Pulmonary manifestation of immunoglobulin G4-related disease in a 7-year-old immunodeficient boy with Epstein-Barr virus infection: a case report.
Immunoglobulin G4-related disease (IgG4-RD) is a multiorgan fibroinflammatory condition with lymphoplasmacytic infiltrates containing abundant IgG4-positive plasma cells. The immunopathogenesis of the disease and the potential role of triggering autoantigens or infectious factors have not been clearly defined. Immunoglobulin G4-related lung disease is a new and emerging condition in pediatric patients and to date, there have been only two reports regarding pulmonary manifestation of IgG4-RD in children recently published. This is the first report of IgG4-related lung disease in an immunodeficient child with Epstein-Barr virus infection. We report on the case of a 7-year old atopic boy who was hospitalized with an initial clinical and radiological diagnosis of pneumonia, positive Epstein-Barr virus (EBV)-DNA in the blood and defective adaptive immunity. The lung CT showed a consolidated mass lesion adjacent to the posterior wall of the chest and the diaphragm. The child underwent surgical resection of the tumor, and the histologic examination of the lung specimens revealed lymphoplasmacytic infiltrates with fibrosis and vasculitis correlating with IgG4-related lung disease. Subsequent monitoring of the patient with lung CT, pulmonary function tests and IgG4 levels did not show signs of active disease. The diagnosis of IgG4-related lung disease in children is challenging because of its rarity, nonspecific symptomatology and heterogeneous morphological manifestations. Further studies are required in children with pulmonary presentation of IgG4-RD to better understand pathogenesis of this condition, possible immunological or infectious triggering factors, and finally, to determine pediatric patient-targeted therapeutic interventions.


Multifaceted Interplay between Hormones, Growth Factors and Hypoxia in the Tumor Microenvironment.
Hormones and growth factors (GFs) are signaling molecules implicated in the regulation of a variety of cellular processes. They play important roles in both healthy and tumor cells, where they function by binding to specific receptors on target cells and activating downstream signaling cascades. The stages of tumor progression are influenced by hormones and GF signaling. Hypoxia, a hallmark of cancer progression, contributes to tumor plasticity and heterogeneity. Most solid tumors contain a hypoxic core due to rapid cellular proliferation that outgrows the blood supply. In these circumstances, hypoxia-inducible factors (HIFs) play a central role in the adaptation of tumor cells to their new environment, dramatically reshaping their transcriptional profile. HIF signaling is modulated by a variety of factors including hormones and GFs, which activate signaling pathways that enhance tumor growth and metastatic potential and impair responses to therapy. In this review, we summarize the role of hormones and GFs during cancer onset and progression with a particular focus on hypoxia and the interplay with HIF proteins. We also discuss how hypoxia influences the efficacy of cancer immunotherapy, considering that a hypoxic environment may act as a determinant of the immune-excluded phenotype and a major hindrance to the success of adoptive cell therapies.


MTDH-stabilized DDX17 promotes tumor initiation and progression through interacting with YB1 to induce EGFR transcription in Hepatocellular Carcinoma.
Metadherin (MTDH) is a well-established oncogene in various cancers including Hepatocellular Carcinoma (HCC). However, the precise mechanism through which MTDH promotes cancer-related signaling pathways in HCC remains unknown. In this study, we identified DDX17 as a novel binding partner of MTDH. Furthermore, MTDH increased the protein level of DDX17 by inhibiting its ubiquitination. We confirmed that DDX17 was a novel oncogene, with dramatically upregulated expression in HCC tissues. The increased expression of DDX17 was closely associated with vascular invasion, TNM stage, BCLC stage, and poor prognosis. In vitro and in vivo tests demonstrated that DDX17, a downstream target of MTDH, played a crucial role in tumor initiation and progression. Mechanistically, DDX17 acted as a transcriptional regulator that interacted with Y-box binding protein 1 (YB1) in the nucleus, which in turn drove the binding of YB1 to its target epidermal growth factor receptor (EGFR) gene promoter to increase its transcription. This in turn increased expression of EGFR and the activation of the downstream MEK/pERK signaling pathway. Our results identify DDX17, stabilized by MTDH, as a powerful oncogene in HCC and suggest that the DDX17/YB1/EGFR axis contributes to tumorigenesis and metastasis of HCC.


Characterization and expression analysis of a chicken interleukin-6 receptor alpha.
Interleukin-6 (IL-6) is a multifunctional cytokine that plays roles in regulating immune responses, acute phase reactions and hematopoiesis. IL-6 signaling is regulated by two receptors, a specific alpha chain (IL-6Ralpha) and a signal transducer, gp130. In this study, cDNA encoding the 445 amino acid propeptide of chicken IL-6Ralpha (chIL-6Ralpha) was identified. The predicted 445 amino acids showed approximately 40% sequence identity with mammalian homologues. In a domain search, chIL-6Ralpha had a signal peptide of 20 residues, an immunoglobulin-like (IG) domain of 71 residues and a fibronectin-type III (FN III) domain of 85 residues. On comparison with mammalian homologues, four conserved cysteine residues and the WSXWS motif were observed in the N- and C-terminal regions of the FN III domain, respectively. Expression analysis revealed that chIL-6Ralpha is strongly expressed in liver and the chicken hepatoma cell line LMH. These findings indicate that the identified chicken cDNA sequence encodes a chIL-6Ralpha homologue.


The complete mitochondrial genome of Sarcoptes scabiei var. nyctereutis from the Japanese raccoon dog: Prediction and detection of two transfer RNAs (tRNA-A and tRNA-Y).
Sarcoptes scabiei (Acari: Sarcoptidae) causes a common contagious skin disease that affects many mammals. Here, the complete mitochondrial genome of a mite, S. scabiei var. nyctereutis, from Japanese wild raccoon dogs was analyzed. The 13,837bp circular genome contained 13 protein-coding genes, two rRNA genes, and 22 tRNA genes. For the first time, two tRNAs (alanine and tyrosine), that were thought to be absent in scabies mites from other animals, were predicted to have short, non-cloverleaf structures by in silico annotation and detected by RT-PCR, sequencing, and northern analysis. The mitochondrial genome structure of S. scabiei is similar to that of Psoroptes cuniculi and Dermatophagoides farinae. While small and unusual tRNA genes seem to be common among acariform mites, further experimental evidence for their presence is needed. Furthermore, through an analysis of the cox1 gene, we have provided new evidence to confirm the transmission of this mite between different animal hosts.


Impairment of glucose tolerance: mechanism of action and impact on the cardiovascular system.
Macrovascular disease, especially coronary heart diseases, have been found to be linked to glucose intolerance. Insulin resistance in respect to glucose uptake in peripheral tissues seems to play an important role in the development of glucose intolerance, since subjects with coronary heart disease mainly are hyperinsulinemic. Insulin resistance may induce not only glucose intolerance but also hypertension, obesity, and dyslipoproteinemia (high very low-density lipoprotein and low high-density lipoprotein values), all variables that add to the risk of coronary heart disease. On the basis of these findings, a new syndrome has been postulated-syndrome X. This syndrome may be caused by inherited insulin resistance in skeletal muscles, and secondary to that arterial hypertension, obesity, and dyslipoproteinemia may develop. Insulin resistance in noninsulin-dependent diabetic persons and in hypertensive subjects is located in skeletal muscles, where insulin's ability to promote nonoxidative glucose metabolism is reduced. The key enzyme in this pathway, glycogen synthase, is proposed as the causal defect responsible for the insulin resistance state, at least in noninsulin-dependent diabetic patients. The pill (sex steroids) may induce a clinical situation that is similar to syndrome X. However, it is important to emphasize that many more studies are needed to substantiate these hypothetical mechanisms behind coronary heart disease.


Hypothyroidism in diabetes mellitus patients in Eastern Nepal.
The coexistence of diabetes mellitus (DM) with hypothyroidism is a known clinical observation. To estimate prevalence and co-relate that of hypothyroidism in patients with DM in relation to the age and sex, the lipid profile, body mass index visiting diabetes clinic and inpatients in B. P. Koirala Institute of Health Sciences. The hospital-based descriptive study. Two hundred and seventy-one known or newly detected cases of DM aged more than 15 years were selected randomly from September 2012 to September 2013 and subjected to evaluation for thyroid function - clinically and biochemically and other relevant investigations were done. For descriptive statistics mean, standard deviation, percentage, proportion were calculated. For inferential statistics following test were carried out at the level of significant 0.05 where confidence interval is 95%. The statistical operations were done through Statistical Package for the Social Sciences version 10. Of 271 subjects, the prevalence of hypothyroidism (clinical and subclinical) in diabetics was, 4.05% (11/271) with females preponderance, of which 7 (30.4%) were clinically hypothyroid and 4 (17.4%) were subclinical hypothyroid. One (4.3%) patient had subclinical hyperthyroidism. The mean age at diagnosis of type 2 DM was 51-60 years. 8.69% of diabetics with primary hypothyroids were having morbid obesity. High-density lipoprotein among different thyroid status were statistically significant (P = 0.042). Hypothyroidism is not uncommon in diabetes, and we found body mass index, mean triglyceride and cholesterol levels were more in those diabetic patients having coexisting hypothyroidism.


Vitamin D supplementation improves bone mineralisation independent of dietary phosphate in male X-linked hypophosphatemic (Hyp) mice.
The disorder of X-linked hypophosphatemia (XLH), results in the supressed renal production of active 1α,25-dihydroxyvitamin D (1,25(OH)2D) due to elevated fibroblast growth factor-23 (FGF23) levels. While adequate 25(OH)D levels are generally associated with improved mineralisation of the skeleton independent of circulating 1,25(OH)2D levels, it is unclear whether raising 25(OH)D to sufficiently high levels through dietary vitamin D3 administration contributes to improving bone mineralisation in the murine homolog for XLH, Hyp mice. Three-week-old male Hyp mice were fed one of four diets containing either 1000 IU (C) or 20,000 IU (D) vitamin D3/kg diet with either 0.35% phosphate or 1.25% phosphate (P) until 12 weeks of age (n = 12/group). When compared to C-fed mice, D-fed mice significantly elevated serum 25(OH)D levels to 72.8 ± 4.9 nmol/L (2-fold, p < 0.001) and increased both cortical bone mineral density (15%, p < 0.01), and vertebral trabecular BV/TV% (80%, p < 0.001), despite persistent hypophosphatemia and normocalcemia. The increase in bone volume was associated with improved Tb.Th (12%, p < 0.01) and Tb.N (63%, p < 0.001). Unlike with D-diet, P-fed mice resulted in increased femoral (15%, p < 0.001) and vertebral (12%, p < 0.001) length, and a 34% increase in vertebral trabecular BV/TV% when compared to control fed animals (p < 0.001). However, the addition of the high P diet to the high D diet did not result in additive effects on bone mineralisation when compared to the effects of D diet alone, despite serum 25(OH)D levels elevated to 118.8 ± 8.6 nmol/L. In D-fed mice, the increase in bone mineral density and volume was associated with reduced osteoid volume, reduced ObS/BS, and a trend for reduced serum PTH levels, suggesting reduced bone turnover in these animals. Thus, elevating serum 25(OH)D levels independently improves bone mineralisation in Hyp mice without causing hypercalcemia, suggesting that further studies are required in XLH patients to establish the role of increasing 25(OH)D levels in improving bone mineralisation.


HRMAS 1H-NMR measured changes of the metabolite profile as mesenchymal stem cells differentiate to targeted fat cells in vitro: implications for non-invasive monitoring of stem cell differentiation in vivo.
Mesenchymal stem cells (MSCs) have shown a great potential for clinical applications in regenerative medicine. However, it remains challenging to follow the transplanted cell grafts in vivo. Nuclear magnetic resonance spectroscopy (NMR or MRS) is capable of determining and quantifying the cellular metabolites in tissue and organs non-invasively, therefore it is an attractive method for monitoring and evaluating the differentiation and functions of transplanted stem cells in vivo. In this study, metabolic changes of MSCs undergoing adipogenic differentiation to targeted fat cells were investigated in vitro, using solid-state high-resolution magic angle spinning (1)H nuclear magnetic resonance spectroscopy. Quantification of metabolite concentrations before and after differentiation of MSCs showed decreased levels of intracellular metabolites, including choline, creatine, glutamate and myo-inositol, and a substantially increased level of fatty acids, when mesenchymal stem cells were differentiated preferentially to fat cells. Intracellular creatine, myo-inositol and choline reduced from 10.4 +/- 0.72, 16.2 +/- 1.2 and 8.22 +/- 0.51 mM to 3.27 +/- 0.34, 6.1 +/- 0.46 and 3.11 +/- 0.32 mM, respectively, while fatty acids increased from 32.6 +/- 1.5 to 91.2 +/- 3.2 mM after undergoing 3 weeks of differentiation. The increase of the fatty acid concentration measured by NMR is confirmed by the observation of 80% fat cells in differentiated cells by cell counting assay, suggesting resonances from fatty acids may be used as metabolite markers for monitoring MSC differentiation to fat cells in vivo, using the magnetic resonance spectroscopic technique readily available on MRI scanners.


Histatin-1 loaded multifunctional, adhesive and conductive biomolecular hydrogel to treat diabetic wound.
Micro-/macroangiopathy, neuropathy and prolonged inflammation are common in diabetic wound, however, traditional wound dressing cannot treat these problems in the same time. Herein, we developed a multifunctional hydrogel with promoted angiogenesis, cell proliferation and anti-inflammation ability to treat diabetic wound. The hydrogel was composed of natural polymers, including gelatin and chitosan, which have excellent biocompatibility. Histatin-1 (His-1) was added into the hydrogel to improve the cell adhesion, proliferation and angiogenesis. Besides, polypyrrole based conductive nanoparticles (G-Ppy) were introduced in the hydrogel to enhance the electrical signal conduction between skin and promote the mechanical strength of the hydrogel. The polypyrrole nanoparticles were growth in the chain of methacryloyl grafted gelatin (Gel-MA), leading to a better biocompatibility and water dispersibility. In vivo wound healing experiment proved that the hydrogel accelerated the wound healing rate, down regulation the expression of pro-inflammation factor TNF-α and upregulation the expression of CD31 and α-SMA, indicating the prospects in the application of diabetic wound healing.


Preventing early, prolonged vitamin E deficiency: an opportunity for better cognitive outcomes via early diagnosis through neonatal screening.
To evaluate whether early diagnosis of cystic fibrosis (CF) through newborn screening (NBS) and early vitamin E status are associated with cognitive function. We assessed cognitive function for 71 children without meconium ileus (ages 7.3-16.9 years) enrolled in the screened (S) or control (C) group of the Wisconsin CF Neonatal Screening Project. The Test of Cognitive Skills, 2nd edition generated the cognitive skills index (CSI; mean = 100, SD = 16). Vitamin E deficiency at diagnosis was defined as plasma alpha-tocopherol (alpha-T) below 300 microg/dL (<300E). Primary analyses evaluated CSI scores across the 4 levels of group (S or C) by using alpha-T status (<300E or >300E) with analysis of covariance. After adjusting for covariates, CSI in the C<300E group was significantly lower than each of the other groups (C>300E, S<300E, and S>300E; P < .05). The highest proportion of CSI scores >84 occurred in the C<300E group (41%). Patients in this group also had the lowest mean head circumference z-scores at diagnosis. Our results show that prolonged alpha-T deficiency in infancy is associated with lower subsequent cognitive performance. Thus, diagnosis via NBS may benefit the cognitive development of children with CF, particularly in those prone to vitamin E deficiency during infancy.


Metabotyping of human colorectal cancer using two-dimensional gas chromatography mass spectrometry.
Colorectal cancer (CRC) is the fourth most common cause of death from cancer in the world. The limitations of the currently available methods and biomarkers for CRC management highlight the necessity of finding novel markers. Metabonomics can be used to search for potential markers that can provide molecular insight into human CRC. The emergence of two-dimensional gas chromatography time of flight mass spectrometry (GC × GC/TOFMS) has comprehensively enhanced the metabolic space coverage of conventional GC/MS. In this study, a GC × GC/TOFMS was developed for the tissue-based global metabonomic profiling of CRC. A Pegasus GC × GC/TOFMS (Leco Corp., St. Joseph, MI, USA) system comprising an Agilent 7890 GC and Pegasus IV TOFMS was used for this purpose. An Agilent DB-1 (30 m × 250 μm × 0.25 μm) fused silica capillary column and a Restek Rxi®-17 (1 m × 100 μm × 0.10 μm) fused silica capillary column were used as the primary and secondary columns, respectively. The method was applied for global metabonomic profiling of matched CRC and normal tissues (n = 63) obtained from 31 CRC patients during surgery. An attempt was also made to compare GC × GC/TOFMS with GC/MS and NMR in similar application. The results showed that the metabotype associated with CRC is distinct from that of normal tissue and led to the identification of chemically diverse marker metabolites. Metabolic pathway mapping suggested deregulation of various biochemical processes such as glycolysis, Krebs cycle, osmoregulation, steroid biosynthesis, eicosanoid biosynthesis, bile acid biosynthesis, lipid, amino acid and nucleotide metabolism.


Potential of targeting host cell calcium dynamics to curtail SARS-CoV-2 infection and COVID-19 pathogenesis.
Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) infection and associated coronavirus disease 2019 (COVID-19) has severely impacted human well-being. Although vaccination programs have helped in reducing the severity of the disease, drug regimens for clinical management of COVID-19 are not well recognized yet. It is therefore important to identify and characterize the molecular pathways that could be therapeutically targeted to halt SARS-CoV-2 infection and COVID-19 pathogenesis. SARS-CoV-2 hijacks host cell molecular machinery for its entry, replication and egress. Interestingly, SARS-CoV-2 interacts with host cell Calcium (Ca2+) handling proteins and perturbs Ca2+ homeostasis. We here systematically review the literature that demonstrates a critical role of host cell Ca2+ dynamics in regulating SARS-CoV-2 infection and COVID-19 pathogenesis. Further, we discuss recent studies, which have reported that SARS-CoV-2 acts on several organelle-specific Ca2+ transport mechanisms. Moreover, we deliberate upon the possibility of curtailing SARS-CoV-2 infection by targeting host cell Ca2+ handling machinery. Importantly, we delve into the clinical trials that are examining the efficacy of FDA-approved small molecules acting on Ca2+ handling machinery for the management of COVID-19. Although an important role of host cell Ca2+ signaling in driving SARS-CoV-2 infection has emerged, the underlying molecular mechanisms remain poorly understood. In future, it would be important to investigate in detail the signaling cascades that connect perturbed Ca2+ dynamics to SARS-CoV-2 infection.


Both N-ras and c-myc are activated in the SHAC human stomach fibrosarcoma cell line.
A transforming N-ras gene was isolated from the SHAC human stomach fibrosarcoma cell line. A single-point mutation resulting in the substitution of histidine for glutamine at codon 61 was found in the SHAC transforming allele. The N-ras gene is overexpressed in the tumor cells and transformant cells. The N-ras p21 product was studied by immunoprecipitation and showed no alteration in mobility as compared to the normal p21 protein. The c-myc gene is amplified and overexpressed in these cells. This report gives evidence that an amplified c-myc and a mutated N-ras gene are both present in this tumor cell line and provides support for the idea that co-operation of at least 2 activated cellular oncogenes is required for carcinogenesis.


Identification of a folate binder in hog kidney.
A macromolecular binder of folic acid (pteroylglutamic acid) and folic acid derivatives has been identified in extracts of hog kidney. With partially purified preparations, binding of [3H]pteroylglutamate was competed for by unlabeled pteroylglutamate, 5-methyltetrahydrofolic acid and its triglutamate derivative, by tetra- and dihydrofolic acid, and by N-10-formyltetrahydrofolic acid. The partially purified extract did not bine [3H]methotrexate nor could methotrexate or 5-formyltetrahydrofolic acid compete for [3H]folic acid-binding sites. The rate of binding of pterolyglutamate at 37 degrees was approximately 3%/s, was independent of pteroylglutamate concentration, and was essentially irreversible between pH 6.0 and 9.0. Below pH 6.0 binding was reversible, and at pH 3.5 the folic acid-binder complex completely disassociated. Based upon Sephadex gel filtration, the molecular weight of the folate-binder complex is 35,000 to 40,000. Binding activity was unaffected by pretreatment with ribonuclease or deoxyribonuclease but was completely destroyed by trypsin. The initial, unfractionated extract showed gamma-glutamyl carboxypeptidase (conjugase) activity which was lost in subsequent steps of purification of the folate binder.


Access to Substituted Thiophenes through Xanthate-Mediated Vinyl C(sp2)-Br Bond Cleavage and Heterocyclization of Bromoenynes.
An environmentally sustainable strategy for the chemoselective heterocyclization of bromoenynes through a transition-metal-free sulfuration/cyclization process is reported. Using inexpensive and safe EtOCS2K as a thiol surrogate and tetrabutylphosphonium bromide and H2O as a mixed solvent, the reaction provided a range of substituted thiophenes in moderate to good yields. In addition, 2,3,4,5-tetrasubstituted thiophenes were able to be prepared under mild reaction conditions by electrophilic heterocyclization with NH4I and EtOCS2K in good yields.


Enteral glutamine does not enhance the effects of hepatocyte growth factor in short bowel syndrome.
This study was designed to determine if luminally administered glutamine alone functions as a growth factor or is synergistic with hepatocyte growth factor (HGF) after massive small bowel resection (MSBR). Twenty Sprague-Dawley rats underwent an 80% small bowel resection and jejunostomy tube placement. Seven days later the rats were divided into four groups: group 1, control, no further treatment (n = 5); group 2 received glutamine (4% of total food intake per day) via an orogastric tube (n = 5); group 3 received intraluminal HGF via a jejunostomy tube at 75 microg/kg/d (n = 5); and group 4 received glutamine and HGF at the same doses, respectively. After a 14-day HGF infusion, glutamine feeding, or both combined, [C14] glycine absorption (micromol/L/cm2 intestine) and mucosal DNA and protein content (microg/mg mucosa) were measured in the remaining small bowel. Glutamine alone had no effect on substrate absorption and protein or DNA content. HGF increased galactose absorption (106% increase over control, P<.01), glycine absorption (95% increase over control, P<.05), protein content (44% increase over control, P<.01), and DNA content (32% increase over control, P<.01). The combination of glutamine and HGF did not prove to be synergistic. These data demonstrate that in this short bowel model, glutamine alone did not enhance intestinal function. Furthermore, glutamine is not synergistic with HGF. This study suggests that glutamine alone may not be useful clinically in patients with inadequate intestinal function.


Activation of Intrinsic Growth State Enhances Host Axonal Regeneration into Neural Progenitor Cell Grafts.
Axonal regeneration after spinal cord injury (SCI) can be enhanced by activation of the intrinsic neuronal growth state and, separately, by placement of growth-enabling neural progenitor cell (NPC) grafts into lesion sites. Indeed, NPC grafts support regeneration of all host axonal projections innervating the normal spinal cord. However, some host axons regenerate only short distances into grafts. We examined whether activation of the growth state of the host injured neuron would elicit greater regeneration into NPC grafts. Rats received NPC grafts into SCI lesions in combination with peripheral "conditioning" lesions. Six weeks later, conditioned host sensory axons exhibited a significant, 9.6-fold increase in regeneration into the lesion/graft site compared with unconditioned axons. Regeneration was further enhanced 1.6-fold by enriching NPC grafts with phenotypically appropriate sensory neuronal targets. Thus, activation of the intrinsic host neuronal growth state and manipulation of the graft environment enhance axonal regeneration after SCI.


Basal cell carcinoma of the prostate gland: a case report and brief review of the basal cell proliferations of the prostate gland.
Basal cell proliferations within the prostate gland encompass a group of benign and malignant entities. Although basal cell hyperplasia is a common finding, basal cell carcinoma of the prostate gland is a rare tumor that can be mistaken by a benign condition and represents a diagnostic problem in genitourinary pathology. We report a case of basal cell carcinoma in a previously healthy 65-year-old man with urinary symptoms and low prostate-specific antigen. The microscopic findings are presented and the use of immunohistochemical markers classifying basal cell lesions of the prostate discussed.


Mitochondrial membrane potential selects hybridomas yielding high viability in fed-batch cultures.
Prior research (Follstad, B. D.; Wang, D. I. C.; Stephanopoulos, G. Mitochondrial membrane potential differentiates cells resistant to apoptosis in hybridoma cultures. Eur. J. Biochem. 2000, 267, 6534-6540.) identified mitochondrial membrane potential (MMP) as a marker of hybridoma subpopulations resistant to apoptosis caused by a variety of apoptosis inducers. In this study, we investigated the viability of hybridoma cell cultures inoculated with cells of varying MMP in regular fed-batch operation. A hybridoma cell population was separated using FACS into subpopulations based on their mean mitochondrial membrane potential (MMP) as measured using the common mitochondrial stain, Rhodamine 123 (Rh123). These subpopulations showed dramatic differences in their apoptotic death kinetics. Fed-batches inoculated with a high MMP subpopulation reached higher viable cell concentrations and viabilities that were maintained for prolonged periods of time relative to fed-batches inoculated with low MMP subpopulations. These results underline the heterogeneous nature of hybridoma cell cultures and suggest that mitochondrial physiology is a critical parameter determining culture performance.


CMKLR1 and GPR1 mediate chemerin signaling through the RhoA/ROCK pathway.
Chemerin is an adipose-derived hormone that regulates immunity and energy homesotasis. To date, all known chemerin functions have been attributed to activation of the G protein-coupled receptor chemokine-like receptor-1 (CMKLR1). Chemerin is also the only known ligand for a second receptor, G protein-coupled receptor-1 (GPR1), whose signaling and function remains unknown. This study investigated the in vitro signal transduction mechanisms of CMKLR1 and GPR1 using a panel of luciferase-reporters and pathway-specific inhibitors. Herein we report the novel finding that chemerin signals through a RhoA and rho-associated protein kinase (ROCK)-dependent pathway for activation of the transcriptional regulator serum-response factor (SRF). Despite similarities in RhoA/ROCK, Gαi/o, and MAPK signaling, we also demonstrate species-specific and receptor-dependent variations in GPR1 and CMKLR1 signaling and expression of the SRF target genes EGR1, FOS and VCL. Moreover, we demonstrate that signaling through p38, Gαi/o, RhoA, and ROCK is required for chemerin-mediated chemotaxis of L1.2 lymphocytes and AGS gastric adenocarcinoma cells. These results provide, to our knowledge, the first empirical evidence that GPR1 is a functional chemerin receptor and identify RhoA/SRF as a novel chemerin-signaling axis via both CMKLR1 and GPR1.


[Understanding child neglect in a gender context: a study performed in a Brazilian city].
To describe the phenomenon of child neglect and understand it in a gender context. Retrospective, quantitative, and exploratory study that analyzed reports of violence by a child and adolescent protection network in a Brazilian city. The theoretical and methodological basis applied was TIPESC (Theory of Nursing Praxical Intervention in Collective Health), with a gender emphasis. Neglect of children under the age of ten represents more than half the reports received over all the years studied; more boys than girls suffered neglect and 41.4% of the reports of neglect involved children under than age of three; women were identified as being solely responsible in 67.9%, and as accessories in 17.3% of the incidents reported. Child neglect is a complex matter, the gender subordinate status inflicted on these children and their mothers who are responsible for their care underscore the social vulnerability of this group.


Norton scale for predicting prognosis in elderly patients undergoing trans-catheter aortic valve implantation: A historical prospective study.
The Norton scale is traditionally used to assess the risk of pressure ulcers. However, recent studies have shown its prognostic utilization in elderly patients with diverse medical conditions. The association between low admission Norton scale scores (ANSS), complications, and mortality in elderly patients following trans-catheter aortic valve implantation (TAVI) has never been studied. We aimed to determine if low ANSS (≤16) is associated with complications and 30-day and 1-year mortality in elderly patients undergoing TAVI. The medical charts of elderly (≥70 years) TAVI patients at the Tel-Aviv Medical Center, a tertiary medical center, were studied for the following measurements: ANSS, demographics, co-morbidities, complications during hospitalization, and 30-day and 1-year mortality. Complications included: an atrio-ventricular block, stroke, and vascular complications. The cohort included 302 elderly patients: 179 (59.3%) were women; the mean age was 83.3±5.1 years. Following TAVI, 112 (37.1%) patients had complications other than pressure ulcers, 10 (3.3%) patients died within 30 days, and 42 (13.9%) patients died within one year. Overall, 36 (11.9%) patients had low ANSS. 1-year mortality rates were almost three times higher in patients with low ANSS relative to patients with high ANSS (27.8% vs. 12.0%; the relative risk 1.1; p=0.018). A stepwise logistic regression analysis showed that ANSS was independently inversely associated with 1-year mortality (p=0.018). Complications and 30-day mortality rates were similar in both groups. Low ANSS are associated with 1-year mortality after TAVI. The Norton scale may therefore be used as an additional tool for elderly patient selection before TAVI.


One-year prospective study on single short (7-mm) implant overdentures in patients with severely resorbed mandibles.
This prospective study aimed to assess the effectiveness of using a single short implant to retain a single-implant mandibular overdenture (SIMO) in participants with severely atrophic edentulous mandibles. The study sample included 18 edentulous participants with severely resorbed mandibular ridges and limited bone height in the symphyseal region sufficient for the insertion of a 7.0-mm-height implant. First, patients received new conventional dentures or had their dentures relined, followed by the insertion of a 3.75 x 7 mm morse-taper implant, and the incorporation of a stud-type attachment and a nylon retentive insert after a 3-month healing period. Implant stability quotient (ISQ) was measured at baseline and after 3 and 12 months. Patient-reported outcomes (satisfaction with the dentures and oral health-related quality of life) were assessed before implant placement and at the 3-, 6-, and 12-month follow-ups. Clinical-radiographic outcomes and the incidence of prosthodontic events were assessed throughout the follow-up period. The implant survival rate was 100%. ISQ increased from baseline (72.7 ± 6.6) to the 3-month (82.0 ± 3.3) and 1-year (85.4 ± 2.9) follow-ups (p < .001). After 1 year, satisfaction with the mandibular prosthesis increased significantly, and the overall OHIP-Edent score decreased by 79.2%. No effects were found for the maxillary denture (p = .420). Due to dissatisfaction with treatment, two participants (11.1%) required additional implants after the 1-year follow-up and were considered as prosthodontic failures. Single-implant mandibular overdenture retained by a short implant showed favorable outcomes after 1 year. Further studies with larger samples and longer follow-up periods are needed to confirm the findings of this single-group prospective study. The present study was registered at the Brazilian Clinical Trials Registry (REBEC), identifier RBR-7p2xmg. The trial registry occurred during the study execution, and its final approval was delayed because of several formatting adequacies required by the platform REBEC.


Risk of colorectal cancer in relation to frequency and total amount of red meat consumption. Systematic review and meta-analysis.
The colon and rectum are common sites of food-related cancer in developed countries. Recent studies strongly suggest that red meat intake is associated with colon cancer, whereas for rectal cancer such an association still needs to be proved. The aim of the study was to assess the role of total amount and frequency of red meat intake in colorectal carcinogenesis based on published data using meta-analysis methods. The literature published until 2009 was selected from: MEDLINE, PubMed, Scopus, Embase, CancerLit, Google Scholar and Cochrane Library databases. The used search terms were: colorectal cancer, colon cancer, rectal cancer, meat intake, red meat intake, red meat consumption, meat consumption, colorectal cancer risk, colon cancer risk, rectal cancer risk and lifestyle. Articles investigating red meat intake of more often than once a day or 50 g per day were reviewed and selected for further analysis. Twenty-two studies fulfilled the established criteria. A meta-analysis confirmed the carcinogenic effect of the consumption of over 50 g of red meat per day for the colon (relative risk 1.21, 1.07-1.37) but not for the rectum (relative risk 1.30, 0.90-1.89). Red meat intake more frequently than once a day can induce both colonic (relative risk 1.37, 1.09-1.71) and rectal cancer (relative risk 1.43, 1.24-1.64). Red meat intake is associated with elevated risk of developing colorectal cancer. The frequency of red meat consumption rather than total amount of consumed meat is associated with a higher risk of colorectal carcinogenesis.


The relationship of age, level of formal education, and duration of employment toward attitudes concerning the use of computers in the workplace.
The introduction of computer technology in the Canadian health care environment necessitates an adaptation by hospital employees in order to perform their jobs. One thousand twenty-two hospital staff were surveyed immediately following their initial orientation to computers. The survey identified attitudes toward change, and the use of computers in the workplace. Demographic data and job classifications of all respondents were tabulated. Hypotheses were tested regarding the relationships between age, formal education, and duration of employment, and attitudes toward computers and change. Results indicated a generally positive response toward the use of computers in the workplace and to change in general. However, statistically significant results were obtained indicating that age, level of education, and length of employment affect the degree of positive response to both computers and change. The results suggest that demographic variables such as age, level of education, and employment duration must be taken into consideration when planning for the implementation of computer systems in the health care setting.


Manufacturing and characterization of a 3D printed lithium disilicate ceramic via robocasting: A pilot study.
The objective of this study was to manufacture and to evaluate the physico-mechanical properties of the Lithium disilicate (Li2O5Si2) ceramic structures fabricated using additive manufacturing (3D printing). Li2O5Si2 samples were divided into (n = 30/group): SM (subtractively manufactured) and AR (additive/robocasting). For the AR group, Li2O5Si2 powder was combined with ammonium polyacrylate, hydroxypropyl methylcellulose, and polyelectrolyte to create a colloidal gel, which was then used for printing. A digital CAD model of a disc was designed, and the G-code transferred to a custom built DIW 3D printer. The control group samples were prepared using pre-crystallized ceramic blocks, which were cut to obtain discs with same dimensions as the AR group. Disc-shaped specimens from both groups were crystallized at 840 °C. Mechanical properties were evaluated using biaxial flexural strength test (BFS) and Vickers hardness test. Representative fractographic images of the specimens were acquired using scanning electron microscopy (SEM) to analyze the fracture origin and crack propagation. Energy-dispersive X-ray spectroscopy (EDS) and attenuated total reflection Fourier transform infrared spectroscopy (FTIR-ATR) were used for chemical analysis, and X-ray diffractometry (XRD) was performed to analyze the crystalline phases. AR group yielded lower values of BFS (120.02 MPa ±33.91) and hardness (4.07 GPa ±0.30), relative to the SM group, (325.09 MPa ±63.98) and (5.63 GPa ±0.14), respectively. For EDS analysis, AR and SM groups showed similar elemental composition. In FTIR-ATR analysis, higher peaks referring to the crystalline structure were found for SM group. XRD analysis indicated a decreased formation of Li2O5Si2 from Lithium metasilicate (Li2O-SiO2) in the AM group. SEM micrographs showed a more porous microstructure associated with the 3D printed samples. The viability of fabricating Li2O5Si2 ceramic constructs using the Robocasting technique was successful. However, the samples prepared using subtractive manufacturing presented higher mechanical properties compared to the 3D printed constructs. The difference in properties between the manufacturing may be correlated to the decreased formation of Li2O5Si2 crystals and higher degrees of porosity.


Red wine acutely induces favorable effects on wave reflections and central pressures in coronary artery disease patients.
To investigate red wine's acute effects on aortic pressures and arterial stiffness in patients with coronary artery disease (CAD). Fifteen patients with CAD were recruited in a double-blind, cross-over study, which was comprised of 2 study days. Each volunteer consumed either 250 mL of regular or 250 mL of dealcoholized red wine. Wave reflections, expressed as augmentation index (AIx), as well as central and peripheral blood pressures (BP) were assessed at fast and 30, 60, and 90 min postprandially. Both regular and dealcoholized red wine caused a significant decrease in AIx by 10.5% +/- 1.4% (P = .001) and 6.1% +/- 1.4% (P = .011), respectively, whereas no significant change was induced in mean BP and timing of wave reflections expressing pulse wave velocity. Peripheral systolic BPs remained unaltered in both beverages, whereas a significant decrease in peripheral and central diastolic BPs was observed after the dealcoholized red wine consumption (P = .03 and P = .035, respectively). Central systolic BP was decreased after the consumption of regular (-7.4 +/- 2.4 mm Hg, P = .05) and dealcoholized red wine (-5.4 +/- 2.7 mm Hg, P = .019). Both types of red wine provoked favorable acute effects on wave reflections and central systolic pressures, whereas no such effect was evident at the brachial artery. Therefore, these findings could be attributed mainly to red wine antioxidant substances, rendering it a possible means of at least acute attenuation of increased wave reflections, arterial stiffness, and central pressures in patients with coronary artery disease.


Acute lung injury with endotoxin or NO2 does not enhance development of airway epithelium from bone marrow.
Adult marrow-derived stem cells can localize to lung and acquire immunophenotypic characteristics of lung epithelial cells. Lung injury increases recruitment of the marrow-derived cells. We speculated that comparing patterns of lung engraftment following different lung injuries would provide insight into potential mechanisms by which marrow-derived cells were recruited to lung. To evaluate this, adult female C57Bl/6 mice irradiated and engrafted with marrow from adult male transgenic GFP mice were exposed to either intranasal inhalation of endotoxin (25 microg/mouse) or 3 days of 25 ppm NO(2) and then compared 1 or 3 months later to transplanted but otherwise uninjured mice. In all cases, the majority of marrow-derived cells recruited to lung were CD45(+) leukocytes. In lungs of transplanted but otherwise uninjured mice, small numbers of CD45(-) donor-derived cells in alveolar septae stained positively for pro-surfactant protein C. Rare donor-derived cells located in the airway epithelium stained positively with cytokeratin. Subsequent exposure of engrafted mice to NO(2) or endotoxin did not significantly increase the number or pattern of donor-derived CD45(-) cells found in recipient lungs. These results suggest that NO(2) or endotoxin lung injury does not result in significant engraftment of marrow-derived cells in lung.


Constitutional risk factors for focal neuropathies in patients referred for electromyography.
It is well established that patient-related constitutional features predispose to focal peripheral neuropathies. Some of these risk factors were investigated in common focal neuropathies encountered in patients referred for electromyography. Gender, age, height and body mass index (BMI) were analysed retrospectively as risk factors for 11 focal neuropathies. In all, 9686 patients (age range 18-96 years; 58% women) were included from three different centres, with identical methods and equipment. High BMI was related to carpal tunnel syndrome (CTS), ulnar neuropathy at the elbow (UNE), combined CTS and UNE, meralgia paraesthetica and lumbar radiculopathy. In women, CTS and Morton's metatarsalgia were more common, whilst long thoracic neuropathies, suprascapular neuropathies and UNE were more common in men. Older age increased the risk for CTS, UNE, Morton's metatarsalgia and radiculopathies. Age, gender and BMI are important risk factors for many common focal neuropathies.


The KEAP1-NRF2 pathway: Targets for therapy and role in cancer.
The KEAP1-NRF2 pathway is the key regulator of cellular defense against both extrinsic and intrinsic oxidative and electrophilic stimuli. Since its discovery in the 1990s, its seminal role in various disease pathologies has become well appreciated, motivating research to elucidate the intricacies of NRF2 signaling and its downstream effects to identify novel targets for therapy. In this graphical review, we present an updated overview of the KEAP1-NRF2 signaling, focusing on the progress made within the past ten years. Specifically, we highlight the advances made in understanding the mechanism of activation of NRF2, resulting in novel discoveries in its therapeutic targeting. Furthermore, we will summarize new findings in the rapidly expanding field of NRF2 in cancer, with important implications for its diagnostics and treatment.


Distributed Fixed-Time Coordination Control for Networked Multiple Euler-Lagrange Systems.
This work investigates the fixed-time distributed coordination control for multiple Euler-Lagrange systems and, in particular, addresses containment control with stationary/dynamic leaders as well as leaderless synchronization control. For the containment control scenario with stationary leaders, the subgraph associated with followers is directed. When dynamic leaders are involved, the information transfer between neighboring followers is bidirectional, for which a novel distributed estimator is developed. For the leaderless synchronization control scenario, the communication network among agents is unidirectional. Three fixed-time distributed control schemes are designed for the aforementioned three cases by applying the fixed-time stability theory. The convergence of the coordination control objectives can be achieved in a fixed time that does not depend on any initial conditions of agents' states, and the settling times are also explicitly derived. Finally, numerical simulations are presented to demonstrate the feasibility of the developed control strategies.


Assessment of mercury presence and exposure in a lighthouse with a mercury drive system.
It is common practice for lighthouses with large Fresnel lenses to use mercury baths as a low-friction rotation mechanism. Some recent acute mercury poisonings and incidents of abnormal behavior in lighthouse keepers have drawn attention to the potential for chronic mercury poisoning in these workplaces. This study evaluated the distribution of mercury in a lighthouse on the Canadian west coast, and the exposure of its keepers and their spouses under two weather conditions. The urine mercury levels found in the lighthouse personnel were all less than would be expected in an occupationally exposed group (less than 4 micrograms/24 hr urine). Air concentrations in the lighthouse ranged from 4.4 to 26.3 micrograms/m3. Swabbing showed considerable accumulation of mercury on surfaces in the area of the light rotation mechanism, as well as transport throughout the lighthouse. The mercury levels in this lighthouse appeared to be under control through effective convective ventilation and employee awareness. The study signals potential problems where precautions have not been taken, especially in situations where the keepers and their families live in the lighthouse.


Complications and survival after subcutaneous ureteral bypass device placement in 24 cats: a retrospective study (2016-2019).
The aim of this study was to document survival, complications and risk factors for the development of complications and mortality prior to discharge after placement of a subcutaneous ureteral bypass (SUB) device in cats. The medical records of cats with SUB placement between January 2016 and August 2019 were retrospectively analysed. The development of complications (overall, intraoperative, perioperative, short- and long-term complications) and risk factors for mortality prior to discharge were statistically assessed with univariate binary logistic regression. All variables with a P value ⩽0.10 in the univariate analysis were assessed in a multivariate model. Variables were significant if P <0.05. Twenty-four cats were included; 12 (50.0%) received a unilateral SUB, 11 (45.8%) a bilateral nephrostomy tube with single cystostomy catheter and the remaining cat (4.2%) two unilateral SUBs. Nearly 80% of the cats developed complications, ranging from mild to fatal, including (partial) SUB obstruction (33.3% of complications), lower urinary tract infection (20.8%), pyelonephritis (20.8%) and sterile cystitis (12.5%). Five cats (20.8%) died prior to discharge. Six cats (25.0%) underwent revision surgery. The overall median survival time (MST) was 274 days (range 1-311 days). Complications were most common in the long-term period (14/16 cats), followed by the short-term (9/18 cats), perioperative (10/23 cats) and intraoperative (4/24 cats) periods. Older cats had an increased risk for developing perioperative complications (P = 0.045) and were less likely to survive to discharge (P = 0.033). An increased haematocrit at presentation was a risk factor for the occurrence of short-term complications (P = 0.03). Although complications similar to those previously described were observed, the complication rate was higher and the MST shorter than previously reported in cats undergoing SUB placement. Despite good short-term survival, the development of complications may necessitate regular and intensive control visits. Owners that consider SUB placement should be informed that follow-up can be strenuous and expensive.


SARS-CoV-2 infection: a possible trigger for the recurrence of IgA nephropathy after kidney transplantation?
Immunoglobulin A nephropathy, the most common primary glomerulonephritis worldwide, is a leading cause of chronic kidney disease and end-stage kidney failure. Several cases of immunoglobulin A nephropathy relapse in native kidneys have been described after COVID-19 vaccination or SARS-CoV-2 infection. Here, we report the case of a 52-year-old kidney transplant recipient who had a stable transplant function for more than 14 years, with a glomerular filtration rate above 30 ml/min/1.73 m2. The patient had been vaccinated against COVID-19 four times with the Pfizer-BioNTech vaccine, most recently in March 2022. Eight weeks after a symptomatic SARS-CoV-2 infection in June 2022, his glomerular filtration rate had decreased by more than 50%, and his proteinuria increased to 17.5 g per day. A renal biopsy indicated highly active immunoglobulin A nephritis. Despite steroid therapy, the function of the transplanted kidney deteriorated, and long-term dialysis became necessary because of recurrence of his underlying renal disease. This case report provides what is, to our knowledge, the first description of recurrent immunoglobulin A nephropathy in a kidney transplant recipient after SARS-CoV-2 infection leading to severe transplant failure and finally graft loss.


Isolation and characterization of novel lipase gene LipHim1 from the DNA isolated from soil samples.
Metagenomics is a magnificent tool to isolate genes from unknown/uncharacterized species and also from organisms that cannot be cultured. In this study, we constructed a metagenomic library from isolated DNA of soil samples collected from Palamuru University campus premises, in Mahabubnagar district of Andhra Pradesh, India. We isolated a novel lipase gene LipHim1, which has an open reading frame of 591 base pairs and encodes ∼23 kDa protein consisting of 196 amino acids. The Lipase LipHim1 showed maximum 32% homology at the protein level with the extracellular Aeromonas hydrophila lipase (Class II, GDSL family) and was significantly different from all other known lipases. The isolated lipase catalyzed the hydrolysis of fatty acid esters of polyoxyethylene sorbitan such as Tween 60. Our results indicate that the isolated lipase gene is novel.


Multimodal imaging of spike propagation: a technical case report.
We report an 11-year-old boy with intractable epilepsy, who had cortical dysplasia in the right superior frontal gyrus. Spatiotemporal source analysis of MEG and EEG spikes demonstrated a similar time course of spike propagation from the superior to inferior frontal gyri, as observed on intracranial EEG. The tractography reconstructed from DTI showed a fiber connection between these areas. Our multimodal approach demonstrates spike propagation and a white matter tract guiding the propagation.


Skeletal muscle fiber characteristics and oxidative capacity in hemiparetic stroke survivors.
Skeletal muscle is changed after stroke, but conflicting data exist concerning muscle morphology and oxidative enzyme capacity. In 36 chronic stroke patients bilateral rectus femoris muscle biopsies were analyzed, and fiber type proportions and cross-sectional areas were determined by ATPase histochemistry. Enzymatic concentrations of citrate synthase (CS) and 3-Hydroxyacyl-coenzymeA-dehydrogenase (HAD) were determined using freeze-dried muscle tissue. Findings were correlated with clinical outcomes. In the paretic muscles the mean fiber area was smaller (P = 0.0004), and a lower proportion of type 1 fibers (P = 0.0016) and a higher proportion of type 2X fibers (P = 0.0002) were observed. The paretic muscle had lower CS (P = 0.013) and HAD concentrations (P = 0.037). Mean fiber area correlated with muscle strength (r = 0.43; P = 0.041), and CS concentration correlated with aerobic capacity (r = 0.47; P = 0.01). In stroke survivors there is a phenotypic shift toward more fatigable muscle fibers with reduced oxidative enzymatic capacity that relates to clinical outcomes.


Bioinformatics Analysis of Programmed Cell Death in Spinal Cord Injury.
Programmed cell death (PCD) in the development of spinal cord injury (SCI) is complicated, including apoptosis, necroptosis, pyroptosis, ferroptosis, cuproptosis, and autophagy. It is necessary to make clear the expression levels of PCD and potential molecular targets after SCI for formulating relevant treatment strategies. We downloaded the rats' SCI expression matrix GSE45006, and the ssGSEA method was used to analyze the PCD after SCI. Then the related differentially expressed genes (DEGs) were identified, and the gene ontology (GO) and pathway analysis, protein-protein interaction (PPI) network construction, and HUB genes were identified. Finally, the correlation between HUB genes and PCD was analyzed. Apoptosis, necroptosis, pyroptosis, ferroptosis, and autophagy increased significantly in acute SCI, and then decreased gradually in the subacute and chronic stages; cuproptosis in acute SCI decreased significantly, and then gradually increased. In addition, we also screened 116 DEGs during the development of SCI. GO and pathway analysis showed that DEGs was related to mitosis and cell cycle. The identified hub genes are closely related to cell apoptosis, necroptosis, pyroptosis, ferroptosis after injury, and autophagy. PCD occurs differently in different stages after SCI. To inhibit apoptosis, necroptosis, pyroptosis, and ferroptosis after injury and induce autophagy may be the therapeutic strategy. In addition, intervention therapy based on related HUB genes may be the therapeutic target of SCI.


A Cost-Effectiveness Analysis of Obtaining Blood Cultures in Children Hospitalized for Community-Acquired Pneumonia.
To determine the clinical utility and cost-effectiveness of universal vs targeted approach to obtaining blood cultures in children hospitalized with community-acquired pneumonia (CAP). We conducted a cost-effectiveness analysis using a decision tree to compare 2 approaches to ordering blood cultures in children hospitalized with CAP: obtaining blood cultures in all children admitted with CAP (universal approach) and obtaining blood cultures in patients identified as high risk for bacteremia (targeted approach). We searched the literature to determine expected proportions of high-risk patients, positive culture rates, and predicted bacteria and susceptibility patterns. Our primary clinical outcome was projected rate of missed bacteremia with associated treatment failure in the targeted approach. Costs per 100 patients and annualized costs on the national level were calculated for each approach. The model predicts that in the targeted approach, there will be 0.07 cases of missed bacteremia with treatment failure per 100 patients, or 133 annually. In the universal approach, 118 blood cultures would need to be drawn to identify 1 patient with bacteremia, in which the result would lead to a meaningful antibiotic change compared with 42 cultures in the targeted approach. The universal approach would cost $5178 per 100 patients or $9,214,238 annually. The targeted approach would cost $1992 per 100 patients or $3,545,460 annually. The laboratory-related cost savings attributed to the targeted approach would be projected to be $5,668,778 annually. This decision analysis model suggests that a targeted approach to obtaining blood cultures in children hospitalized with CAP may be clinically effective, cost-saving, and reduce unnecessary testing.


Peroxidase catalyzed formation of azine pigments--a convenient and sensitive method for the identification of human cells with positive myeloperoxidase reactivity.
A new and efficient method for the identification of human cells with positive myeloperoxidase reactivity was developed. This method is based on the myeloperoxidase catalyzed formation of p-naphthoquinone-thiazol-2-on-azine pigments and offers a number of advantages compared to standard methods. In addition, we report the horseradish peroxidase (HRP) catalyzed oxidative coupling of 2-hydrazono-4-thiazolines with alpha-naphthol. This transformation allows a new and efficient synthesis of p-naphthoquinone-thiazol-2-on-azines under mild conditions.


Single-molecule imaging of DNA polymerase I (Klenow fragment) activity by atomic force microscopy.
We report a DNA origami-facilitated single-molecule platform that exploits atomic force microscopy to study DNA replication. We imaged several functional activities of the Klenow fragment of E. coli DNA polymerase I (KF) including binding, moving, and dissociation from the template DNA. Upon completion of these actions, a double-stranded DNA molecule was formed. Furthermore, the direction of KF activities was captured and then confirmed by shifting the KF binding sites on the template DNA.


Immediate hip spica is as effective as, but more efficient than, flexible intramedullary nailing for femoral shaft fractures in pre-school children.
Flexible intramedullary nailing (FIN) is the gold standard treatment for femur fracture in school-aged children. It has been performed successfully in younger children, although Spica cast immobilisation (SCI) has been the most widely used strategy to date. A retrospective analysis was performed between two comparable groups of children aged 1-4 years with a femoral shaft fracture. Two University hospitals, each using specific treatment guidelines, participated in the study: SCI in Group I (Basel, Switzerland) and FIN in Group II (Lausanne, Switzerland). Group I included 19 children with a median age of 26 months (range 12-46 months). Median hospital stay was 1 day (range 0-5 days) and casts were retained for a median duration of 21 days (range 12-29 days). General anaesthesia was used in six children and sedation in four. Skin breakdown secondary to cast irritation occurred in two children (10.5%). The median follow-up was 114 months (range 37-171 months). No significant malunion was noted. Group II included 27 children with a median age of 38.4 months (range 18.7-46.7 months). Median hospital stay was 4 days (range 1-13 days). All children required general anaesthesia for insertion and removal of the nails. Free mobilisation and full weight bearing were allowed at a median of 2 days (range 1-10 days) and 7 days (range 1-30 days), respectively, postoperatively. Nail exteriorisation was noted in three children (11%). The median follow-up was 16.5 months (range 8-172 months). No significant malunion was reported. Young children with a femoral shaft fracture treated by SCI or FIN had similarly favourable outcomes and complication rates. FIN allowed earlier mobilisation and full weight bearing. Compared to SCI, a greater number of children required general anaesthesia. In a pre-school child with a femoral shaft fracture, immediate SCI applied by a paediatric orthopaedic team following specific guidelines allowed early discharge from hospital with few complications.


Prebiotic synthesis of orotic acid parallel to the biosynthetic pathway.
By heating an aqueous solution of aspartic acid and urea, carbamylaspartic acid is first formed and then the molecule is cyclized to dihydroorotic acid (DHO) with loss of water. Irradiation of an aqueous solution of DHO with a tungsten lamp yields orotic acid by photo-dehydrogenation of the molecule. This pathway of orotic acid formation is quite similar to that of biosynthesis of the molecule.


[Looking back on 50 years of stone treatment].
Urolithiasis is an urologist's daily business. This article provides an overview of the developments in the management of urinary stone disease since the 1970s. While conventional KUB X-rays and intravenous pyelography were standard imaging procedures in the past, computed tomography is the first choice today. Conservative treatments such as medical expulsive therapy were established to facilitate ureteral stone passage, but have come into discussion in the past few years. Fifty years ago, open stone surgery was the standard procedure to access renal stones. This has been superseded by minimally-invasive procedures ranging from percutaneous nephrolithotomy (PCNL) to extracorporeal shock-wave lithotripsy (ESWL) and ureteroscopy (URS), which is today's standard for most upper urinary tract stones. It would have been desirable if such enormous efforts and progress had been made in the evaluation of stone pathogenesis and the improvement of preventative measures. Unfortunately, the knowledge on stone formation has hardly improved compared to 1970.


Frontiers in endocrine disruption: Impacts of organotin on the hypothalamus-pituitary-thyroid axis.
Endocrine disruptors (EDs), chemical substances widely used in industry and ubiquitously distributed in the environment, are able to interfere with the synthesis, release, transport, metabolism, receptor binding, action, or elimination of endogenous hormones. EDs affect homeostasis mainly by acting on nuclear and nonnuclear steroid receptors but also on serotonin, dopamine, norepinephrine and orphan receptors in addition to thyroid hormone receptors. Tributyltin (TBT), an ED widely used as a pesticide and biocide in antifouling paints, has well-documented actions that include inhibiting aromatase and affecting the nuclear receptors PPARγ and RXR. TBT exposure in humans and experimental models has been shown to mainly affect reproductive function and adipocyte differentiation. Since thyroid hormones play a fundamental role in regulating the basal metabolic rate and energy homeostasis, it is crucial to clarify the effects of TBT on the hypothalamus-pituitary-thyroid axis. Therefore, we review herein the main effects of TBT on important metabolic pathways, with emphasis on disruption of the thyroid axis that could contribute to the development of endocrine and metabolic disorders, such as insulin resistance and obesity.


Symbiosis-inspired de novo synthesis of ultrahigh MOF growth mixed matrix membranes for sustainable carbon capture.
Mixed matrix membranes (MMMs) are one of the most promising solutions for energy-efficient gas separation. However, conventional MMM synthesis methods inevitably lead to poor filler-polymer interfacial compatibility, filler agglomeration, and limited loading. Herein, inspired by symbiotic relationships in nature, we designed a universal bottom-up method for in situ nanosized metal organic framework (MOF) assembly within polymer matrices. Consequently, our method eliminating the traditional postsynthetic step significantly enhanced MOF dispersion, interfacial compatibility, and loading to an unprecedented 67.2 wt % in synthesized MMMs. Utilizing experimental techniques and complementary density functional theory (DFT) simulation, we validated that these enhancements synergistically ameliorated CO2 solubility, which was significantly different from other works where MOF typically promoted gas diffusion. Our approach simultaneously improves CO2 permeability and selectivity, and superior carbon capture performance is maintained even during long-term tests; the mechanical strength is retained even with ultrahigh MOF loadings. This symbiosis-inspired de novo strategy can potentially pave the way for next-generation MMMs that can fully exploit the unique characteristics of both MOFs and matrices.


Kinetics of EGFR expression during fractionated irradiation varies between different human squamous cell carcinoma lines in nude mice.
Preclinical and clinical data indicate that high pretherapeutic EGFR expression is associated with poor local tumour control, possibly caused by a high repopulation rate of clonogenic cells during radiotherapy in these tumours. Previous data reported from our laboratory showed a correlation between EGFR expression and acceleration of repopulation in poorly differentiated FaDu human squamous cell carcinoma (SCC) during fractionated irradiation. To test whether this is a general phenomenon, two further SCC were investigated in the present study. GL and UT-SCC-14, two moderately well differentiated and keratinising hSCC, were grown as xenografts in nude mice. Functional data on the repopulation kinetics during fractionated irradiation for these tumour models have been previously determined. The expression of EGFR during fractionation was analysed by immunohistochemistry. Endpoints were the membrane-staining score and the proportion of EGFR-positive cells (EGFR labelling index). Different kinetics of EGFR expression during fractionated RT were found. In UT-SCC-14, EGFR staining score and labelling index increased significantly during radiotherapy. In GL SCC, the EGFR expression was unchanged. Both tumours are characterized by a small but significant repopulation rate during radiotherapy. The expression of EGFR may change significantly during fractionated irradiation. No clear correlation between EGFR expression and the repopulation kinetics of clonogenic tumour cells during fractionated irradiation was found. The changes in EGFR expression during irradiation warrant further investigation on their prognostic implications and on their importance for therapeutic interventions.


Graphene bilayers with a twist.
Near a magic twist angle, bilayer graphene transforms from a weakly correlated Fermi liquid to a strongly correlated two-dimensional electron system with properties that are extraordinarily sensitive to carrier density and to controllable environmental factors such as the proximity of nearby gates and twist-angle variation. Among other phenomena, magic-angle twisted bilayer graphene hosts superconductivity, interaction-induced insulating states, magnetism, electronic nematicity, linear-in-temperature low-temperature resistivity and quantized anomalous Hall states. We highlight some key research results in this field, point to important questions that remain open and comment on the place of magic-angle twisted bilayer graphene in the strongly correlated quantum matter world.


Up-regulation of thioredoxin system by puerarin inhibits lipid uptake in macrophages.
Cellular oxidative stress promotes lipid accumulation in macrophages during atherogenesis. Puerarin is a natural isoflavone with beneficial effects against oxidation and atherosclerosis. In this study, we investigated the effects of puerarin on lipid uptake and explored the underlying molecular regulation. We found puerarin up-regulated thioredoxin-1 (Trx1) and Trx reductase-1 (TrxR1) expression; it increased TrxR1 activity, cellular thiols contents and decreased oxidized form of Trx1, thus inhibiting cellular ROS generation. Confocal microscope and flow cytometry analysis showed fluorescence labeled Dil-oxLDL uptake was dramatically inhibited by puerarin in RAW264.7 cells as well as in primary bone marrow derived macrophages and peritoneal macrophages. The effects were reversed when Trx1 was inhibited by treatment with Trx1 inhibitor PX-12 or Trx1 siRNA. We also found scavenger receptors such as SR-A and Lox-1, but not CD36 were involved in the Trx1-mediated lipid uptake inhibition. Moreover, measurements of foam cell accumulation and ROS production in sections of aortic roots showed those were reduced by puerarin but raised when additional treatment with PX-12 or Trx1 siRNA in apoE-/- mice, which demonstrates the lipid uptake reduction by puerarin requires Trx1 inhibition in vivo. In addition, we analyzed the upstream regulation and found puerarin induced Nrf2 activity; cooperation between Nrf2 and ATF4 facilitated the puerarin effects. PERK phosphorylation was detected to be increased by puerarin, while PERK inhibition reduced cellular Trx1, TrxR1, nuclear Nrf2 and ATF4. Altogether, puerarin modulates PERK/Nrf2 that coordinates with ATF4 to active Trx1, which causes SR-A and Lox-1 reduction and lipid uptake inhibition in macrophages. This suggests Trx1 could be an effective target by puerarin in the prevention of atherosclerosis.


Clinical Outcomes in Metabolically Healthy and Unhealthy Obese and Overweight Patients With Atrial Fibrillation: Findings From the GLORIA-AF Registry.
To explore the association between metabolic status, body mass index (BMI), and natural history of patients with atrial fibrillation (AF). The global, prospective GLORIA-AF Registry Phase II and III included patients with recent diagnosis of AF between November 2011 and December 2014 for Phase II and between January 2014 and December 2016 for Phase III. With this analysis, we categorized patients with AF according to BMI (normal weight [18.5 to 24.9 kg/m2], overweight [25.0 to 29.9 kg/m2], obese [30.0 to 60.0 kg/m2]) and metabolic status (presence of hypertension, diabetes, and hyperlipidemia). We analyzed risk of major outcomes using multivariable Cox regression analyses; the primary outcome was the composite of all-cause death and major adverse cardiovascular events. There were 24,828 (mean age, 70.1±10.3 years; 44.6% female) patients with AF included. Higher BMI was associated with metabolically unhealthy status and higher odds of receiving oral anticoagulants and other treatments. Normal-weight unhealthy patients showed a higher risk of the primary composite outcome (adjusted hazard ratio [aHR], 1.20; 95% CI, 1.01 to 1.42) and thromboembolism, whereas a lower risk of cardiovascular death (aHR, 0.35; 95% CI, 0.14 to 0.88) and major adverse cardiovascular events (aHR, 0.56; 95% CI, 0.33 to 0.93) was observed in metabolically healthy obese individuals. Unhealthy metabolic groups were also associated with increased risk of major bleeding (aHR, 1.51 [95% CI, 1.04 to 2.20] and aHR, 1.96 [95% CI, 1.34 to 2.85] in overweight and obese groups, respectively). Increasing BMI was associated with poor metabolic status and with more intensive treatment. Prognosis was heterogeneous between BMI groups, with metabolically unhealthy patients showing higher risk of adverse events.


Impact of genetic testing on complex diseases.
Wide applications for genetic testing in the clinical care of complex diseases have been discussed. However, it has never been quantified to what extent genetic testing could eventually be useful in the clinical care of common disorders. We aimed to quantify the theoretically possible utilization in common ischemic stroke, an example of a complex disease. We computed the possible impact of genetic testing on risk assessment, secondary prevention and prognosis of ischemic stroke. This was done for hypothetical genotypes with different frequencies and effects. To apply pharmacogenetic secondary prevention based on a genotype with a frequency of 10% and 33% risk reduction, 204 subjects would have to be screened and 110 given genotype specific treatment for 1 year, in order to prevent one recurrent stroke, compared with standard therapy. The application of genetic testing seems to be less promising in risk assessment and the assessment of prognosis of common ischemic stroke. The highest impact of genetic testing on clinical practice of stroke will be in the areas of secondary prevention. Given the weak effects of reported susceptibility genotypes, it is theoretically unlikely that genetic screening will be used for the assessment of risk or prognosis of a complex disorder, except for Mendelian types of disease.


Implications of threonine hydrogen bonding in the glycophorin A transmembrane helix dimer.
The transmembrane helix of glycophorin A contains a seven-residue motif, LIxxGVxxGVxxT, that mediates protein dimerization. Threonine is the only polar amino acid in this motif with the potential to stabilize the dimer through hydrogen-bonding interactions. Polarized Fourier transform infrared spectroscopy is used to establish a robust protocol for incorporating glycophorin A transmembrane peptides into membrane bilayers. Analysis of the dichroic ratio of the 1655-cm(-1) amide I vibration indicates that peptides reconstituted by detergent dialysis have a transmembrane orientation with a helix crossing angle of <35 degrees. Solid-state nuclear magnetic resonance spectroscopy is used to establish high resolution structural restraints on the conformation and packing of Thr-87 in the dimer interface. Rotational resonance measurement of a 2.9-A distance between the gamma-methyl and backbone carbonyl carbons of Thr-87 is consistent with a gauche- conformation for the chi1 torsion angle. Rotational-echo double-resonance measurements demonstrate close packing (4.0 +/- 0.2 A) of the Thr-87 gamma-methyl group with the backbone nitrogen of Ile-88 across the dimer interface. The short interhelical distance places the beta-hydroxyl of Thr-87 within hydrogen-bonding range of the backbone carbonyl of Val-84 on the opposing helix. These results refine the structure of the glycophorin A dimer in membrane bilayers and highlight the complementary role of small and polar residues in the tight association of transmembrane helices in membrane proteins.


Endothelial GATA4 controls liver fibrosis and regeneration by preventing a pathogenic switch in angiocrine signaling.
Angiocrine signaling by liver sinusoidal endothelial cells (LSECs) regulates hepatic functions such as growth, metabolic maturation, and regeneration. Recently, we identified GATA4 as the master regulator of LSEC specification during development. Herein, we studied the role of endothelial GATA4 in the adult liver and in hepatic pathogenesis. We generated adult Clec4g-icretg/0xGata4fl/fl (Gata4LSEC-KO) mice with LSEC-specific depletion of Gata4. Livers were analyzed by histology, electron microscopy, immunohistochemistry/immunofluorescence, in situ hybridization, and LSECs were isolated for gene expression profiling, ChIP- and ATAC-sequencing. Partial hepatectomy was performed to assess regeneration. We used choline-deficient, l-amino acid-defined (CDAA) diet and chronic carbon tetrachloride exposure to model liver fibrosis. Human single cell RNA-seq data sets were analyzed for endothelial alterations in healthy and cirrhotic livers. Genetic Gata4 deficiency in LSECs of adult mice caused perisinusoidal liver fibrosis, hepatopathy and impaired liver regeneration. Sinusoidal capillarization and LSEC-to-continuous endothelial transdifferentiation were accompanied by a profibrotic angiocrine switch involving de novo endothelial expression of hepatic stellate cell-activating cytokine PDGFB. Increased chromatin accessibility and amplification by activated MYC mediated angiocrine Pdgfb expression. As observed in Gata4LSEC-KO livers, CDAA diet-induced perisinusoidal liver fibrosis was associated with GATA4 repression, MYC activation and a profibrotic angiocrine switch in LSECs. Comparison of CDAA-fed Gata4LSEC-KO and control mice demonstrated that endothelial GATA4 indeed protects against dietary-induced perisinusoidal liver fibrosis. In human cirrhotic livers, GATA4-positive LSECs and endothelial GATA4 target genes were reduced, while non-LSEC endothelial cells and MYC target genes including PDGFB were enriched. Endothelial GATA4 protects against perisinusoidal liver fibrosis by repressing MYC activation and profibrotic angiocrine signaling at the chromatin level. Therapies targeting the GATA4/MYC/PDGFB/PDGFRβ axis offer a promising strategy for prevention and treatment of liver fibrosis. The liver vasculature is supposed to play a major role in the development of liver fibrosis and cirrhosis, which can lead to liver failure and liver cancer. Herein, we discovered that structural and transcriptional changes induced by genetic deletion of the transcription factor GATA4 in the hepatic endothelium were sufficient to cause liver fibrosis. Activation of the transcription factor MYC and de novo expression of the "angiocrine" growth factor PDGFB were identified as downstream drivers of fibrosis and as potential therapeutic targets for this potentially fatal disease.


Therapies for cardiac light chain amyloidosis: An update.
Light-chain (AL) amyloidosis is the most common type of systemic amyloidosis, affecting around 10 people per million per year. This serious disorder is characterized by the presence of a clone of bone marrow plasma cells that produces monoclonal light chains (LCs) of the κ or predominantly λ type. These amyloidogenic LCs undergo extracellular misfolding and aggregation into proteotoxic soluble oligomers and amyloid fibrils that deposit within tissues. The lethal consequences of AL amyloidosis are due to the toxic products (the LCs) and not to the malignant behaviour of the plasma cell clone. Almost 80% of patients with AL amyloidosis have some degree of cardiac involvement, manifesting as heart failure (HF), and carrying a particularly poor prognosis. The past decade has seen major advances in the treatment of AL amyloidosis, and a rapidly fatal disease has become a treatable and possibly curable condition. The number of therapeutic options is rapidly expanding, offering hope to address currently unmet needs (most notably, the treatment of frail patients). The treatment of AL amyloidosis consists in a combination of agents targeting multiple steps of the amyloid cascade, associated with effective HF management, and there is ground for hope for dramatically improving the outcome in the near future. In the present review we will summarize our current knowledge on therapy for cardiac AL amyloidosis, targeting clinical cardiologists involved in the care of this serious disorder.


Glycaemic control with add-on thiazolidinedione or a sodium-glucose co-transporter-2 inhibitor in patients with type 2 diabetes after the failure of an oral triple antidiabetic regimen: A 24-week, randomized controlled trial.
To evaluate the effectiveness and safety of adding either a sodium-glucose co-transporter-2 inhibitor (SGLT2i) or thiazolidinedione (TZD) in patients with type 2 diabetes (T2D) inadequately controlled with triple therapy. In this prospective, open-label, multicentre, 24-week clinical trial, we randomly assigned 119 patients with T2D who failed to achieve glycaemic control (7% < HbA1c ≤ 10%) with conventional triple oral antidiabetic agents (OADs; metformin, sulphonylurea and dipeptidyl peptide-4 [DPP-4] inhibitor) into two groups who received either an SGLT2i or TZD. The primary endpoint was mean change in HbA1c level between the two groups at 24 weeks. In total, 119 patients were enrolled in the SGLT2i (n = 60) and TZD (n = 59) groups. Mean age of the study subjects was 61.86 years, and the mean duration of T2D was 13.89 years. After 24 weeks, both groups showed significant reductions in HbA1c (from 7.94% ± 0.74% to 6.97% ± 0.84% in the SGLT2i group and from 8.00% ± 0.78% to 7.18% ± 0.98% in the TZD group), without a significant between-group difference (P = .235). A significant body mass index (BMI) reduction was noted in the SGLT2i group, whereas an increase in BMI was noted in the TZD group (-0.79 ± 1.37 vs. 0.92 ± 0.86 kg/m2 , P < .001). Other safety profiles were favourable in both groups. The current study shows that an SGLT2i or TZD could be a valid option as a fourth OAD for treatment of patients with T2D inadequately controlled with a triple combination of OADs.


Developmental studies on an apparent cell-lethal mutant gene-ut-in the Mexican axolotl, Ambystoma mexicanum.
The discovery of a new mutant gene in stocks of the Mexican axolotl derived from breeding stock of the Hubrecht Laboratory, the Netherlands, is described. The gene appears to be a simple recessive and displays complete penetrance. ut/ut larvae develop normally through hatching, but begin to lag in growth and display characteristics defects in gill and limb formation shortly thereafter. The results of parabiosis of normal and mutant embryos, as well as embryological transplants of mutant limb and branchial rudiments, support the conclusion that the gene ut is expressed as an 'autonomous-cell lethal'. Despite gross morphological defects in ut/ut larvae, comparisons between normal and mutant animals of the protein spectra of various tissues and organs revealed no substantial differences. A subtle change in the metabolism of ut/ut larvae apparently, therefore, leads to developmental arrest.


Study on the emergency capacity of coal mine enterprises in Longdong Area based on D-FAHP method.
Emergency capability assessment is a complex system with multiple factors, variables and levels. Incomplete and uncertain assess information often occurs during assessment. Based on this, a method combining D-number theory and fuzzy analytic Hierarchy process (FAHP) is proposed to study the emergency capacity of coal enterprises in Longdong area. On the basis of analyzing the limitation of D-S evidence theory, the D-number theory was optimized and improved. According to the principles of systematicness, feasibility, scientificity and timeliness, a hierarchical structure model of enterprise emergency capability assessment was constructed from the perspective of pre-incident, mid-incident and post-incident, which consisted of 4 first-level indicators and 18 s-level indicators. The weight and importance of the assessment index of emergency response capability are calculated by organically integrating the D-number preference relation with the hierarchy structure. Combined with the assessment results of experts, a quantitative analysis and evaluation of the emergency response capacity of a coal enterprise was conducted by using FAHP. The comprehensive score of the enterprise's emergency response capability was 80.45, and the level of emergency response capacity was "good". The research results show that the D-FAHP method has high reliability in evaluating the emergency response capability of coal enterprises, avoiding the impact of uncertain and incomplete information on the assessment results. This can not only effectively identify the weak links in emergency management, but also meet the emergency decision-making needs of enterprises in the emergency state, which has important guiding significance to improve the ability and level of enterprise emergency management.


The RAM classification: a novel, systematic approach to the adult-acquired flatfoot.
In summary, prior classifications have provided broad guidelines for treating the AAFF without accounting for case-specific variables in determining a treatment plan. The current system breaks down the deformity into three independent levels of involvement: the rearfoot, the ankle, and the midfoot. Via a simple, easy to remember, and reproducible schema based off the original Johnson and Strom classification, each level can be independently evaluated and a patient-specific surgical treatment plan can be formulated based on our most current understanding of the AAFF.


Impact of an intrahospital mobile thrombolysis team on 3-month clinical outcomes in patients benefiting from intravenous thrombolysis for acute ischemic stroke.
The benefits of thrombolysis in patients presenting with acute ischemic stroke (IS) are highly time-dependent. The aim of our study was to evaluate the clinical benefit, after 3 months, of an intrahospital mobile thrombolysis team (MTT) for thrombolysis in IS. A total of 95 consecutive patients treated with IV tPA for acute IS at the neurology department of Rouen University Hospital between 1 January and 31 December 2015 were retrospectively identified. Patients who had benefited from mechanical thrombectomy or hemicraniectomy were excluded. The study compared 33 patients who had benefited from our MTT (thrombolysis whatever the location and as soon as possible by a specific nurse) with 62 patients treated in the usual way (thrombolysis only at the stroke unit). Management timescales, inhospital and 3-month clinical outcomes, and imaging data were also compared between the two groups. Demographic data and factors known to influence the clinical course after thrombolysis were similar between the two groups (P>0.05). However, use of the MTT allowed significant decreases in the median onset-to-treatment (OTT) time of 26min and median door-to-needle (DTN) time of 27min (P<0.001). The proportion of patients with a DTN time<60min was higher in the MTT group than in the usual care (UC) group: 64% vs. 14%, respectively (P<0.001), according to American Heart Association/American Stroke Association guidelines. Although there was a smaller proportion of negative 3-month outcomes (modified Rankin Scale score: 6; 6% vs. 16%) and a larger proportion of highly favorable 3-month outcomes (mRS score: 0-1; 79% vs. 64%) in the MTT vs. UC groups, respectively, these differences were not statistically significant (P>0.05). Use of an MTT is a simple way to reduce thrombolysis delays, and the present results encourage us to improve the system to make it even more effective and available for all patients.


Prolactin secretion by cultured anterior pituitary cells: influence of culture conditions and endocrine status of the pituitary donor.
The characteristics of prolactin (PRL) secretion by cultured anterior pituitary cells in the presence and absence of catecholamines were studied. PRL secretion was markedly influenced by culture conditions such as cell density, culture duration and length of short-term incubation. Dopamine (DA) inhibited PRL release in a dose-dependent manner within a physiological range, and this inhibition was reversed by the stereospecific DA receptor antagonist (+)-butaclamol. In contrast, the inhibition of PRL secretion by norepinephrine (NE) required much higher doses and lacked specificity. Cells obtained from male donors had the lowest basal PRL secretion and were the least responsive to DA inhibition, whereas those obtained from females in late pregnancy had the highest basal PRL secretion and were the most sensitive to DA.


The expression of specific proteins in cultured renal collecting duct cells.
It is still uncertain whether cell cultures attain the functional maturity of corresponding in vivo cells. The degree of differentiation of cultured collecting-duct (CD) epithelium cells was therefore examined using immunohistochemical procedures. Three monoclonal antibodies (mabs CD1, CD2, and CD3) were raised against proteins (PCD) isolated from the renal papilla. At Western-blot analysis, each of these antibodies reacted with a specific protein that was distinguishable according to its molecular weight [PCD1, 190 kilodaltons (kDa); PCD2, 210 kDa; PCD3, 50 kDa]. Using immunofluorescence, these proteins were found to be localized exclusively in the renal CD system. Other renal structures, such as the proximal or distal tubular portions, the glomeruli and the interstitial network, were not reactive. The mabs, CD2 and CD3, labeled both the cortical and medullary CD in a uniform way, whereas mab CD1 produced heterogeneous immunolabeling along the length of the cortical, medullary, and papillary CD. As revealed by immunohistochemistry, the mabs revealed differences with respect to the expression of the specific renal proteins in cultured CD cells. In polar-differentiated epithelium cultured for 5 days on a specific renal support, mab CD1 was unreactive, whereas mabs CD2 and CD3 were positive. This demonstrated the biochemical immaturity of this cultured epithelium with respect to CD1 reactivity. In morphologically dedifferentiated CD monolayer cells grown on the bottom of a culture dish, only a weak reaction for mab CD3 was observed. The loss of epithelial polarization in CD monolayer cells obviously coincides with the absence of the renal proteins PCD1 and PCD2.(ABSTRACT TRUNCATED AT 250 WORDS)


High-dose-rate endorectal brachytherapy in the treatment of locally advanced rectal carcinoma: technical aspects.
In this era of new radiation technologies and tumor imaging, a high-dose-rate endorectal brachytherapy has been developed and tested in a phase I/II study of advanced rectal tumors. In this article, we report technical aspects of the treatment modality. Forty-nine patients underwent staging with endoscopic endorectal ultrasound, and the tumor dimensions were determined with MRI of the pelvis. Patients with resectable rectal cancer (staged T2, T3, or early T4) were treated with preoperative high-dose-rate endorectal brachytherapy followed by surgery 6-8 weeks later. Under direct rectoscopy, radio-opaque clips were used to mark the tumor margins. The treatment planning was done with the use of a CT simulator, and the treatment was delivered using a flexible endorectal applicator with eight catheters arranged around the circumference of the applicator and a high-dose-rate brachytherapy remote afterloading system with an Iridium-192 source. Isodose distributions were generated by the Plato planning system (Nucletron B.V., Veenendaal, The Netherlands) and digitally reconstructed radiographs were used as references for daily treatment. A tumor dose of 26 Gy in four fractions was prescribed, and intramesorectal deposits were documented on the magnetic resonance images. Forty-nine patients received planned treatment, and all but 2 patients underwent planned surgery. The pathology specimens showed a complete macroscopic response in 64% of the patients and tumor downstaging in 67% of the patients. Advances in tumor imaging and 3D treatment planning systems allow for better tumor mapping and dose planning. The use of a multichannel flexible endorectal applicator leads to tumor downstaging before surgery in patients with resectable locally advanced rectal carcinomas. The technique used in our center was practical and validated by this study.


Sensory processing and detection thresholds of burn-injured patients: A comparison to normative data.
Emerging evidence suggests that individual levels of sensory sensitivity may impact treatment outcomes for people recovering from burn injuries. For example, individuals with higher levels of sensory sensitivity were less adherent with compression garment wear, often used for scar management. The purpose of this study was to characterise sensory patterns for a sample of burn-injured patients as a cohort, using normative data as the reference. As different patterns of sensory processing can have implications clinically, understanding this at the cohort level may provide valuable insight for therapy. This was a secondary analysis of data collected during a cross-sectional study. Adults (N = 117) attending the Professor Stuart Pegg Adult Burns Unit outpatient clinic completed the Adolescent/Adult Sensory Profile and the following quantitative sensory tests: two-point discrimination; mechanical detection threshold; and pressure pain threshold. Compared to matched normative data, burn-injured patients reported higher levels of sensory sensitive and avoiding patterns, and experienced lower detection thresholds for touch and pain. Higher reports of sensory sensitivity and sensory avoiding, and lower thresholds for touch and pain, have been correlated with tactile defensiveness. Tactile defensiveness has been associated with social withdrawal and isolation, all of which could contribute to decreased engagement in therapy. The ways in which these sensory characteristics impact on burn-related treatments, such as compression garment adherence, warrant further investigation.


Predicting crystal structure by merging data mining with quantum mechanics.
Modern methods of quantum mechanics have proved to be effective tools to understand and even predict materials properties. An essential element of the materials design process, relevant to both new materials and the optimization of existing ones, is knowing which crystal structures will form in an alloy system. Crystal structure can only be predicted effectively with quantum mechanics if an algorithm to direct the search through the large space of possible structures is found. We present a new approach to the prediction of structure that rigorously mines correlations embodied within experimental data and uses them to direct quantum mechanical techniques efficiently towards the stable crystal structure of materials.


Dependence of knee range of motion on the alignment of femoral and tibial components after medial unicompartmental knee arthroplasty.
This study evaluated the relationship between postoperative knee flexion angles and the position of femoral and tibial components in unicompartmental knee arthroplasty (UKA). Eighteen patients (a total of 22 knees: three men, four knees; 15 females, 18 knees) who underwent navigation-assisted UKA were included. Pre- and postoperative computed tomography images were applied on 3D software, which were matched and used to calculate the position of femoral and tibial components. Correspondingly, we investigated the relationship between the knee range of motion (ROM) at 1-year postoperative follow-up and the position of femoral and tibial components. At 1-year post-UKA, the knee flexion angle was associated with the posterior flexion angle of tibial components. This particular angle was significantly greater in the group with equal or greater postoperative knee ROM compared to preoperative ROM (5.2 ± 2.1°) than in the group with less postoperative knee ROM compared to preoperative ROM (2.6 ± 1.6°, p < 0.01). There was no significant difference between both groups in the femoral component position, preoperative posterior slope of the medial tibial plateau, change in the pre- to postoperative posterior tibial slope, and postoperative knee society score. The posterior flexion angle of the tibial component affected the improvement/deterioration of the postsurgery knee flexion angle in navigation-assisted UKA. For improved outcomes after UKA using navigation systems, surgeons should aim to achieve a 5° to 8° posterior flexion angle of the tibial component.


Investigating neuropsychological and reward-related deficits in a chronic corticosterone-induced model of depression.
Chronic stress is a known risk factor for the development of major depression (MDD) and is commonly used to induce a depression-like phenotype in rodents. Similar phenotypic effects are also observed in rodents when treated chronically with the stress hormone corticosterone. In this study, we investigated the neuropsychological consequences of chronic corticosterone treatment in male rats using two translational rodent assays of affective bias, the judgement bias task (JBT) and affective bias test (ABT). We also used the reward learning assay (RLA) and sucrose preference test (SPT) to quantify reward-related behaviours. Negative biases in decision-making were observed in the chronic corticosterone-treated group but only when the treatment was given shortly before each behavioural session. The same dose of corticosterone, when given daily after completion of the behavioural session had no effects. Chronic corticosterone treatment did not potentiate negative affective biases in the ABT induced by either an acute pharmacological or stress manipulation but both reward learning and reward sensitivity were blunted. Analysis of the brain tissue from animals receiving chronic corticosterone found reduced hippocampal neurogenesis consistent with previous studies suggesting corticosterone-induced neurotrophic deficits. Taken together, these data suggest chronic corticosterone treatment induces neuropsychological effects related to changes in reward learning, memory and negative biases in decision making, but these decision-making biases depend on whether rewarding outcomes were experienced during the acute effects of the drug. These findings suggest an important interaction between psychological and biological factors resulting in negative biases in decision-making in this model.