[Calcinosis of the scrotum. Report of a case associated with epidermoid cysts].
Clinical and histological features of a case of the so-called idiopathic calcinosis of the scrotum in a 22 year-old otherwise healthy man are presented. In a first biopsy we did not observe any evidence of squamous epithelial lining around calcified masses, but in a second one we observed some intact epidermoid cysts and granulomas of foreign-body type around keratinous material. These findings suggest that scrotal calcinosis is not idiopathic, but rather the end stage of dystrophic calcification of scrotal epidermoid cysts.


Dual-wavelength excitation for fluorescence-based quantification of zinc protoporphyrin IX and protoporphyrin IX in whole blood.
Quantification of erythrocyte zinc protoporphyrin IX (ZnPP) and protoporphyrin IX (PPIX), individually or jointly, is useful for the diagnostic evaluation of iron deficiency, iron-restricted erythropoiesis, lead exposure, and porphyrias. A method for simultaneous quantification of ZnPP and PPIX in unwashed blood samples is described, using dual-wavelength excitation to effectively eliminate background fluorescence from other blood constituents. In blood samples from 35 subjects, the results of the dual-wavelength excitation method and a reference high performance liquid chromatography (HPLC) assay were closely correlated both for ZnPP (rs = 0.943, p < 0.0001; range 37-689 μmol ZnPP/mol heme, 84-1238 nmol/L) and for PPIX (rs = 0.959, p < 0.0001; range 42-4212 μmol PPIX/mol heme, 93-5394 nmol/L). In addition, for ZnPP, the proposed method is compared with conventional single-wavelength excitation and with commercial front-face fluorimetry of washed erythrocytes and whole blood. We hypothesize that dual-wavelength excitation fluorimetry will provide a new approach to the suppression of background fluorescence in blood and tissue measurements of ZnPP and PPIX.


Hepatic insulin resistance directly promotes formation of cholesterol gallstones.
Despite the well-documented association between gallstones and the metabolic syndrome, the mechanistic links between these two disorders remain unknown. Here we show that mice solely with hepatic insulin resistance, created by liver-specific disruption of the insulin receptor (LIRKO mice) are markedly predisposed toward cholesterol gallstone formation due to at least two distinct mechanisms. Disinhibition of the forkhead transcription factor FoxO1, increases expression of the biliary cholesterol transporters Abcg5 and Abcg8, resulting in an increase in biliary cholesterol secretion. Hepatic insulin resistance also decreases expression of the bile acid synthetic enzymes, particularly Cyp7b1, and produces partial resistance to the farnesoid X receptor, leading to a lithogenic bile salt profile. As a result, after twelve weeks on a lithogenic diet, all of the LIRKO mice develop gallstones. Thus, hepatic insulin resistance provides a crucial link between the metabolic syndrome and increased cholesterol gallstone susceptibility.


Suspected appendicitis in children: US and CT--a prospective randomized study.
To evaluate the accuracy of ultrasonography (US) and of abdominal computed tomography (CT) performed in addition to US in the diagnosis of childhood appendicitis. Six hundred children with suspected appendicitis were included in a prospective randomized study. After clinical examination, the patients were randomly assigned to undergo US only (283 patients) or US with abdominal CT (317 patients). Radiologic findings were correlated with surgical, histopathologic, and clinical follow-up findings. Two hundred forty-four (40.7%) of all patients had appendicitis. In the US only group, US had a sensitivity of 86%, specificity of 95%, positive predictive value of 91%, negative predictive value of 92%, and diagnostic accuracy of 92%. The combination of US and CT performed in the other group yielded a sensitivity of 99%, specificity of 89%, positive predictive value of 87%, negative predictive value of 99%, and diagnostic accuracy of 93%. Analysis of data on US performed in all 600 patients and on CT performed in 317 patients revealed, respectively, sensitivities of 80% and 97%, specificities of 94% and 93%, positive predictive values of 91% and 92%, negative predictive values of 88% and 98%, and diagnostic accuracies of 89% and 95%. The overall negative appendectomy rate was 3.7%; and the perforation rate, 21%. US is valuable in the diagnosis of appendicitis in children. In inconclusive cases, performing additional abdominal CT can improve diagnostic accuracy and thereby decrease the negative appendectomy rate without an increase in the perforation rate.


Prostratin and bortezomib are novel inducers of latent Kaposi's sarcoma-associated herpesvirus.
Kaposi's sarcoma-associated herpesvirus (KSHV) establishes latent infections in lymphocytes and endothelial cells, and latent infection is closely linked to tumorigenesis. As few viral markers are expressed during latency, compounds that can safely and efficiently increase lytic gene expression in vivo have been sought. We have found that the non-tumour-promoting phorbol ester prostratin and the proteasome inhibitor bortezomib induce immediate-early, early and late KSHV gene expression from two lymphoma cell lines in vitro. Their ability to induce lytic gene expression supports a role for phorbol-ester and proteasome-regulated signalling pathways in KSHV reactivation and prompts further investigation of prostratin and bortezomib as therapeutic agents for KSHV-associated malignancies.


Pulse Oximetry Waveform Represents an Earlier Ventricular Contraction in Relation to the Intraarterial Blood Pressure Tracing and Electrocardiogram on Multichannel Monitors: A Case Series.
Multichannel patient monitors apparently display the electrocardiogram, intra-arterial blood pressure, and pulse oximetry waveforms in synchrony during sinus cardiac rhythm. We recorded 20 events of a premature cardiac contraction on multichannel monitors from 10 patients. Interestingly, during premature cardiac contraction, the low-amplitude arterial trace was aligned with the premature electrocardiogram, but the low-amplitude pulse oximetry plethysmograph was aligned with the next normal electrocardiogram and arterial tracing. In conclusion, the pulse oximetry plethysmograph tracing is offset by 1 ventricular depolarization on the electrocardiogram (QRS) and the arterial tracing on multichannel monitors.


Lysosomal cystine export regulates mTORC1 signaling to guide kidney epithelial cell fate specialization.
Differentiation is critical for cell fate decisions, but the signals involved remain unclear. The kidney proximal tubule (PT) cells reabsorb disulphide-rich proteins through endocytosis, generating cystine via lysosomal proteolysis. Here we report that defective cystine mobilization from lysosomes through cystinosin (CTNS), which is mutated in cystinosis, diverts PT cells towards growth and proliferation, disrupting their functions. Mechanistically, cystine storage stimulates Ragulator-Rag GTPase-dependent recruitment of mechanistic target of rapamycin complex 1 (mTORC1) and its constitutive activation. Re-introduction of CTNS restores nutrient-dependent regulation of mTORC1 in knockout cells, whereas cell-permeant analogues of L-cystine, accumulating within lysosomes, render wild-type cells resistant to nutrient withdrawal. Therapeutic mTORC1 inhibition corrects lysosome and differentiation downstream of cystine storage, and phenotypes in preclinical models of cystinosis. Thus, cystine serves as a lysosomal signal that tailors mTORC1 and metabolism to direct epithelial cell fate decisions. These results identify mechanisms and therapeutic targets for dysregulated homeostasis in cystinosis.


[Efficacy of oxaliplatin-based chemotherapy in Stage IV colorectal cancer patients with peritoneal carcinomatosis].
This retrospective study evaluated the outcome of oxaliplatin-based chemotherapy for peritoneal metastasis from colorectal cancer. The study included 49 patients with peritoneal metastasis from colorectal cancer who underwent oxaliplatin-based chemotherapy between January 2006 and November 2012 and 25 patients who underwent systematic chemotherapy before oxaliplatin-based chemotherapy. Patient background characteristics, overall survival(l OS), and prognostic outcomes were examined. OS was significantly longer in patients treated with oxaliplatin -based chemotherapy than in those treated with prior chemotherapy( median, 20.5 months vs 11.7 months, p= 0.04). Multivariate analysis showed that treatment with oxaliplatin-based chemotherapy, age less than 70 years(p=0.03), and primary tumor resection( p=0.02) were significant independent factors affecting OS. Oxaliplatin-based chemotherapy improved the prognosis of patients with peritoneal metastasis from colorectal cancer. In the treatment of such patients, initiation of chemotherapy after primary tumor resection appears to be important, especially in patients aged <70 years.


"Renal dose" dopamine in surgical patients: dogma or science?
"Renal dose" dopamine is widely used in the perioperative period to provide renal protection. A comprehensive review of the literature was performed to determine whether dopamine does in fact confer protection on the kidneys of surgical patients. Studies in healthy animals and human volunteers reveal that dopamine causes diuresis and natriuresis, as well as some degree of renal vasodilatation. Studies of the perioperative use of dopamine fail to demonstrate any benefit of dopamine in preventing renal failure. Studies in congestive heart failure, critical illness, and sepsis also fail to show any benefit of dopamine other than diuresis. Further, dopamine administration is not completely without risk, because of dopamine's catecholamine and neuroendocrine functions. Routine use of prophylactic "renal dose" dopamine in surgical patients is not recommended.


[Theoretical and applied aspects of methanogenesis].
The basic aspects of organic matter breakdown through methanogenesis are dealt with. The process is referred to as a method of obtaining energy-- biogas --and a way to solve a number of problems associated with the protection of the environment from waste products of the agricultural industry. Both traditional and modern technical means and basic parameters of the technology of biogas production are presented. Stated are also the important practical aspects of the biochemistry and kinetics of the process.


The relation between plasminogen activator inhibitor activity and disease activation in acute myeloblastic leukaemia patients.
Coagulation and fibrinolytic abnormalities are common in patients with acute myeloblastic leukaemia (AML) like other forms of leukaemias. In this study, we investigated if total plasminogen activator inhibitor (PAI) activity, which is believed to increase in initial diagnosis and relapse in AML patients could be accepted as a relapse criterion or not. Total of 34 AML patients and 18 healthy volunteers were included in this study. The patients' diagnosis were based on clinical criteria as well as morphological, cytochemical, immunuphenotypic examinations of peripheral blood and bone marrow specimens. Total PAI activity was measured with Dade Behring Bericrom PAI reagent in BCS system. Total PAI activity was higher than 3.5 U/ml in 11 AML patients while it was normal (0.3-3.5 U/ml) in control group (P < 0.01). There was no significant difference in total PAI activity between AML subgroups (P > 0.05). We found significant difference in total PAI activity between patients who have active disease and remission. In conclusion, the total PAI activity could be accepted as a relapse and an initial diagnosis criterion of AML patients during follow up.


Early detection and successful treatment of cerebral venous thrombosis associated with minimal change nephrotic syndrome.
Although venous thrombosis is a major complication in nephrotic syndrome, cerebral venous thrombosis (CVT) is rarely reported. We describe a 29-year-old male with nephrotic syndrome who suddenly developed headache and nausea. Although computed tomography scan and magnetic resonance imaging detected no abnormal lesions, phase-contrast magnetic resonance venography (PC MRV) demonstrated extensive thrombosis of the superior sagittal sinus. After receiving systemic anticoagulant therapy and oral prednisolone, his neurological symptoms improved dramatically, and complete remission from nephrotic syndrome was achieved. Follow-up PC MRV demonstrated recanalization of the superior sagittal sinus, and renal biopsy confirmed the diagnosis of minimal change nephrotic syndrome. Although CVT is difficult to detect with conventional diagnostic methods, PC MRV may help the establishment of an early diagnosis and prompt treatment for a successful outcome.


Burkholderia cepacia Induced Occipital Subcutaneous Abscess and Fracture in a Brain-Dead Woman.
Burkholderia cepacia is an aerobic opportunistic Gram-negative pathogen that mainly infects immunodeficiency patients. However, soft tissue infections with fractures due to Burkholderia cepacia are rare with no cases reported. Here, we report a rare case of 3 years of brain death in a 43-year-old woman with Burkholderia cepacia resulting in an occipital subcutaneous abscess and fracture. Through the second-generation sequencing of the whole genome of this strain, it was found that there were no high virulence genes and virulence factors. The patient received targeted antibiotic therapy and showed improvement in clinical symptoms and radiological signs. Bone destruction because of Burkholderia cepacia is easily overlooked due to the lack of characteristic symptoms and limited clinical examination. This case reminds us that Burkholderia cepacia without major virulence factors could damage the bone in immune-compromised patients.


[Complex treatment of patients with pyo-necrotic complications of the neuropathic form of diabetic foot syndrome].
Of study is improving the results of treatment of patients with pyo-necrotic complications of diabetic foot syndrome by including the method of negative pressure wound treatment in the complex treatment program in combination with using of the combined antibacterial drug Cifran ST and immunocorrective therapy. The results of examination and treatment of 184 patients with pyo-necrotic complications of the neuropathic form of diabetic foot syndrome were analyzed. According to choice of treatment methods in the postoperative period all patients were divided into two groups. In 95 patients (group I), iodine-containing ointments based on polyethylene glycol were used for local treatment of purulent foot wounds and standard systemic antibacterial therapy was performed. In 89 patients (group II), negative pressure wound treatment (NPWT) was used to treat wounds in the postoperative period. In addition to standard parenteral antimicrobial therapy, these patients also received an oral combined antibacterial drug Cifran ST and immunocorrective cytokine therapy (Leukinferon). The analysis of the dynamics of the wound process was carried out based on the clinical picture and the results of cytological, bacteriological and immunological studies of the wound exudate. The presented strategy of complex treatment of pyo-necrotic complications of the neuropathic form of diabetic foot syndrome allowed group II patients to significantly reduce the degree of microbial contamination of wounds, to achieve a faster regression of the content of proinflammatory and inflammatory cytokines in the wound exudate, as well as to reduce the time of wound cleansing and the transition of the pyo-necrotic process to the reparative stage in comparison with group I patients. This allowed group II patients to reduce the time of plastic closure of the wound from 24.3±0.5 to 15.6±1.7 days, to avoid generalization of infection, death and high level amputation of the limb. At the same time, 11.6% of patients in group I had high level limb amputation due to generalization of infection. The mortality rate in group I was 5.3%. Adding of vacuum therapy of wounds, systemic antimicrobial therapy using the combined antibacterial drug Cifran ST and immunocorrective cytokine therapy in the complex treatment program for patients with neuropathic form of diabetic foot syndrome after radical surgical treatment of the pyo-necrotic lesion allows reducing the time of wound cleansing and the transition of the pyo-necrotic process to the reparative stage. On the other hand, this makes it possible for this category of patients to perform plastic closure of the wound at an earlier date, avoid generalization of infection and high level amputation of the limb.


Calcium transients and calcium signalling during early neurogenesis in the amphibian embryo Xenopus laevis.
Development of the vertebrate embryonic nervous system is characterized by a cascade of signalling events. In Xenopus, the initial step in this cascade results from signals emanating from the dorsal mesoderm that divert the fate of the ectoderm from an epidermal to a neural lineage. These signals include extracellular antagonists of the bone morphogenetic protein (BMP). Experiments performed with isolated ectoderm suggest that epidermis is induced by BMP, whereas neural fates arise by default following BMP inhibition; however, we show that this mechanism is not sufficient for neural determination. Ca2+ imaging of intact Xenopus embryos reveals patterns of Ca2+ transients in the dorsal ectoderm but not in the ventral ectoderm. These increases in intracellular calcium concentration ([Ca2+](i)), which occur via the activation of dihydropyridine (DHP)-sensitive Ca2+ channels, are necessary and sufficient to orientate the ectodermal cells toward a neural fate. On the one hand, the treatments that antagonize the increase in [Ca2+](i), inhibit neuralization, while on the other hand, an artificial increase in [Ca2+](i), whatever its origin, neuralizes the ectoderm. Using these properties, we have constructed a subtractive cDNA library between untreated ectoderm and caffeine-treated ectoderm. The caffeine stimulates an increase in [Ca2+](i) and thus orientates the cells towards the neural pathway. We have identified early Ca2+ target genes expressed in neural territories. One of these genes, an arginine methyl transferase, controls the expression of the early proneural gene, Zic3. Here, we discuss an alternative model where Ca2+ plays a central regulatory role in early neurogenesis. This model integrates the activation of a Ca2+ -dependent signalling pathway due to an influx of Ca2+ through DHP-Ca2+ channels. While Ca2+ is required for neural determination, epidermal determination occurs when Ca2+ -dependent signalling pathways are inactive.


Evidence for Narrow Transfer after Short-Term Cognitive Training in Older Adults.
The degree to which "brain training" can improve general cognition, resulting in improved performance on tasks dissimilar from the trained tasks (transfer of training), is a controversial topic. Here, we tested the degree to which cognitive training, in the form of gamified training activities that have demonstrated some degree of success in the past, might result in broad transfer. Sixty older adults were randomly assigned to a gamified cognitive training intervention or to an active control condition that involved playing word and number puzzle games. Participants were provided with tablet computers and asked to engage in their assigned training for 30 45-min training sessions over the course of 1 month. Although intervention adherence was acceptable, little evidence for transfer was observed except for the performance of one task that most resembled the gamified cognitive training: There was a trend for greater improvement on a version of the corsi block tapping task for the cognitive training group relative to the control group. This task was very similar to one of the training games. Results suggest that participants were learning specific skills and strategies from game training that influenced their performance on a similar task. However, even this near-transfer effect was weak. Although the results were not positive with respect to broad transfer of training, longer duration studies with larger samples and the addition of a retention period are necessary before the benefit of this specific intervention can be ruled out.


Elevated pretreatment serum levels of interferon-inducible protein-10 (CXCL10) predict disease relapse and prognosis in diffuse large B-cell lymphoma patients.
Although standard clinical prognostic factors predict outcome in diffuse large B-cell lymphoma (DLBCL), predicting the outcome of patients might be further refined using biological factors. We tested whether serum cytokines could provide prognostic information in DLBCL patients. Thirty cytokines were measured in pretreatment samples from newly diagnosed DLBCL patients using a multiplex ELISA. Sixty-nine patients treated with R-CHOP plus epratuzumab were used in an initial cohort and 185 patients treated with standard R-CHOP served as a subsequent validation cohort. In the initial cohort, elevated serum interleukin-10 [IL-10; hazard ratio (HR) = 6.6, P = 0.022], granulocyte macrophage colony-stimulating factor (HR = 10.8, P= 0.027) and IP-10 (interferon-inducible protein-10, CXCL10; HR = 3.32, P = 0.015) were associated with event-free survival (EFS). An identical analysis of the subsequent validation cohort confirmed that elevated serum levels of IP-10 were strongly associated with a poor EFS (HR = 2.42, P = 0.0007); and also identified interleukin-8 (IL-8; HR = 3.40, P = 0.00002) and interleukin-2 receptor (IL-2R, CD25; HR = 2.59, P = 0.0012) as significantly associated with prognosis. The prognostic significance of elevated IP-10 remained significant after adjustment for the International Prognostic Index (EFS - HR 1.99, P = 0.009, overall survival-HR 1.93, P = 0.021). Elevated pretreatment serum IP-10 levels are therefore associated with an increased likelihood of disease relapse and an inferior survival in patients with DLBCL.


Molecular Phylogenetics of Centrocestus formosanus (Digenea: Heterophyidae) Originated from Freshwater Fish from Chiang Mai Province, Thailand.
This study aimed to investigate the morphology and reconstruct the phylogenetic relationships of Centrocestus formosanus originating from 5 species of freshwater fish, i.e., Esomus metallicus, Puntius brevis, Anabas testudineus, Parambassis siamensis, and Carassius auratus, in Chiang Mai province, Thailand. Sequence-related amplified polymorphism (SRAP) and phylogeny based on internal transcribed spacer 2 (ITS2) and mitochondrial cytochrome c oxidase subunit 1 (CO1) were performed. The results showed similar morphologies of adult C. formosanus from day 5 after infection in chicks. C. formosanus originated from 4 species of freshwater fish had the same number of circumoral spines on the oral sucker, except for those from C. auratus which revealed 34 circumoral spines. The phylogenetic tree obtained from SRAP profile and the combination of ITS2 and CO1 sequence showed similar results that were correlated with the number of circumoral spines in adult worms. Genetic variability of C. formosanus also occurred in different species of freshwater fish hosts. However, more details of adult worm morphologies and more sensitive genetic markers are needed to confirm the species validity of C. formosanus with 34 circumoral spines originating from C. auratus in the future.


Multifunctional Coatings on Sulfide-Based Solid Electrolyte Powders with Enhanced Processability, Stability, and Performance for Solid-State Batteries.
Sulfide-based solid-state electrolytes (SSEs) exhibit many tantalizing properties including high ionic conductivity and favorable mechanical properties for next-generation solid-state batteries. Widespread adoption of these materials is hindered by their intrinsic instability under ambient conditions, which makes them difficult to process at scale, and instability at the Li||SSE and cathode||SSE interfaces, which limits cell performance and lifetime. Atomic layer deposition is leveraged to grow thin Al2 O3 coatings on Li6 PS5 Cl powders to address both issues simultaneously. These coatings can be directly grown onto Li6 PS5 Cl particles with negligible chemical modification of the underlying material and enable exposure of powders to pure and H2 O-saturated oxygen environments for ≥4 h with minimal reactivity, compared with significant degradation of the uncoated powder. Pellets fabricated from coated powders exhibit ionic conductivities up to 2× higher than those made from uncoated material, with a simultaneous decrease in electronic conductivity and significant suppression of chemical reactivity at the Li-SSE interface. These benefits result in significantly improved room temperature cycle life at high capacity and current density. It is hypothesized that this enhanced performance derives from improved intergranular properties and improved Li metal adhesion. This work points to a completely new framework for designing active, stable, and scalable materials for next-generation solid-state batteries.


Chemical Constituents of the Deep-sea Gammarid Shrimp-Derived Fungus Penicillium citrinum XIA-16.
One new alkaloid, (S)-2-acetamido-4-(2-(methylamino)phenyl)-4-oxobutanoic acid (1), was isolated from the deep-sea-derived Penicillium citrinum XIA-16, together with 25 known compounds including ten polyketones (2-11), eight alkaloids (12-19), six steroids (20-25), and a fatty acid (26). Their planar and relative structures were determined by an analysis of 1D and 2D nuclear magnetic resonance (NMR) as well as high resolution electrospray ionization mass spectroscopy (HR-ESI-MS) data. The absolute configuration of 1 was determined by comparison of the experimental and calculated electronic circular dichroism (ECD) spectra. Penicitrinol B (6) significantly inhibited RSL3-induced ferroptosis (EC50 =2.0 μM) by reducing lipid peroxidation and heme oxygenase 1 (HMOX1) expression. Under the concentration of 10 μM, penicitrinol A (7) was able to inhibit cuproptosis with the cell viabilities of 68.2 % compared to the negative control (copper and elesclomol) with the cell viabilities of 14.8 %.


Promote health, not nuclear weapons: ethical duty of medical professionals.
Despite ongoing tensions in various parts of the world, the year 2017 ended on a positive note. The Treaty Prohibiting Nuclear Weapons (TPNW) was passed by the UN General Assembly on July 7, 2017, which will always be a red-letter day in history. It has raised many hopes for a future world without nuclear weapons and staved off the impending humanitarian catastrophe. Good health is a basic need of every individual. Therefore, each person yearns for a life free of violence and free of man-made catastrophes like the ones at Hiroshima and Nagasaki in 1945, which killed over two hundred thousand people and resulted in genetic mutations affecting generations thereafter. Unfortunately, instead of working for nuclear disarmament, the world moved towards an unending nuclear arms race, costing billions which could have been used for healing millions of people living in despair and sickness. This is why on December 10, 2017, Oslo, the capital of Norway, was filled with excitement when the Nobel Peace Prize for this year was bestowed upon the International Campaign to Abolish Nuclear Weapons (ICAN). Large numbers of medical professionals from around the globe had gathered there to affirm their commitment to a healthy future through diversion of wasteful expenditure from the nuclear arms race towards universal health.


When Traditional Offloading is not an Option, Could an External Fixator be a Solution?: A Case Report.
Offloading is a cornerstone in managing diabetic plantar foot ulcers; however, it often represents one of the most challenging aspects of treatment for clinician and patient alike. The authors present a case of a 61-year-old African American man with type 2 diabetes and a limb-threatening plantar foot ulcer that required aggressive wound and surgical management. Due to the heavy drainage and patient adherence issues, traditional offloading techniques such as total contact cast, DH Pressure Relief Walker (Össur, Foothill Ranch, CA), and wedge shoes, among others, were not viable options. Without offloading, healing will be difficult to achieve and will take a long time, carrying a higher risk of limb loss. The decision was made to apply an Ilizarov circular frame with footplate to facilitate offloading and weight bearing in tandem with negative pressure therapy. Although this is still considered an unusual use of this device, the results were positive and the wound progressed to complete reepithelialization.


A case of IMP-4-, OXA-421-, OXA-96-, and CARB-2-producing Acinetobacter pittii sequence type 119 in Australia.
An IMP-4-producing Acinetobacter pittii strain coproducing oxacillinases was isolated from a leg wound of a 67-year-old female patient. Identification to the species level by rpoB and gyrB sequencing and multiplex-PCR-based analysis revealed that the isolate was A. pittii. Whole-genome sequencing of this A. pittii isolate determined the presence of blaOXA-96, blaCARB-2, and a novel blaOXA-421 gene. The position of this novel blaOXA-421 gene was similar to that of blaOXA-51 in A. baumannii, downstream of the phosphinothricin N-acetyltransferase gene and upstream of fxsA in the chromosome. This A. pittii isolate was found to belong to sequence type 119 (ST119). Here, we report the first isolation of IMP-4-producing A. pittii ST119 with a novel blaOXA-421 gene from a patient in Australia and characterize its draft genome.


[A critical note on bioceramics].
In this note I, the authors point out the uncertainty and confusion which exists in the bioceramics field. They then list the calcium phosphate biomaterials today in use.


Plasma protein binding affinity and its relationship to molecular structure: an in-silico analysis.
In-silico plasma protein binding (PPB) models have been generated on human and rat in-house datasets, and on a human dataset sourced from the literature. From the results reported herein, it is apparent that models built on datasets relevant to the chemotypes under investigation in lead optimization programs will perform considerably better in this role than those generated on diverse compounds sourced from the literature. The in-house human and rat partial least-squares regression (PLS) models have cross-validated q2 values of 0.53 and 0.42 on the training sets, respectively. On the independent test and validation sets, they display similar predictive ability, with logK prediction errors of approximately 0.5 log units. This compares to approximately 0.25 log units variability expected for experiment. Given the considerable interspecies PPB differences, the prediction of PPB in one species using measurements in the other is no better than a prediction from an in-silico model generated on that species.


Efficacy and safety of vertebroplasty for treatment of painful osteoporotic vertebral fractures: a randomised controlled trial [ACTRN012605000079640].
Vertebroplasty is a promising but as yet unproven treatment for painful osteoporotic vertebral fractures. It involves radiographic-guided injection of various types of bone cement directly into the vertebral fracture site. Uncontrolled studies and two controlled quasi-experimental before-after studies comparing volunteers who were offered treatment to those who refused it, have suggested an early benefit including rapid pain relief and improved function. Conversely, several uncontrolled studies and one of the controlled before-after studies have also suggested that vertebroplasty may increase the risk of subsequent vertebral fractures, particularly in vertebrae adjacent to treated levels or if cement leakage into the adjacent disc has occurred. As yet, there are no completed randomised controlled trials of vertebroplasty for osteoporotic vertebral fractures. The aims of this participant and outcome assessor-blinded randomised placebo-controlled trial are to i) determine the short-term efficacy and safety (3 months) of vertebroplasty for alleviating pain and improving function for painful osteoporotic vertebral fractures; and ii) determine its medium to longer-term efficacy and safety, particularly the risk of further fracture over 2 years. A double-blind randomised controlled trial of 200 participants with one or two recent painful osteoporotic vertebral fractures. Participants will be stratified by duration of symptoms (< and >or= 6 weeks), gender and treating radiologist and randomly allocated to either the treatment or placebo. Outcomes will be assessed at baseline, 1 week, 1, 3, 6, 12 and 24 months. Outcome measures include overall, night and rest pain on 10 cm visual analogue scales, quality of life measured by the Assessment of Quality of Life, Osteoporosis Quality of Life and EQ-5D questionnaires; participant perceived recovery on a 7-point ordinal scale ranging from 'a great deal worse' to 'a great deal better'; disability measured by the Roland-Morris Disability Questionnaire; timed 'Up and Go' test; and adverse effects. The presence of new fractures will be assessed by radiographs of the thoracic and lumbar spine performed at 12 and 24 months. The results of this trial will be of major international importance and findings will be immediately translatable into clinical practice.


Command performance. CMS ready to debut pay-for-performance plan, but regardless of whether it's a hit, quality comparisons could be a rising star.
Whether the CMS' pay-for-performance plan ever materializes is inconsequential. Executives say quality performance will undoubtedly play a larger role in reimbursement. Opal Reinbold, left, from Palomar Pomerado Health says the system participated in a quality incentive demonstration project because "it prepared us for how this evidence-based practice was going to be applied."


Interleukin 1 release by human alveolar macrophages and blood monocytes.
These studies utilized a sensitive and specific radioimmunoassay for interleukin 1 beta (IL-1 beta) to compare release of IL-1 by human alveolar macrophages and blood monocytes. The studies demonstrate that alveolar macrophages release amounts of antigenic IL-1 beta that are similar to that of blood monocytes. The amounts of IL-1 released were similar for both cell types at early (4 h) and late (24 h) time points and with differing amounts of stimuli [endotoxin; lipopolysaccharide (LPS)]. In addition, alveolar macrophages actually produced more total IL-1 (intracellular IL-1 plus released IL-1) than did blood monocytes. Alveolar macrophages that were stimulated with LPS released significantly more prostaglandin E2 (PGE2, an inhibitor of IL-1) than did blood monocytes. These studies demonstrate that human alveolar macrophages are not defective in their capacity to release IL-1.


A negative elongation factor for human RNA polymerase II inhibits the anti-arrest transcript-cleavage factor TFIIS.
Formation of productive transcription complexes after promoter escape by RNA polymerase II is a major event in eukaryotic gene regulation. Both negative and positive factors control this step. The principal negative elongation factor (NELF) contains four polypeptides and requires for activity the two-polypeptide 5,6-dichloro-1-beta-D-ribobenzimidazole-sensitivity inducing factor (DSIF). DSIF/NELF inhibits early transcript elongation until it is counteracted by the positive elongation factor P-TEFb. We report a previously undescribed activity of DSIF/NELF, namely inhibition of the transcript cleavage factor TFIIS. These two activities of DSIF/NELF appear to be mechanistically distinct. Inhibition of nucleotide addition requires > or = 18 nt of nascent RNA, whereas inhibition of TFIIS occurs at all transcript lengths. Because TFIIS promotes escape from promoter-proximal pauses by stimulating cleavage of back-tracked nascent RNA, TFIIS inhibition may help DSIF/NELF negatively regulate productive transcription.


Recurrent attacks of fear and visual hallucinations in a child.
The case of a 7-year-old boy suffering from recurrent nocturnal and occasional daytime attacks with intense fear and complex visual hallucinations is presented. His state was otherwise normal, as were routine electroencephalographic (EEG) and magnetic resonance imaging (MRI) investigations in the past. Several differential diagnoses such as panic disorder, pavor nocturnus, and nightmares were considered but could not be definitely established or excluded. Since the attacks appeared after the divorce of his parents, an adjustment disorder was suspected, and the patient received psychotherapy for more than 2 years without an effect on the attacks. Only when long-term video-EEG recorded two typical attacks with left temporal ictal seizure patterns was focal epilepsy diagnosed and successfully treated with antiepileptic medication. A suspected origin of seizures in the amygdala was supported by a high-resolution MRI showing a cortical dysplasia extending from the left anteromedial temporal lobe to the amygdala. The case exemplifies difficulties in the differential diagnosis of panic-like attacks and underlines the value of long-term video-EEG, which may be necessary to establish the correct diagnosis and to prevent ineffective therapeutical approaches.


Simplification of the representation of the organic component of atmospheric particulates.
We present an analysis of recent field data to investigate the variation in the organic component of atmospheric aerosol and its behaviour in the moist environment. In all locations the degree of oxygenation of the organic material increases with photochemical age, as does the particulate hygroscopicity. These changes will generally occur in spatial scales comparable to a single cell in global models at representative boundary layer wind speeds. Using ADDEM, a new model of the equilibrium state of multicomponent aerosol, we show that inorganic component changes must be responsible for the increase in particulate hygroscopicity with photochemical age. It is suggested that a common representation of nearfield and background organic aerosol composition is sufficient to describe the behaviour of organic components in a variety of field experiments; nearfield small mode organics being dominated by a combustion-derived unoxidised signature, whilst the background accumulation mode is more oxygenated and dominates in air masses with a photochemical age of more than a couple of days. This representation may be used within the sub-saturated regime to predict the behaviour of ambient particulates in the moist atmosphere. Whether a similar common representation can be used for cloud activation prediction in supersaturated environments, or for investigation of gas-to-particle partitioning, should be investigated.


Variation in cellulase-constituting components from Trichoderma reesei with agitation intensity.
Cellulase producing activity of Trichoderma reesei QM9414 was examined under various agitation intensities and at the dissolved oxygen concentration above 3 ppm. The producing activity greatly depended upon the agitation intensity, and the dependence on the agitation was different for each cellulase-constituting component. The maximum producing activities of FPA, CM Case, and beta-glucosidase were obtained under different agitation conditions, 1.0, 0.7, and 1.4 m/s in tip velocity, respectively. Intensive agitation brought about remarkable reduction in all cellulase components. The mycelial transformation through agitation intensity was also observed. Comparatively mild agitation of 0.3-1.0 m/s caused pellet formation as the culture progressed, although the pelletization was delayed with increasing agitation intensity. The behavior of the pelletization did not occur at 1.3 and 1.7 m/s throughout the course of cultivation, and under the latter agitation condition hyphae were broken up into short fragments. The cellulase producing activity is discussed in relation to such morphological changes.


Operant matching is a generic outcome of synaptic plasticity based on the covariance between reward and neural activity.
The probability of choosing an alternative in a long sequence of repeated choices is proportional to the total reward derived from that alternative, a phenomenon known as Herrnstein's matching law. This behavior is remarkably conserved across species and experimental conditions, but its underlying neural mechanisms still are unknown. Here, we propose a neural explanation of this empirical law of behavior. We hypothesize that there are forms of synaptic plasticity driven by the covariance between reward and neural activity and prove mathematically that matching is a generic outcome of such plasticity. Two hypothetical types of synaptic plasticity, embedded in decision-making neural network models, are shown to yield matching behavior in numerical simulations, in accord with our general theorem. We show how this class of models can be tested experimentally by making reward not only contingent on the choices of the subject but also directly contingent on fluctuations in neural activity. Maximization is shown to be a generic outcome of synaptic plasticity driven by the sum of the covariances between reward and all past neural activities.


Lipoblastomas and liposarcomas in paediatric patients: A case series.
Lipoblastomas and liposarcomas are rare causes of soft tissue masses in paediatric patients. In this retrospective clinical case series we identified 11 patients from our paediatric database (10 with a lipoblastoma and one with a liposarcoma) who had attended our hospital between 1998 and 2019. The median age of patients with lipoblastoma was 29 months. All lipoblastoma cases were managed with surgical excision and histological examination. The 18-year old patient with liposarcoma presented with a metastatic and unresectable tumour that was unresponsive to chemotherapy and radiation. Our experience demonstrates the importance of differentiating the type of soft tissue mass in children.


Sustained effects of interprofessional shared learning on student attitudes to communication and team working depend on shared learning opportunities on clinical placement as well as in the classroom.
Delivering high quality healthcare increasingly requires effective team working, and interprofessional shared learning (SL) is crucial to this. This study compares the attitudes, 1 year after experience of an undergraduate SL programme, of students who had participated in the programme with their peers who had not. 207 students were invited to complete a questionnaire to assess the impact of SL on attitudes to clinical competence and behaviour. Responses were received from 171 students (83%) who had either had no experience of SL, SL in lectures only, or SL in lectures and clinical placement. Significantly different responses were found between the three groups for a number of the statements, and these were further developed in responses to the open-ended questions. Only group 3 had developed and sustained a less exclusive attitude and were better able to appreciate that SL can make an important contribution to learning communication skills and understanding patient problems. This raises important questions about the approach taken to undergraduate SL if it is to have a contributory effect to attitudes about professional identity, and a significant effect in improving the quality of care provided by the doctors and nurses of tomorrow.


Acute renal failure associated with inhaled tobramycin.
A case of nephrotoxicity possibly caused by tobramycin inhalation solution is presented. A 62-year-old Caucasian woman was admitted for treatment of decreased urine output and sepsis secondary to Pseudomonas aeruginosa. Her past medical history was significant for multiple diseases, including chronic renal insufficiency (baseline serum creatinine concentration [SCr] 2 mg/dL). One month postadmission, the patient was diagnosed with health care-associated pneumonia. The patient was initiated on piperacillin-tazobactam and tobramycin 2 mg/kg i.v. She was changed to imipenem-cilastatin with continuation of i.v. tobramycin. A month after discontinuation of her antibiotic regimen, the patient was diagnosed with P. aeruginosa pneumonia. The patient received imipenem-cilastatin, vancomycin, and inhaled tobramycin 300 mg twice daily. At that time, her SCr was 2 mg/dL. Inhaled tobramycin was continued for four weeks, and the patient's SCr steadily rose to a peak of 4.5 mg/dL. During week 1 of treatment, multidrug-resistant P. aeruginosa and methicillin-resistant Staphylococcus aureus were diagnosed. The patient continued to receive i.v. imipenem-cilastatin, vancomycin, and inhaled tobramycin with an SCr of 1.9 mg/dL. However, at the end of week 2, the patient's SCr began to slowly rise (2.3 mg/dL). At week 3, imipenem-cilastatin was discontinued; inhaled tobramycin was continued. The patient's SCr continued to rise (3.2 mg/dL). At week 4, her SCr rose to 4.5 mg/dL, resulting in initiation of hemodialysis and discontinuation of inhaled tobramycin. The patient's SCr never returned to baseline, and renal function was never regained. Acute renal failure requiring dialysis occurred in a high-risk patient receiving an extended course of treatment with inhaled tobramycin.


Congenital nuclear cataracts in the Morgan horse.
Nuclear cataracts were found in 2 groups of related Morgan horses. The cataracts were finely reticulated central spherical translucencies that sometimes extended to the region of the posterior "Y" suture. The cataracts were not associated with other ocular defects and did not impair vision. In 1 group of 8 horses, 5 were affected; in the other group, 6 of 8 were affected. Although a pattern of inheritance could not be determined, the familial distribution of the cataracts supported the conclusion that the defect was a heritable disorder.


Haloperidol plasma and red blood cell levels and clinical antipsychotic response.
Two investigators have recently suggested therapeutic ranges for plasma haloperidol in the treatment of schizophrenia. An apparent optimal therapeutic range of red blood cell haloperidol as early as day 7 of the drug trial is described in this article. With continued treatment, an optimal plasma haloperidol range for response could be observed by day 14 of treatment. The previously described correlation between response at day 7 and plasma/red blood cell haloperidol ratio was confirmed but was found not to predict response at day 14 of drug treatment in this cohort of DSM-III schizophrenic patients.


TGF-β2 and collagen play pivotal roles in the spheroid formation and anti-aging of human dermal papilla cells.
Dermal papilla cells (DPCs) tend to aggregate both in vitro and in vivo, which increases the hair inductivity of DPCs. However, the underlying mechanism of spheroid formation is unknown. We investigated whether collagen expression in human DPCs (hDPCs) is involved in the spheroid formation and hair inductivity of hDPCs and further examined the underlying molecular mechanism of collagen upregulation. The expression of diverse collagens, such as COL13A1 and COL15A1, was upregulated in three dimensional (3D)-cultured or intact DPCs, compared to 2D-cultured hDPCs. This collagen expression was a downregulated in aged hair follicle, and aged DPCs were difficult to aggregate. Blocking of COL13A1 and COL15A1 by small interfering RNA reduced aggregation, while induced senescence of hDPCs in vitro. Further, transforming growth factor-β2 (TGF-β2) expression decreases with aging, and is involved in regulating the expression of COL13A1 and COL15A1. Addition of recombinant TGF-β2 delayed cellular senescence, and recovered spheroid formation in aged hDPCs by upregulating collagen levels. On the contrary, knock-out of TGF-β2 induced the aging of DPCs, and inhibited spheroid formation. These results suggested that COL13A1 and COL15A1 expression is downregulated with aging in DPCs, and upregulation of collagen by TGF-β2 induces the spheroid formation of DPCs. Therefore, TGF-β2 supplement in DPC culture medium could enhance the maintenance and hair inductivity of DPCs.


Jeotgalicoccus schoeneichii sp. nov. isolated from exhaust air of a pig barn.
A Gram-staining-positive, non-motile, non-spore-forming, coccus (strain 140805-STR-02T) was isolated from exhaust air of a pig barn on Columbia Blood Agar Base (Oxoid) supplemented with 5 % defibrinated horse blood, Streptococcus selective supplement and 0.5 mg erythromycin l-1. The strains shared high 16S rRNA gene sequence similarity to Jeotgalicoccus pinnipedialis (98.6 %) but only a maximum of 94 % sequence similarity to all other species of the genus Jeotgalicoccus. DNA-DNA hybridisation values between strain 140805-STR-02T and J. pinnipedialis CIP 107946T were 60.3 % (reciprocal, 51.2 %). The quinone system of 140805-STR-02T contained predominantly menaquinone MK-7 and minor amounts of MK-6. The polar lipid profile of strain 140805-STR-02T contained the major compounds diphosphatidylglycerol and phosphatidylglycerol and four unidentified lipids present in minor to moderate amounts. In the polyamine pattern spermidine and spermine were predominant. The fatty acid profile comprising iso-C15 : 0 and anteiso-C15 : 0 as major fatty acids, and was in congruence with those reported for other species of the genus Jeotgalicoccus and thus supported the affiliation of strain 140805-STR-02T to this genus. The results of physiological and biochemical tests allowed a clear phenotypic differentiation of strain 140805-STR-02T from the most closely related species. Strain 140805-STR-02T represents a novel species, for which the names Jeotgalicoccus schoeneichii sp. nov. is proposed, with the type strain 140805-STR-02T (=LMG 29445T=CCM 8667T).


Mobilization of endothelial precursor cells: systemic vascular response to musculoskeletal trauma.
Postnatal vasculogenesis, the process by which vascular committed bone marrow stem cells or endothelial precursor cells (EPC) migrate, differentiate, and incorporate into the nacent endothelium contributing to physiological and pathological neovascularization, has stimulated much interest. Its contribution to tumor nonvascularization, wound healing, and revascularization associated with skeletal and cardiac muscles ischaemia is established. We evaluated the mobilization of EPCs in response to musculoskeletal trauma. Blood from patients (n = 15) following AO type 42a1 closed diaphyseal tibial fractures was analyzed for CD34 and AC133 cell surface marker expression. Immunomagnetically enriched CD34+ mononuclear cell (MNC(CD34+)) populations were cultured and examined for phenotypic and functional vascular endothelial differentiation. Circulating MNC(CD34+) levels increased sevenfold by day 3 postinjury. Circulating MNC(AC133+) increased 2.5-fold. Enriched MNC(CD34+) populations from day 3 samples in culture exhibited cell cluster formation with sprouting spindles. These cells bound UEA-1 and incorporated fluorescent DiI-Ac-LDL intracellularily. Our findings suggest a systemic provascular response is initiated in response to musculoskeletal trauma. Its therapeutic manipulation may have implications for the potential enhancement of fracture healing.


Early stage primary gastric diffuse large B-cell lymphoma in a young HIV-positive patient.
HIV infection is known to be associated with the development of a wide range of neoplasia. About 25 to 40% of HIV-positive patients will present some kind of malignancy in the course of the disease; among them 10% are non-Hodgkin lymphomas (NHL) and 20% of these are represented by the diffuse large B-cell lymphoma. HIV-positive patients have a relative risk of 110 times higher to develop neoplasia, than the non-infected population. The gastrointestinal (GI) tract is the most frequent extranodal site of involvement. However, the primary GI lymphoma is rare. The authors present a case of a 31-year-old male patient with a 16-year history of HIV infection, who deliberately withdrew the Highly Active Antiretroviral Therapy (HAART) regimen and was hospitalized because of a respiratory infection. Because of a long-term complaint of dyspepsia, an upper gastrointestinal endoscopy was performed disclosing a large elevated and ulcerated gastric lesion, which biopsy revealed a diffuse large B-cell lymphoma. Clinical, imaging and laboratory tests showed an early stage diagnosis: Lugano stage I. Although not frequent, the authors alert to considering this neoplasia in all HIV-positive patients with dyspeptic symptoms.


Scorpions and Scorpionism in Roudan County, Southern Iran.
Every year, thousands of cases and many deaths from scorpion sting are reported in tropical areas of South and Southwestern parts of Iran. The aim of this study was to identify the fauna and dangerous species of scorpions in Roudan County, southern Iran. This descriptive study was conducted in 10 stations in Hormozgan Province. Scorpion sampling was done randomly by searching for their shelter and digging out their nests during day, and with the use of UV light during night from February 2013 to October 2014. Data of scorpion stings were obtained from health center of Hormozgan Province during 2014-2016. Overall, 155 scorpions were collected on a set of eight species belonging to Buthidae and Hemiscorpiidae families. These species were identified as Mesobuthus persicus, Mesobuthus phillipsi, Hottentotta schach, Odontobuthus doriae, Compsobuthus persicus, Orthochirus farzanpayi, Androctonus crassicauda and Hemiscorpius acanthocercus. One thousand and twenty-seven cases of scorpion sting were recorded during 2014-2016 with a peak period in summer. Most of cases were <44yr old. Five out of six medically important scorpions in Iran were actively identified in the study area. Results of this study would greatly help to identify risk factors of scorpion sting in high-risk areas for planning, management and treatment of patients with scorpion sting in these areas.


Dynamics of a periodic stoichiometric model with application in predicting and controlling algal bloom in Bohai Sea off China.
We develop a nonautonomous stoichiometric algal growth model incorporating a season-driven light intensity. We characterize the model dynamics by showing positive invariance, dissipativity, boundary dynamics, and internal dynamics. We use numerical simulations to uncover the impacts of the seasonal light intensity and the nutrient availability on the algal dynamics. We discuss two control methods, removing algae (RA) periodically and blocking nutrient (BN) input from rivers constantly, via ourmodeling approach. By comparison, the BNmethod is amore effective way to terminate algal bloom in Yellow Sea off China. The model dynamics can fit the Bohai Sea data well. Our model and analysis provide a possible explanation of seasonal algal bloom and give some measurements for controlling algal bloom in China's coastal regions.


A severe case of Lemierre Syndrome with Streptococcus constellatus Infection.
A 76-year-old Japanese male presenting with high fever, headache, and disturbance of consciousness was hospitalized. Contrast computed tomography revealed thrombophlebitis in the internal jugular vein and abscesses in the posterior neck region, pharynx, and pterygoid muscle. Streptococcus constellatus infection was confirmed by culture of blood samples, and the patient was diagnosed with Lemierre syndrome. In addition to the administration of antibiotics and anticoagulants, abscess drainage was performed. S. constellatus should be considered as a causative bacterium in elderly patients with Lemierre syndrome.


Selected developmental factors of cleft palate children between two and thirty-three months of age.
The literature suggests that cleft palate children and adults perform below their peers on both linguistic and nonlinguistic tasks. The purpose of this investigation was to ascertain if developmental delay could be observed in 24 cleft palate children below the age of three years when compared to matched controls. Three developmental scales were administered to all infants, and complete medical histories and current weights were obtained. Results indicated that the performance of the cleft palate subjects paralleled but fell below their peers on all developmental subtests. The differences were significant (p is less than .05) on three language subtests and on one personal-social and one motor subtest. The mean percentile weight also differed significantly (p is less than .05). Multivariate analysis explored the relationship of 1) extent of cleft, 2) sex, 3) hearing loss, 4) length of hospitalization, and 5) weight to the developmental scores. The extent of the cleft proved to be significant.


An examination of current stroke rehabilitation practice in Peru: Implications for interprofessional education.
This study aimed to better understand current clinical practice of rehabilitation professionals in Lima, Peru, and to explore the existence of and potential for interprofessional collaboration. A secondary purpose was to assess rehabilitation professionals' agreement with evidence-based stroke rehabilitation statements and confidence performing stroke rehabilitation tasks prior to and following an interprofessional stroke rehabilitation training. Current clinical practice for rehabilitation professionals in Peru differs from high-income counties like the United States, as physical therapists work with dysphagia and feeding, prosthetist orthotists serve a strictly technical role, and nurses have a limited role in rehabilitation. Additionally, while opportunity for future interprofessional collaboration within stroke rehabilitation exists, it appears to be discouraged by current health system policies. Pre- and post-training surveys were conducted with a convenience sample of 107 rehabilitation professionals in Peru. Survey response options included endorsement of professionals for rehabilitation tasks and a Likert scale of agreement and confidence. Training participants largely agreed with evidence-based stroke rehabilitation statements. Differences in opinion remained regarding the prevalence of dysphagia and optimal frequency of therapy post-stroke. Substantially increased agreement post-training was seen in favour of early initiation of stroke rehabilitation and ankle foot orthosis use. Participants were generally confident performing traditional profession-specific interventions and educating patients and families. Substantial increases were seen in respondents' confidence to safely and independently conduct bed to chair transfers and determine physiological stability. Identification of key differences in rehabilitation professionals' clinical practice in Peru is a first step toward strengthening the development of sustainable rehabilitation systems and interprofessional collaboration.


Controlled shear affinity filtration (CSAF): a new technology for integration of cell separation and protein isolation from mammalian cell cultures.
Controlled shear affinity filtration (CSAF) integrates animal cell separation and product isolation in a single unit operation through the use of a specifically designed rotating disk filter with incorporated membrane chromatography column. Because of the decoupling of shear force and pressure generation and the specific hydrodynamics of the system, shear rates can be easily optimized and precisely controlled to maximize filtration performance while viability of the shear sensitive animal cells is maintained. In this study, the general methodology is demonstrated using the integration of Chinese hamster ovary cell separation and isolation of recombinant tissue plasminogen activator (t-PA) as a model example. Direct capture of t-PA from cell culture broth was realized by using custom-made affinity membranes with lysine as a robust, small molecular weight affinity ligand. Small-scale t-PA adsorption experiments, as well as microfiltration experiments, were used to design the integrated CSAF process. A Chinese hamster ovary batch culture was processed with a lab-scale prototype, yielding 86% of the t-PA in the concentrated, particle-free eluate, whereas 95% of the bulk protein was removed. Because the viability of the cells is not significantly affected and high specific flux rates can be achieved, the CSAF technology should also be well suited for continuous perfusion with integrated product isolation. A truly continuous operation could be realized with two systems in tandem configuration.


Mitochondria regulate intestinal stem cell proliferation and epithelial homeostasis through FOXO.
A metabolic transition from glycolysis to oxidative phosphorylation is often associated with differentiation of many types of stem cells. However, the link between mitochondrial respiration and stem cells' behavior is not fully understood. We genetically disrupted electron transport chain (ETC) complexes in the intestinal stem cells (ISCs) of Drosophila. We found that ISCs carrying impaired ETC proliferated much more slowly than normal and produced very few enteroblasts, which failed to further differentiate into enterocytes. One of the main impediments to ISC proliferation and lineage specification appeared to be abnormally elevated forkhead box O (FOXO) signaling in the ETC-deficient ISCs, as genetically suppressing the signaling pathway partially restored the number of enterocytes. Contrary to common belief, reactive oxygen species (ROS) accumulation did not appear to mediate the ETC mutant phenotype. Our results demonstrate that mitochondrial respiration is essential for Drosophila ISC proliferation and lineage specification in vivo and acts at least partially by repressing endogenous FOXO signaling.


Hepatitis C treatment uptake and adherence among injecting drug users in the Czech Republic.
Injecting drug users (IDUs) represent a major subpopulation of hepatitis C virus (HCV)-infected people in developed countries. Yet their uptake to treatment is generally low despite well-documented effectiveness of HCV treatment among former and active IDUs. The present study represents the first attempt to describe the HCV treatment coverage among IDUs and identify factors that affect treatment uptake in the Czech Republic. From January to March 2011, a questionnaire survey was conducted among viral hepatitis treatment centres in the Czech Republic. From a total of 76 identified hepatitis treatment centres existing in the country, 39 provided HCV treatment to (mainly former or abstaining) IDUs in 2010. Most clinicians reported being cautious in initiating HCV treatment in IDUs. Abstinence, a screening phase before treatment initiation, opioid substitution treatment and an external evaluation by a specialist were often prerequisites for skrting treatment. However, HCV treatment centres rarely provided drug-use specific services. Financial constraints were also reported, further limiting the inclusion of IDUs into treatment, as non-users are widely preferred to active drug users. Clinicians reported no difference in treatment uptake and adherence between drug users and non-users, nor between opioid and methamphetamine users. A number of system- and provider-related factors limit HCV treatment in IDUs in the Czech Republic, despite permissive national clinical guidelines. Targeting these factors is crucial to reduce HCV prevalence at population level.


Examination and characterisation of burst spinal cord stimulation on cerebrospinal fluid cellular and protein constituents in patient responders with chronic neuropathic pain - A Pilot Study.
Patients with neuropathic pain have altered proteomic and neuropeptide constituents in cerebrospinal fluid (CSF) compared to controls. Tonic spinal cord stimulation (SCS) has demonstrated differential expression of neuropeptides in CSF before and after treatment suggesting potential mechanisms of action. Burst-SCS is an evidence-based paraesthesia free waveform utilised for neuropathic pain with a potentially different mechanistic action to tonic SCS. This study examines the dynamic biological changes of CSF at a cellular and proteome level after Burst-SCS. Patients with neuropathic pain selected for SCS had CSF sampled prior to implant of SCS and following 8 weeks of continuous Burst-SCS. Baseline and 8-week pain scores with demographics were recorded. T cell frequencies were analysed by flow cytometry, proteome analysis was performed using mass spectrometry and secreted cytokines, chemokines and neurotrophins were measured by enzyme-linked immunosorbent assay (ELISA). 4 patients (2 females, 2 males) with a mean age of 51 years (+/-SEM 2.74, SD 5.48) achieved a reduction in pain of >50% following 8 weeks of Burst-SCS. Analysis of the CSF proteome indicated a significant alteration in protein expression most related to synapse assembly and immune regulators. There was significantly lower expression of the proteins: growth hormone A1 (PRL), somatostatin (SST), nucleobindin-2 (NUCB2), Calbindin (CALB1), acyl-CoA binding protein (DBI), proSAAS (PCSK1N), endothelin-3 (END3) and cholecystokinin (CCK) after Burst-SCS. The concentrations of secreted chemokines and cytokines and the frequencies of T cells were not significantly changed following Burst-SCS. This study characterised the alteration in the CSF proteome in response to burst SCS in vivo. Functional analysis indicated that the alterations in the CSF proteome is predominately linked to synapse assembly and immune effectors. Individual protein analysis also suggests potential supraspinal mechanisms.


Expression and characterization of biologically active human hepatocyte growth factor (HGF) by insect cells infected with HGF-recombinant baculovirus.
A cDNA containing the entire coding sequence of human hepatocyte growth factor (HGF) [also known as scatter factor (SF)] was inserted into the genome of Autographa california nuclear polyhedrosis virus (baculovirus) adjacent to the polyhedrin promoter by homologous recombination. Insect cells (Spodoptera frugiperda) infected with the recombinant virus secrete relatively high levels (3-8 mg/L) of biologically active HGF into the culture medium. The recombinant HGF induces pronounced morphological changes and scattering of primary cultures of rat, mouse, and human hepatocytes within 24 h after plating and stimulates DNA synthesis in these cells with the same magnitude as native HGF derived from human placenta or rabbit serum. The human recombinant HGF produced by the insect cells is N-glycosylated, binds to heparin like native HGF, and is recognized by polyclonal antiserums raised against human or rabbit HGF as assessed by immunoblot, ELISA, and immunoneutralization experiments. Metabolic radiolabeling with L-[35S]methionine (pulse-chase experiments) as well as Western blot analysis indicates that the recombinant HGF is synthesized and secreted by the infected insect cells as the unprocessed single-chain form (pro-HGF) when the cells are cultured in serum-free medium. However, when the infected insect cells are cultured in insect culture medium (Grace's medium) containing fetal bovine serum, the secreted HGF is present mainly in the mature heterodimeric form. Addition of serum to the baculovirus-expressed single-chain [125I]HGF in a cell-free system results in conversion to the heterodimeric two-chain form, and the activation is prevented by the serine protease inhibitor PMSF. Incubation of 125I-labeled pro-HGF with rat liver or spleen extracts resulted in conversion of pro-HGF to the heterodimeric two-chain form. A truncated form of HGF containing the N-terminal portion of HGF (kringles 1-3) was also produced in the same expression system. This deleted HGF, by itself, did not have any detectable biological activity; however, it abrogated the stimulatory effects of full-length HGF on hepatocytes. This is the first successful production of bioactive recombinant HGF in large quantities, which will allow purification on the milligram scale of pro-HGF and will permit future studies to elucidate pathways involved in HGF activation by its target tissues.


Time-resolved and mechanistic study of the photochemical uncaging reaction of the o-hydroxycinnamic caged compound.
The o-hydroxycinnamic derivatives represent efficient caged compounds that can realize quantification of delivery upon uncaging, but there has been lack of time-resolved and mechanistic studies. We used time-resolved infrared (TRIR) spectroscopy to investigate the photochemical uncaging dynamics of the prototype o-hydroxycinnamic compound, (E)-3-(2-hydroxyphenyl)-acrylic acid ethyl ester (HAAEE), leading to coumarin and ethanol upon uncaging. Taking advantage of the specific vibrational marker bands and the IR discerning capability, we have identified and distinguished two key intermediate species, the cis-isomers of HAAEE and the tetrahedral intermediate, in the transient infrared spectra, thus providing clear spectral evidence to support the intramolecular nucleophilic addition mechanism following the trans-cis photoisomerization. Moreover, the product yields of coumarin upon uncaging were observed to be greatly affected by the solvent polarity, suppressed in CH2Cl2 but enhanced in D2O/CH3CN with the increasing volume ratio of D2O. The highly solvent-dependent behavior indicates E1 elimination of the tetrahedral intermediate to give rise to the final uncaging product coumarin. The photorelease rate of coumarin was directly characterized from TRIR (3.6 × 10(6) s(-1)), revealing the promising application of such o-hydroxycinnamic compound in producing fast alcohol jumps. The TRIR results provide the first time-resolved detection and thus offer direct dynamical information about this photochemical uncaging reaction.


LED phototherapy to prevent mucositis: a case report.
The purpose of this case report was to evaluate the efficacy of phototherapy using light-emitting diodes (LEDs) to prevent oral mucositis in a Hodgkin's disease patient treated with the ABVD (doxorubicin [Adriamycin], bleomycin, vinblastine, and dacarbazine) chemotherapy regimen. Mucositis is a common dose-limiting complication of cancer treatment, and if severe it can lead to alterations in treatment planning or suspension of cancer therapy, with serious consequences for tumor response and survival. Therefore, low-power lasers and more recently LEDs, have been used for oral mucositis prevention and management, with good results. In this study, a 34-year-old man received intraoral irradiation with an infrared LED array (880 nm, 3.6 J/cm2, 74 mW) for five consecutive days, starting on chemotherapy day 1. In each chemotherapy cycle, he received the ABVD protocol on days 1 and 15, and received LED treatment for 5 d during each cycle. To analyze the results, the World Health Organization (WHO) scale was used to grade his mucositis, and a visual analogue scale (VAS) was used for pain evaluation, on days 1, 3, 7, 10, and 13 post-chemotherapy. The results showed that the patient did not develop oral mucositis during the five chemotherapy cycles, and he had no pain symptoms. LED therapy was a safe and effective method for preventing oral mucositis in this case report. However, further randomized studies with more patients are needed to prove the efficacy of this method.


Resting B cells from autoimmune lupus-prone New Zealand Black and (New Zealand Black x New Zealand White)F1 mice are hyper-responsive to T cell-derived stimuli.
To determine whether B cells from New Zealand Black (NZB) and (New Zealand Black x New Zealand White)F1 (NZB/W) mice possess intrinsic defects that lead to altered immune responsiveness, we purified resting B cells from these mice and compared their surface phenotype and function with those of resting B cells isolated from BALB/c and DBA/2 nonautoimmune mouse strains. Flow cytometric analysis of freshly isolated resting B cells revealed that NZB and NZB/W resting B cells are conventional B2-type cells similar to their nonautoimmune counterparts. Despite this, resting B cells from young NZB and NZB/W mice express lower levels of CD23 on their surface and aberrant levels of intracellular IgM. Upon stimulation, resting B cells from young NZB and NZB/W mice demonstrate increased proliferation, IgM secretion, or enhanced expression of costimulatory molecules in response to a variety of different T cell-derived stimuli, including cytokines and signals generated through CD40. Therefore, B cell hyper-responsiveness to T cell stimuli is immunodominant or codominant in NZB/W mice. Taken together, our results suggest that intrinsic B cell hyper-responsiveness may play a role in the pathogenesis of autoimmune disease in NZB and NZB/W mice. The increased clonal expansion of these B cells together with increased Ig production and enhanced costimulatory capacity serve to amplify the immune response. In the context of normal but incomplete T cell tolerance, B cell hyperresponsiveness to the limited signals provided by partially tolerant T cells may be sufficient to yield an autoantibody response.


Design, Synthesis, and Pharmacological Evaluation of Novel Multisubstituted Pyridin-3-amine Derivatives as Multitargeted Protein Kinase Inhibitors for the Treatment of Non-Small Cell Lung Cancer.
A novel series of pyridin-3-amine derivatives were designed, synthesized, and evaluated as multitargeted protein kinase inhibitors for the treatment of non-small cell lung cancer (NSCLC). Hit 1 was first disclosed by in silico screening against fibroblast growth factor receptors (FGFR), which was subsequently validated by in vitro experiments. The structure-activity relationship (SAR) of its analogues was then explored to afford novel FGFR inhibitors 2a-2p and 3a-3q. Among them, 3m showed potent inhibition against FGFR1, 2, and 3. Interestingly, compound 3m not only inhibited various phosphorylation and downstream signaling across different oncogenic forms in FGFR-overactivated cancer cells but also showed nanomolar level inhibition against several other NSCLC-related oncogene kinases, including RET, EGFR, EGFR/T790M/L858R, DDR2, and ALK. Finally, in vivo pharmacology evaluations of 3m showed significant antitumor activity (TGI = 66.1%) in NCI-H1581 NSCLC xenografts with a good pharmacokinetic profile.


[Psychophysical stress among students of secondary schools during the years of education].
A group of students attended in successive classes of secondary schools were examined in the school-years 1984/85--1987/88. The studied material comprised 165 students (116 girls and 42 boys) beginning education in the first classes. In second classes were 161 students, in third--160 students and in four classes--149 students. The psychophysical stress of the students was assessed by a standardized 77 questions grouped so that it was possible to assess the psychophysical strain connected with school-education, work outside school, day timetable, situation in the class and in the family and physical well-being. The sum of the points obtained in the answers provided information about the general psychophysical stress the answers to certain questions described objectively the stressful situations and the personal experiencing of the troublesomeness++ of them. It was found that with longer time spent in school psychophysical situation of girls and boys was deteriorating because the psychophysical stress from classes to classes was increasing. The psychophysical stress of students in compared classes differed for both sexes. It was greater in girls in successive classes. The objective stressful situations and the personal experiencing of this stress increased among girls from year to year. The psychophysical stress connected with objective situation deteriorated in the fourth classes among boys, nevertheless the subjective experiencing of this stress was raised to its highest value. The final examination (diploma from a European gymnasium) determined a great emotional stress of students, especially for boys.


Diabetes mellitus and its complications in an African-American community: project DIRECT.
Project DIRECT (Diabetes Intervention Reaching and Educating Communities Together) is a multilevel community-based intervention project designed to address diabetes and its complications in an African-American community. This article presents results of the Project DIRECT pilot study and describes risk factors for diabetes, diabetes prevalence, complications, and care practices. During 1993, a pilot study was conducted among persons 20 to 74 years of age in Wake County, North Carolina. The study involved household interviews and examinations, and more extensive health center interviews and examinations based on the race of the head of the household, previous diagnosis of diabetes, and results of capillary glucose tests done in the household. Of the black population aged 20 to 74 years, 52 +/- 3% reported being inactive and 51 +/- 3% were overweight; the prevalence of diagnosed diabetes was 5.2 +/- 0.9%; the prevalence of undiagnosed diabetes was 5.7 +/- 2.7%; and the prevalence of impaired glucose tolerance was 11.4 +/- 7.5%. Blacks with diabetes were significantly more likely than nonblacks with diabetes to have uncontrolled hypertension and to smoke cigarettes. Blacks with diabetes were significantly less likely to report having health insurance or to have a private health-care provider. Diabetes mellitus is a major public health problem in the African-American community of Wake County. Modifiable risk factors for diabetes and undiagnosed diabetes are common. Project DIRECT is attempting to improve the health-related quality of life of this population by reducing the burden of diabetes and its complications through a multilevel, community-based intervention.


Shikonin Inhibits Tumor Growth of ESCC by suppressing PKM2 mediated Aerobic Glycolysis and STAT3 Phosphorylation.
Background: Shikonin, a small molecule inhibitor of pyruvate kinase 2 (PKM2), has been demonstrated to play the antitumor effect in various cancers. However, the specific effects and related regulatory mechanism of Shikonin in esophageal squamous cell carcinoma (ESCC) have not been clearly declared. Materials and methods: Cell viability was valued through 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Glucose consumption, lactate production, glycolytic intermediates and pyruvate kinase enzymatic activity were measured using corresponding assay kits. Patient-derived xenograft (PDX) models were constructed to observe the anti-ESCC effect of Shikonin in vivo. PKM2, p-PKM2, signal transducer and activator of transcription 3 (STAT3), p-STAT3, glucose transporter 1 (GLUT1) and hexokinase 2 (HK2) in ESCC tissues were assessed by western blot. The expression of PKM2, p-PKM2, p-STAT3, GLUT1 and HK2 was assessed by immunohistochemistry (IHC) in ESCC tissue based on PDXs. Results: Shikonin effectively inhibited cell proliferation in dose-dependent and time-dependent manner compared with the control group. The detection of glycolysis showed that Shikonin suppressed the glucose consumption, lactate production, glycolytic intermediates and pyruvate kinase enzymatic activity. Furthermore, Shikonin not only inhibited the growth of ESCC, but also decreased the expression of p-PKM2 and p-STAT3 in vivo. Finally, Shikonin suppressed the expression of GLUT1 and HK2 proteins which are related to glycolysis. Conclusion: Shikonin has a significant antitumor effect in the ESCC by suppressing PKM2 mediated aerobic glycolysis and regulating PKM2/STAT3 signal pathway.


Meniscal Tear at Knee: Repair or Resect? An Early Experience from a University Hospital in Nepal.
Background The meniscus plays an important role in maintaining healthy articular cartilage. Meniscus tear, one of the common intra-articular knee lesions, is treated by either debridement or repair. Objective This study aims at identifying the early outcome of meniscus tears treated by debridement or repair. This study also elaborates on the spectrum of meniscal injuries presented in a tertiary care hospital in Nepal. Method A retrospective descriptive cross-sectional study was conducted at Orthopedic Department of Dhulikhel Hospital from February 2018 until January 2020 among patients who underwent knee arthroscopies for meniscal tears treated either by debridement or repair. Patients having intra articular fractures, osteochondral injuries and multi-ligament injuries were excluded. The meniscal tears were classified according to location and type of tear. Those patients who had at least one-year of follow up were evaluated with Lysholm score for functional outcome. Data were compiled and analyzed with Microsoft Excel 2011. Result One hundred and ten cases of meniscal tears were managed over the study period. Ninty-three cases could be traced for outcome evaluation, which included 50 cases of meniscal debridement and 43 cases of meniscal repair. The mean Lysholm score of the patients who received debridement was 81.5 (SD 10.4) and those who received meniscal repair was 84.9 (SD 9.1) (p=0.105). The population distribution was found to be similar in both the groups according to age and sex distribution and associated ligamentous injuries. Conclusion Good functional outcome was seen for meniscal tears managed with debridement or repair in at least one year follow up and could not establish one modality of management better than the other.


On-board generation of a highly volatile starting fuel to reduce automobile cold-start emissions.
The on-board distillation system (OBDS) was developed to extract, from gasoline, a high-volatility fuel for exclusive use during the starting and warm-up periods. The use of OBDS distillate fuel results in much improved mixture preparation, allowing combinations of air/fuel ratio and ignition timing that are not possible with gasoline, even with a fully warm engine. The volatility of the distillate is a function of the parent fuel volatility; however, the variability in distillate quality can be diminished via manipulation of the OBDS operating conditions. Thus, it is possible to develop aggressive starting calibrations that are relatively immune to variations in pump gasoline volatility. The key benefits provided bythe OBDS fuel relative to standard gasoline were found to be (1) improved mixture preparation allowing a 70% reduction of cranking fuel requirements, elimination of air-fuel mixture enrichment during the warm-up period, and significant extension of warm-up ignition timing retard; (2) a 57% decrease in catalyst light-off time, (3) emissions reductions over the FTP drive cycle of 81% for regulated hydrocarbons (NMOG); (4) emissions index (NMOG) approaching that of SULEV/PZEV vehicles; and (5) an apparent 1% increase in fuel economy over the FTP drive cycle.


Comparisons of pharmacokinetics and NO-releasing of nitrofibriate and fenofibrate after oral administration in rats.
Nitrofibriate, a new compound of hypolipidemic, is modified based on fenofibrate. Both of them are used for prevention and treatment of cardiovascular diseases. In this study, an accurate and sensitive analytical method of reversed-phase high-performance liquid chromatography was developed to determine fenofibric acid, which is an active metabolite of both nitrofibriate and fenofibrate in rat plasma. This method was validated and successfully applied to pharmacokinetic study of nitrofibriate and fenofibrate after oral administration. The results suggested that the pharmacokinetic behavior of nitrofibriate followed a nonlinear process, while fenofibrate was linear, demonstrating that the two drugs were different in pharmacokinetic behaviors. Moreover, the effect of fenofibrate and nitrofibriate on releasing NO in rat serum was explored. This study showed that nitrofibriate, as a nitric oxide donor, could slowly release nitric oxide in vivo. This study provided a biopharmaceutical basis for further study of nitrofibriate.


The application of low shear modeled microgravity to 3-D cell biology and tissue engineering.
The practice of cell culture has been virtually unchanged for 100 years. Until recently, life scientists have had to content themselves with two-dimensional cell culture technology. Clearly, living creatures are not constructed in two dimensions and thus it has become widely recognized that in vitro culture systems must become three dimensional to correctly model in vivo biology. Attempts to modify conventional 2-D culture technology to accommodate 3-D cell growth such as embedding cells in extracellular matrix have demonstrated the superiority of concept. Nevertheless, there are serious drawbacks to this approach including limited mass transport and lack of scalability. Recently, a new cell culture technology developed at NASA to study the effects of microgravity on cells has emerged to solve many of the problems of 3-D cell culture. The technology, the Rotating Wall Vessel (RWV) is a single axis clinostat consisting of a fluid-filled, cylindrical, horizontally rotating culture vessel. Cells placed in this environment are suspended by the resolution of the gravitational, centrifugal and Coriolis forces with extremely low mechanical shear. These conditions, which have been called "low shear modeled microgravity", enable cells to assemble into tissue-like aggregates with high mass transport of nutrients, oxygen and wastes. Examples of the use of the RWV for basic cell biology research and tissue engineering applications are discussed.


Development and evaluation of a multiple-plate fraction collector for sample processing: application to radioprofiling in drug metabolism studies.
Microplate scintillation counters are utilized routinely in drug metabolism laboratories for the off-line radioanalysis of fractions collected during HPLC radioprofiling. In this process, the current fraction collection technology is limited by the number of plates that can be used per injection as well as the potential for sample loss due to dripping or spraying as the fraction collector head moves from well to well or between plates. More importantly, sample throughput is limited in the conventional process, since the collection plates must be manually exchanged after each injection. The Collect PAL, an innovative multiple-plate fraction collector, was developed to address these deficiencies and improve overall sample throughput. It employs a zero-loss design and has sub-ambient temperature control. Operation of the system is completely controlled with software and up to 24 (96- or 384-well) fraction collection plates can be loaded in a completely automated run. The system may also be configured for collection into various-sized tubes or vials. At flow rates of 0.5 or 1.0 mL/min and at collection times of 10 or 15s, the system precisely delivered 83-μL fractions (within 4.1% CV) and 250-μL fractions (within 1.4% CV), respectively, of three different mobile phases into 12 mm × 32 mm vials. Similarly, at a flow rate of 1 mL/min and 10s collection times, the system precisely dispensed mobile phase containing a [(14)C]-radiolabeled compound across an entire 96-well plate (% CV was within 5.3%). Triplicate analyses of metabolism test samples containing [(14)C]buspirone and its metabolites, derived from three different matrices (plasma, urine and bile), indicated that the Collect PAL produced radioprofiles that were reproducible and comparable to the current technology; the % CV for 9 selected peaks in the radioprofiles generated with the Collect PAL were within 9.3%. Radioprofiles generated by collecting into 96- and 384-well plates were qualitatively comparable; however, the peak resolution was greater in the profiles that were collected in 384-well plates due to the collection of a larger number of fractions per minute. In conclusion, this new and innovative fraction collector generated radioprofile results that were comparable to current technology and should provide a major improvement in capacity and throughput for radioprofiling studies.


Enkephalin in the goldfish retina.
Enkephalin-like immunoreactive amacrine cells were visualized using the highly sensitive avidin-biotin method. The somas of these cells were situated in the inner nuclear and ganglion cell layers. Enkephalin-stained processes were observed in layers 1, 3, and 5 of the inner plexiform layer. The biosynthesis of sulfur-containing compounds in the goldfish retina was studied by means of a pulse-chase incubation with 35S-methionine. A 35S-labeled compound, which comigrated with authentic Met5-enkephalin on high-performance liquid chromatography (HPLC), was synthesized and was bound competitively by antibodies to enkephalin and by opiate receptors. This compound was tentatively identified as "Met5-enkephalin." The newly synthesized 35S-Met5-enkephalin was released upon depolarization of the retina with a high K+ concentration. This K+-stimulated release was greatly suppressed by 5 mM Co2+, suggesting that the release was Ca2+ dependent. Using a double-label technique, enkephalin immunoreactivity and gamma-aminobutyric acid (GABA) uptake were colocalized to some amacrine cells, whereas others labeled only for enkephalin or GABA. The possible significance of enkephalin-GABA interactions is also discussed.


miR‑132 inhibits high glucose‑induced vascular smooth muscle cell proliferation and migration by targeting E2F5.
The dysregulated behavior of vascular smooth muscle cells (VSMCs) serves an important role in the pathogenesis of cardiovascular diseases in diabetes. The present study aimed to investigate the effects of microRNA (miR)‑132 on the proliferation and migration of VSMCs under high glucose conditions to mimic diabetes. We observed that the expression of miR‑132 was significantly decreased and that of E2F transcription factor 5 (E2F5) was upregulated in high glucose (HG)‑treated VSMCs or those obtained from diabetic rats. A dual luciferase reporter gene assay revealed that miR‑132 could specifically bind to the 3'‑untranslated region of E2F5 and significantly suppress the luciferase activity. The proliferation and migration of diabetic rat or HG‑treated VSMCs were increased compared with non‑diabetic rat VSMCs and those under normal glucose conditions. Upregulation of miR‑132 significantly inhibited the proliferation and migration of diabetic rat VSMCs; similar effects were observed following E2F5 downregulation. The inhibitory effects of miR‑132 on the proliferation and migration of HG‑treated VSMCs could be reversed by E2F5 overexpression. In conclusion, miR‑132 was proposed to inhibit the proliferation and migration of diabetic rat or high‑glucose‑treated VSMCs by targeting E2F5. The findings of the present study suggested that increasing the expression of miR‑132 may serve as a novel therapeutic approach to inhibit the progression of cardiovascular disease in diabetes.


Plant cystoliths: a complex functional biocomposite of four distinct silica and amorphous calcium carbonate phases.
Plant cystoliths are mineralized objects that are formed by specialized cells in the leaves of certain plants. The main mineral component of cystoliths by volume is amorphous calcium carbonate (ACC) and the minor component is silica. We show that the silica stalk is formed first and is essential for ACC formation. Furthermore, the cystolith is shown to be composed of four distinct mineral phases with different chemical properties: an almost pure silica phase grades into a Mg-rich silica phase. This Mg-rich silica is overlaid by a relatively stable ACC phase. A bulky and less stable ACC phase encapsulates the first ACC phase. This architecture poses interesting questions about the role of Mg in the silica phase and suggests a strategy for ACC stabilization that takes advantage of a precise regulation of the mineral-growth microenvironment.


Identification of contaminant interferences which cause positive urine reagent test strip reactions in a cage setting for the laboratory-housed nonhuman primate, Beagle dog, and Sprague-Dawley rat.
A high incidence of unexplained positive urine reagent test strip reactions was observed in healthy, untreated laboratory-housed nonhuman primates, Beagle dogs, and Sprague-Dawley rats. Exposure of urine to cage pan contaminants was the suspected cause of the positive reactions. The objective of this study was to identify cage pan contaminants which could cause positive reagent test strip reactions. Contaminated urine was simulated by exposing water samples to cage pan contaminants, including cleaning solutions, feces from nonhuman primates, Beagle dogs, and Sprague-Dawley rats, certified laboratory animal diets, and dietary enrichments (vegetables, fruits, and food treats). Ten samples were prepared for each contaminant and analyzed for blood, glucose, bilirubin, ketones, pH, protein, urobilinogen, nitrite, and leukocyte esterase using commercially available urine reagent test strips and an automated urine chemistry analyzer. Positive reactions were common for all but one analyte and frequently associated with multiple contaminants. Blood, glucose, and protein reactions had the highest incidence and/or strongest positive reactions. Positive reactions for other reagent test strip analytes were observed, but generally of lower incidence and magnitude. We identified a high incidence of contaminant interferences in a water matrix causing positive reagent test strip reactions, primarily for the blood, glucose, and protein reactions. These findings highlight the potential limited value of urine reagent test strip assays as reliable biomarkers for detecting kidney toxicity in nonclinical studies, and imply that urine collection methods that minimize exposure to contaminants will likely improve the diagnostic validity of reagent test strip assays.


Quantum mass acquisition in spinor Bose-Einstein condensates.
Quantum mass acquisition, in which a massless (quasi)particle becomes massive due to quantum corrections, is predicted to occur in several subfields of physics. However, its experimental observation remains elusive since the emergent energy gap is too small. We show that a spinor Bose-Einstein condensate is an excellent candidate for the observation of such a peculiar phenomenon as the energy gap turns out to be 2 orders of magnitude larger than the zero-point energy. This extraordinarily large energy gap is a consequence of the dynamical instability. The propagation velocity of the resultant massive excitation mode is found to be decreased by the quantum corrections as opposed to phonons.


Structures of acrolein-guanine adducts: a semi-empirical self-consistent field and nuclear magnetic resonance spectral study.
The structures and conformations of adducts formed by reaction of guanosine with several mutagenic alpha,beta-unsaturated carbonyl compounds have been investigated by semi-empirical molecular orbital calculations and compared with NMR spectral results. Two cyclization processes taking place on the pyrimidine ring of guanine leading to two sets of regioisomers, 11-hydroxy- and 13-hydroxytetrahydropyrimidinoguanines (THPG), were considered. Relative stabilities and geometries of all configurations and conformations of adducts with acrolein, crotonaldehyde, and alpha-chloroacrolein were calculated by PM3, AM1, and MNDO methods. PM3 results were the most compatible with experimental structures based on 400-MHz 1H NMR spectroscopy. The most stable structures for the 11-hydroxy and 13-hydroxy THPG isomers from acrolein are predicted to have chair-like structures for the tetrahydropyrimidine ring and axial hydroxyl groups, as suggested by the NMR spectra of the isolated adducts. Of the possible isomers from guanine and crotonaldehyde, cis-11-hydroxy-13-methyl THPG with methyl and hydroxyl groups axial is predicted to be the most stable. The only isolated adduct is the trans-13-hydroxy-11-methyl THPG with methyl shown to be equatorial and hydroxyl axial by 1H NMR. This is completely consistent with the geometry predicted by PM3 for the 13-hydroxy regioisomer of crotonaldehyde. In the case of adducts of alpha-chloroacrolein, one stereoisomer predominates for each of the two possible regioisomers. For the 12-chloro-11-hydroxy isomer, the cis configuration with chlorine axial and hydroxyl quasi-axial is calculated to have the most stable geometry. In contrast, the 1H NMR spectrum supports a trans diaxial orientation, although the cis computed structure could also be accommodated by the spectrum. The 12-chloro-13-hydroxy regioisomer is unambiguously assigned as trans diaxial by PM3 calculations and 1H NMR spectroscopy.


In vitro culture of tetrathyridia of Mesocestoides corti using a gel based diphasic medium.
Tetrathyridia of Mesocestoides corti were cultured in vitro in a diphasic medium consisting of a liquid medium (CMRL Sigma) and a thixotropic nutrient gel (Oxoid). Tests demonstrated that a 50% medium/gel mixture produced optimum conditions for the survival and development of tetrathyridia. Established anthelminthic drugs were inoculated into the gel which demonstrated that this system can be used for preliminary anthelminthic drug screening. The development and survival of the tetrathyridia were influenced by the addition of pepsin, trypsin and liver peptone to the culture media. The development and maturation of proglottids were observed in addition to asexual reproduction by the process of budding. Tetrathyridia maintained in vitro and reinfected into both mouse and rat hosts retained their viability.


Alcohol, high blood pressure, and serum gamma-glutamyl transpeptidase level.
The influence of the level of serum gamma-glutamyl transpeptidase, a biological marker of alcohol consumption, on elevations of blood pressure and on the development of hypertension related to increases in alcohol consumption was determined in a cross-sectional study of 1,492 middle-aged male workers and in a subsequent 5-year follow-up study of 1,393 workers. Blood pressure levels, as well as the prevalence and incidence of hypertension, were higher in the subjects with serum gamma-glutamyl transpeptidase levels above 50 units/l than in those with normal levels. These differences were more marked in drinkers who consumed 30 ml or more of alcohol per day. Thus, elevated serum gamma-glutamyl transpeptidase activity may identify drinkers at higher risk for the development of alcohol-related hypertension.