Efficacy of ultrasound-guided testicle-sparing surgery for small testicular masses.
The aim of the present study was to evaluate evolution of conservative echo-guided surgery for testicular tumours ≤1.5 cm and to assess the safety of this surgical procedure. 49 consecutive patients diagnosed with ultrasonography testicular lesions (mono or bilateral) ≤1.5 cm and treated with conservative echo-guided testicular surgery were examined. The parameters considered in this retrospective analysis included case history, physical examination, scrotal and abdominal ultrasonography, computed tomography scan, size of the nodule (maximum diameter), tumour markers, chest radiography, frozen section examination, histologic size of the tumour, overall survival (OS) and findings on follow-up. Mean age of patients was 33 years (range 18-62). Patients who presented with a palpable testicular nodule were 32.7 %, gynecomastia 10.2 %, precocious pseudopuberty 4 %, and scrotal pain 12.2 %. Permanent section examination confirmed the FSE data in all cases of certainty malignant lesion and definitive histological types were: 22 seminoma, 13 non-seminomatous or mixed germ cell tumours, 4 Leydig tumours, 2 hamartoma, 1 epidermoid cyst, 2 sertoli cell tumours, and 5 fibrous pseudotumour. No complications intra- and postoperative were observed. Overall survival was 100 % and scrotal US showed evidence of local tumour recurrence in 6 patients (12.2 %) after a mean follow-up of 34.7 months. The OS, the low rate of local recurrence, and absence of complications, tend to demonstrate the safety of the procedure. The benefits to testicular-sparing surgery include improving the patient's overall quality of life, fertility, endocrine function and negative cosmetic effects of radical orchiectomy.


The temporal dynamics of plasma fractalkine levels in ischemic stroke: association with clinical severity and outcome.
The chemokine fractalkine (CX3CL1, FKN) is involved in neural-microglial interactions and is regarded as neuroprotective according to several in vivo studies of inflammatory and degenerative states of the brain. Recently, an association with outcome in human ischemic stroke has been proposed. In this study, we aimed to investigate the temporal pattern of FKN levels in acute ischemic stroke in relation to stroke severity and outcome. FKN levels were measured in plasma specimens of fifty-five patients with acute ischemic stroke. Blood was available for time points 6 hours (h), 12 h, 3 days (d), 7 d and 90 d after stroke onset. Clinical outcome was evaluated using the modified Rankin Scale (mRS) at 7 d and 90 d. The time course of FKN significantly differs depending on stroke severity, with higher FKN levels linked to a lower severity. FKN levels in patients with moderate to severe strokes differ significantly from controls. In outcome analysis, we found an association of dynamics of FKN with clinical outcome. Decrease of FKN is pronounced in patients with worse outcome. Multivariate analysis including stroke severity and stroke etiology revealed that deltaFKN between 6 h and 3 d is independently associated with mRS at 90 d. In addition deltaFKN is inversely correlated with the extent of brain damage, as measured by S100B. FKN dynamics are independently associated with stroke outcome. Further studies might give insight on whether FKN is actively involved in the inflammatory cascade after acute ischemic stroke.


Design of the FRESH-DOSE study: A randomized controlled noninferiority trial evaluating a guided self-help family-based treatment program for children with overweight or obesity.
Overweight and obesity affect 45% of children and increases the risk for several negative health sequelae. Family-Based Behavioral Treatment (FBT) is the most efficacious treatment for child weight management and consists of nutrition and physical activity education, behavior change skills and parenting skills training. FBT is time and staff intensive and can include 20, 60-min separate groups for parents and children, as well as 20-min behavior coaching sessions to help problem solve barriers to implementing the skills learned and individualize the program. Guided self-help (GSH) therapies involve providing families a manual to review independently and brief coaching sessions by an interventionist to facilitate adherence. We developed a GSH version of FBT (gshFBT) which provides a manual to both parents and children and includes 14, 20-min coaching sessions over 6-months. The current study randomized 150 children (mean age = 10.1 years (SD = 1.38); mean BMI% = 97.3% (SD = 2.84); mean BMIz = 2.09 (SD = 0.40); 49% female; 43% Hispanic) and one of their parents (mean age = 41.8 years (SD = 6.52); mean BMI = 32.0 (SD = 7.24); 87.3% female; 43% Hispanic) to either a group-based FBT program or a gshFBT program. Assessments are conducted at baseline, post-treatment (6 months), 6-month follow-up (12 months) and 12-month follow-up (18 months). Primary outcomes are child weight change (BMIz) and cost effectiveness. Recruitment occurred between May 2017 and October 2021 and follow-up assessments are underway. Given the public health concern for children with obesity and the low level of access to FBT, gshFBT could prove extremely useful to provide intervention to a greater proportion of the population.


Preimplantation genetic testing for aneuploidy (PGT-A): The biology, the technology and the clinical outcomes.
Preimplantation genetic testing for aneuploidy (PGT-A) seeks to identify preimplantation embryos with a normal chromosome complement (euploid) during in vitro fertilisation (IVF). By sifting out embryos with abnormal chromosome numbers (aneuploid), PGT-A should theoretically improve pregnancy success. However, earlier versions of PGT-A were ineffective, and in some cases, detrimental, due to biopsy-induced trauma and because the technology at the time could analyse only a fraction of all chromosomes. More recently, the emergence of technologies enabling all chromosomes to be analysed and a switch to less traumatic blastocyst-stage biopsy have seen widespread uptake of PGT-A. Assessing the full impact of blastocyst biopsy PGT-A requires consideration of multiple factors, including embryonic mosaicism, sensitivity of the technological platform used, embryo loss during long-term in vitro culture, embryo cryopreservation and inter-clinic variability in expertise. Significantly, there hasnt yet been an appropriately designed randomised controlled trial (RCT) of blastocyst biopsy PGT-A analysed by intention-to-treat that accounts for all these parameters on a per-cycle basis. The three RCTs reporting benefits studied outcomes on a per-embryo transfer basis were small and underpowered and demonstrated benefits for a very select sub-group of good prognosis patients. The liberal use of this very expensive IVF add-on for other patient populations has not yet been shown to be effective, or indeed, without harm.


Age-related changes in the immune system of mice of eight medium and long-lived strains and hybrids. II. Short- and long-term effects of natural infection with parainfluenza type 1 virus (Sendai).
Studies on the effects of natural infection with parainfluenza type 1 virus (Sendai) on the immune system of 8 strains and hybrids of aging mice revealed that (a) 55 of 63 indices tested were abnormal as late as 8 months after the disappearance of clinical systoms, (b) thymus weight and cell indices and lymph node T cell activity still exhibited a statistically significant negative correlation with age following infection, and (c) immunologically immature young and immunodeficient old mice suffered more severe sequelae following viral infection than did adult or middle-aged mice as determined by accelerated decline of thymus indices, mortality, IgM Coombs' titer, and in the case of old mice, anemia. These findings suggest that chronic viral infection can accelerate immunologic aging and that viral infection in utero can suppress or retard the development of the T cell component of the immune system. In support of our previous study, they also suggest that the thymus regulates immunologic aging and that genetic factors regulating immunologic aging are not easily influenced by environmental factors.


Exercise capacity in children with bronchopulmonary dysplasia at school age.
To assess the exercise capacity, exercise habits, and lung function of preterm born children with bronchopulmonary dysplasia (BPD) compared to term born controls at school age. Cardiopulmonary exercise test (CPET) by cycle ergometer and pulmonary function test were performed in children with BPD (n = 42) and compared with a term born control group (n = 42). Daily activity, participation in sports and respiratory symptoms were assessed by questionnaire. Children with BPD versus controls had significantly lower values for oxygen consumption (V̇O2 [mL/min] 1442 ± 417 vs. 1766 ± 541), minute ventilation (VE [L/min] 48 ± 14.92 vs. 60 ± 18.33), and workload (W [watt] 96.1 ± 16.7 vs. 110.6 ± 17.2) at peak exercise and a lower anaerobic threshold (VO2 AT [mL/min] 1183 ± 345 vs. 1382 ± 398). When corrected for weight, only for the workload (2.7 ± 0.5 vs. 3.1 ± 0.5, p = 0.0013) did significant differences persist. The forced expiratory volume in 1 s and forced expiratory flow between 25 and 75% of expired forced vital capacity were significantly reduced in the BPD group (p < 0.0001). Children with BPD have a higher risk of reporting difficulties in physical activity (OR 2.5) and of suffering from wheezing or shortness of breath while exercising (OR 2.5). Compared to term born controls, children with BPD at school age show airflow obstruction, a lower workload in CPET, and more respiratory symptoms related to physical activity. The comparable oxygen consumption based on weight suggests a functionally normal alveolar compartment.


Qualitative and quantitative preservation of the fine structure of absorptive cells in cultured biopsies of human small-intestine.
The fine structure of the absorptive cells in human small-intestinal biopsies cultured for 6, 24, and 48 h was analyzed qualitatively and quantitatively. The findings show generally good preservation of the cultured absorptive cells and a normal distribution, size, and relative volume of their cell organelles, but there was a systematic decrease in the apical cell surface and an increase in the number of apical vesicles and tubules after culturing. Since the apical vesicles and tubules are thought to have a function in the transport of cell-coat material from the Golgi apparatus to the cell surface, these findings raise the question of whether a delayed transport or extrusion of cell surface material occurs. The diminished relative volume of the mitochondria and the increased signs of autophagy in some poorly preserved absorptive cells, are assumed to be an adaption to less favourable culture conditions.


Arteriovenous malformation of the pancreas: assessment of clinical and multislice CT features.
This study was undertaken to analyze the clinical and CT features of arteriovenous malformation (AVM) of the pancreas. Seven lesions in six consecutive patients (one woman and five men, mean age 51 years) with AVM of the pancreas who underwent multiphase contrast-enhanced CT with 0.5 or 1-mm collimation were retrospectively studied. CT images were evaluated and correlated with angiographic findings by two radiologists. In four patients, the lesions were incidental findings. Two patients presented with abdominal pain and gastrointestinal bleeding, respectively. Serum amylase levels were within normal limits in all patients. The mean size of the lesions was 32.1 mm (3.0-97.3 mm). Conglomeration of strong nodular stains and early enhancement of the portal venous system were observed for all the lesions. The diagnosis of AVM of the pancreas was retrospectively established for all lesions, but was prospectively established for four lesions. The feeding arteries of all lesions were depicted, showing agreement with those observed by angiography performed for four lesions. One lesion showed pseudocyst formation, haziness around the lesion, and strong enhancement of the duodenal wall. Multiphase CT examination using multislice CT is useful for diagnosis of AVM of the pancreas, including identification of the feeding arteries.


Apical junctions and growth control in Drosophila.
Recent studies have revealed unexpected links between cell polarity and proliferation, suggesting that the polarized organization of cells is necessary to regulate growth. Drosophila melanogaster is a genetically simple model that is especially suited for the study of polarity and growth control, as polarized tissues undergo a well-defined pattern of proliferation and differentiation during the development. In addition, genetic studies have identified a number of tumor suppressor genes, which later studies have shown to be associated with junctions, or in the regulation of junctional proteins. We will explore in this review the links between growth and apical junction proteins in the regulation of growth control in Drosophila.


Novel dual LSD1/HDAC6 inhibitors for the treatment of multiple myeloma.
Lysine specific demethylase 1 (LSD1) and HDAC6 are epigenetic proteins associated with several diseases, including cancer and combined inhibition of these proteins could be highly beneficial in treating some cancers such as AML, MM and solid tumors. Multiple myeloma (MM) is a challenging cancer with fast relapse rate where novel treatment options are the need of the hour. We have designed and developed novel, LSD1 and HDAC6 selective dual inhibitors to target MM. Our dual inhibitor compound 1 shows superior potency in multiple MM cell lines. In MM.1S xenograft model compound 1 shows superior efficacy compared to single agent LSD1 and HDAC6 inhibitors by oral administration and is well tolerated. Further evaluation of the molecule in other cancers is in progress.


Routine staging laparoscopy has no place in the management of colorectal liver metastases.
Staging laparoscopy has been recommended in the management of patients with colorectal liver metastases prior to hepatectomy in order to reduce the incidence and associated morbidity of futile laparotomies. The utility of staging laparoscopy has not been assessed in patients undergoing CT, PET-CT and MRI as standard preoperative staging. All patients undergoing attempted open hepatectomy for colorectal liver metastases between 1/4/2008 and 31/3/2012 were identified from a prospectively maintained research database. All patients who underwent futile laparotomy were identified, with demographics and operative notes subsequently analysed. A total of 274 patients underwent attempted open hepatectomy during the study period. At laparotomy 12 (4.4%) patients were found to have irresectable disease. There were no unifying demographic factors within the patients undergoing futile laparotomy. With modern imaging, the potential yield of staging laparoscopy is low. Staging laparoscopy should not be used routinely, but may have a role in the case of specific clinical concerns.


Groundwater conditions the effectiveness of surface water diversion in the remediation of the eutrophicated volcanic lake of Furnas, Azores archipelago.
Volcanic lakes in oceanic islands represent extremely important areas for biodiversity and offer exceptional conditions for nature-based tourism as one of the main pillars of economic growth in these regions. Nitrogen and phosphorus fertilizers are being used extensively at the Azores archipelago, similarly to other places in the world, to increase agricultural production and is causing severe pollution and eutrophication of surface freshwater reservoirs. This work concentrates on the evaluation of the efficiency of surface water diversion as a remediation measure to reduce nutrient loading and reverse eutrophication of Furnas crater lake on the island of São Miguel. Nutrient loading was monitored using an extensive water quality monitoring program in the main watershed of the eutrophicated lake that, together with watershed-scale mass balance methods and groundwater and solute transport models, allowed us to identify an average 98% efficiency in the reduction of nitrate loading. However, phosphorus total load in the discharged water to Furnas lake was observed to only be decreased by 33% due to the groundwater origin of phosphorus. Results from modelling suggested that nutrients were emitted from both point (nitrates) and diffuse (phosphorus) sources as surface runoff and as groundwater seepage, respectively. The results obtained recognized a partially successful surface water diversion, since groundwater path was not initially identified, thus highlighting the importance of groundwater flow regime in the design of such remediation measures. This work also provides a perspective on surface water diversion to revert eutrophication under a volcanic formation, where lakes can be naturally more nutrient rich.


Longitudinal changes in angiogenesis biomarkers within 72 h of diagnosis and time-to-delivery in early-onset preeclampsia.
To evaluate the association between the increment of the sFlt-1/PlGF ratio within the first 72 h after the diagnosis of early-onset preeclampsia (PE) and the time-to-delivery. Secondarily we aimed to test its predictive value for maternal adverse outcomes. Retrospective cohort study of 155 women with early-onset PE and measurement of sFlt-1/PlGF at diagnosis and delivery from which the expected distributions of the daily increment (Δ) of sFlt-1/PlGF ratio, sFlt-1 and PlGF were obtained. Of them, in 110 a short-term evaluation at 72 +/- 24 h was available and Δ72h were calculated and compared to the expected distributions. The high-risk groups were those with Δ72h sFlt-1/PlGF and Δ72h sFlt-1 > 3rd expected quartile or Δ72h PlGF < 1st expected quartile. The low-risk groups were those with Δ72h ≤ 0 for sFlt-1/PlGF and sFlt-1 or Δ72h PlGF ≥ 0. The rest were considered intermediate risk. Time-to-delivery and maternal adverse outcomes were compared between the three groups. Δ72h sFlt-1/PlGF and sFlt-1 > 3rd quartile had a significant reduction of time-to-delivery when compared to increments < 3rd quartile or ≤ 0 (5 vs 11 vs 14 days, p < 0.01) and (6 vs 8 vs 15 days, p < 0.01), respectively. Both were limited for the prediction of maternal adverse outcomes. Δ72h PlGF showed no significant relation with time-to-deliver but all abruptio placentae had PlGF < 70 pg/mL at diagnosis. High Δ72h sFlt-1/PlGF and sFlt-1 are associated to a shorter time-to-delivery while low PlGF at diagnosis is associated to abruptio placentae.


Reducing Postoperative Length of Stay for Idiopathic Scoliosis Patients using Quality Improvement Methodology.
Approximately 1%-3% of the US population is diagnosed with scoliosis. In addition, 80% of those diagnosed have idiopathic scoliosis, with about 10% requiring surgical intervention. This Quality Improvement initiative aimed to reduce the length of stay (LOS) after posterior spinal fusion for these patients. According to the Pediatric Health Information System, our institution had a poorer performance, with an actual LOS greater than or equal to the expected LOS compared with peer institutions. The aim was to increase the percentage of idiopathic scoliosis patients with a procedure to discharge LOS of less than or equal to 4 days after posterior spinal fusion from 39.13% to 90%. Interventions included implementing a new pain management protocol, a daily checklist, education on expectations of postoperative pain, and updated order sets. Interventions improved patients discharged in less than 4 days from 39.13% to 93.48% (P ≤ 0.001), reducing the average postprocedure LOS from 4.93 to 2.59 (P ≤ 0.001) days. A key process measure tracked was the percentage of patients off the patient-control analgesia pump by postoperative day 2, which increased from 13% to 97.75% (P ≤ 0.001). These improvements did not affect the balancing measure of readmissions or Emergency Department visits for pain. By implementing a more standardized pathway, including a patient-focused daily checklist for providers and families, we established expectations for LOS and pain. This checklist and updates to the pain management protocol successfully reduced the length of stay in idiopathic scoliosis patients after posterior spinal fusion.


Production of active recombinant human aldehyde oxidase (AOX) in the baculovirus expression vector system (BEVS) and deployment in a pre-clinical fraction-of-control AOX compound exposure assay.
Human aldehyde oxidase (AOX) has emerged as a key enzyme activity for consideration in modern drug discovery. The enzyme catalyzes the oxidation of a wide variety of compounds, most notably azaheterocyclics that often form the building blocks of small molecule therapeutics. Failure to consider and assess AOX drug exposure early in the drug development cycle can have catastrophic consequences for novel compounds entering the clinic. AOX is a complex molybdopterin-containing iron-sulfur flavoprotein comprised of two identical 150 kDa subunits that has proven difficult to produce in recombinant form, and a commercial source of the purified human enzyme is currently unavailable. Thus, the potential exposure of novel drug development candidates to human AOX metabolism is usually assessed by using extracts of pooled human liver cytosol as a source of the enzyme. This can complicate the assignment of AOX-specific compound exposure due to its low activity and the presence of contaminating enzymes that may have overlapping substrate specificities. Herein is described a two-step process for the isolation of recombinant human AOX dimers to near homogeneity following production in the baculovirus expression vector system (BEVS). The deployment of this BEVS-produced recombinant human AOX as a substitute for human liver extracts in a fraction-of-control AOX compound-exposure screening assay is described. The ability to generate this key enzyme activity readily in a purified recombinant form provides for a more accurate and convenient approach to the assessment of new compound exposure to bona fide AOX drug metabolism.


High resolution Fourier domain optical coherence tomography in the 2 μm wavelength range using a broadband supercontinuum source: erratum.
We correct a mistake in the OCT system sensitivity given in our recent paper [Opt. Express23(3), 1992 (2015)].


[Mechanical property of tooth-like yttria-stabilized tetragonal zirconia polycrystal by adding rare earth oxide].
To evaluate the influence of mechanical property of tooth-like yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) by adding rare earth oxide as colorants. Six kinds of tooth-like Y-TZP were made by introducing internal coloration technology. The colorants included rare earth oxide (Pr6O11, CeO2, Er2O3) and transition element oxide (MnO2). Mechanical properties (flexural strength, vickers hardness and fracture toughness) were tested. Microstructure was examined by scanning electron microscope(SEM), and the fracture model was analyzed. The range of flexural strength of the six kinds of tooth-like Y-TZP were (792 +/- 20)-(960 +/- 17) MPa, the fracture toughness were (4.72 +/- 0.31)-(5.64 +/- 0.38) MPam(1/2), and the vickers hardness were (1332 +/- 19)-(1380 +/- 17) MPa. SEM observation on the cross section of the six kinds of sintered composites showed a relatively dense polycrystal structure, and the fracture models was mixed type. Tooth-like Y-TZP is acquired with better mechanical properties (fracture toughness and vickers hardness) by adding rare earth oxide as colorants. It is available for clinical application.


Low aspartate aminotransferase/alanine aminotransferase ratio is a predictor of diabetes incidence in Japanese people: Population-based Panasonic cohort study 5.
The aim of this study was to calculate the cut-off values of liver enzymes to identify the risk of incident type 2 diabetes (DM) and to investigate the association between liver enzymes and incident DM in participants with or without obesity. The long-term cohort study included 70,688 subjects who underwent medical health checkups in 2008. The cut-off values of alanine aminotransferase (ALT) and the aminotransferase (AST)/ALT ratio for incident DM were evaluated using the time-dependent receiver operating characteristic curves. The risk of incident type 2 DM was examined according to cut-off values of liver enzymes and the group with body mass index (BMI) ≥25 kg/m2 using Cox regression analyses. In total, 4181 of 70,688 subjects developed DM within 10 years. The area under the curve and cut-off values for the ALT and the AST/ALT ratio for incident type 2 DM at 10 years were 0.707 and 23 IU/L and 0.694 and 0.875, respectively. The risk of incident DM was higher in subjects with ALT ≥23 or AST/ALT ≤0.875 and BMI <25 kg/m2 than in those with ALT <23 IU/L or AST/ALT >0.875 and BMI ≥25 kg/m2 , respectively. The cut-off values of ALT and the AST/ALT ratio associated with the risk of incident type 2 DM were determined. Non-obese individuals with AST/ALT ≤0.875 had a higher risk of incident type 2 DM than obese individuals with AST/ALT >0.875.


Author Correction: Neutrophil to lymphocyte ratio and breast cancer risk: analysis by subtype and potential interactions.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Forced to communicate: Integration of mechanical and biochemical signaling in morphogenesis.
Morphogenesis is a physical process that requires the generation of mechanical forces to achieve dynamic changes in cell position, tissue shape, and size as well as biochemical signals to coordinate these events. Mechanical forces are also used by the embryo to transmit detailed information across space and detected by target cells, leading to downstream changes in cellular properties and behaviors. Indeed, forces provide signaling information of complementary quality that can both synergize and diversify the functional outputs of biochemical signaling. Here, we discuss recent findings that reveal how mechanical signaling and biochemical signaling are integrated during morphogenesis and the possible context-specific advantages conferred by the interactions between these signaling mechanisms.


Long-term safety and effectiveness of adalimumab in 462 patients with intestinal Behçet's disease: results from a large real-world observational study
The safety and effectiveness of adalimumab was demonstrated in a phase 3 trial in Japanese patients with intestinal Behçet's disease. The aim of this study was to evaluate the long-term safety and effectiveness of adalimumab in Japanese patients with intestinal Behçet's disease. This prospective, all-case, post-marketing study was conducted at 254 centers in Japanese patients with intestinal Behçet's disease receiving adalimumab. The primary endpoint was incidence of adverse drug reactions. Effectiveness endpoints included global improvement rating and change in C-reactive protein levels. Of the 473 registered patients, 462 and 383 included in the safety and effectiveness populations were administered adalimumab for a mean of 515.3 and 579.5 days, respectively. Overall, 395 patients (85.5%) received adalimumab at the recommended dose. Adverse drug reactions and serious adverse drug reactions were reported in 120 (25.97%) and 51 (11.04%) patients, respectively. The incidence of adverse drug reactions was significantly higher in patients with comorbidities (P< 0.0001), patients taking concomitant oral corticosteroids (P< 0.0001), and those not self-administering adalimumab (P= 0.0257). At study end, global improvement rating was "effective" (n = 156, 40.7%) or "markedly effective" (n = 168, 43.9%) in 324 patients (overall effective, 84.6%). Mean C-reactive protein levels (mg/dL) decreased from 1.96 at baseline (n = 324) to 0.58 at week 24 (n = 208) and 0.25 at week 156 (n = 37). This large real-world study confirmed the long-term safety and effectiveness of adalimumab in patients with intestinal Behçet's disease. No new safety concerns were identified. (Clinical trial registration number: NCT01960790).


Investigational study of iodine-124 with a positron camera.
A case is presented where I-124 produced by a clinical cyclotron was used with a positron emission tomography camera for clinical usage. This represents the first report of the utilization of this modality with this radionuclide. We feel the increased spatial resolution of PET should be of value in looking at thyroid disease.


Chemical probes reveal no evidence of Hoogsteen base pairing in complexes formed between echinomycin and DNA in solution.
Five different DNA fragments have been treated with a range of conformationally sensitive reagents in an effort to probe structural changes in DNA associated with binding of the bis-intercalating antibiotic echinomycin. For each probe, the intensity and pattern of its reactivity with DNA have been analyzed in order to elucidate the effect of antibiotic binding on the accessibility of a specific site or sites to chemical attack. It was found that in one of the DNA fragments, pTyr2 DNA, several purine residues exhibit enhanced reactivity to diethyl pyrocarbonate (DEPC) in the absence of bound antibiotic, and that this strongly sequence specific reaction is enhanced in the presence of quite low echinomycin concentrations. The echinomycin-dependent reactivities towards DEPC of three homologous DNA fragments, chosen for their subtly different antibiotic binding characteristics, were also investigated. It was found that small changes in base sequence generate striking changes in susceptibility to modification by DEPC. The abolition of one antibiotic binding site leads to the creation of a new, intense DEPC-reactive site. In the presence of moderate concentrations of echinomycin, specific thymidine residues exhibit enhanced reactivity towards osmium tetroxide. No differences in the reactivities of the DNA fragments towards bromoacetaldehyde, S1 nuclease, dimethyl sulphate or potassium tetrachloropalladinate were observed in the presence of the antibiotic. DEPC reactions were performed on tubercidin (7-deaza-adenosine) to determine the DEPC reactive positions in situation where N-7 is inaccessible. Tubercidin was found to be generally resistant to attack by DEPC followed by treatment with base. We conclude that the bulk of structural changes induced by the binding of echinomycin to DNA do not involve Hoogsteen base pairing, but rather are due to sequence-specific unwinding of the helix in a manner which is strongly dependent on the nature of surrounding nucleotide sequences.


Trypanosomes of Australian Mammals: Knowledge Gaps Regarding Transmission and Biosecurity.
Trypanosomes infect humans, domestic animals, and wildlife, and are transmitted by haematophagous invertebrate vectors. Eight native trypanosome species have been described from Australian indigenous mammals, along with other unnamed isolates and genotypes. Associated difficulties relating to the confirmation of cyclical and mechanical vector candidates has hindered vector identification in Australia, with no successful experimental transmission documented for any of these native trypanosomes to indigenous mammals. We discuss pending biosecurity issues, with significant importance placed on the close phylogenetic and phenotypic relationship shared between Trypanosoma cruzi and some Australian trypanosomes. With such a dearth of information, we highlight the importance of keeping an open mind, which considers all possibilities during future investigations of vectors and their associated biosecurity issues in Australia.


E3-ubiquitin ligase Nedd4 determines the fate of AID-associated RNA polymerase II in B cells.
Programmed mutagenesis of the immunoglobulin locus of B lymphocytes during class switch recombination (CSR) and somatic hypermutation requires RNA polymerase II (polII) transcription complex-dependent targeting of the DNA mutator activation-induced cytidine deaminase (AID). AID deaminates cytidine residues on substrate sequences in the immunoglobulin (Ig) locus via a transcription-dependent mechanism, and this activity is stimulated by the RNA polII stalling cofactor Spt5 and the 11-subunit cellular noncoding RNA 3'-5' exonucleolytic processing complex RNA exosome. The mechanism by which the RNA exosome recognizes immunoglobulin locus RNA substrates to stimulate AID DNA deamination activity on its in vivo substrate sequences is an important question. Here we report that E3-ubiquitin ligase Nedd4 destabilizes AID-associated RNA polII by a ubiquitination event, leading to generation of 3' end free RNA exosome RNA substrates at the Ig locus and other AID target sequences genome-wide. We found that lack of Nedd4 activity in B cells leads to accumulation of RNA exosome substrates at AID target genes and defective CSR. Taken together, our study links noncoding RNA processing following RNA polII pausing with regulation of the mutator AID protein. Our study also identifies Nedd4 as a regulator of noncoding RNAs that are generated by stalled RNA polII genome-wide.


Calcium(II)(3) (3,5-Diisopropylsalicylate)(6)(H(2)O)(6) Activates Nitric Oxide Synthase: An Accounting for its Action in Decreasing Platelet Aggregation.
Purposes of these studies were first; to determine whether or not Calcium(II)(3) (3,5- diisopropylsalicylate)(6)(H(2)O)(6) [Ca(II)(3)(3,5-DIPS)(6)], a lipophilic calcium complex, could decrease activated-platelet aggregation, and second; to determine whether or not it is plausible that Ca(II)(3)(3,5-DIPS)(6) decreases activated-platelet aggregation by facilitating the synthesis of Nitric Oxide (NO) by Nitric Oxide Synthase (NOS). The influence of Ca(II)(3)(3,5-DIPS)(6) on the initial rate of activated-platelet aggregation was determined by measuring the decrease in rate of increase in transmission at 550 nm for a suspension of Thrombin-CaCl(2) activated platelets following the addition of 0, 50, 100, 250, or 500 muM Ca(II)(3)(3,5-DIPS)(6). To establish that the Ca(lI)(3)(3,5- DIPS)(6)-mediated decrease in aggregation was due to activation of NOS, the effect of L-NMMA, an inhibitor of NOS, on the inhibition of platelet aggregation by Ca(II)(3)(3,5-DIPS)(6) was determined using a suspension of activated platelets contaimng 0 or 250 muM Ca(II)(3)(3,5-DIPS)(6) without or with 1 mM L-NMMA. An in vitro Bovine Brain NOS reaction mixture, containing CaCl(2) for the activation of Phosphodiesterase-3' ,5'-Cyclic Nucleotide Activator required for the activation of NOS, was used to determine whether or not Ca(II)(3)(3,5-DIPS)(6) could be used as a substitute for the addition of Ca. The decrease in absorbance at 340 nm, lambda maximum for NADPH, was measured to determine NOS activity following the addition of NOS to the complete reaction mixture containing either CaCl(2), Ca(II)(3)(3,5-DIPS)(6), or neither Ca compound. Increasing the concentration of Ca(II)(3)(3,5-DIPS)(6) caused a concentration related decrease in activated platelet aggregation. The addition of L-NMMA to activated platelets, in the absence of Ca(II)(3)(3,5-DIPS)(6), caused a 129% increase in initial rate of platelet aggregation. The initial rate of platelet aggregation decreased 74% with the addition of 250 muM Ca(II)(3)(3,5-DIPS)(6) and the addition of L-NMMA plus 250 muM Ca(II)(3)(3,5-DIPS)(6) caused a 197% decrease in initial rate of aggregation compared to the initial rate observed width the presence of 1 mM L-NMMA alone. There was only a small, 27%, increase in initial rate of 0.4 mM NADPH oxidation when 0.9 mM CaCl(2) was added to the NOS reaction mixture in comparison to the initial rate of NADPH oxidation with no addition of CaCI(2). Addition of an equivalent amount of Ca in the form of Ca(II)(3)(3,5-DIPS)(6), 333 muM, caused a 37% increase in initial rate of NADPH oxidation compared to the addition of 0.9 mM CaCl(2). Addition of increasing concentrations of L-NMMA plus 0.9 mM CaCl(2) or 333 muM Ca(II)(3)(3,5-DIPS)(6) to the NOS reaction mixture caused a concentration related increase in initial rate of NADPH oxidation. Addition of L-NMMA while expected to decrease NADPH oxidation actually increased the rate of NADPH oxidation. Additions of 133 muM or 267 muM Ca(II)(3)(3,5- DIPS)(6) also caused concentration related increases in initial rate of NADPH oxidation in the presence of 113 muM L-NMMA. However, the addition of 533 muM Ca(II)(3)(3,5-DIPS)(6) caused a dramatic decrease in initial rate of NADPH oxidation by NOS. It is concluded that: 1) Ca(II)(3)(3,5- DIPS)(6) activates platelet NOS in preventing platelet aggregation, 2) in vitro NOS activity can be observed spectrophotometrically by following the consumption of NADPH as a decrease in absorbance at 340 nm, 3) Ca(II)(3)(3,5-DIPS)(6) plays a role in enhancing Bovine Brain NOS activity resulting in an increased rate of NADPH oxidation by NOS, 4) Ca(II)(3)(3,5-DIPS)(6) is a useful form of Ca in activating NOS and superior to CaCl(2) with regard to the facilitation of a NADPH oxidation, and 5) L-NMMA stimulates Bovine Brain NOS activity rather than causing an inhibition of this enzyme and must serve as a reducible substrate for Bovine Brain NOS.


Percutaneous Treatment of an Aorto-Right Ventricular Fistula Following Balloon-Expandable Transcatheter Aortic Valve Replacement.
We present the case of a 71-year-old man who experienced congestive cardiac failure after transcatheter aortic valve replacement with a balloon-expandable transcatheter heart valve. Echocardiography and cardiac computed tomography demonstrated an aorto-right ventricular fistula, and successful percutaneous closure was performed with a vascular plug. (Level of Difficulty: Advanced.).


Heart rate variability during sleep and subsequent sleepiness in patients with chronic fatigue syndrome.
We determined whether alterations in heart rate dynamics during sleep in patients with chronic fatigue syndrome (CFS) differed from controls and/or correlated with changes of sleepiness before and after a night in the sleep laboratory. We compared beat-to-beat RR intervals (RRI) during nocturnal sleep, sleep structure, and subjective scores on visual analog scale for sleepiness in 18 CFS patients with 19 healthy controls aged 25-55 after excluding subjects with sleep disorders. A short-term fractal scaling exponent (α1) of RRI dynamics, analyzed by the detrended fluctuation analysis (DFA) method, was assessed after stratifying patients into those who reported more or less sleepiness after the night's sleep (a.m. sleepier or a.m. less sleepy, respectively). Patients in the a.m. sleepier group showed significantly (p<0.05) higher fractal scaling index α1 during non-rapid eye movement (non-REM) sleep (Stages 1, 2, and 3 sleep) than healthy controls, although standard polysomnographic measures did not differ between the groups. The fractal scaling index α1 during non-REM sleep was significantly (p<0.05) higher than that during awake periods after sleep onset for healthy controls and patients in the a.m. less sleepy group, but did not differ between sleep stages for patients in the a.m. sleepier group. For patients, changes in self-reported sleepiness before and after the night correlated positively with the fractal scaling index α1 during non-REM sleep (p<0.05). These results suggest that RRI dynamics or autonomic nervous system activity during non-REM sleep might be associated with disrupted sleep in patients with CFS.


Single-cell transcriptome sequencing of rumen ciliates provides insight into their molecular adaptations to the anaerobic and carbohydrate-rich rumen microenvironment.
Rumen ciliates are a specialized group of ciliates exclusively found in the anaerobic, carbohydrate-rich rumen microenvironment. However, the molecular and mechanistic basis of the physiological and behavioral adaptation of ciliates to the rumen microenvironment is undefined. We used single-cell transcriptome sequencing to explore the adaptive evolution of three rumen ciliates: two entodiniomorphids, Entodinium furca and Diplodinium dentatum; and one vestibuliferid, Isotricha intestinalis. We found that all three species are members of monophyletic orders within the class Litostomatea, with E. furca and D. dentatum in Entodiniomorphida and I. intestinalis in Vestibuliferida. The two entodiniomorphids might use H2-producing mitochondria and the vestibuliferid might use anaerobic mitochondria to survive under strictly anaerobic conditions. Moreover, carbohydrate-active enzyme (CAZyme) genes were identified in all three species, including cellulases, hemicellulases, and pectinases. The evidence that all three species have acquired prokaryote-derived genes by horizontal gene transfer (HGT) to digest plant biomass includes a significant enrichment of gene ontology categories such as cell wall macromolecule catabolic process and carbohydrate catabolic process and the identification of genes in common between CAZyme and HGT groups. These findings suggest that HGT might be an important mechanism in the adaptive evolution of ciliates to the rumen microenvironment.


Diagnostic accuracy of a uniform research case definition for TBM in children: a prospective study.
Bacteriological confirmation of tuberculous meningitis (TBM) is problematic, and rarely guides initial clinical management. A uniform TBM case definition has been proposed for research purposes. We prospectively enrolled patients aged 3 months to 13 years with meningitis confirmed using cerebrospinal fluid analysis at Tygerberg Hospital, Cape Town, South Africa. Criteria that differentiated TBM from other causes were explored and the accuracy of a probable TBM score assessed by comparing bacteriologically confirmed cases to 'non-TBM' controls. Of 139 meningitis patients, 79 were diagnosed with TBM (35 bacteriologically confirmed), 10 with bacterial meningitis and 50 with viral meningitis. Among those with bacteriologically confirmed TBM, 15 were Mycobacterium tuberculosis culture-positive and 20 were culture-negative but positive on GenoType(®) MTBDRplus or Xpert(®) MTB/RIF; 18 were positive on only a single commercial nucleic acid amplification test. A probable TBM score provided a sensitivity of 74% (95%CI 57-88) and a specificity of 97% (95%CI 86-99) compared to bacteriologically confirmed TBM. A probable TBM score demonstrated excellent specificity compared to bacteriological confirmation. However, 26% of children with TBM would be missed due to the limited accuracy of the case definition. Further prospective testing of an algorithm-based approach to TBM is advisable before recommendation for general clinical practice.


Human allospecific T cell proliferative clones: different T cell clones expressing an apparently identical HLA class II specificity are heterogeneous in regard to their proliferative inhibition by a panel of monoclonal antibodies against human Ia-like antigens.
Four independent PLT clones displaying an apparently identical class II specificity (i.e., Dw/DR3) were found to give a heterogeneous pattern of inhibition in relation to 15 anti-class II mAb and an anti-beta 2m mAb used as a control. Some clones were inhibited by all anti-class II mAbs, irrespective of the cluster of molecular subsets with which they reacted. Such clones were also inhibited by the control anti-beta 2m mAb. Other clones were inhibited by only a few of the mAb tested. Within this group of T clones following the addition of a limiting amount of conditioned medium the inhibitory data of all independent clones with an identical specificity were inhibited by the same mAb; under these conditions, it was possible to relate the mAb inhibition patterns with the specificity of the T cell clones and these T cell specificities with the epitopic cluster/molecular subsets defined by these mAbs. A new level of T cell subset heterogeneity within T cell clones with apparent identical proliferative specificities is demonstrated, in relation to the T cell clone "susceptibility" or "resistance" to the effects of anti-class II mAbs directed towards their own Ia-like antigens.


Effects of alternate wetting and drying on oxyanion-forming and cationic trace elements in rice paddy soils: impacts on arsenic, cadmium, and micronutrients in rice.
Rice is a global dietary staple and its traditional cultivation under flooded soil conditions leads to accumulation of arsenic (As) in rice grains. Alternate wetting and drying (AWD) is a widely advocated water management practice to achieve lower As concentrations in rice, water savings, and decreased methane emissions. It is not yet clear whether AWD leads to tradeoffs between concentrations of As and micronutrient elements (e.g., zinc, manganese, molybdenum) in rice grain. We analyzed pore water chemistry and rice grain composition data from a field experiment conducted in Arkansas, USA, in 2017 and 2018 to test the hypothesis that AWD will have diverging effects on oxyanion-forming (arsenic, molybdenum) vs. cationic (cadmium, zinc, manganese, copper) trace elements. This was hypothesized to occur via decreases in soil pH and/or precipitation of iron oxide minerals during oxidizing conditions under AWD. Solubility of all trace elements, except zinc, increased in more reducing conditions. Consistent with our hypothesis, AWD tended to increase grain concentrations of cationic elements while decreasing grain concentrations of oxyanionic elements. Decreases in total As in rice grains under AWD were mainly driven by changes in dimethylarsinic concentrations, with negligible changes in inorganic As. Linear mixed-effects modeling showed that effects of AWD on grain composition were more significant in 2017 compared to 2018. These differences may be related to the timing of dry-downs in the developmental stage of rice plants, with dry-downs during the heading stage of rice development leading to larger impacts on grain composition of certain elements. We also observed significant interannual variability in grain elemental composition from continuously-flooded fields and postulate the warmer temperatures in 2018 may have played a role in these differences.


Social learning and memory.
The adaptability of human populations to changing environments is often attributed to the human capacity for social learning, innovation, and culture. In rapidly changing environments, it has been shown that maintaining high levels of cultural variation is beneficial because it allows for efficient adaptation. However, in many theoretical models, a high level of cultural variation also implies that a large amount of useless and perhaps detrimental information must be maintained and used, leading to lower population fitness in general. Here, we begin to investigate this often conflicting relationship between adaptation and cultural variation. We explicitly allow for the interplay between social learning and environmental variability, alongside the capacity for "memory," i.e., the storage, retrieval, and forgetting of information. Here, memory allows individuals to retain unexpressed cultural variation, which does not directly impact adaptation. We show that this capacity for memory facilitates the evolution of social learning across a broader range of circumstances than previously thought. Results from this analysis may help to establish whether and when memory should be incorporated into cultural evolutionary models focused on questions of adaptation.


[Current concepts in a systemic-informatics study of feedback and blood volume homeostasis. Proposal of a mathematical reference model].
Attention is drawn to the relationship between information and biological models in the study of problems of human physiology and physiopathology. The biomathematical principles, to which reference must be made for the elaboration of the expert systems, necessary in evaluating biological feedback in particular are analyzed.


Reduction of hexavalent chromium by cell-free extract of Bacillus sphaericus AND 303 isolated from serpentine soil.
Cell-free extracts (CFEs) of chromium-resistant bacterium Bacillus sphaericus AND 303 isolated from serpentine soil of Andaman, India reduced Cr(VI) in in vitro condition, and the reductase activity was solely localized in the soluble cell-fractions (S12, S32, and S150). The enzyme was constitutive as the CFEs from cells grown in Cr(VI)-free and Cr(VI)-containing media reduced a more or less equal amount of Cr(VI). Optimum Cr(VI) reductase activity was obtained at an enzyme (S150) concentration equivalent to 4.56 mg protein/mL, 300 microM: Cr(VI) and pH 6.0 after 30 min incubation at 30 degrees C. The enzyme was heat labile; 80% of its activity was lost when exposed at 70 degrees C for 15 min. Kinetics of Cr(VI) reductase activity fit well with the linearized Lineweaver-Burk plot and showed a V(max) of 1.432 micromol Cr(VI)/mg protein/min and K(m) of 158.12 microM: Cr(VI). The presence of additional electron donors accelerated Cr(VI) reductase activity of CFE, and an increase of 28% activity over control was recorded with 1.0 microM: NADH. Heavy metal ions such as Ni(II), Cu(II), and Cd(II) were strong inhibitors of Cr(VI) reductase unlike that of 100 microM: Co(II), which retained 93% activity over control.


Failure properties of vena cava tissue due to deep penetration during filter insertion.
In this work, we use an in-vitro mechanical test to explore the resistance of biaxially stretched vena cava tissue against deep perforation and a methodology which integrates experimental and numerical modeling to identify constitutive fracture properties of the vena cava. Six sheep vena cava were harvested just after killing, and cyclic uniaxial tension tests in longitudinal and circumferential directions and biaxial deep penetration tests were performed. After that, we use a nonlinear finite element model to simulate in vitro penetration of the cava tissue in order to fit the fracture properties under penetration of the vena cava by defining a cohesive fracture zone. An iterative process was developed in order to fit the fracture properties of the vena cava using the previously obtained experimental results. The proposed solutions were obtained with fracture energy of 0.22 or 0.33 N/mm. In comparison with the experimental data, the simulation using [Formula: see text], [Formula: see text], and [Formula: see text] parameters ([Formula: see text]) is in good agreement with results from penetration experiments of cava tissue. It is noticeable that the parameter estimation process of the fracture behavior is more accurate than the estimation process of the elastic behavior for the toe region of the curve.


Therapeutic Potential of BAY-117082, a Selective NLRP3 Inflammasome Inhibitor, on Metastatic Evolution in Human Oral Squamous Cell Carcinoma (OSCC).
Oral squamous cell carcinoma (OSCC) is a commonly occurring head and neck cancer and it is characterized by a high metastasis grade. The aim of this study was to evaluate for the first time the effect of BAY-117082, a selective NLRP3 inflammasome inhibitor, in an in vivo orthotopic model of OSCC and its role in the invasiveness and metastasis processes in neighbor organs such as lymph node, lung, and spleen tissues. Our results demonstrated that BAY-117082 treatment, at doses of 2.5 mg/kg and 5 mg/kg, was able to significantly reduce the presence of microscopic tumor islands and nuclear pleomorphism in tongue tissues and modulate the NLRP3 inflammasome pathway activation in tongue tissues, as well as in metastatic organs such as lung and spleen. Additionally, BAY-117082 treatment modulated the epithelial-mesenchymal transition (EMT) process in tongue tissue as well as in metastatic organs such as lymph node, lung, and spleen, also reducing the expression of matrix metalloproteinases (MMPs), particularly MMP2 and MMP9, markers of cell invasion and migration. In conclusion, the obtained data demonstrated that BAY-117082 at doses of 2.5 mg/kg and 5 mg/kg were able to reduce the tongue tumor area as well as the degree of metastasis in lymph node, lung, and spleen tissues through the NLRP3 inflammasome pathway inhibition.


Role of connexin 43 in the mechanism of action of alendronate: dissociation of anti-apoptotic and proliferative signaling pathways.
Bisphosphonates (BPs) inhibit osteocyte and osteoblast apoptosis via opening of connexin (Cx) 43 hemichannels and activating the extracellular signal regulated kinases ERKs. Previously, we hypothesized that intracellular survival signaling is initiated by interaction of BPs with Cx43. However, using whole cell binding assays with [(3)H]-alendronate, herein we demonstrated the presence of saturable, specific and high affinity binding sites in the Cx43-expressing ROS 17/2.8 osteoblastic cells, authentic osteoblasts and MLO-Y4 cells expressing Cx43 or not, as well as in HeLa cells lacking Cx43 expression and ROS 17/2.8 cells pretreated with agents that disassemble Cx channels. In addition, both BPs and the PTP inhibitor Na(3)VO(4) increased proliferation of cells expressing Cx43 or not. Furthermore, although BPs are internalized and inhibit intracellular enzymes in osteoclasts, whether the drugs penetrate non-resorptive bone cells is not known. To clarify this, we evaluated the osteoblastic uptake of AF-ALN, a fluorescently labeled analog of alendronate. AF-ALN was rapidly internalized in cells expressing Cx43 or not indicating that this process is not mediated via Cx43 hemichannels. Altogether, these findings suggest that although required for triggering intracellular survival signaling by BPs, Cx43 is dispensable for cellular BP binding, its uptake, as well as the proliferative effects of these agents.


Complement Factor I Gene Variant as a Treatable Cause of Recurrent Aseptic Neutrophilic Meningitis: A Case Report.
Mutations in the complement factor I (CFI) gene have previously been identified as causes of recurrent CNS inflammation. We present a case of a 26-year-old man with 18 episodes of recurrent meningitis, who had a variant in CFI(c.859G>A,p.Gly287Arg) not previously associated with neurologic manifestations. He achieved remission with canakinumab, a human monoclonal antibody targeted at interleukin-1 beta.


Vaccination with heat-shocked mononuclear cells as a strategy for treating neurodegenerative disorders driven by microglial inflammation.
Naturally occurring T regulatory cells targeting epitopes derived from various heat shock proteins escape thymic negative selection and can be activated by vaccination with heat shock proteins; hence, vaccination with such proteins has exerted favorable effects on rodent models of autoimmune disorders. A more elegant way to achieve such vaccination, first evaluated clinically by Al-Harbi in the early 1990s, is to subject mononuclear cells to survivable heat shock ex vivo, incubate them at physiological temperature for a further 24-48 h, and then inject them subcutaneously; anecdotally, beneficial effects were observed with this strategy in a wide range of autoimmune and inflammatory conditions. There is growing evidence that M1-activated microglia play a primary or secondary role in the pathogenesis of numerous neurodegenerative diseases, as well as in major depression. T regulatory cells, by polarizing microglial toward a reparative M2 phenotype, have the potential to aid control of such disorders. It would be appropriate to test the heat-shocked mononuclear cell vaccination strategy in animal models of neurodegeneration and major depression, and to evaluate this approach clinically if such studies yield encouraging results.


[Tyrosine kinase receptor gene fusion: A series of four cases of infantile-type hemispheric glioma].
Infant-type hemispheric gliomas belong to pediatric-type diffuse high-grade gliomas according to the 2021 WHO classification of central nervous system tumors. They are characterized by tyrosine kinase gene rearrangements (NTRK1/2/3, ALK, ROS1, MET). The aim of the study was to describe the clinical, histopathologic, and molecular characteristics of such tumors, and to provide a review of the literature. This retrospective series comprises four cases of infant-type hemispheric glioma diagnosed at Angers University Hospital between 2020 and 2022. The diagnosis was suspected based on morphology and immunohistochemistry and was confirmed by molecular biology techniques. The most common clinical sign was raised intracranial pressure. Imaging showed a large cerebral hemispheric tumor with contrast enhancement. Microscopic examination revealed diffuse astrocytoma with high-grade features, sometimes with neuronal or pseudo-ependymal differentiation. Identification of a gene fusion involving a tyrosine kinase gene allowed to make a definitive diagnosis of infant-type hemispheric glioma. Infant-type hemispheric gliomas are rare and present as large cerebral hemispheric tumors in very young children. Searching for a tyrosine kinase gene fusion should be systematic when dealing with a high-grade glioma in an infant. Importantly, these gene fusions are therapeutic targets. The impact of targeted therapies on patient survival should be evaluated in future prospective studies.


Stability and singularity analysis of the cosmologies with different scenarios for deceleration parameter in the presence of torsion.
In the current paper, path deviation equations in absolutely parametric parallel geometries are derived. It is considered as a geodesic deviation equation. Additionally, it is modified by a torsion term. It proposes the deviation path equation that describes the trajectory deviation of a particle under the influence of the gravitational field. To examine the singularity of the Cosmological models, the modified version of the Raychaudhuri equation is utilized. The generalized law of the variation of Hubble's parameter is utilized to achieve some Cosmological models.


Clinical course of diffuse invasive carcinoma of the tongue excised after Bleomycin treatment.
According to the mode of invasion across the tumour-host boundary which was described and graded by Jacobsson et al. (1973), the clinical course of diffuse invasive squamous cell carcinoma of the tongue (grade 4) was investigated comparing it with the other grade groups (grades 1-3). Metastasis to regional lymph nodes and local recurrence after surgery were extremely common in grade 4. These local recurrences led to death in most grade 4 cases while a better clinical outcome was achieved in the other grades. Grade 4 was further subclassified into a cord-like type (grade 4 C) and a diffuse type (grade 4 D) from the histological features of the tumour-host interface, with the latter being more malignant than the former.


[Morphological properties of erythrocytes and their deformability in cerebral circulatory disorders in patients with arterial hypertension].
Using a scanning electron microscope, the authors examined changes in the shape of erythrocytes (E) in 85 patients with impaired cerebral circulation (ICC). Parallelly 48 patients were studied for the deformity and osmotic resistance of E. The control group consisted of 17 healthy individuals. The data obtained suggest significant alterations in the shape and functional properties of E in patients with ICC which correlated with parameters of blood viscosity and E aggregation. The degree of changes was determined by the duration of the underlying vascular disease, recurrent episodes of ICC and their severity.


[Phloem, transport between organs and long-distance signalling].
Phloem plays a major role in carbohydrate partitioning in the plant. It also controls the redistribution of various metabolites such as amino acids, vitamins, hormones, and ions. The molecular mechanisms responsible for phloem loading and unloading have been particularly well characterised, with the identification of sucrose and polyol transporters. The discovery of the role of phloem in the long-distance translocation of macromolecules, proteins, mRNA and small RNA has modified our understanding of the regulation of the coordination of some developmental and adaptation processes. This review details recent results concerning the transport and long-distance signalling that take place in the phloem.


Mossy cells and different subpopulations of pyramidal neurons are immunoreactive for cocaine- and amphetamine-regulated transcript peptide in the hippocampal formation of non-human primates and tree shrew (Tupaia belangeri).
Cocaine- and amphetamine-regulated transcript peptide mRNA was discovered in the rat striatum following cocaine and amphetamine administration. Since both psychostimulants elicit memory-related effects, localization of cocaine- and amphetamine-regulated transcript peptide in the hippocampal formation may have functional importance. Previous studies demonstrated different cellular localizations of cocaine- and amphetamine-regulated transcript peptide in humans and in rodents. Mossy cells were cocaine- and amphetamine-regulated transcript-positive in the human dentate gyrus, whereas granule cells contained this peptide in the rat. In the present study, the localization of cocaine- and amphetamine-regulated transcript peptide was examined using immunohistochemistry in the hippocampal formation of the rhesus monkey (Macaca mulatta), the common marmoset monkey (Callithrix jacchus) and in the tree shrew (Tupaia belangeri). In these species principal neurons of the hippocampal formation were cocaine- and amphetamine-regulated transcript-immunoreactive. In both monkeys and tree shrews, mossy cells of the hilus were cocaine- and amphetamine-regulated transcript-positive whereas granule cells of the dentate gyrus were cocaine- and amphetamine-regulated transcript-negative. The dense cocaine- and amphetamine-regulated transcript-immunoreactive axonal plexus of the associational pathway outlined the inner one-third of the dentate molecular layer. In the hippocampus of the tree shrew and marmoset monkey, a subset of CA3 pyramidal cells were cocaine- and amphetamine-regulated transcript-immunoreactive. In the marmoset monkey, cocaine- and amphetamine-regulated transcript labeling was found only in layer V pyramidal cells of the entorhinal cortex, while in the rhesus monkey, pyramidal cells of layers II and III were cocaine- and amphetamine-regulated transcript-immunopositive. Our results show that cocaine- and amphetamine-regulated transcript positive neurons in the dentate gyrus of non-human primates are similar to that of the human. Furthermore, in the hippocampal formation of the tree shrew similar cocaine- and amphetamine-regulated transcript-immunoreactive cell-types were observed as in monkeys, supporting their evolutionary relationship with primates. Mossy cells and granule cells are members of a mutual excitatory intrahippocampal circuitry, therefore cocaine- and amphetamine-regulated transcript-immunoreactivity of these neurons in primates and rodents suggests that psychostimulants cocaine and amphetamine may induce memory-related effects at different points of the same excitatory circuitry in the hippocampal formation.


Cytotoxicity of electrophilic iron(II)-clathrochelates in human promyelocytic leukemia cell line.
We observed that electrophilic iron(II)-clathrochelates exhibit significant cytotoxicity in human promyelocytic leukemia cells (IC50=6.5±4.6μM), which correlates with the enhancement of intracellular oxidative stress (17-fold increase with respect to the cells treated with the solvent only). Based on in vitro studies we suggested that this effect is caused by alkylation of glutathione leading to inhibition of the cellular antioxidative system and by catalytic generation of reactive oxygen species by products of the alkylation reaction.


Effect of novel synthetic evodiamine analogue on sexual behavior in male rats.
Currently phosphodiestrase5 (PDE5) inhibitors are the first-line treatment for erectile dysfunction. Drugs such as sildenafil and tadalafil are available as PDE5 inhibitors which are potent and reversible but lack selectivity with side effects such as headache, facial flushing, dyspepsia, and visual disturbances. We herein report for the first time novel condensed thienopyrimidines as evodiamine analogue and their effect on sexual behavior in male rats hitherto unreported. Novel synthetic evodiamine significantly showed improvement in male rat copulatory behavior. The test compound MKAC9 could be of promising importance in the treatment of sexual disorders like desire disorder or erectile dysfunction.FigureEvodiamine analogue on sexual behavior in male rats.


[Study of blood viscosity and fibrinogen as a guide to the therapy of atherosclerotic peripheral arteriopathy].
Of the many blood parameters now regarded as useful in the study of arteriopathy particular importance is attached to viscosity and fibrinogen levels. Changes in viscosity lead to changes in flow and the supply of O2 to the tissues, while fibrinogen alterations frequently accompany or precede atherosclerosis, since they both encourage the intravascular deposition of fibrin and help to modify viscosity. The pharmacological correction of such changes is discussed.


Exploring the Pattern of Metabolic Alterations Causing Energy Imbalance via PPARα Dysregulation in Cardiac Muscle During Doxorubicin Treatment.
Cardiotoxicity by anthracycline antineoplastic drug doxorubicin is one of the systemic toxicity of the cardiovascular system. The mechanism responsible for doxorubicin cardiotoxicity and lipid metabolism remains elusive. The current study tested the hypotheses that the role of peroxisome proliferator-activated receptor α (PPARα) in the progress of doxorubicin-induced cardiomyopathy and its mechanism behind lipid metabolism. In the present study, male rats were subjected to intraperitoneal injection (5-week period) of doxorubicin with different dosages such as low dosage (1.5 mg/kg body weight) and high dosage (15 mg/kg body weight) to induce doxorubicin cardiomyopathy. Myocardial PPARα was impaired in both low dosage and high dosage of doxorubicin-treated rats in a dose-dependent manner. The attenuated level of PPARα impairs the expression of the genes involved in mitochondrial transporter, fatty acid transportation, lipolysis, lipid metabolism, and fatty acid oxidation. Moreover, it disturbs the reverse triacylglycerol transporter apolipoprotein B-100 (APOB) in the myocardium. Doxorubicin elevates the circulatory lipid profile and glucose. Further aggravated lipid profile in circulation impedes the metabolism of lipid in cardiac tissue, which causes a lipotoxic condition in the heart and subsequently associated disease for the period of doxorubicin treatment. Elevated lipids in the circulation translocate into the heart dysregulates lipid metabolism in the heart, which causes augmented oxidative stress and necro-apoptosis and mediates lipotoxic conditions. This finding determines the mechanistic role of doxorubicin-disturbed lipid metabolism via PPARα, which leads to cardiac dysfunction.


Hydroxychloroquine-associated, photo-induced toxic epidermal necrolysis.
A patient developed toxic epidermal necrolysis (TEN), which was triggered by sun exposure while the patient was on long-term hydroxychloroquine. Phototoxic and photoallergic reactions are known to occur with hydroxychloroquine, but, to our knowledge, this is the first reported case of photo-induced TEN associated with the drug.


Inferior alveolar nerve function after sagittal split osteotomy of the mandible: correlation with degree of intraoperative nerve encounter and other variables in 496 operations.
Dysfunction of the inferior alveolar nerve indicated by various degrees of numbness of the lower lip and chin is one of the few drawbacks of sagittal split osteotomy (SSO) of the mandible. Although it has been recorded throughout the history of this technique its true aetiology is poorly understood. In this study of 496 SSOs, we have evaluated possible correlations between neurosensory dysfunction and several variables that have been implicated, such as the age of the patient, mandibular movement, type of split technique and osteosynthesis, degree of intraoperative nerve encounter, and surgical skill. Nerve dysfunction developed after 200/496 SSOs (40%). The patient's age had a significant influence on the recovery of the neurosensory function. Intraoperative nerve encounter such as nerve manipulation correlated with dysfunction to a much lesser degree than expected. Surgical skill seemed to influence the recovery of neurosensory function after SSO, which is often referred to as a technique-sensitive procedure. We suggest that the dissection of the soft tissue on the medial aspect of the mandibular ramus might be partly responsible for nerve dysfunction of the lower lip and chin after SSO of the mandible.


Inhibition of lipid peroxidation by prostaglandin oligomeric derivatives.
The inhibition of lipid peroxidation by oligomeric derivatives synthesized from prostaglandin E1 (PGE1) and PGB2 was studied using two rat models. In an in vitro model, the brain was exposed to decapitation-ischemia, the cortex was removed and homogenized, and the formation of thiobarbituric acid reactive substances (TBAR) was measured after exposing the homogenate to in vitro reoxygenation either in the presence or absence of oligomers. It was found that these oligomers could inhibit lipid peroxidation, and that their activities were higher than that of superoxide dismutase (SOD). In an in vivo administration model, either the oligomer or the vehicle was injected i.p. 30 min before decapitation. The brain was exposed to decapitation-ischemia, the cortex was homogenized and exposed to 'in vitro' reoxygenation, after which TBAR value was determined. Ester-type compounds had a greater activity than free-acid type compounds in inhibiting lipid peroxidation. A possible mechanism of the protective effect of these oligomers in ischemia/reperfusion injury may be to scavenge oxygen free radicals.


Photo-polymerizable ferrous sulfate liposomes as vehicles for iron fortification of food.
The development of photo-polymerizable ferrous sulfate liposomes has been perused for iron fortification of food. Iron fortification is accompanied by several limitations that reduce its quality as it provokes sensory problems due to the oxidation of Fe2+ into Fe3+. To overcome such undesirable effects and improve the bioavailability of iron, photo-polymerizable 1,2-bis(10,12-tricosadiynoyl)-sn-glycero-3-phosphocholine (DC8,9PC) phospholipids have been employed as iron delivery system. DC8,9PC possesses diacetylene groups that undergo intramolecular cross-linking following UV treatment, resulting in enhanced stability, high encapsulation efficiency (~91%) without inducing sensory changes during milk fortification, along with less oxidation and con-tent leakage after long term storage in ethanol. Furthermore, polymeric Fe-DC8,9PC liposomes present high in vivo iron bioavailability in hemoglobin (Hb)-repletion study in rats, with no indications of toxicity examined by hematoxylin-eosin test. This methodology offers great promise for a simple, reliable, cost-effective, and robust platform for treating iron deficiency anemia, seeking to bring its application toward market.


Statistical modelling of roadside and urban background ultrafine and accumulation mode particle number concentrations using generalized additive models.
Quantification of the exposure of urban residents to ultrafine particle number concentrations (UFP) is challenging due to its high spatial and temporal variability. Hence, statistical models, e.g. generalized additive models (GAM), may be used to estimate time series or spatial characteristics of UFP. The GAM approach allows the representation of non-linear relations of a response variable with explanatory variables without the need to pre-define model functions. Up to now, GAMs were usually fitted to UFP data from a single site or from mobile measurement campaigns with limited temporal coverage. In this study, GAMs were used to determine UFP, accumulation mode particle (ACC) and total number concentration (TNC) at five urban sites in the cities of Leipzig and Dresden, Germany for the period 2011-2013. As explanatory variables, reanalysis data sets of meteorological quantities, urban geometry and traffic volume data were evaluated. Variables causing concurvity, which is the equivalent to collinearity in non-linear model approaches, were neglected to guarantee the interpretability of the final models. The models were then validated in a ten-fold cross-validation approach. The final models contained smooth functions for the building surface fraction, planetary boundary layer height, traffic volume, air temperature, wind direction, atmospheric pressure, relative humidity, global radiation and precipitation. Adjusted coefficients of determination (R2adj.) for the final models were R2adj. = 0.44 for UFP, R2adj. = 0.51 for ACC and R2adj. = 0.48 for TNC. Coefficients of determination of the cross-validation were in a similar range (0.44 for UFP, 0.51 for ACC, 0.49 for TNC). Finally, our study shows that GAMs are able to represent important processes that contribute to the particle number concentration from the smooth functions, i.e. emission, dilution, nucleation, deposition and long-range transport.


A circulating sodium transport inhibitor and essential hypertension.
Evidence for the hypothesis that the rise in peripheral resistance that occurs in inherited hypertension is due largely to the observed rise in the circulating level of a sodium transport inhibitor is reviewed. A sequence of events is proposed linking a postulated genetic defect in the kidney's ability to excrete sodium, salt intake, and the rise in the circulating concentration of a sodium transport inhibitor and thereby the rise in peripheral resistance in essential hypertension.


Surface dual redox cycles of Mn(III)/Mn(IV) and Cu(I)/Cu(II) for heterogeneous peroxymonosulfate activation to degrade diclofenac: Performance, mechanism and toxicity assessment.
Advanced oxidation processes (AOPs) based on heterogeneous catalytic activated peroxymonosulfate (PMS) have been becoming alternatives to conventional wastewater treatment technologies to directly degrade chemical contaminants. To build dual/multi redox cycles of different metal ions may be an effective means for better PMS activation. Herein, this study designed Mn3O4/CuBi2O4 with dual redox cycles of Mn(III)/Mn(IV) and Cu(I)/Cu(II) to activate PMS for efficiently decomposing and mineralizing diclofenac sodium (DCF). Under optimal reaction conditions, DCF (50 mg/L) was degraded totally within 10 min, and TOC removal rate reached up to 74.3%. The possible mechanism of PMS activation by Mn3O4/CuBi2O4 was proposed, wherein dual redox cycles of Mn(III)/Mn(IV) and Cu(I)/Cu(II) on Mn3O4/CuBi2O4 effectively facilitated PMS activation to generate ·O2-, 1O2, SO4·- and ·OH, which was responsible for DCF degradation. Moreover, combined with degraded products detected by high resolution liquid chromatography coupled to mass spectrometry and corresponding toxic assessment results, the possible degradation pathways of DCF were proposed and the relative toxicity of degraded products was evaluated. This work may be useful for developing stronger heterogeneous activators of PMS to construct more efficient AOPs for purifying wastewater.


Epidemiology of pneumonia in rural underfives.
Rural underfives (5335) were followed for a period of one year from January to December 1987 for acute respiratory infections (ARI). Those affected with pneumonias were studied in detail to know the epidemiology. The children reported an attack rate of 0.29/child/year. Severe cases constitute 0.5%. Infants had higher attack rate (0.59/child/year), 47.7% of episodes occurred in infants and 87.7% occurred in children below 3 years. Males had a higher attack rate (0.32 vs 0.27). Mean duration of attack was 5.45 +/- 1.95 days. The attack rate was higher in winter. The case fatality rate was 1.26%. It was highest in neonates (10.7%) Females had higher case fatality rate than males (1.5% Vs 1.1%). Mortality was very high in severe cases. Case fatalities were high in seasons when the climatic change occurred, the highest being in autumn.


Discriminatory proteomic biomarker analysis identifies free hemoglobin in the cerebrospinal fluid of women with severe preeclampsia.
Preeclampsia is an idiopathic multisystem disorder specific to human pregnancy. This study used proteomic analysis of cerebrospinal fluid (CSF) to identify protein biomarkers characteristic of preeclampsia and related to its severity. CSF was collected from women diagnosed clinically with severe preeclampsia (sPE: n = 7), mild preeclampsia (mPE: n = 8), and normotensive controls (CRL: n = 8). Samples were subjected to proteomic analysis using surface-enhanced laser desorption/ionization time-of-flight (SELDI-TOF) mass spectroscopy. A discriminative proteomic biomarker profile was extracted by applying Mass Restricted analysis, and a Preeclampsia Proteomic Biomarker (PPB) score developed based on the presence or absence of four discriminatory protein peaks in individual CSF SELDI tracings. In-gel tryptic digests, Western blot analysis, on-chip immunoassays, ELISA, and spectral analysis were used to identify the biomarkers composing the PPB score. PPB score distinguished patients with a clinical diagnosis of sPE from mPE and CRLs. (PPB median [range]: sPE: 4 [0-4] vs mPE: 1 [0-1] vs CRL: 0 [0-0]; P < 0.001). PPB scores were unaffected by parity, magnesium seizure prophylaxis, CSF leukocyte counts, and total protein content. Proteomic identification techniques matched the discriminatory protein peaks to the alpha- and beta-hemoglobin chains. ELISA confirmed that women diagnosed clinically with sPE had significantly higher CSF hemoglobin concentrations than women with mPE or CRL (median [range]: sPE: 6.6 [0.0-10.3] microg/mL vs mPE: 0 [0-1.3] microg/mL vs CRL: 0 [0-0] microg/mL; P < 0.001). Proteomic analysis of CSF can accurately distinguish sPE from both mPE and CRL. Patients with sPE have nanomolar amounts of free hemoglobin in their CSF. Further studies are needed to confirm these observations and determine their physiologic implications.


Molecular Evaluation of HIF-1α Gene Variation and Determination of Its Frequency and Association with Breast Cancer Susceptibility in Saudi Arabia.
Hypoxia-inducible factor 1 (HIF-1α) is responsible in regulating oxygen homeostasis in tissues and is a central effector of the hypoxic response besides its protein overexpression has been shown to have prognostic relevance in several cancers including breast cancer. Several reports indicated that HIF-1α gene variation C1772T (Pro582Ser) is associated with increased breast susceptibility but results remained controversial. Therefore, we performed the molecular evaluation of HIF-1α gene variation and determined its frequency and association with Breast Cancer susceptibility in Saudi Arabia. This study was conducted on histologically confirmed Breast cancer patients and gender matched healthy women. HIF-1α C1772T (Pro582Ser) genotyping was done by Amplification refractory mutation system PCR method. The HIF-1α gene genotypes were correlated with different clinicopathological characteristics of breast cancer patients. A significant difference was observed in genotype distribution of HIF-1α gene variation C1772T (Pro582Ser) between breast cancer cases and gender matched healthy controls (P=0.010). Our findings showed that the HIF- 1α variant was associated with an increased risk of Breast cancer for HIF-1α CC vs CT genotype OR = 2.20, 95% CI = (1.28 -3.77), P = 0.004) in codominant inheritance model. The significant association was reported for HIF1A for genotypes CC vs (CT+ TT) OR = 1.98, 95% CI = (1.17-3.34), P = 0.010) in dominant inheritance model tested. In case of recessive inheritance model, a non-significant association of HIF-1 alpha gene variants was reported for (CC+ CT) vs TT) OR = 1.03, 95% CI = (0. 064-16.79), P = 0.97). During the allelic comparison, a non-significant association was reported between A vs C allele among Breast cancer patients. A significant association of HIF- 1α polymorphism was reported with stage as well as distant metastasis of the disease. A significant difference was observed in genotype distribution of HIF-1α gene variation C1772T (Pro>Ser) between breast cancer cases and gene matched healthy controls (P=0.010). HIF-1α- CT heterozygosity and CC genotype increased the susceptibility .The HIF-1α polymorphism was reported to be significantly associated with the distant metastasis of Breast cancer. Further studies with larger data set and well-designed models are required to validate our findings.


Eye movements in patients with absent voluntary horizontal gaze.
Despite the complete absence of horizontal saccades, two patients with pontine gliomas had horizontal reflex eye movements within a range of +/- 20 degrees. The gain (peak eye velocity/peak stimulus velocity) and phase of the vestibulo-ocular reflex were normal, but the optokinetic gain was decreased. The latency, accuracy, and peak velocity of vertical saccades were normal. Apparently the voluntary gaze centers in the pontine reticular formation are not crucial for generating horizontal vestibular or vertical saccadic eye movements.


Insulin receptors in the brain: structural and physiological characterization.
The present study was conducted to characterize insulin receptors and to determine the effects of insulin in synaptosomes prepared from adult rat brains. Binding of 125I-insulin to synaptosome insulin receptors was highly specific and time dependent: equilibrium binding was obtained within 60 minutes, and a t1/2 of dissociation of 26 minutes. Cross-linking of 125I-insulin to its receptor followed by SDS-PAGE demonstrated that the apparent molecular weight of the alpha subunit of the receptor was 122,000 compared with 134,000 for the liver insulin receptor. In addition, insulin stimulated the dose-dependent phosphorylation of exogenous tyrosine containing substrate and a 95,000 MW plasma membrane associated protein, in a lectin-purified insulin receptor preparation. The membrane associated protein was determined to be the beta subunit of the insulin receptor. Incubation of synaptosomes with insulin caused a dose-dependent inhibition of specific sodium-sensitive [3H]norepinephrine uptake. Insulin inhibition of [3H]norepinephrine uptake was mediated by a decrease in active uptake sites without any effects in the Km, and was specific for insulin since related and unrelated peptides influenced the uptake in proportion to their structural similarity with insulin. These observations indicate that synaptosomes prepared from the adult rat brain possess specific insulin receptors and insulin has inhibitory effects on norepinephrine uptake in the preparation.


Upper thermal limits of the hearts of Arctic cod Boreogadus saida: adults compared with larvae.
Wild adult and reared larval Boreogadus saida were acclimated to 3·5° C before testing their cardiac response to acute warming. Heart rate transition temperatures during warming were similar for adult and larval hearts, except that the maximum temperature for heart rate was 3° C warmer for adults. Thus, in a rapidly warming Arctic Ocean, the upper temperature limit for larval rather than adult B. saida appears more likely to dictate the southern range of the species.


rac-5-(1-Methyl-eth-yl)-2-sulfanylidene-imidazolidin-4-one.
In the title compound, C6H10N2OS, the 2-sulfanylideneimidazolidin-4-one fragment is essentially planar (r.m.s. deviation = 0.0033 Å). In the crystal, one amino group is involved in N-H⋯O hydrogen bonding, which links pairs of mol-ecules into inversion dimers, while the other amino group generates weak N-H⋯S hydrogen bonds, which link these dimers into chains in [10-1]. The chains are further aggregated into layers parallel to the ac plane through weak C-H⋯O inter-actions.


MicroRNAs: potential diagnostic markers and therapeutic targets for EBV-associated nasopharyngeal carcinoma.
Nasopharyngeal carcinoma (NPC) is a highly malignant cancer with local invasion and early distant metastasis. NPC is highly prevalent in the Southern China and South-eastern Asia. The genetic susceptibility, endemic environment factors, and Epstein-Barr virus (EBV) infection are believed to be the major etiologic factors of NPC. Once metastasis occurs, the prognosis is very poor. It is urgently needed to develop biomarkers for early clinical diagnosis/prognosis, and novel effective therapies for nasopharyngeal carcinoma. In this paper, we systematically reviewed the current progress of miRNA studies in NPC. It has been shown that both host encoded miRNAs and EBV encoded miRNAs play key roles in almost all the steps of epithelia cell carcinogenesis, including epithelial-mesenchymal to stem-like transition, cell growth, migration, invasion, and tumorigenesis. More importantly, some miRNAs could be secreted out and play a role in the microenvironments. The level of sera miRNAs is correlated with the copy numbers of host miRNAs in tumor biopsies. Promising results of gene therapy have been also achieved by lentiviral delivered miRNAs. Taken together, cell free miRNAs would be potential biomarkers of early clinical diagnosis/prognosis; while some miRNAs could be further developed into therapeutic agents in the future.


Memory effect of spider major ampullate silk in loading-unloading cycles and the structural connotations.
Spider silk is repeatedly stretched while performing biological functions. There is a close relationship between the shape change of the fibre materials and their mechanical properties. However, the effect of the deformation and interval time on the structure and tensile behaviour properties of spider silk after repeatedly stretching by given strain value has been rarely reported. Here we found that major ampullate silk (MAS) can revert its tensile behaviour independent of its previous loading history via intervals of approximately 8 s to 5 min with constant and increased elongation, respectively, after being subjected to yield and hardening regions. The true stress-true strain curve beyond a given value of true strain is independent from the previous loading history of the sample. Even after longer intervals (≥1 h), MAS can reproduce the last tensile behaviour via one stretched. Despite recognizing the development of irreversible deformations in the material when tested in air, the reversible change in tensile behaviour outside the spider silk's elastic region has rarely been observed before. MAS has at least one proper ground state that allows it to present good shape and mechanical behaviour memory in terms of longitudinal stretching, functioning as a new strategy to achieve certain tensile properties. The analysis of the true stress-true strain curves was performed from a series of loading‒unloading tests to evaluate the evolution of those mechanical parameters with the cycle number. The elastic modulus measured in the loading steps increases monotonously with increasing values of true strain reached in the cycles. In contrast, a marginal variation is found in the values of the yield stress measured in the different cycles. The memory and variation in the mechanical behaviour and performance of MAS can be accounted for through the irreversible and reversible deformation micromechanisms and its combination in which the viscoelasticity of the material plays a leading role. These findings may be helpful to guide the biomimetic design of novel fibre materials such as spider silk gut via artificially stretching spider silk glands.


A rare combination of closed fracture of right talar body Sneppen 2 with associated medial malleolus: A case report.
Ankle fractures are common, with the incidence of up to 174 cases per 100,000 adults per year. The talus, the second largest tarsal bone, has distinctive imaging characteristics and injury patterns. The combination of talar body fracture in sagittal plane along with medial malleolus fracture is an unusual pattern of injury and rarely reported in the literature. We presented a case of nineteen-year old male with pain on the right ankle as a result from motorcycle accident twelve hours before admitted to Cipto Mangunkusumo Hospital. After the patient underwent close reduction and immobilization using back slab, patient then underwent open reduction surgery of the right ankle then fixated using plate and screw combined by tension band wiring (TBW) at the right medial malleolus. Talar and Medial malleolus fracture was initially evaluated by physical examination and radiographic imaging and classified by Sneppen classification. After the diagnosis was established the patient underwent ORIF plate, screw and tension band wiring. In post-operative x-ray examination, there was a significant improvement in talocrural angle and medial clear space but same result in tibiofibular overlap and tibiofibular. Open reduction and internal fixation (ORIF) plate and screw combined by tension band wiring (TBW) is the right surgical treatment for patient with closed fracture of right talar body Sneppen 2 with associated medial malleolus fracture. The outcome of the treatment is excellent because the patient presented was able to fully recover to normal activity.


Pathogenic mechanisms and regulatory factors involved in alcoholic liver disease.
Alcoholism is a widespread and damaging behaviour of people throughout the world. Long-term alcohol consumption has resulted in alcoholic liver disease (ALD) being the leading cause of chronic liver disease. Many metabolic enzymes, including alcohol dehydrogenases such as ADH, CYP2E1, and CATacetaldehyde dehydrogenases ALDHsand nonoxidative metabolizing enzymes such as SULT, UGT, and FAEES, are involved in the metabolism of ethanol, the main component in alcoholic beverages. Ethanol consumption changes the functional or expression profiles of various regulatory factors, such as kinases, transcription factors, and microRNAs. Therefore, the underlying mechanisms of ALD are complex, involving inflammation, mitochondrial damage, endoplasmic reticulum stress, nitrification, and oxidative stress. Moreover, recent evidence has demonstrated that the gut-liver axis plays a critical role in ALD pathogenesis. For example, ethanol damages the intestinal barrier, resulting in the release of endotoxins and alterations in intestinal flora content and bile acid metabolism. However, ALD therapies show low effectiveness. Therefore, this review summarizes ethanol metabolism pathways and highly influential pathogenic mechanisms and regulatory factors involved in ALD pathology with the aim of new therapeutic insights.


Competitive MS binding assays for dopamine D2 receptors employing spiperone as a native marker.
A competitive MS binding assay employing spiperone as a native marker and a porcine striatal membrane fraction as a source for dopamine D2 receptors in a nonvolatile buffer has been established. Binding of the test compounds to the target was monitored by mass-spectrometric quantification of the nonbound marker, spiperone, in the supernatant of the binding samples obtained by centrifugation. A solid-phase extraction procedure was used for separating spiperone from ESI-MS-incompatible supernatant matrix components. Subsequently, the marker was reliably quantified by LC-ESI-MS-MS by using haloperidol as an internal standard. The affinities of the test compounds, the dopamine receptor antagonists (+)-butaclamol, chlorpromazine and (S)-sulpiride obtained from the competitive MS binding assay were verified by corresponding radioligand binding experiments with [3H]spiperone. The results of this study demonstrate that competitive MS binding assays represent a universally applicable alternative to conventional radioligand binding assays.


Complexity of the microRNA transcriptome of cow milk and milk-derived extracellular vesicles isolated via differential ultracentrifugation.
MicroRNAs (miRNAs) are small gene-regulatory noncoding RNA that are highly enriched in cow milk. They are encapsulated in different extracellular vesicle (EV) subsets that protect them from the extracellular milieu and the harsh conditions of the gastrointestinal tract during digestion. Here, we isolated pellets enriched in 4 different EV subsets, via differential ultracentrifugation of commercial cow milk: 12,000 × g (P12K), 35,000 × g (P35K), 70,000 × g (P70K), and 100,000 × g (P100K). Small RNA sequencing (sRNA-Seq) analyses revealed an unprecedented level of diversity in the complete miRNA repertoire and features of unfractionated cow milk and derived EV subsets. Although 5 miRNA sequences represented more than 50% of all miRNAs, milk EV exhibited heterogeneous content of miRNAs and isomeric variants (termed isomiR): P100K EV were enriched in reference miRNA sequences, and P12K and P35K EV in related isomiR. Incubation of milk EV with human cultured HeLa cells led to cellular enrichment in miRNA miR-223, which was concomitant with decreased expression of a reporter gene placed under the control of miR-223, thereby demonstrating the functionality of miR-223. These results suggest that cow milk EV may transfer their miRNAs to human cells and regulate recipient cell gene expression programming in a manner as complex as that of their miRNA transcriptome. The biological activity and relevance of the different milk EV subsets and bioactive mediators, including small noncoding RNA, in health and disease, warrants further investigation.


BK virus regulatory region sequence deletions in a case of human polyomavirus associated nephropathy (PVAN) after kidney transplantation.
The pathogenesis of polyomavirus-associated interstitial nephropathy (PVAN) in kidney transplant recipients is likely to depend upon multiple risk factors, which may include viral characteristics. We report a case of interstitial nephropathy in a young kidney allograft recipient, associated with a BK virus (BKV) strain with a rearranged transcription control region (TCR). BKV strains with deletions and nucleotide substitutions in the TCR were present in a kidney biopsy and urine samples. After retransplantation, following loss of renal function, a BKV strain with an archetypal TCR was detected in association with asymptomatic reactivation.


Different Pathways to the Lysosome: Sorting out Alternatives.
Considerable research supports a model in which hydrolytic enzymes of mammalian lysosomes are sorted to their destinations in a receptor-dependent mechanism. The ligand for the mammalian sorting receptors is mannose 6-phosphate (M6P). Two M6P receptors have been defined in mammals. Here, we review the foundational evidence supporting this mechanism and highlight the remaining gaps in our understanding of the mammalian mechanism, including evidence for M6P-independent sorting, and its relevance to lysosomal enzyme sorting in metazoa.


There's more in a number than you think: new validity data for the Global Assessment Scale.
Holcomb and Otto (1988) recently suggested that the widely used Global Assessment Scale may have ". . . questionable validity as an index of psychopathology." The present paper points out procedural and conceptual problems in their study and presents extensive evidence supporting the concurrent and predictive validity of the scale.


Outcome after ipsilateral breast tumor recurrence in patients who receive accelerated partial breast irradiation.
The objective of this study was to examine clinical outcomes and patterns of failure in patients with early stage breast cancer who developed an ipsilateral breast tumor recurrence (IBTR) after breast-conserving therapy (BCT) using accelerated partial breast irradiation (APBI). In total, 1440 patients (1449 tumors) with early stage breast cancer who underwent BCT were treated with the MammoSite device to deliver APBI (34 Gray [Gy] in 3.4-Gy fractions). One thousand two hundred fifty-five patients (87%) had invasive breast cancer (IBC) (median tumor size, 10 mm), and 194 patients (13%) had ductal carcinoma in situ (DCIS) (median tumor size, 8 mm). The median follow-up was 60 months. Fifty patients (3.5%) developed an IBTR for a 5-year actuarial rate of 3.61% (3.65% for IBC and 3.36% for DCIS). It was determined that 36 recurrences (72%) represented new primary cancers, and 14 recurrences (28%) represented recurrences of the index lesion. Of the 32 recurrences with known histology, 78% were IBC, and 22% were DCIS. After IBTR, 28 of 38 patients (74%) underwent salvage mastectomy, and 9 of 38 patients (26%) had a second attempt at BCT. Adjuvant therapies included tamoxifen in 8 patients (16%) and systemic chemotherapy in 6 patients (12%). The 3-year rates of disease-free survival, cause-specific survival, and overall survival after IBTR were 58.7%, 92.1%, and 80.5%, respectively. With 5 years of follow-up, APBI produced clinical outcomes and patterns of failure comparable to those achieved with whole breast irradiation. Patients who developed an IBTR after APBI had excellent 3-year survival outcomes after salvage treatments.


Epac and phospholipase Cepsilon regulate Ca2+ release in the heart by activation of protein kinase Cepsilon and calcium-calmodulin kinase II.
Recently, we identified a novel signaling pathway involving Epac, Rap, and phospholipase C (PLC)epsilon that plays a critical role in maximal beta-adrenergic receptor (betaAR) stimulation of Ca2+-induced Ca2+ release (CICR) in cardiac myocytes. Here we demonstrate that PLCepsilon phosphatidylinositol 4,5-bisphosphate hydrolytic activity and PLCepsilon-stimulated Rap1 GEF activity are both required for PLCepsilon-mediated enhancement of sarcoplasmic reticulum Ca2+ release and that PLCepsilon significantly enhances Rap activation in response to betaAR stimulation in the heart. Downstream of PLCepsilon hydrolytic activity, pharmacological inhibition of PKC significantly inhibited both betaAR- and Epac-stimulated increases in CICR in PLCepsilon+/+ myocytes but had no effect in PLCepsilon-/- myocytes. betaAR and Epac activation caused membrane translocation of PKCepsilon in PLCepsilon+/+ but not PLCepsilon-/- myocytes and small interfering RNA-mediated PKCepsilon knockdown significantly inhibited both betaAR and Epac-mediated CICR enhancement. Further downstream, the Ca2+/calmodulin-dependent protein kinase II (CamKII) inhibitor, KN93, inhibited betaAR- and Epac-mediated CICR in PLCepsilon+/+ but not PLCepsilon-/- myocytes. Epac activation increased CamKII Thr286 phosphorylation and enhanced phosphorylation at CamKII phosphorylation sites on the ryanodine receptor (RyR2) (Ser2815) and phospholamban (Thr17) in a PKC-dependent manner. Perforated patch clamp experiments revealed that basal and betaAR-stimulated peak L-type current density are similar in PLCepsilon+/+ and PLCepsilon-/- myocytes suggesting that control of sarcoplasmic reticulum Ca2+ release, rather than Ca2+ influx through L-type Ca2+ channels, is the target of regulation of a novel signal transduction pathway involving sequential activation of Epac, PLCepsilon, PKCepsilon, and CamKII downstream of betaAR activation.