (SD) show somatosensory impairments similar in scope to other forms of focal dystonia. Proprioceptive acuity (ability to discriminate between two stimuli) for forearm position and motion sense was assessed in 14 spasmodic dystonia subjects and 28 age-matched controls using a passive motion apparatus. Psychophysical thresholds, uncertainty area (UA), and a proprioceptive acuity index (AI) were computed based on the subjects' verbal responses. The main findings are as follows: first, the SD group showed significantly elevated thresholds and UAs for forearm position sense compared with the control group. Second, 9 of 14 dystonia subjects (64%) exhibited an AI for position sense above the control group maximum. Three SD subjects had a motion sense AI above the control group maximum. The results indicate that impaired limb proprioception is a common feature of SD. Like other forms of focal dystonia, spasmodic dystonia does affect the somatosensation of nondystonic muscle systems. That is, SD is associated with a generalized somatosensory deficit.


Myocardial perfusion imaging and infarct characterization using multidetector cardiac computed tomography.
Until recently, computed tomography coronary angiography was restricted to the anatomical assessment of coronary stenosis, whereas the functional significance of coronary lesions remained outside of its scope. Nevertheless, the kinetics of iodinated contrast is similar to gadolinium-diethylenetriamine pentaacetic acid used in contrast-enhanced magnetic resonance imaging, allowing assessment of myocardial perfusion and viability by cardiac computed tomography.


Placebo effect in surgery for Ménière's disease. A double-blind, placebo-controlled study on endolymphatic sac shunt surgery.
To investigate the placebo effect in surgery for Ménière's disease, a double-blind controlled study was undertaken, comparing effects of a regular endolymphatic shunt with those of regular mastoidectomy. Thirty patients with typical Ménière's diseases, selected because of unsuccessful medical treatment, participated. Patients completed daily dizziness questionnaires three months before and 12 months after surgery, with registration of nausea, vomiting, vertigo, tinnitus, hearing impairment, and pressure in the ears. Patients were operated on at two universities, and the patients operated on at one underwent controlled study each month at the other. At termination of the trial, both investigators and patients gave their opinions of the efficacy of the operations. Minor differences were seen between active and placebo groups, but the greatest difference in symptoms was found when preoperative and postoperative scores were compared: both groups improved significantly


Phase III trial comparing protracted intravenous fluorouracil infusion alone or with yttrium-90 resin microspheres radioembolization for liver-limited metastatic colorectal cancer refractory to standard chemotherapy.
Liver dissemination is a major cause of mortality among patients with advanced colorectal cancer. Hepatic intra-arterial injection of the beta-emitting isotope yttrium-90 ((90)Y) bound to resin microspheres (radioembolization) delivers therapeutic radiation doses to liver metastases with minimal damage to adjacent tissues. We conducted a prospective, multicenter, randomized phase III trial in patients with unresectable, chemotherapy-refractory liver-limited metastatic CRC (mCRC) comparing arm A (fluorouracil [FU] protracted intravenous infusion 300 mg/m(2) days 1 through 14 every 3 weeks) and arm B (radioembolization plus intravenous FU 225 mg/m(2) days 1 through 14 then 300 mg/m(2) days 1 through 14 every 3 weeks) until hepatic progression. The primary end point was time to liver progression (TTLP). Cross-over to radioembolization was permitted after progression in arm A. Forty-six patients were randomly assigned and 44 were eligible for analysis (arm A, n = 23; arm B, n = 21). Median follow-up was 24.8 months. Median TTLP was 2.1 and 5.5 months in arms A and B, respectively (hazard ratio [HR] = 0.38; 95% CI, 0.20 to 0.72; P = .003). Median time to tumor progression (TTP) was 2.1 and 4.5 months, respectively (HR = 0.51; 95% CI, 0.28 to 0.94; P = .03). Grade 3 or 4 toxicities were recorded in six patients after FU monotherapy and in one patient after radioembolization plus FU treatment (P = .10). Twenty-five of 44 patients received further treatment after progression, including 10 patients in arm A who received radioembolization. Median overall survival was 7.3 and 10.0 months in arms A and B, respectively (HR = 0.92; 95% CI, 0.47 to 1.78; P = .80). Radioembolization with (90)Y-resin microspheres plus FU is well tolerated and significantly improves TTLP and TTP compared with FU alone. This procedure is a valid therapeutic option for chemotherapy-refractory liver-limited mCRC.


Osteoarthropathy in thalassemia minor.
Osteoarthropathy associated with thalassemia minor is rare. The arthritic changes are usually thought to be secondary to bone infarction or erythropoiesis, with decreased trabeculation of the femoral head followed by microfracture and arthritic changes. This report presents the radiographic changes in the hip of a patient with thalassemia minor and discusses the etiology.


Variation in GRM3 affects cognition, prefrontal glutamate, and risk for schizophrenia.
GRM3, a metabotropic glutamate receptor-modulating synaptic glutamate, is a promising schizophrenia candidate gene. In a family-based association study, a common GRM3 haplotype was strongly associated with schizophrenia (P = 0.0001). Within this haplotype, the A allele of single-nucleotide polymorphism (SNP) 4 (hCV11245618) in intron 2 was slightly overtransmitted to probands (P = 0.02). We studied the effects of this SNP on neurobiological traits related to risk for schizophrenia and glutamate neurotransmission. The SNP4 A allele was associated with poorer performance on several cognitive tests of prefrontal and hippocampal function. The physiological basis of this effect was assessed with functional MRI, which showed relatively deleterious activation patterns in both cortical regions in control subjects homozygous for the SNP4 A allele. We next looked at SNP4's effects on two indirect measures of prefrontal glutamate neurotransmission. Prefrontal N-acetylaspartate, an in vivo MRI measure related to synaptic activity and closely correlated with tissue glutamate, was lower in SNP4 AA homozygotes. In postmortem human prefrontal cortex, AA homozygotes had lower mRNA levels of the glial glutamate transporter EAAT2, a protein regulated by GRM3 that critically modulates synaptic glutamate. Effects of SNP4 on prefrontal GRM3 mRNA and protein levels were marginal. Resequencing revealed no missense or splice-site SNPs, suggesting that the intronic SNP4 or related haplotypes may exert subtle regulatory effects on GRM3 transcription. These convergent data point to a specific molecular pathway by which GRM3 genotype alters glutamate neurotransmission, prefrontal and hippocampal physiology and cognition, and thereby increased risk for schizophrenia.


Characteristics and mental health of Hispanic dementia caregivers in New York City.
Dementia prevalence and related caregiving burden are increasing, particularly among Hispanics. We studied the characteristics and mental health of Hispanic caregivers in New York City. We recruited 139 Hispanic family caregivers. We collected data on sociodemographic characteristics and predictors of caregiver burden, measured with the Zarit Caregiver Burden Scale, and depressive symptoms, measured with the Geriatric Depression Scale. The mean age was 59.3 ± 10.4 years. The majority of caregivers were daughters and earned less than US$30 000 a year. In multivariate analyses with linear regression, lower satisfaction with social networks was associated with higher caregiver burden and a greater number of depressive symptoms. Higher dementia severity was associated with higher caregiver burden, while higher caregiver comorbidities were associated with higher depressive symptoms. Caregiver comorbidities and satisfaction with social support may be targets for intervention that could improve caregiver burden and depressive symptoms among Hispanic caregivers.


Human monocytes are associated with the formation of fibrin.
Purified populations of human monocytes (greater than 99% monocytic) contain a subpopulation of cells (8-20%) that will surround themselves with a radial array of needles within 15 s after exposure to plasma. The needles are composed of fibrin as demonstrated by gel electrophoresis of isolated needles and by their reaction with a monoclonal antibody, UC45, that is specific for the alpha chain of fibrin. The addition of UC45 antibody to plasma inhibits the formation of the fibrin needles by monocytes, suggesting that the recognized antigen is important for polymerization. In contrast, UC45 binds weakly to conventional plasma fibrin and does not prevent clotting of plasma although the process is prolonged. Thus, the UC45 antigen is more prominently represented or more accessible on monocyte fibrin. At least 1% of the monocytes were able to form fibers at the earliest time that they could be tested. A greater proportion developed this ability between 2 and 6 h in culture. Macrophages from lung, thymus, and breast milk, when incubated for similar periods of time, do not form fibrin needles. However, phagocytes from tonsils were extremely active in this respect. It is speculated that monocytes may be induced to express fibrin as part of their activities in inflammatory lesions.


Differences between human subjects in the composition of the faecal bacterial community and faecal metabolism of linoleic acid.
Conjugated linoleic acid (CLA) is formed from linoleic acid (LA; cis-9,cis-12-18:2) by intestinal bacteria. Different CLA isomers have different implications for human health. The aim of this study was to investigate LA metabolism and the CLA isomers formed in two individuals (V1 and V2) with different faecal metabolic characteristics, and to compare fatty acid metabolism with the microbial community composition. LA incubated with faecal samples was metabolized at similar rates with both subjects, but the products were different. LA was metabolized extensively to stearic acid (SA; 18:0) in V1, with minor accumulation of CLA and more rapid accumulation of vaccenic acid (VA; trans-11-18:1). CLA accumulation at 4 h was almost tenfold higher with V2, and little SA was formed. At least 12 different isomers of CLA were produced from LA by the colonic bacteria from the two individuals. The predominant (>75%) CLA isomer in V1 was rumenic acid (RA; cis-9,trans-11-18:2), whereas the concentrations of RA and trans-10,cis-12-18:2 were similar with V2. Propionate and butyrate proportions in short-chain fatty acids were higher in V1. A 16S rRNA clone library from V1 contained mainly Bacteroidetes (54% of clones), whereas Firmicutes (66% of clones) predominated in V2. Both samples were devoid of bacteria related to Clostridium proteoclasticum, the only gut bacterium known to metabolize VA to SA. Thus, the CLA formed in the intestine of different individuals may differ according to their resident microbiota, with possibly important implications with respect to gut health.


Changes in the Fitness Fatness Index following reduced exertion high-intensity interval training versus moderate-intensity continuous training in physically inactive adults.
Many adults do not reach the recommended exercise participation guidelines, often citing lack of time as a barrier. Reduced exertion high-intensity training (REHIT) is a mode of exercise that takes as few as 10 min and has been shown to be as effective as other modalities. The Fitness Fatness Index (FFI) is a recently developed index that is used to predict cardiovascular disease (CVD) risk. The aim of this study was to determine the efficacy of a REHIT vs. a traditional moderate-intensity continuous training (MICT) on FFI in physically inactive adults. Thirty-two participants were randomized into one of two 8-week exercise intervention groups: (i) REHIT (n = 16); (ii) MICT (n = 16). The REHIT group performed 10 min of individualized cycling intervals on 2-4 days of the week. The MICT group were prescribed aerobic exercise at 50-65% of their heart rate reserve (HRR) on 3-5 days of the week. FFI was recorded at baseline and post 8-weeks, with FFI being calculated as cardiorespiratory fitness (CRF) (expressed as metabolic equivalents) divided by waist to height ratio (WtHR). A 1-unit increase in FFI was recognized as a clinically significant change in FFI. The REHIT group showed significantly greater (+1.95, ±0.63) improvements in FFI compared to those in the MICT (+0.99, ±0.47) group (between group difference, p < 0.001). Furthermore, there was a greater proportion of participants who achieved a clinically significant change in FFI in the REHIT group (12/12, 100%) than in the MICT group (8/15, 53%) (between group difference, p = 0.01). This study suggests that REHIT may be a more efficacious exercise modality to increase FFI than MICT. This outcome is beneficial as the clinician can prescribe REHIT to physically inactive adults who cite lack of time as a barrier to physical activity participation and achieve significant reductions in CVD risk.


Environmental hypothermia in porcine polytrauma and hemorrhagic shock is safe.
We have previously demonstrated survival benefit to induced hypothermia in a porcine model of controlled hemorrhagic shock simulating an associated delay to definitive care. In the current study, we wished to evaluate the effects of environmental hypothermia in a porcine model of hemorrhagic shock with the addition of polytrauma. Sixteen pigs were randomized to normothermic (39°C, n = 7) or hypothermic (34°C, n = 9) groups. The model included instrumentation, chest injury (captive bolt device), hemorrhage to systolic blood pressure (SBP) of ∼50 mmHg, and crush liver injury. Animals received limited fluid resuscitation for a 1-h period with goal SBP of greater than 80 mmHg and ice packs or warming blankets to achieve goal temperatures, followed by full resuscitation with goal SBP of greater than 90 mmHg, adequate urine output, and hemoglobin by protocol for 20 h. Survivors were observed for an additional 24 h with end points including mortality, markers of organ injury, and neurologic function. There were no differences in survival between the groups (mortality = 1/9, hypothermia group vs. 2/7, normothermia group, P = 0.39). Markers of organ injury were elevated in the hypothermia group at 24 h after injury but were identical between groups at the end of the experimental protocol (48 h after injury). There were no noted differences in neurologic function between the two groups. Environmental hypothermia in a model of polytrauma and hemorrhagic shock was not associated with worse outcomes.


World's first report of sigmoidectomy for sigmoid cancer using the Saroa surgical system with tactile feedback.
Increasing evidence based on the safety and benefits of robot-assisted surgery indicates the disadvantage of the lack of tactile feedback. A lack of tactile feedback increases the risk of intraoperative complications, prolongs operative times, and delays the learning curve. A 40-year-old female patient presented to our hospital with a positive fecal occult blood test. A colonoscopy revealed type 2 advanced cancer of the sigmoid colon, and histological examination showed a well-differentiated adenocarcinoma. Furthermore, abdominal contrast-enhanced computed tomography revealed a tumor in the sigmoid colon and several swollen lymph nodes in the colonic mesentery without distant metastases. The patient was diagnosed with cStage IIIb (cT3N1bM0) sigmoid cancer and underwent sigmoidectomy using the Saroa Surgical System, which was developed by RIVERFIELD, a venture company at the Tokyo Medical and Dental University, and the Tokyo Institute of Technology. Based on adequate simulation, surgery was safely performed with appropriate port placement and arm base-angle adjustment. The operating time was 176 min, with a console time of 116 min and 0 ml blood loss. The patient was discharged 6 days postoperatively without complications. The pathological diagnosis was adenocarcinoma, tub1, tub2, pT2N1bM0, and pStage IIIa. Herein, we report the world's first surgery for sigmoid cancer using the Saroa Surgical System with tactile feedback in which a safe and appropriate oncological surgery was performed.


Dammarane-type triterpenes from the Brazilian medicinal plant Cordia multispicata.
From the Brazilian medicinal plant Carucaá (Cordia multispicata), oleanane- and ursane-type triterpenoids were previously reported as anti-androgenic constituents of the plant. In this study, purification of the polar elements of the EtOAc-soluble fraction of the plant revealed nine novel dammarane-type triterpenes, named cordianols A-I (1-9) along with the known compound cordialin A (10). The structures of these new compounds were elucidated by means of spectral methods including HRFABMS, (1)H NMR, (13)C NMR, and 2D NMR (HMQC, HMBC, NOESY). Absolute configuration at C-23 of compound 7 was determined by an excitone chirality method. Some of these new compounds revealed a hemiketal structure on the A ring and a hydroxylated or epoxidated 20(22)-(E)-ene side chain and showed weak anti-androgenic activity.


Connectivity analysis as a novel approach to motor decoding for prosthesis control.
The use of neural signals for prosthesis control is an emerging frontier of research to restore lost function to amputees and the paralyzed. Electrocorticography (ECoG) brain-machine interfaces (BMI) are an alternative to EEG and neural spiking and local field potential BMI approaches. Conventional ECoG BMIs rely on spectral analysis at specific electrode sites to extract signals for controlling prostheses. We compare traditional features with information about the connectivity of an ECoG electrode network. We use time-varying dynamic Bayesian networks (TV-DBN) to determine connectivity between ECoG channels in humans during a motor task. We show that, on average, TV-DBN connectivity decreases from baseline preceding movement and then becomes negative, indicating an alteration in the phase relationship between electrode pairs. In some subjects, this change occurs preceding and during movement, before changes in low or high frequency power. We tested TV-DBN output in a hand kinematic decoder and obtained an average correlation coefficient (r(2)) between actual and predicted joint angle of 0.40, and as high as 0.66 in one subject. This result compares favorably with spectral feature decoders, for which the average correlation coefficient was 0.13. This work introduces a new feature set based on connectivity and demonstrates its potential to improve ECoG BMI accuracy.


The youth-centricity of three university-associated general practices.
Young people are poor attenders of mainstream general practice; however, university general practices may be an exception, given the age demographic of the population from which they draw patients. The objective of this article was to explore the youth-centricity of 12 months of encounter data from three university-associated general practices: Queensland University of Technology (QUT) in Brisbane, University of New South Wales (UNSW) in Sydney and the University of Melbourne (UniMelb). Encounter data from January to December 2016 were extracted from information management software and analysed using SPSS Statistics 24. The highest percentages of encounters were with patients aged 20-24 years at QUT (39.8%), UNSW (40.6%) and UniMelb (40%). The age encounter data from the general practices associated with QUT, UNSW and UniMelb demonstrate their youth-centricity, with almost 50% of encounters with patients aged 15-24 years. This result is unsurprising given the age demographic from which these practices draw patients, but a proportional under-representation of younger students suggests possible access issues that require further research. Questions regarding the role of these and other Australian university-associated general practices in the health of young people attending university are signposted for future enquiry.


Psychological flexibility and inflexibility as sources of resiliency and risk during a pandemic: Modeling the cascade of COVID-19 stress on family systems with a contextual behavioral science lens.
The coronavirus disease (COVID-19) pandemic and the historic economic shutdown and stay-at-home efforts to slow its spread have radically impacted the lives of families across the world, completely disrupting routines and challenging them to adjust to new health risks as well as to new work and family demands. The current study applied a contextual behavioral science lens to the spillover hypothesis of Family Systems Theory to develop a multi-stage mechanistic model for how COVID-19 stress could impact family and child functioning and how parents' psychological flexibility could shape those processes. A total of 742 coparents (71% female; 84% Caucasian, 85% married, M = 41 years old) of children (ages 5-18, M = 9.4 years old, 50% male) completed an online survey from March 27th to the end of April 2020. Path analyses highlighted robust links from parent inflexibility to all components of the model, predicting: greater COVID-19 stress, greater coparenting discord and family discord, greater caustic parenting, and greater parent and child distress. Parent flexibility was associated with greater family cohesion, lower family discord and greater use of constructive parenting strategies (inductive, democratic/autonomy supportive, positive). Results further suggested that COVID-19 stressors predicted greater family and coparent discord, which in turn predicted greater use of caustic parenting (reactive, inconsistent, aggressive), which in turn predicted greater child and parent distress. The current results highlight parental flexibility and inflexibility as key points of intervention for helping families navigate the current global health crisis, highlighting the crucial role they play in the lives of families.


Developing a decision-making model based on an interdisciplinary oncological care group for the management of colorectal cancer.
To report our experience on implementation and preliminary results of a decision-making model based on the recommendations of an Interdisciplinary Oncological Care Group developed for the management of colorectal cancer. The multidisciplinary team identified a reference guideline using appraisal of guidelines for research and evaluation (AGREE) tool based on a sequential assessment of the guideline quality. Thereafter, internal guidelines with diagnostic and therapeutic management for early, locally advanced and metastatic colonic and rectal cancer were drafted; organizational aspects, responsibility matrices, protocol actions for each area of specialty involved and indicators for performing audits were also defined. The National Institute for Health and Care Excellence (NICE) UK guideline was the reference for drafting the internal guideline document; from February to November 2013, 125 patients with colorectal cancer were discussed by and taken under the care of the Interdisciplinary Oncological Care Group. The first audit performed in December 2013 revealed optimal adherence to the internal guideline, mainly in terms of uniformity and accuracy of perioperative staging, coordination and timing of multi-modal therapies. To date, all patients under observation are within the diagnostic and therapeutic course, no patient came out from the multidisciplinary "path" and only in 14% of cases have the first recommendations proposed been changed. The selected indicators appear effective and reliable, while at the moment, it is not yet possible to assess the impact of the multidisciplinary team on clinical outcome. Although having a short observation period, our model seems capable of determining optimal uniformity of diagnostic and therapeutic management, to a high degree of patient satisfaction. A longer observation period is necessary in order to confirm these observations and for assessing the impact on clinical outcome.


Enhanced hit-to-lead process using bioanalogous lead evolution and chemogenomics: application in designing selective matrix metalloprotease inhibitors.
The authors describe an innovative approach for designing novel inhibitors. This approach effectively integrates the emerging chemogenomics concept of target-family-based drug discovery with bioanalogous design strategies, including privileged structures, molecular frameworks as well as bioisosteric and bioanalogous/isofunctional modifications. The authors applied this method in the design of selective inhibitors of matrix metalloproteases (MMPs), also referred to as matrixins, on the basis of a unique analysis of the ligand-target knowledge base, the 'matrixinome'. For this analysis, the authors created an annotated MMP database containing ∼ 300 inhibitors with their published activity profile. The ligand space was then arranged into a lead evolution tree, where the substructural transformations in each virtual step led to marked changes in the activity pattern. This allowed subtype-specific privileged fragments to be extracted as well as modifications, which improve activity and/or selectivity. Furthermore, the compounds with the preferred activity profile were correlated with sequence homology as well as binding site similarity within the target family, thereby leading to the identification of substructural modifications that turn non-selective, biohomologous structures into selective inhibitors. The matrixinomic application of the authors' approach, therefore, provides an example of how the combination of ligand space knowledge with sequence-related data can radically improve the outcome of the lead optimisation process to achieve higher selectivity within a given target family.


High Heterogeneity of Virus-Neutralizing and RBD-Binding Activities of COVID-19 Convalescent Sera.
The parameters of the humoral response are an important immunological characteristic of donors who recovered from COVID-19 and vaccinated individuals. Analysis of the level of virus-binding antibodies has become widespread. The most accurate predictor of effective immune protection against symptomatic SARS-CoV-2 infection is the activity of virus-neutralizing antibodies. We determined virus-neutralizing activities in plasma samples of individuals (n = 111) who had COVID-19 from April to September 2020. Three independent methods were used: conventional with live virus, with virus-like particles pseudotyped with spike protein, and a surrogate virus-neutralization test (cVNT, pVNT and sVNT, respectively). For comparison, the levels of IgG, IgA and IgM antibodies against the receptor-binding domain of the SARS-CoV-2 spike protein were also evaluated. The levels of virus-binding as well as virus-neutralizing antibodies in cVNT and pVNT showed high heterogeneity. A comparison of cVNT and pVNT results showed a high correlation, sVNT results also correlated well with both cVNT and pVNT. To the greatest extent, the level of IgG antibodies correlated with the results of cVNT, pVNT and sVNT. These results can be used in the selection of plasmas that are best suited for transfusion and treatment of acute COVID-19. In addition, data on the virus-neutralizing activity of plasma are important for the selection of potential donors, for the isolation of SARS-CoV-2-specific B-lymphocytes, in order to further generate monoclonal virus-neutralizing antibodies.


Blood monocytes consist of two principal subsets with distinct migratory properties.
Peripheral blood monocytes are a heterogeneous population of circulating leukocytes. Using a murine adoptive transfer system to probe monocyte homing and differentiation in vivo, we identified two functional subsets among murine blood monocytes: a short-lived CX(3)CR1(lo)CCR2(+)Gr1(+) subset that is actively recruited to inflamed tissues and a CX(3)CR1(hi)CCR2(-)Gr1(-) subset characterized by CX(3)CR1-dependent recruitment to noninflamed tissues. Both subsets have the potential to differentiate into dendritic cells in vivo. The level of CX(3)CR1 expression also defines the two major human monocyte subsets, the CD14(+)CD16(-) and CD14(lo)CD16(+) monocytes, which share phenotype and homing potential with the mouse subsets. These findings raise the potential for novel therapeutic strategies in inflammatory diseases.


Application of Contrast-Enhanced Ultrasound in Cystic Pancreatic Lesions Using a Simplified Classification Diagnostic Criterion.
Objective. Classification diagnosis was performed for cystic pancreatic lesions using ultrasound (US) and contrast-enhanced ultrasound (CEUS) to explore the diagnostic value of CEUS by comparison with enhanced CT. Methods. Sixty-four cases with cystic pancreatic lesions were included in this study. The cystic lesions of pancreas were classified into four types by US, CEUS, and CT: type I unilocular cysts; type II microcystic lesions; type III macrocystic lesions; and type IV cystic lesions with solid components or irregular thickening of the cystic wall or septa. Results. Eighteen type I, 7 type II, 10 type III, and 29 type IV cases were diagnosed by CT. The classification results by US were as follows: 6 type I; 5 type II; 4 type III; and 49 type IV cases. Compared with the results by enhanced CT, the kappa value was 0.36. Using CEUS, 15, 6, 12, and 31 cases were diagnosed as types I-IV, respectively. The kappa value was 0.77. Conclusion. CEUS has obvious superiority over US in the classification diagnostic accuracy in cystic pancreatic lesions and CEUS results showed substantial agreement with enhanced CT. CEUS could contribute to the differential diagnosis of cystic pancreatic diseases.


Spin-paired solvated electron couples in alkali-ammonia systems.
Alkali metals form deep blue solutions of solvated electrons in liquid ammonia. To explain the diamagnetism of more concentrated solutions, DFT and ab initio computations have been used to show that spin-paired couples of electrons can overcome Coulomb repulsion, occupying a cavity formed by solvent molecules, comparable in size to that of single solvated electrons. Sluggish hydrogen evolution from the solutions is rationalized by a high activation barrier for reaction of the electron pair with an ammonia molecule.


[Evolution of demand for childbirth services from rural cities to Sergipe, Aracaju, Brazil, 1970-1996].
This study analyzes trends in the migration of childbirth from rural areas of Sergipe State, Brazil, to the capital city, Aracaju, from 1970 to 1996. Data on "mother's place of residence" were obtained from mothers whose children were born in maternity hospitals in Aracaju in 1970, 1976, 1986, and 1996. Significant differences occurred in the proportion of mothers who resided outside of Aracaju but came there to give birth, especially from 1976 to 1986. This shift was stimulated by improvement of State highways and the granting of personal political favors in exchange for votes. Changes in the State hospital structure did not appear to have influenced the childbirth shift to Aracaju. There was a 134.6% increase in deliveries occurring in the State capital. There were also different trends among the various regions of the State, with some regional health districts displaying a downward trend in the number of deliveries from 1986 to 1996. It is urgent that more regional and hierarchical support be provided for childbirth and neonatal care in order to ensure quality assistance for pregnant women and their infants.


A transmembrane intracellular estrogen receptor mediates rapid cell signaling.
The steroid hormone estrogen regulates many functionally unrelated processes in numerous tissues. Although it is traditionally thought to control transcriptional activation through the classical nuclear estrogen receptors, it also initiates many rapid nongenomic signaling events. We found that of all G protein-coupled receptors characterized to date, GPR30 is uniquely localized to the endoplasmic reticulum, where it specifically binds estrogen and fluorescent estrogen derivatives. Activating GPR30 by estrogen resulted in intracellular calcium mobilization and synthesis of phosphatidylinositol 3,4,5-trisphosphate in the nucleus. Thus, GPR30 represents an intracellular transmembrane estrogen receptor that may contribute to normal estrogen physiology as well as pathophysiology.


Multiple cullin-associated E3 ligases regulate cyclin D1 protein stability.
Cyclin D1 is a key regulator of cell cycle progression, which forms a complex with CDK4/6 to regulate G1/S transition during cell cycle progression. Cyclin D1 has been recognized as an oncogene since it was upregulated in several different types of cancers. It is known that the post-translational regulation of cyclin D1 is controlled by ubiquitination/proteasome degradation system in a phosphorylation-dependent manner. Several cullin-associated F-box E3 ligases have been shown to regulate cyclin D1 degradation; however, it is not known if additional cullin-associated E3 ligases participate in the regulation of cyclin D1 protein stability. In this study, we have screened an siRNA library containing siRNAs specific for 154 ligase subunits, including F-box, SOCS, BTB-containing proteins, and DDB proteins. We found that multiple cullin-associated E3 ligases regulate cyclin D1 activity, including Keap1, DDB2, and WSB2. We found that these E3 ligases interact with cyclin D1, regulate cyclin D1 ubiquitination and proteasome degradation in a phosphorylation-dependent manner. These E3 ligases also control cell cycle progression and cell proliferation through regulation of cyclin D1 protein stability. Our study provides novel insights into the regulatory mechanisms of cyclin D1 protein stability and function.


[Effect of heme oxygenase-1 expression and activity on the heart function in ischemia-reperfusion].
The purpose of this study was to determine the protective effects of heme oxygenase-1 (HO-1) expression against postischemic myocardial dysfunction. We also investigated HO-1 expression in cardiac tissue from the left and right ventricles of myocardium. Rat hearts were isolated and perfused according to Langendorff technique to evaluate the recovery of myocardial function after 20 min of global ischemia and 40 min of reperfusion. We found that HO-1 expression was more expressed in left ventricles of myocardium in basic conditions and after ischemia/reperfusion as well as after its previous induction by hemin. Upregulation of the inducible isoform of HO-1 and increase its activity after treatment of animals with hemin 24 h before ischemia ameliorated myocardial function (raised left ventricular developed pressure, decreased end-diastolic pressure, attenuated vasoconstriction) and reduced oxydative stress in cardiac tissue during reperfusion of isolated hearts. Zinc protoporphyrin IX, an inhibitor of heme oxygenase activity, completely abolished the HO-1 expression in left ventricles of myocardium and increased postischemic myocardial dysfunction. Likewise, cardiac tissue injury was exacerbated by treatment with zinc protoporphyrin IX through significant inhibition of HO activity and increasing of hydroxyl radical production on reperfusion. The treatment of animals with hemin and following ischemia/reperfusion resulted in 5-6-times increase of HO-1 expression in the left ventricle of myocardium whereas in right ventricle only in 3-times. Our data provide strong evidence for a primary role of HO-1 in cardioprotection against reperfusion injury and show different HO-1 expression in left and right ventricles of myocardium.


Ablation of the Cl-/HCO3- Exchanger Pendrin Enhances Hydrochlorothiazide-Induced Diuresis.
The Cl-/HCO3- exchanger pendrin and the thiazide-sensitive Na-Cl cotransporter NCC are expressed in the kidney distal nephron and mediate salt absorption. We hypothesized that deletion of pendrin leaves NCC as the major salt absorbing transporter in the distal nephron and therefore enhances salt excretion by hydrochlorothiazide (HCTZ). Metabolic cage studies were performed in wild type, pendrin KO and NCC KO mice at baseline and following HCTZ treatment. In parallel studies, systemic blood pressure was measured in mice treated with HCTZ with the tail cuff method. Urine output, salt excretion and water intake were comparable in all groups under baseline condition. Urine output and water intake increased significantly only in pendrin KO mice in response to HCTZ, but not in WT or NCC KO mice. Sodium and chloride excretion increased in HCTZ-treated pendrin KO mice, but they remained unchanged in WT or NCC KO mice. Pendrin KO mice treated with HCTZ developed volume depletion, as determined by increased expression of renin mRNA and protein. The expression of ENaC and pendrin increased in HCTZ-treated WT mice. HCTZ treatment did not significantly modify blood pressure in any of the experimental group. The ablation of the Cl-/HCO3- exchanger Pendrin enhances the magnitude of salt wasting by HCTZ.


Differential roles of unsaturated and saturated fatty acids on autophagy and apoptosis in hepatocytes.
Fatty acid-induced lipotoxicity plays a critical role in the pathogenesis of nonalcoholic liver disease. Saturated fatty acids and unsaturated fatty acids have differential effects on cell death and steatosis, but the mechanisms responsible for these differences are not known. Using cultured HepG2 cells and primary mouse hepatocytes, we found that unsaturated and saturated fatty acids differentially regulate autophagy and apoptosis. The unsaturated fatty acid, oleic acid, promoted the formation of triglyceride-enriched lipid droplets and induced autophagy but had a minimal effect on apoptosis. In contrast, the saturated fatty acid, palmitic acid, was poorly converted into triglyceride-enriched lipid droplets, suppressed autophagy, and significantly induced apoptosis. Subsequent studies revealed that palmitic acid-induced apoptosis suppressed autophagy by inducing caspase-dependent Beclin 1 cleavage, indicating cross-talk between apoptosis and autophagy. Moreover, our data suggest that the formation of triglyceride-enriched lipid droplets and induction of autophagy are protective mechanisms against fatty acid-induced lipotoxicity. In line with our in vitro findings, we found that high-fat diet-induced hepatic steatosis was associated with autophagy in the mouse liver. Potential modulation of autophagy may be a novel approach that has therapeutic benefits for obesity-induced steatosis and liver injury.


A Distinct Mechanism of Temporal Integration for Motion through Depth.
Temporal integration of visual motion has been studied extensively within the frontoparallel plane (i.e., 2D). However, the majority of motion occurs within a 3D environment, and it is unknown whether the principles from 2D motion processing generalize to more realistic 3D motion. We therefore characterized and compared temporal integration underlying 2D (left/right) and 3D (toward/away) direction discrimination in human observers, varying motion coherence across a range of viewing durations. The resulting discrimination-versus-duration functions followed three stages, as follows: (1) a steep improvement during the first ∼150 ms, likely reflecting early sensory processing; (2) a subsequent, more gradual benefit of increasing duration over several hundreds of milliseconds, consistent with some form of temporal integration underlying decision formation; and (3) a final stage in which performance ceased to improve with duration over ∼1 s, which is consistent with an upper limit on integration. As previously found, improvements in 2D direction discrimination with time were consistent with near-perfect integration. In contrast, 3D motion sensitivity was lower overall and exhibited a substantial departure from perfect integration. These results confirm that there are overall differences in sensitivity for 2D and 3D motion that are consistent with a sensory difference between binocular and dichoptic sensory mechanisms. They also reveal a difference at the integration stage, in which 3D motion is not accumulated as perfectly as in the 2D motion model system.


VP2 virus-like particles elicit protective immunity against duckling short beak and dwarfism syndrome in ducks.
Duckling short beak and dwarfism syndrome virus (SBDSV), an emerging goose parvovirus, has caused short beak and dwarfism syndrome (SBDS) in Chinese duck flocks since 2015. Presently, there is no commercial vaccine against SBDS. In the present study, a virus-like particle (VLP)-based candidate vaccine was developed against this disease. A baculovirus expression system was used to express the SBDSV VP2 protein in Sf9 cells. Immunofluorescence assay, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting were used to confirm protein expression. Furthermore, transmission electron microscopy was used to observe the formation of VLPs. VLPs were formulated into an oil-adjuvanted maternal vaccine to evaluate humoral responses in breeding ducks via latex particle agglutination inhibition assay (LPAI) and microneutralization assay. The offspring were challenged with SBDSV to test the protective efficacy. A single dose of SBDSV was able to induce the high level of LPAI antibodies in ducks, with LPAI and neutralization peak titres of 4.9 ± 1.20 log2 and 7.1 ± 1.20 log2, respectively, at 4 weeks post-vaccination (wpv). The average LPAI titre of yolk antibodies in duck eggs receiving 2 doses (first and boost doses) of the vaccine was 5.3 ± 1.09 log2 at 4 weeks post-boost. The protective efficacy of the maternal vaccine was 87.5%-100%. These results indicate that SBDSV VLPs can be a promising vaccine candidate for controlling SBDS.


Microrheology of interphase chromosomes with spatial constraints: a computational study.
Chromatin fibers within the interior of the nucleus of the cell make stable interactions with the nucleoskeleton, an ensemble of 'extra-chromatin' structures which help ensuring genome stability. Although the role of these interactions appears crucial to the correct behavior of the cell, their impact on chromatin structure and dynamics remains to be elucidated. In order to tackle this important issue, in this work we introduce a simple polymer model for chromatin fibers in interphase which takes into account the two generic properties of chain-versus-chain mutual uncrossability and the presence of stable binding interactions to an extra-chromatin nuclear matrix. To study how these constraints affect chromatin structure from small to large scales, we employ extensive molecular dynamics computer simulations and we monitor the motion of nanoprobes of different sizes embedded within the polymer medium. Our results demonstrate that nanoprobes show hampered motion whenever their linear size becomes larger than chromatin stiffness. This transition is also displaying features which usually belong to the realm of glassy systems, namely long-tail correlations in the distribution functions of nanoprobe spatial displacements and heterogeneous behavior accompanied by ergodicity breaking.


Research progress of targeted therapy combined with immunotherapy for hepatocellular carcinoma.
Hepatocellular carcinoma is a common gastrointestinal malignancy with a high mortality rate and limited treatment options. Molecularly targeted drugs combined with immune checkpoint inhibitors have shown unique advantages over single-agent applications, significantly prolonging patient survival. This paper reviews the research progress of molecular-targeted drugs combined with immune checkpoint inhibitors in the treatment of hepatocellular carcinoma and discusses the effectiveness and safety of the combination of the two drugs to provide a reference for the further application of molecular-targeted drugs combined with immune checkpoint inhibitors in clinical practice.


The modulation of leaf metabolism plays a role in salt tolerance of Cymodocea nodosa exposed to hypersaline stress in mesocosms.
Applying proteomics, we tested the physiological responses of the euryhaline seagrass Cymodocea nodosa to deliberate manipulation of salinity in a mesocosm system. Plants were subjected to a chronic hypersaline condition (43 psu) to compare protein expression and plant photochemistry responses after 15 and 30 days of exposure with those of plants cultured under normal/ambient saline conditions (37 psu). Results showed a general decline in the expression level of leaf proteins in hypersaline stressed plants, with more intense reductions after long-lasting exposure. Specifically, the carbon-fixing enzyme RuBisCo displayed a lower accumulation level in stressed plants relative to controls. In contrast, the key enzymes involved in the regulation of glycolysis, cytosolic glyceraldehyde-3-phosphate dehydrogenase, enolase 2 and triose-phosphate isomerase, showed significantly higher accumulation levels. These responses suggested a shift in carbon metabolism in stressed plants. Hypersaline stress also induced a significant alteration of the photosynthetic physiology of C. nodosa by means of a down-regulation in structural proteins and enzymes of both PSII and PSI. However we found an over-expression of the cytochrome b559 alpha subunit of the PSII initial complex, which is a receptor for the PSII core proteins involved in biogenesis or repair processes and therefore potentially involved in the absence of effects at the photochemical level of stressed plants. As expected hypersalinity also affects vacuolar metabolism by increasing the leaf cell turgor pressure and enhancing the up-take of Na(+) by over-accumulating the tonoplast specific intrinsic protein pyrophosphate-energized inorganic pyrophosphatase (H(+)-PPase) coupled to the Na(+)/H(+)-antiporter. The modulation of carbon metabolism and the enhancement of vacuole capacity in Na(+) sequestration and osmolarity changes are discussed in relation to salt tolerance of C. nodosa.


Impact of different management measures on the colonization of broiler chickens with ESBL- and pAmpC- producing Escherichia coli in an experimental seeder-bird model.
The colonization of broilers with extended-spectrum β-lactamase- (ESBL-) and plasmid-mediated AmpC β-lactamase- (pAmpC-) producing Enterobacteriaceae has been extensively studied. However, only limited data on intervention strategies to reduce the colonization throughout the fattening period are available. To investigate practically relevant management measures for their potential to reduce colonization, a recently published seeder-bird colonization model was used. Groups of 90 broilers (breed Ross 308) were housed in pens under conventional conditions (stocking of 39 kg/m2, no enrichment, water and feed ad libitum). Tested measures were investigated in separate trials and included (I) an increased amount of litter in the pen, (II) the reduction of stocking density to 25 kg/m2, and (III) the use of an alternative broiler breed (Rowan x Ranger). One-fifth of ESBL- and pAmpC- negative broilers (n = 18) per group were orally co-inoculated with two E. coli strains on the third day of the trial (seeder). One CTX-M-15-positive E. coli strain (ST410) and one CMY-2 and mcr-1-positive E. coli strain (ST10) were simultaneously administered in a dosage of 102 cfu. Colonization of all seeders and 28 non-inoculated broilers (sentinel) was assessed via cloacal swabs during the trials and a final necropsy at a target weight of two kilograms (= d 36 (control, I-II), d 47 (III)). None of the applied intervention measures reduced the colonization of the broilers with both the ESBL- and the pAmpC- producing E. coli strains. A strain-dependent reduction of colonization for the ESBL- producing E. coli strain of ST410 by 2 log units was apparent by the reduction of stocking density to 25 kg/m2. Consequently, the tested management measures had a negligible effect on the ESBL- and pAmpC- colonization of broilers. Therefore, intervention strategies should focus on the prevention of ESBL- and pAmpC- colonization, rather than an attempt to reduce an already existing colonization.


New-onset pediatric nephrotic syndrome following Pfizer-BioNTech SARS-CoV-2 vaccination: a case report and literature review.
Various new vaccines against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been rapidly developed. The new onset and recurrence of nephrotic syndrome triggered by some vaccines have been documented and several adult cases of minimal change nephrotic syndrome newly developing after SARS-CoV-2 vaccination have been reported. However, no reports of pediatric cases have been published. Indications for SARS-CoV-2 vaccines have been expanded to those as young as 12 years old and vaccination of children has just started in Japan. We encountered a 15-year-old boy without underlying disease who newly developed nephrotic syndrome after SARS-CoV-2 vaccination with BNT162b2 (Pfizer-BioNTech). He developed eyelid edema 4 days after vaccination and peripheral edema of the lower extremities a further 4 days later. Twenty-one days after vaccination, 60 mg of oral daily prednisolone was started. He achieved complete remission in 12 days without complications such as hypertension or acute kidney injury. We clinicians should be aware of the possibility of nephrotic syndrome developing after SARS-CoV-2 vaccination, not only in adults, but also in children.


[Two cases of stroke associated with the use of finasteride, an approved drug for male-pattern hair loss in Japan].
We report two cases of stroke associated with the use of finasteride at 1 mg/day, which is approved in Japan for the treatment of male-pattern hair loss. The first case involved a 35-year-old male taking 1 mg of finasteride daily for 6 months to prevent male-pattern hair loss. He was taken to a hospital and later admitted to our hospital owing to headache and seizures. Brain computed tomography (CT) images showed a low-density area in the right frontal lobe. CT venography (CTV) revealed sinus thrombosis and he was treated with an anticoagulant. As the headache gradually subsided, medications were tapered and terminated 10 months later when venous flow to the sagittal sinus and left transverse sinus was confirmed to be recanalized. The second case involved a 41-year-old male taking 1 mg of finasteride and 6 mg of minoxidil daily for 1 year for male-pattern hair loss. He started having headaches and was admitted to our hospital when diffusion-weighted images of brain magnetic resonance imaging (MRI) showed a high-intensity area in the left parietotemporal lobe. He was treated with antiplatelet and anticoagulation medicines. The Japan Pharmaceutical and Medical Devices Agency (PMDA) has reported 14 cases of thrombosis in patients taking finasteride in Japan; 4 cases of stroke (our 2 cases and 2 reported by PMDA), 6 cases of myocardial infarction, and 4 cases of other thrombotic diseases. Increases in estrone and estradiol levels in prostate cancer patients and controls receiving 5 mg of finasteride have been reported. Gynecomastia has also been reported as one of the adverse effects of finasteride at 1 mg or 5 mg daily. Taken together, we assume that the increases in estrone and estradiol levels induced by finasteride lead to thrombosis development.


Genotype-phenotype correlations in children with congenital hyperinsulinism due to recessive mutations of the adenosine triphosphate-sensitive potassium channel genes.
Congenital hyperinsulinism (HI) is most commonly caused by recessive mutations of the pancreatic beta-cell ATP-sensitive potassium channel (K(ATP)), encoded by two genes on chromosome 11p, SUR1 and Kir6.2. The two mutations that have been best studied, SUR1 g3992-9a and SUR1 delF1388, are null mutations yielding nonfunctional channels and are characterized by nonresponsiveness to diazoxide, a channel agonist, and absence of acute insulin responses (AIRs) to tolbutamide, a channel antagonist, or leucine. To examine phenotypes of other K(ATP) mutations, we measured AIRs to calcium, leucine, glucose, and tolbutamide in infants with recessive SUR1 or Kir6.2 mutations expressed as diffuse HI (n = 8) or focal HI (n = 14). Of the 24 total mutations, at least seven showed evidence of residual K(ATP) channel function. This included positive AIR to both tolbutamide and leucine in diffuse HI cases or positive AIR to leucine in focal HI cases. One patient with partial K(ATP) function also responded to treatment with the channel agonist, diazoxide. Six of the seven patients with partial defects had amino acid substitutions or insertions; whereas, the other patient was compound heterozygous for two premature stop codons. These results indicate that some K(ATP) mutations can yield partially functioning channels, including cases of hyperinsulinism that are fully responsive to diazoxide therapy.


The chemokine CXCL12 generates costimulatory signals in T cells to enhance phosphorylation and clustering of the adaptor protein SLP-76.
The CXC chemokine CXCL12 mediates the chemoattraction of T cells and enhances the stimulation of T cells through the T cell receptor (TCR). The adaptor SLP-76 [Src homology 2 (SH2) domain-containing leukocyte protein of 76 kD] has two key tyrosine residues, Tyr(113) and Tyr(128), that mediate signaling downstream of the TCR. We investigated the effect of CXCL12 on SLP-76 phosphorylation and the TCR-dependent formation of SLP-76 microclusters. Although CXCL12 alone failed to induce SLP-76 cluster formation, it enhanced the number, stability, and phosphorylation of SLP-76 microclusters formed in response to stimulation of the TCR by an activating antibody against CD3, a component of the TCR complex. Addition of CXCL12 to anti-CD3-stimulated cells resulted in F-actin polymerization that stabilized SLP-76 microclusters in the cells' periphery at the interface with antibody-coated coverslips and increased the interaction between SLP-76 clusters and those containing ZAP-70, the TCR-associated kinase that phosphorylates SLP-76, as well as increased TCR-dependent gene expression. Costimulation with CXCL12 and anti-CD3 increased the extent of phosphorylation of SLP-76 at Tyr(113) and Tyr(128), but not that of other TCR-proximal components, and mutation of either one of these residues impaired the CXCL12-dependent effect on SLP-76 microcluster formation, F-actin polymerization, and TCR-dependent gene expression. The effects of CXCL12 on SLP-76 microcluster formation were dependent on the coupling of its receptor CXCR4 to G(i)-family G proteins (heterotrimeric guanine nucleotide-binding proteins). Thus, we identified a costimulatory mechanism by which CXCL12 and antigen converge at SLP-76 microcluster formation to enhance T cell responses.


Vasopressin withdrawal produces hypotension in the spontaneously hypertensive rat.
In spontaneously hypertensive rats (SHR) and their normotensive Wistar-Kyoto controls (WKY), prolonged intravenous infusions of either arginine vasopressin (AVP, 8 mU X kg-1 X min-1) or phenylephrine (PE, 20 nmol X kg-1 X min-1) resulted in similar rises in arterial pressure. Heart rate fell greatly in the WKY but not in the SHR. Withdrawal of the PE infusion resulted in moderate decreases in blood pressure and increases in heart rate; these responses were similar in SHR and WKY. At 5 h after PE withdrawal, blood pressure and heart rate returned to basal values. In contrast, withdrawal of the AVP infusion was associated with greater falls in blood pressure and rises in heart rate. Blood pressure and heart rate in both the SHR and the WKY at 5 h after AVP were significantly different from their respective basal values. The effects of AVP withdrawal on either blood pressure or heart rate were significantly greater in the SHR than in the WKY. At 5 h after the withdrawal of AVP, blood pressure in the SHR was reduced to normotensive levels. These results suggest that the withdrawal effect was specific to AVP, was more marked in the SHR, and might not result from only the rise in blood pressure seen during the intravenous infusion of the pressor agent.


Modular approach to triazole-linked 1,6-α-D-oligomannosides to the discovery of inhibitors of Mycobacterium tuberculosis cell wall synthetase.
Aiming at developing inhibitors of mannosyltransferases, the enzymes that participate in the biosynthesis of the cell envelope of Mycobacterium tuberculosis, the synthesis of a range of designed triazole-linked 1,6-oligomannosides up to a hexadecamer has been accomplished by a modular approach centered on the Cu(I)-catalyzed azide-alkyne cycloaddition as key process. The efficiency and fidelity of the cycloaddition are substantiated by high yields (76-96%) and exclusive formation of the expected 1,4-disubstituted triazole ring in all oligomer assembling reactions. Key features of oligomers thus prepared are the anomeric carbon-carbon bond of all mannoside residues and the 6-deoxymannoside capping residue. Suitable bioassays with dimer, tetramer, hexamer, octamer, decamer, and hexadecamer showed variable inhibitor activity against mycobacterial α-(1,6)-mannosyltransferases, the highest activity (IC(50) = 0.14-0.22 mM) being registered with the hexamannoside and octamannoside.


Induction of H2O2 in transgenic rice leads to cell death and enhanced resistance to both bacterial and fungal pathogens.
Oxidative burst, mediated by hydrogen peroxide (H2O2), has been recognized as a key component of plant defense response during an incompatible interaction. To determine if elevated levels of H2O2 lead to cell death, activation of defense genes and enhanced resistance to diverse pathogens, transgenic rice plants expressing a fungal glucose oxidase gene (GOX) were generated using both constitutive and inducible expression systems. Constitutive or wound/pathogen-induced expression of GOX also allowed us to determine the effectiveness of these systems in conferring long lasting resistance to various pathogens. Both constitutive and wound/pathogen-induced expression of GOX lead to increases in the endogenous levels of H2O2, which in turn caused cell death. Elevated levels of H2O2 also activated the expression of several defense genes and these transgenic plants showed enhanced resistance to both bacterial and fungal pathogens. In comparison to inducible expression, constitutive expression of GOX resulted in 3-10-fold higher levels of the GOX transcript and the corresponding enzymatic activity. Such increased levels of GOX, which would result in elevated levels of H2O2, caused improper seed set and decreased seed viability in transgenic plants constitutively expressing GOX. Our results suggest that pathogen inducible expression of heterologous genes may be a practical and robust way of generating broad spectrum disease resistance.


Ikaros-Notch axis in host hematopoietic cells regulates experimental graft-versus-host disease.
Host hematopoietically derived APCs play a vital role in the initiation of GVH responses. However, the APC autonomous molecular mechanisms that are critical for the induction of GVHD are not known. We report here that the Ikaros-Notch axis in host hematopoietically derived APCs regulates the severity of acute GVHD across multiple clinically relevant murine models of experimental bone marrow transplantation. In the present study, Ikaros deficiency (Ik(-/-)) limited to host hematopoietically derived APCs enhanced donor T-cell expansion and intensified acute GVHD, as determined by survival and other GVHD-specific parameters. The Ik(-/-) conventional CD8(+) and CD8(-)CD11c(+) dendritic cells (DCs), the most potent APCs, showed no increase in the expression of activation markers or in response to TLR stimulation compared with wild-type controls. However, Ik(-/-) DCs demonstrated an enhanced stimulation of allogeneic T cells. Deficiency of Ikaros in the conventional CD8(+) and CD8(-)CD11c(+) DCs was associated with an increase in Notch signaling, the blockade of which mitigated the enhanced in vitro and in vivo allostimulatory capacity. Therefore, the Ikaros-Notch axis is a novel pathway that modulates DC biology in general, and targeting this pathway in host hematopoietically derived APCs may reduce GVHD.


[Prevalence of caries, fissure sealants and filling materials among German children and children of migrants].
The aim of this study was to collect information on dental health and dental care of German children and children of migrants for planning oral health promotion in the Rems-Murr-district. To meet this aim all first and fourth degree children in primary and special schools were examined. The dmft- and DMFT-Index, the number of fissure sealants, the filling material used and the children's nationality were recorded. The dmft of 6- and 7-year-olds was 1.50 among Germans, 4.61 among German migrants from Russia, 4.02 among Turks, 4.05 among children from former Yugoslavia, 2.35 among Italians, 1.95 among Greeks and 3.76 among children of other nationalities. At the age of 9 to 10 years Germans had an average of 0.31, German migrants from Russia 0.77, Turks 1.19, children from former Yugoslavia 1.32, Italians 0.64, Greeks 0.69 and children of other nationalities 0.57 DMF-teeth. In both age groups the proportion of caries-free children was highest among Germans and lowest among Turks. Also, more migrants than Germans were referred to a dentist for caries treatment. Compared to the Germans fewer migrants had at least one tooth sealed. The proportion of amalgam fillings to the total number of fillings was higher among 9- and 10-year-olds of Turkish, Italian or Yugoslavian origin than among other nationalities. The results of this study show that further development of oral health promotion programmes for children and parents with Turkish and Russian cultural background has the top-most priority.


Effects of forest disturbance on the fitness of an endemic rodent in a biodiversity hotspot.
Praomys delectorum occurs abundantly in both disturbed and intact forests in the Ukaguru Mountains within the Eastern Arc Mountains (EAM), Morogoro, Tanzania. While previous studies have reported that anthropogenic disturbances such as grazing, wood cutting, and harvesting have a positive effect on the population density of P. delectorum, the impact of habitat disturbance on its demographic traits is still unknown. We performed a capture-mark-recapture study in both disturbed and intact forests from June 2018 to February 2020 in order to investigate the effects of habitat disturbance on abundance and two demographic traits: survival and maturation of P. delectorum in the Ukaguru Mountains. We found no variation in abundance or maturation between intact and disturbed forests, but habitat type did affect survival. However, this effect was sex-dependent since female survival was higher in disturbed forests, while male survival remained similar across the two forest types potentially due to differences in predation pressure or food availability between the two habitats. Continuous demographic monitoring of P. delectorum in EAM is necessary given that the increasing human population surrounding the landscape is leading to higher deforestation rates and expansion of the pine plantation in the forest reserve.


Run to the hills: exotic fish invasions and water quality degradation drive native fish to higher altitudes.
While the significance of anthropogenic pressures in shaping species distributions and abundances is undeniable, some ambiguity still remains on their relative magnitude and interplay with natural environmental factors. In our study, we examined 91 late-invasion-stage river locations in Northern Italy using ordination methods and variance partitioning (partial-CCA), as well as an assessment of environmental thresholds (TITAN), to attempt to disentangle the effects of eutrophication and exotic species on native species. We found that exotic species, jointly with water quality (primarily eutrophication) and geomorphology, are the main drivers of the distribution of native species and that native species suffer more joint effects than exotic species. We also found that water temperature clearly separates species distributions and that some native species, like Italian bleak (Alburnus alborella) and Italian rudd (Scardinius hesperidicus), seem to be the most resilient to exotic fish species. We also analyzed the dataset for nestedness (BINMATNEST) to identify priority targets of conservation. As a result, we confirmed that altitude correlated negatively with eutrophication and nestedness of exotic species and positively with native species. Overall, our analysis was able to detect the effects of species invasions even at a late invasion stage, although reciprocal effects seemed comparable at this stage. Exotic species have pushed most native species on the edge of local extinction in several sites and displaced most of them on the rim of their natural distribution. Any potential site- and species-specific conservation action aimed at improving this situation could benefit from a carefully considered prioritization to yield the highest results-per-effort and success rate. However, we encourage future research to update the information available before singling out specific sites for conservation or outlining conservation actions.


Dietary lysophosphatidylcholine-EPA enriches both EPA and DHA in the brain: potential treatment for depression.
EPA and DHA protect against multiple metabolic and neurologic disorders. Although DHA appears more effective for neuroinflammatory conditions, EPA is more beneficial for depression. However, the brain contains negligible amounts of EPA, and dietary supplements fail to increase it appreciably. We tested the hypothesis that this failure is due to absorption of EPA as triacylglycerol, whereas the transporter at the blood-brain barrier requires EPA as lysophosphatidylcholine (LPC). We compared tissue uptake in normal mice gavaged with equal amounts (3.3 μmol/day) of either LPC-EPA or free EPA (surrogate for current supplements) for 15 days and also measured target gene expression. Compared with the no-EPA control, LPC-EPA increased brain EPA >100-fold (from 0.03 to 4 μmol/g); free EPA had little effect. Furthermore, LPC-EPA, but not free EPA, increased brain DHA 2-fold. Free EPA increased EPA in adipose tissue, and both supplements increased EPA and DHA in the liver and heart. Only LPC-EPA increased EPA and DHA in the retina, and expression of brain-derived neurotrophic factor, cyclic AMP response element binding protein, and 5-hydroxy tryptamine (serotonin) receptor 1A in the brain. These novel results show that brain EPA can be increased through diet. Because LPC-EPA increased both EPA and DHA in the brain, it may help in the treatment of depression as well as neuroinflammatory diseases, such as Alzheimer's disease.


Emergence of resistance to atovaquone-proguanil in malaria parasites: insights from computational modeling and clinical case reports.
The usefulness of atovaquone-proguanil (AP) as an antimalarial treatment is compromised by the emergence of atovaquone resistance during therapy. However, the origin of the parasite mitochondrial DNA (mtDNA) mutation conferring atovaquone resistance remains elusive. Here, we report a patient-based stochastic model that tracks the intrahost emergence of mutations in the multicopy mtDNA during the first erythrocytic parasite cycles leading to the malaria febrile episode. The effect of mtDNA copy number, mutation rate, mutation cost, and total parasite load on the mutant parasite load per patient was evaluated. Computer simulations showed that almost any infected patient carried, after four to seven erythrocytic cycles, de novo mutant parasites at low frequency, with varied frequencies of parasites carrying varied numbers of mutant mtDNA copies. A large interpatient variability in the size of this mutant reservoir was found; this variability was due to the different parameters tested but also to the relaxed replication and partitioning of mtDNA copies during mitosis. We also report seven clinical cases in which AP-resistant infections were treated by AP. These provided evidence that parasiticidal drug concentrations against AP-resistant parasites were transiently obtained within days after treatment initiation. Altogether, these results suggest that each patient carries new mtDNA mutant parasites that emerge before treatment but are killed by high starting drug concentrations. However, because the size of this mutant reservoir is highly variable from patient to patient, we propose that some patients fail to eliminate all of the mutant parasites, repeatedly producing de novo AP treatment failures.


Lifting capacity is associated with central sensitization and non-organic signs in patients with chronic back pain.
To analyze the associations between lifting capacity, and central sensitization (CS) and non-organic signs (NOS) in patients with chronic back pain (CBP) attending vocational rehabilitation. Cross-sectional observational multicenter study among patients with CBP undergoing a return to work assessment within care as usual. Main analyses: step 1: partial correlation between lifting capacity, and CS, NOS, and additional variables; step 2: multiple regression in stepwise forward method for dependent variable lifting capacity, and for independent variables CS and NOS, and additional variables significant (p < 0.05) at step 1. All analyses were controlled for sex. Fifty-six patients of mean age 42.5 years and 59% women participated in the study. Correlations between lifting capacity and CS and NOS were r = -0.53 and r = -0.50, respectively. CS and NOS, as well as age and sex, contributed significantly to the final regression model, which explained 57.6% of variance. After controlling for confounders, CS and NOS were negatively associated with lifting capacity in patients with CBP. Explained variance was substantially higher than previously reported studies.Implications for RehabilitationThe identification of central sensitization and non-organic signs (NOS) in patients with chronic back pain can alert clinicians about central nervous system being in a hypersensitive state and about pain behavior.Central sensitization and NOS are relevant determinants of lifting capacity.Better understanding of the factors affecting lifting capacity lead to better design and tailoring of interventions, resulting in optimized vocational rehabilitation programs and faster return to work.


The emergence of carbapenemase-producing Enterobacterales in hospitals: a major challenge for a debilitated healthcare system in Lebanon.
Carbapenem- and extended-spectrum cephalosporin-resistant Enterobacterales (CR-E and ESCR-E, respectively) are increasingly isolated worldwide. Information about these bacteria is sporadic in Lebanon and generally relies on conventional diagnostic methods, which is detrimental for a country that is struggling with an unprecedented economic crisis and a collapsing public health system. Here, CR-E isolates from different Lebanese hospitals were characterized. Non-duplicate clinical ESCR-E or CR-E isolates (N = 188) were collected from three hospitals from June 2019 to December 2020. Isolates were identified by MALDI-TOF, and their antibiotic susceptibility by Kirby-Bauer disk diffusion assay. CR-E isolates (n = 33/188) were further analyzed using Illumina-based WGS to identify resistome, MLST, and plasmid types. Additionally, the genetic relatedness of the CR-E isolates was evaluated using an Infrared Biotyper system and compared to WGS. Using the Kirby-Bauer disk diffusion assay, only 90 isolates out of the 188 isolates that were collected based on their initial routine susceptibility profile by the three participating hospitals could be confirmed as ESCR-E or CR-E isolates and were included in this study. This collection comprised E. coli (n = 70; 77.8%), K. pneumoniae (n = 13; 14.4%), Enterobacter spp. (n = 6; 6.7%), and Proteus mirabilis (n = 1; 1.1%). While 57 were only ESBL producers the remaining 33 isolates (i.e., 26 E. coli, five K. pneumoniae, one E. cloacae, and one Enterobacter hormaechei) were resistant to at least one carbapenem, of which 20 were also ESBL-producers. Among the 33 CR-E, five different carbapenemase determinants were identified: blaNDM-5 (14/33), blaOXA-244 (10/33), blaOXA-48 (5/33), blaNDM-1 (3/33), and blaOXA-181 (1/33) genes. Notably, 20 CR-E isolates were also ESBL-producers. The analysis of the genetic relatedness revealed a substantial genetic diversity among CR-E isolates, suggesting evolution and transmission from various sources. This study highlighted the emergence and broad dissemination of blaNDM-5 and blaOXA-244 genes in Lebanese clinical settings. The weak AMR awareness in the Lebanese community and the ongoing economic and healthcare challenges have spurred self-medication practices. Our findings highlight an urgent need for transformative approaches to combat antimicrobial resistance in both community and hospital settings.


Subconjunctival and external hemorrhage secondary to oral anticoagulation.
Recurrent, bilateral, and severe conjunctival hemorrhages mandate the search for an underlying etiology, such as a blood dyscrasia, blood clotting disorder, or recurrent increased intrathoracic pressure caused by repetitive vomiting or coughing spells. Medical and ophthalmic management is discussed in a case of subconjunctival and external hemorrhage from the eye secondary to prolonged prothrombin time in oral anticoagulation therapy.


Coronavirus disease 2019: a bibliometric analysis and review.
On December 8, 2019, many cases of pneumonia with unknown etiology were first reported in Wuhan, China, subsequently identified as a novel coronavirus infection aroused worldwide concern. As the outbreak is ongoing, more and more researchers focused interest on the COVID-19. Therefore, we retrospectively analyzed the publications about COVID-19 to summarize the research hotspots and make a review, to provide reference for researchers in the world. We conducted a search in PubMed using the keywords "COVID-19" from inception to March 1, 2020. Identified and analyzed the data included title, corresponding author, language, publication time, publication type, research focus. 183 publications published from 2020 January 14 to 2020 February 29 were included in the study. The first corresponding authors of the publications were from 20 different countries. Among them, 78 (42.6%) from the hospital, 64 (35%) from the university and 39 (21.3%) from the research institution. All the publications were published in 80 different journals. Journal of Medical Virology published most of them (n=25). 60 (32.8%) were original research, 29 (15.8%) were review, 20 (10.9%) were short communications. 68 (37.2%) epidemiology, 49 (26.8%) virology and 26 (14.2%) clinical features. According to our review, China has provided a large number of research data for various research fields, during the outbreak of COVID-19. Most of the findings play an important role in preventing and controlling the epidemic around the world. With research on the COVID-19 still booming, new vaccine and effective medicine for COVID-19 will be expected to come out in the near future with the joint efforts of researchers worldwide.


[Role of the individual sensitivity to various cytostatic agents in children with neuroblastomas].
Primary plasmic cultures of 33 tumors of the sympathic nervous system of children (27 neuroblastomas and 6 ganglioneuromas) were used for determination of their individual sensitivity to vincristin, vinblastin, dactinomycin, mithramycin, olivomycin, cyclophosphant bruneomycin and adriamycin. Most of the tumors were sensitive to vincristin, rubomycin an adriamycin. The data obtained in vitro and in the clinic were compared with respect to 12 cases (treatment with vincristin, combination of olivomycin with cyclophosphan, vincristin with rubomycin or cyclophosphan). Coinsidence of the results was observed in 8 cases. The method of primary plasmic cultures may be successfully used for determination of individual sensitivity of children with neuroblastomas to various cytostatics.