Delayed recanalization reduced neuronal apoptosis and neurological deficits by enhancing liver-derived trefoil factor 3-mediated neuroprotection via LINGO2/EGFR/Src signaling pathway after middle cerebral artery occlusion in rats.
Delayed recanalization at days or weeks beyond the therapeutic window was shown to improve functional outcomes in acute ischemic stroke (AIS) patients. However, the underlying mechanisms remain unclear. Previous preclinical study reported that trefoil factor 3 (TFF3) was secreted by liver after cerebral ischemia and acted a distant neuroprotective factor. Here, we investigated the liver-derived TFF3-mediated neuroprotective mechanism enhanced by delayed recanalization after AIS. A total of 327 male Sprague-Dawley rats and the model of middle cerebral artery occlusion (MCAO) with permanent occlusion (pMCAO) or with delayed recanalization at 3 d post-occlusion (rMCAO) were used. Partial hepatectomy was performed within 5 min after MCAO. Leucine-rich repeat and immunoglobulin-like domain-containing nogo receptor-interacting protein 2 (LINGO2) siRNA was administered intracerebroventricularly at 48 h after MCAO. Recombinant rat TFF3 (rr-TFF3, 30 μg/Kg) or recombinant rat epidermal growth factor (rr-EGF, 100 μg/Kg) was administered intranasally at 1 h after recanalization, and EGFR inhibitor Gefitinib (75 mg/Kg) was administered intranasally at 30 min before recanalization. The evaluation of outcomes included neurobehavior, ELISA, western blot and immunofluorescence staining. TFF3 in hepatocytes and serum were upregulated in a similar time-dependent manner after MCAO. Compared to pMCAO, delayed recanalization increased brain TFF3 levels and attenuated brain damage with the reduction in neuronal apoptosis, infarct volume and neurological deficits. Partial hepatectomy reduced TFF3 levels in serum and ipsilateral brain hemisphere, and abolished the benefits of delayed recanalization on neuronal apoptosis and neurobehavioral deficits in rMCAO rats. Intranasal rrTFF3 treatment reversed the changes associated with partial hepatectomy. Delayed recanalization after MCAO increased the co-immunoprecipitation of TFF3 and LINGO2, as well as expressions of p-EGFR, p-Src and Bcl-2 in the brain. LINGO2 siRNA knockdown or EGFR inhibitor reversed the effects of delayed recanalization on apoptosis and brain expressions of LINGO2, p-EGFR, p-Src and Bcl-2 in rMCAO rats. EGFR activator abolished the deleterious effects of LINGO2 siRNA. In conclusion, our investigation demonstrated for the first time that delayed recanalization may enhance the entry of liver-derived TFF3 into ischemic brain upon restoring blood flow after MCAO, which attenuated neuronal apoptosis and neurological deficits at least in part via activating LINGO2/EGFR/Src pathway.


A novel androgen-regulated isoform of the TSC2 tumour suppressor gene increases cell proliferation.
TSC2 (Tuberous sclerosis complex 2) is an important tumour suppressor gene, mutations within which are linked to the development of tuberous sclerosis and implicated in multiple tumour types. TSC2 protein complexes with TSC1 and blocks the ability of the Rheb (Ras homolog enriched in brain) GTPase to activate mTOR (mammalian target of rapamycin), a crucial signal transducer which regulates protein synthesis and cell growth. Here, we report the characterisation of a novel isoform of TSC2 which is under direct control of the ligand-activated androgen receptor. TSC2 isoform A (TSC2A) is derived from an internal androgen-regulated alternative promoter and encodes a 508-amino acid cytoplasmic protein corresponding to the C-terminal region of full-length TSC2, lacking the interaction domain for TSC1 and containing an incomplete interaction domain required for Rheb inactivation. Expression of TSC2A is induced in response to androgens and full-length TSC2 is co-ordinately down-regulated, indicating an androgen-driven switch in TSC2 protein isoforms. In contrast to the well-characterised suppressive effect on cell proliferation of full-length TSC2 protein, both LNCaP and HEK293 cells over-expressing TSC2 isoform A proliferate more rapidly (measured by MTT assays) and have increased levels of cells in S-phase (measured by both Edu staining and FACS analysis). Our work indicates, for the first time, a novel role for this well-known tumour suppressor gene, which encodes an activator of cell proliferation in response to androgen stimulation.


Development of tolerance to ethanol in cultured adrenal chromaffin cells.
Dissociated bovine adrenal chromaffin cells in culture were utilized to study the mechanisms for development of cellular tolerance to ethanol. Three days after plating, cells were grown in either control medium or medium containing 200 mM ethanol for periods up to 6 days. Catecholamine release induced by carbachol, 80 mM K+, BAY K 8644, and veratrine was studied in ethanol-treated and untreated cultures. The presence of ethanol in vitro was inhibitory to all these stimuli in untreated cell cultures, carbachol-induced release being most sensitive (IC50 approximately 30 mM). After growth in ethanol for 6 days, carbachol-induced catecholamine release was completely resistant to the inhibitory effects of ethanol. The resistance to ethanol of carbachol-induced catecholamine release from cells grown in medium containing ethanol also extended to the inhibitory effects of butanol. The results show some similarities with changes observed in the central nervous system during the development of ethanol tolerance, and it is suggested that this model system may be useful in examining the mechanisms by which tolerance occurs.


The neuronal ceroid-lipofuscinoses.
The neuronal ceroid-lipofuscinoses (NCL's, Batten disease) represent a group of severe neurodegenerative diseases, which mostly present in childhood. The phenotypes are similar and include visual loss, seizures, loss of motor and cognitive function, and early death. At autopsy, there is massive neuronal loss with characteristic storage in remaining neurons. Neurons appear to die because of increased rates of apoptosis and altered autophagy. Ten genes have been identified so far that result in an NCL (CLN1-10). The most common forms are CLN1, CLN2, and CLN3, which were previously known as Infantile, Late-Infantile, and Juvenile NCL's, respectively. CLN1 and CLN2 result from mutations in soluble lysosomal enzymes palmitoyl-protein thioesterase (PPT) and tripeptidyl peptidase 1 (TPP1), which can be measured in white blood cells for clinical diagnosis. Molecular diagnostic testing is routinely available for CLN1, CLN2, and CLN3. Sequencing of other NCL genes may be required to establish a diagnosis when the common forms are ruled out. The pathogenesis of NCL neuronal loss resulting from loss of function of any of the NCL gene products remains unknown and no treatment options are presently available.


KELVIN: a software package for rigorous measurement of statistical evidence in human genetics.
This paper describes the software package KELVIN, which supports the PPL (posterior probability of linkage) framework for the measurement of statistical evidence in human (or more generally, diploid) genetic studies. In terms of scope, KELVIN supports two-point (trait-marker or marker-marker) and multipoint linkage analysis, based on either sex-averaged or sex-specific genetic maps, with an option to allow for imprinting; trait-marker linkage disequilibrium (LD), or association analysis, in case-control data, trio data, and/or multiplex family data, with options for joint linkage and trait-marker LD or conditional LD given linkage; dichotomous trait, quantitative trait and quantitative trait threshold models; and certain types of gene-gene interactions and covariate effects. Features and data (pedigree) structures can be freely mixed and matched within analyses. The statistical framework is specifically tailored to accumulate evidence in a mathematically rigorous way across multiple data sets or data subsets while allowing for multiple sources of heterogeneity, and KELVIN itself utilizes sophisticated software engineering to provide a powerful and robust platform for studying the genetics of complex disorders.


Antioxidant vitamin supplementation in cardiovascular diseases.
Cardiovascular disease is the most important adult health problem in wealthy countries, where biological factors such as obesity, hypertension, dyslipidemia, diabetes, inappropriate diet, cigarette smoking, and sedentary life-style have contributed to its dissemination. Research concerning nutritional regimens has shown that persons who consume large amounts of fruit and vegetables have lower incidences of cardiovascular diseases, stroke, and tumors, although the precise mechanisms for this protective effect are elusive. Possible explanations include (a) increased consumption of dietary fiber, (b) reduced consumption of dietary cholesterol and other lipids, and (c) increased intake of the antioxidant vitamins (A, C, and E). Numerous studies have raised the question whether vitamin supplements help to prevent cardiovascular diseases. Results of randomized controlled trials of antioxidant vitamin supplements in large numbers of participants has been ambiguous or contradictory. This minireview examines the relevant clinical reports on dietary supplements of vitamins A, C, and E to determine whether they support the premise that patients at risk of cardiovascular disease may be candidates for this therapeutic option.


Virus isolations from pet birds submitted for importation into the United States.
From October 1973 to September 1981, 2,882,111 birds were offered for importation into the United States. All were quarantined for 30 days, and specimens were submitted to the laboratory for virus-isolation studies. Viruses were isolated from specimens from 26.3% (598/2,274) of the quarantined lots of birds. Viscerotropic velogenic Newcastle disease virus (VVNDV) was isolated from 141 lots. Nonviscerotropic velogenic Newcastle disease virus (VNDV) was isolated from six lots. All VVNDV- and VNDV-positive lots were refused entry. The percentage of lots refused entry declined from 31.6% in 1974 to 2.9% in 1981. Mesogenic Newcastle disease virus (NDV) was isolated from three lots, and lentogenic NDV was isolated from 23 lots. Lots positive for mesogenic and lentogenic NDV were allowed to enter the United States. Hemagglutinating (HA) viruses other than NDV were isolated from 24.5% (373/1,521) of the lots imported through privately owned quarantine facilities. Of the 8,563 HA viruses isolated, 1,558 were selected for identification. Forty-six percent were identified as paramyxovirus (PMV)-2, 34% were PMV-3, and 20% were influenza A viruses possessing the hemagglutinin subtypes H3, H4, H7, and H10 and the neuraminidase subtypes N1, N6, N7, and N8. The frequency of PMV-2 and PMV-3 isolations fluctuated from year to year, whereas the frequency of isolations of influenza A viruses decreased from 64% in 1974 to 0.2% in 1981. Viruses that did not agglutinate chicken red blood cells were isolated from 52 lots. Psittacine herpesvirus (Pacheco's disease) was isolated from 25 lots of psittacines. Viruses identified by electron microscopy as reoviruses were isolated from 24 lots.(ABSTRACT TRUNCATED AT 250 WORDS)


Bedside troubleshooting during venovenous extracorporeal membrane oxygenation (ECMO).
In this review, we discuss common difficulties that clinicians may encounter while managing patients treated with venovenous (VV) extracorporeal membrane oxygenation (ECMO). ECMO is an increasingly important tool for managing severe respiratory failure that is refractory to conventional therapies. Its overall goal is to manage respiratory failure-induced hypoxemia and hypercarbia to allow "lung rest" and promote recovery. Typically, by the time VV-ECMO is initiated, the patient's pulmonary condition requires conventional ventilator settings that are detrimental to lung recovery or that exceed the remaining functional lung's ability to maintain acceptable physiological conditions. Standard mechanical ventilation can activate inflammation and worsen the pulmonary damage caused by the underlying disease, leading to ventilator-induced lung injury. In contrast, VV-ECMO facilitates lung-protective ventilation, decreasing further ventilator-induced lung injury and allowing lung recovery. Such lung-protective ventilation seeks to avoid barotrauma (by monitoring transpulmonary pressure), volutrauma (by reducing excessive tidal volume to promote lung rest), atelectotrauma [by maintaining adequate positive end-expiratory pressure (PEEP)], and oxygen toxicity (by decreasing ventilator oxygen levels when PEEP is adequate). ECMO for adult respiratory failure was associated with overall survival of 62% in 2018, according to the Extracorporeal Life Support Organization (ELSO) January 2019 registry report. Difficulties that may arise during VV-ECMO require timely diagnosis and optimal management to achieve the most favorable outcomes. These difficulties include ventilation issues, hypoxemia (especially as related to recirculation or low ECMO-flow-to-cardiac-output ratio), sepsis, malfunctioning critical circuit components, lack of clarity regarding optimal hemoglobin levels, hematological/anticoagulation complications, and right ventricular (RV) dysfunction. A culture of safety should be emphasized to optimize patient outcomes. A properly functioning team-not only the bedside clinician, but also nurses, perfusionists, respiratory therapists, physical therapists, pharmacists, nutritionists, and other medical specialists and allied health personnel-is vital for therapeutic success.


[Long-term intermittent renal replacement therapy at an intensive care unit].
Standard intermittent hemodialysis (IHD) used for the treatment of acute renal failure (ARF) at an intensive care unit has significant biochemical and physiological drawbacks. In the past 20 years, these drawbacks have stimulated the development of continuous renal replacement therapy (CRRT) and its ever-increasing use. However, CRRT is technically complicated and requires 24-hour monitoring. In some clinics, the use of CRRT leads to that each patient is under his/her nurse's surveillance, instead 1 nurse per 2 patients as before; this change has economic consequences and may limit nursing accessibility to other patients. The procedures prolonging intermittent therapy do not require 24-hour monitoring may benefit the treatment of ARF at the intensive care therapy. In this paper the authors call such procedures for continuous intermittent renal replacement therapy. They are characterized by a number of basic principles: (1) the use of modified or standard dialysis apparatuses; (2) the application of diffuse, convection, or both; (3) a certain reduction in the rate of elimination of dissolved substances as compared with IHD; (4) more prolonged treatment: above usual 3 or 4 hours of IHD, but not more than 8-12 hours (hence the term "intermittent"); (5) the use of on-line generation dialysate or substituting fluid. Information on the effectiveness and safety of this procedure is being now compiled.


Hypoxia protects HepG2 cells against etoposide-induced apoptosis via a HIF-1-independent pathway.
Tumor hypoxia has been described to increase the resistance of cancer cells to radiation therapy and chemotherapy. It also supports the invasiveness and metastatic potential of the tumor. However, few data are available on the transduction pathway set up under hypoxia and leading to this resistance against anti-cancer therapies. HIF-1, the main transcription factor activated by hypoxia, has been recently shown to participate to this process although its role as an anti- or a pro-apoptotic protein is still controversy. In this study, we showed that hypoxia protected HepG2 cells against etoposide-induced apoptosis. The effect of hypoxia on cell death was assayed by measuring different parameters of the apoptotic pathway, like DNA fragmentation, caspase activity and PARP-1 cleavage. The possible implication of HIF-1 in the anti-apoptotic role of hypoxia was investigated using HIF-1alpha siRNA. Our results indicated that HIF-1 is not involved in the hypoxia-induced anti-apoptotic pathway. Another transcription factor, AP-1, was studied for its potential role in the hypoxia-induced protection against apoptosis. Specific inhibition of AP-1 decreased the protection effect of hypoxia against etoposide-induced apoptosis. Together, all these data underline that hypoxia could mediate its anti-apoptotic role via different transcription factors depending on the cellular context and pro-apoptotic stimuli.


From intestinal metaplasia to gastric cancer: Witnessing the rise of evil over time and space.
Intestinal metaplasia (IM) is a precancerous lesion associated with increased gastric cancer (GC) risk. However, the molecular characteristics and heterogeneity distinguishing the two stages remain unclear. Huang et al. provide a spatiotemporal insight into the transition from IM to GC, offering the potential for tailored precision prevention strategies for GC.


Hibernating myocardium.
According to estimates, up to 50% of patients with coronary artery disease and impaired left ventricular function have areas of viable myocardium. This dysfunctional, yet viable myocardial tissue, which can improve functionally after myocardial oxygen supply is reestablished, has been called hibernating myocardium. The possible pathophysiological mechanism that leads to hibernating myocardium is controversial: is the phenomenon due to persistent ischemia or is it the result of repetitive episodes of ischemia and reperfusion, such as myocardial stunning? Regardless of the mechanism, the presence of viable myocardial tissue indicates that structural and biochemical cellular changes occur, and the recovery of left ventricular function after revascularization depends on the severity and extent of these changes. Whether these changes reflect a long-lasting state of cellular dedifferentiation, an adaptive process that is reversible, or eventually lead to cellular degeneration has not been determined. Perhaps early detection of hibernating myocardial tissue via noninvasive imaging techniques used to assess contractile response, integrity of the cellular membrane, myocardial metabolism, and myocardial blood flow and subsequent early coronary revascularization may prevent infarction and deterioration in left ventricular function. Knowledge that reversible changes and areas of viable myocardium can occur in patients with left ventricular dysfunction will assist healthcare providers in the care and management of patients with hibernating myocardium.


Management of residual pleural space after lung resection: fully controllable paralysis of the diaphragm through continuous phrenic nerve block.
Residual pleural space after lung resection associated with air leak is a challenging issue, potentially causing serious complications. We report a new, postoperative technique to reduce the pleural space, inducing a controlled and reversible paralysis of the diaphragm. Ten patients were enrolled (7 lobectomies, 2 bilobectomy, 1 wedge resection). Inclusion criteria were: digitally detected air flow >200 mL/min at post-op day 3, presence of empty pleural space at chest x-ray, absence of restrictive lung disease, absence of known arrhythmias. A 22G nerve-block catheter was place under ultrasound guidance in proximity to the phrenic nerve, between the sternocleidomastoid muscle and the anterior scalene muscle at the level of 6th cervical vertebra. Continuous infusion of ropivacaine 0.2% 3 mL/h was started. Fluoroscopy was used to confirm significant reduction in hemidiaphragm movements. Monitoring of vital signs and intense respiratory physiotherapy were enhanced. The infusion was stopped at air leak cessation and the catheter was removed along with the chest drain. No peri- and post-procedural complications occurred. In all patients, we observed an immediate reduction of the empty pleural space and resolution of the air leak within few days (3±1.16 days). After suspension of local anaesthetic, complete restoration of the hemidiaphragm function has been documented. This is an effective and minimally invasive method to reduce the residual pleural space after lung resections. Narrowing of the pleural space facilitates the contact between the lung and the chest wall promoting the resolution of the air leak. Diaphragm paralysis is controlled and temporary with no residual disabilities.


Neuronal and physiological correlation to hemodynamic resting-state fluctuations in health and disease.
Low-frequency, spatially coherent fluctuations present in functional magnetic resonance imaging time series have had a tremendous impact on brain connectomics. This work aims to explore the degree with which hemodynamic connectivity is associated with neuronal, metabolic, and vascular connectivity measures. For this purpose, GCaMP and nontransgenic mice were used to image neuronal activity and oxidative metabolism activity, respectively, along with blood-oxygenation- and cerebral blood volume (CBV)-sensitive hemodynamic changes from the same animals. Although network clusters calculated using either GCaMP (neuronal activity) or optical imaging of intrinsic signal (OIS)-BOLD (blood oxygenation) data did not exhibit strong spatial similarity, the strengths of node-to-node connectivity measured with these modalities were strongly correlated with one another. This finding suggests that hemodynamic connectivity as measured by blood oxygenation measurements, such as functional connectivity magnetic resonance imaging, is a valuable surrogate for the underlying neuronal connectivity. In nontransgenic animals, greater connectivity correlation was observed between tissue oxidative metabolism (flavoprotein autofluorescence imaging [FAI]) and blood oxygenation measurements, suggesting that metabolic contributions to hemodynamic signals are likely responsible for its significant correlation with neuronal connectivity. Lastly, a mouse model of Alzheimer's disease was used to explore the source of decreases in connectivity reported in these mice, a finding that is thought to be associated with amyloid load-driven metabolic decline. The intercluster connectivity measured by metabolic-sensitive measurements (FAI and OIS-BOLD) was maintained while vascular-only signals (OIS-CBV) provided negligible correlation. Therefore, metabolism-sensitive measurements as used in this work are better positioned to capture changes in neuronal connectivity, such that decreases in hemodynamic connectivity likely reflect decreases in oxidative metabolic function.


Optical chromatography using a photonic crystal fiber with on-chip fluorescence excitation.
We describe the realization of integrated optical chromatography, in conjunction with on-chip fluorescence excitation, in a monolithically fabricated poly-dimethylsiloxane (PDMS) microfluidic chip. The unique endlessly-single-mode guiding property of the Photonic Crystal Fiber (PCF) facilitates simultaneous on-chip delivery of beams to perform optical sorting in conjunction with fluorescence excitation. We use soft lithography to define the chip and insert the specially capped PCF into it through a predefined fiber channel that is intrinsically aligned with the sorting channel. We compare the performance of the system to a standard ray optics model and use the system to demonstrate both size-driven and refractive index-driven separations of colloids. Finally we demonstrate a new technique of enhanced optofluidic separation of biological particles, by sorting of human kidney embryonic cells (HEK-293), internally tagged with fluorescing microspheres through phagocytocis, from those without microspheres and the separation purity is monitored using fluorescence imaging.


Volumetric integration model of the Stiles-Crawford effect of the first kind and its experimental verification.
The integrated Stiles-Crawford function is commonly used as apodization model for vision through the natural eye pupil. However, this method does not account for possible effects related to the retinal thickness, the large length-to-diameter aspect ratio of the photoreceptors, or the use of nonMaxwellian illumination. Here, we introduce a geometrical optics model to calculate the fraction of overlap between light at the retina and the photoreceptor outer segments where absorption triggers vision. The model, which does not account for photoreceptor waveguiding, is discussed for both Maxwellian and nonMaxwellian illumination. The integrated Stiles-Crawford effect is analyzed experimentally with a uniaxial pupil-size flicker methodology and we find that the psychophysical measurements match better to the geometrical optics predictions than direct integration of a Stiles-Crawford function.


What affects the quality of economic analysis for life-saving investments?
Economic analysis of life-saving investments in both the public and private sectors has the potential to dramatically improve longevity and the quality of life, but only if the analyses on which decisions are based are done well. In this article, we analyze a data set that provides information on the content and quality of journal articles that measure the cost-effectiveness of life-saving investments. Our study is the first to provide a detailed multivariate analysis of factors affecting objective measures of quality. We also explore whether a series of recommendations by an expert panel convened by the U.S. Public Health Service affect the way analyses of specific life-saving investments are done. Our results suggest that four factors are positively correlated with an index we construct to measure analytical quality: (1) having at least one author affiliated with a university, (2) publication in a journal that has experience in publishing these analyses, (3) if the life-saving investment is located in the United States, and (4) if the analysis considers a measure of social costs or benefits. Somewhat surprisingly, a study's funding source and whether it is affiliated with industry are not significantly correlated with the quality index. Finally, neither time nor the panel guidelines had an impact on the index.


High-dynamic-range single-shot cross-correlator based on an optical pulse replicator.
The operation of a single-shot cross-correlator based on a pulse replicator is described. The correlator uses a discrete sequence of sampling pulses that are nonlinearly mixed with the pulse under test. The combination of a high reflector and partial reflector replicates an optical pulse by multiple internal reflections and generates a sequence of spatially displaced and temporally delayed sampling pulses. This principle is used in a cross-correlator characterizing optical pulses at 1053 nm. A dynamic range higher than 60 dB is obtained over a temporal range larger than 200 ps.


Molecular architecture of the inner membrane of mitochondria from rat liver: a combined biochemical and stereological study.
The molecular structure of mitochondria and their inner membrane has been studied using a combined approach of stereology and biochemistry. The amount of mitochondrial structures (volume, number, surface area of inner membrane) in a purified preparation of mitochondria from rat liver was estimated by stereological procedures. In the same preparation, the oxidative activity of the respiratory chain with different substrates and the concentration of the redox complexes were measured by biochemical means. By relating the stereological and biochemical data, it was estimated that the individual mitochondrion isolated from rat liver has a volume of 0.27 micron 3, an inner membrane area of 6.5 micron 2, and contains between 2,600 (complex I) and 15,600 (aa3) redox complexes which produce an electron flow of over 100,000 electrons per second with pyruvate as substrate. The individual redox complexes and the H+-ATPase together occur at a density of approximately 7,500/micron 2 and occupy approximately 40% of the inner membrane area. From the respective densities it was concluded that the mean nearest distance between reaction partners is small enough (70-200 A) to cause the formation of micro-aggregates. The meaning of these results for the mechanism of mitochondrial energy transduction is discussed.


Improved motor function with Bowen therapy for rehabilitation in chronic stroke: a pilot study.
Bowen therapy is an established complementary therapy with anecdotal reports of effectiveness. However, there is limited published research to substantiate safety or effectiveness. This is a pilot study to explore the potential impact of Bowen therapy in chronic stroke. A case series of 14 people with chronic stroke were offered 13 sessions of Bowen therapy over a three-month period. Motor assessments of the 13 people who participated showed improvements--gross motor function trended to improvement; SF-36 role-physical, physical health summary scale and total SF-36 scores showed statistically significantly improvements. However, grip strength reduced. In this pilot study, Bowen therapy was associated with improvements in neuromuscular function in people with chronic stroke. At this stage of study, it is not possible to conclude that there is definite benefit; however the results suggest that exploration through further research is appropriate.


Inhaled Insulin: A Clinical and Historical Review.
Insulin is the most effective blood glucose lowering agent and remains one of the cornerstones of diabetes management. However, many individuals with diabetes are either reluctant to initiate or are nonadherent to their insulin therapy for various reasons, including fear of frequent injections. Technosphere Insulin (TI) is a novel inhaled insulin powder that is approved by the United States Food and Drug Administration for the management of diabetes. The results from 2 phase III clinical trials have shown that TI was noninferior to subcutaneous insulin aspart and superior to inhaled placebo in lowering HbA1c in patients with diabetes mellitus types 1 and 2, respectively. Across both studies, TI appears to be generally well tolerated, with the most common adverse events being hypoglycemia and cough. However, long-term pulmonary safety concerns have not been addressed and additional studies are needed. Overall, TI appears to be a promising noninvasive prandial insulin alternative for individuals with diabetes who are at risk for medication nonadherence due to aversion to frequent injections. This article provides a review of the historical development of TI, its safety and efficacy data, and its advantages and disadvantages over traditional injectable insulins.


Cell death during corneal storage at 4 degrees C.
To evaluate cell death in human donor corneas stored at 4 degrees C, to determine whether terminal deoxynucleotidyl transferase-mediated dUTP-fluorescein nick-end labeling (TUNEL) discriminates between apoptosis and necrosis in corneas stored at 4 degrees C. Ten human corneas were stored in Optisol (Chiron Ophthalmics, Irvine, CA) at 4 degrees C for periods ranging from 0 to 21 days and then fixed for histologic examination. Central corneal sections from each cornea were examined by transmission electron microscopy (TEM) and by the TUNEL assay. Electron micrographs of at least 15 keratocytes each from the anterior, middle, and posterior stroma were examined by three masked observers who graded each cell as normal, apoptotic, or necrotic. Central sections from the same corneas were processed by the TUNEL assay and evaluated with a laser scanning confocal microscope to determine the percentage of apoptotic cells. By TEM, apoptosis occurred in 23% of the keratocytes and necrosis in 12%. By TUNEL assay, apoptosis occurred in 11% of the keratocytes, with the results in individual corneas being similar to the findings by TEM for apoptosis, rather than for necrosis. By TUNEL assay, apoptosis occurred in 13% of the epithelial cells and in 8% of the endothelial cells. The percentage of apoptotic cells and storage time correlated significantly for the epithelium, but not for the keratocytes or endothelium in this small sample. Both apoptosis and necrosis occur in cells during corneal storage at 4 degrees C, with apoptosis appearing to predominate. The TUNEL assay identifies cells undergoing apoptosis, but not necrosis, in corneal tissue. Inhibition of apoptosis in corneas stored at 4 degrees C may prolong acceptable storage times.


Eating disorder symptoms and obesity at the intersections of gender, ethnicity, and sexual orientation in US high school students.
We examined purging for weight control, diet pill use, and obesity across sexual orientation identity and ethnicity groups. Anonymous survey data were analyzed from 24 591 high school students of diverse ethnicities in the federal Youth Risk Behavioral Surveillance System Survey in 2005 and 2007. Self-reported data were gathered on gender, ethnicity, sexual orientation identity, height, weight, and purging and diet pill use in the past 30 days. We used multivariable logistic regression to estimate odds of purging, diet pill use, and obesity associated with sexual orientation identity in gender-stratified models and examined for the presence of interactions between ethnicity and sexual orientation. Lesbian, gay, and bisexual (LGB) identity was associated with substantially elevated odds of purging and diet pill use in both girls and boys (odds ratios [OR] range = 1.9-6.8). Bisexual girls and boys were also at elevated odds of obesity compared to same-gender heterosexuals (OR = 2.3 and 2.1, respectively). Interventions to reduce eating disorders and obesity that are appropriate for LGB youths of diverse ethnicities are urgently needed.


Cardiovascular effects of matrine isolated from the Chinese herb Shan-dou-gen.
Matrine, a pure compound isolated from the Chinese herb Shan-don-Gen (Sophora subprostrata), was studied for its effects on the cardiovascular system of the rat. Intravenous injections of matrine at doses from 5 mg to 20 mg/kg body weight exhibited dose-dependent hypotensive and bradycardiac effects. These effects lasted only 1 to 3 min. In the isolated atria and ventricle preparations, matrine at doses from 50 micrograms to 200 micrograms/ml increased the amplitudes of spontaneous contraction of the atria and electrically induced contraction of the ventricle, whereas the frequency of the spontaneous beating of the atria was reduced. The dose-dependent effects of matrine on the isolated preparations persisted as long as the compound was present. Tachyphylaxis was not observed with repeated applications of this compound to the isolated preparations. The positive inotropic effects on both atria and ventricle and the negative chronotropic effect on spontaneous beating of the atria by matrine were not blocked by diphenhydramine, atropine, phenoxybenzamine, propranolol, trifluoperazine, or methysergide. In contrast, verapamil significantly reduced the positive inotropic effect of matrine on the ventricle. In the isolated aortic strip preparation, matrine at a dose of 200 micrograms/ml led to a slight increase in muscle tone. The same dose of matrine induced a 35% increase of perfusion pressure in the hindleg perfusion model. These results suggest that the in vivo transient hypotensive effect of matrine is likely associated with a decrease in heart rate itself, since positive inotropic effects on both the atria and the ventricle, and vasoconstriction of some vascular beds could not be the cause of hypotension.(ABSTRACT TRUNCATED AT 250 WORDS)


Religiosity, values, and horizontal and vertical individualism-collectivism: a study of Turkey, the United States, and the Philippines.
The authors examined the links between two dimensions that have been useful in understanding cross-cultural differences and similarities, namely, individualism-collectivism (I-C) and value orientations. The authors examined the relations and parallels between the two variables by directly relating them and examining the patterns of relations that both have with a third variable, religiosity. Participants were 475 college students from the Philippines, the United States, and Turkey who responded to measures of horizontal and vertical I-C, value orientations, and religiosity. The authors found partial support for the parallels between I-C and value types, particularly for collectivism and conservative values. Moreover, religiosity was associated positively with conservative values and collectivism, across all three cultures. The authors found individualism to also relate to openness-to-change values, though the patterns were not as consistent as those that they found between collectivism and conservation. Differences and similarities emerged in links of I-C-values to religiosity across the three samples.


A position effect on TRPS1 is associated with Ambras syndrome in humans and the Koala phenotype in mice.
Ambras syndrome (AS) is a rare form of congenital hypertrichosis with excessive hair on the shoulders, face and ears. Cytogenetic studies have previously implicated an association with rearrangements of chromosome 8. Here we define an 11.5 Mb candidate interval for AS on chromosome 8q based on cytogenetic breakpoints in three patients. TRPS1, a gene within this interval, was deleted in a patient with an 8q23 chromosomal rearrangement, while its expression was significantly downregulated in another patient with an inversion breakpoint 7.3 Mb downstream of TRPS1. Here, we describe the first potential long-range position effect on the expression of TRPS1. To gain insight into the mechanisms by which Trps1 affects the hair follicle, we performed a detailed analysis of the hair abnormalities in Koa mice, a mouse model of hypertrichosis. We found that the proximal breakpoint of the Koa inversion is located 791 kb upstream of Trps1. Quantitative real-time polymerase chain reaction, in situ hybridization and immunofluorescence analysis revealed that Trps1 expression levels are reduced in Koa mutant mice at the sites of pathology for the phenotype. We determined that the Koa inversion creates a new Sp1 binding site and translocates additional Sp1 binding sites within a highly conserved stretch spanning the proximal breakpoint, providing a potential mechanism for the position effect. Collectively, these results describe a position effect that downregulates TRPS1 expression as the probable cause of hypertrichosis in AS in humans and the Koa phenotype in mice.


The epidemiology of hepatitis C infection in the United States.
The prevalence of hepatitis C virus (HCV) infection in the United States has remained constant from 1988 through 2002, although the peak age of infection has increased. While the number of new HCV cases is declining, the rates of HCV-associated morbidity and mortality are increasing. We reviewed the risk factors for HCV infection, the laboratory methods used to diagnose it, the dynamics of disease progression, and the natural history of HCV infection. Medline searches were performed using the key word HCV, together with incidence, risk factors, demographics, diagnostic methods, disease progression, natural history, normal alanine aminotransferase (ALT), fibrosis, and hepatocellular carcinoma (HCC). Three characteristics-abnormal serum ALT, history of injection drug use, and blood transfusion before 1992-identified 85% of HCV-positive individuals 20-59 years old. About 75%-85% of acutely infected individuals progress to chronic infection, with up to 20% developing liver cirrhosis over 20-25 years, putting them at increased risk for end-stage liver disease and/or HCC. HCV-associated cirrhosis is the leading cause of liver transplantation in the United States. Rates of infection are higher in non-Hispanic blacks than in non-Hispanic whites and Mexican Americans and higher in men than in women. In the United States, over 70% of HCV-infected individuals are infected with genotype 1. HCV infection is more prevalent than human immunodeficiency virus or hepatitis B virus infection and is particularly common among certain demographic groups. Individual rates of fibrosis progression vary, but identification of host and viral characteristics associated with disease progression may reveal the mechanisms of HCV-associated hepatic fibrosis/cirrhosis.


Vorapaxar expands antiplatelet options. Which patients may benefit from thrombin receptor antagonism?
Vorapaxar is the first substance of a new class of antiplatelet drugs that has been tested in large clinical trials. The protease-activated receptor 1 (PAR-1) antagonist inhibits thrombin-induced platelet activation to prevent atherothrombosis. In the phase 3 trials TRACER (acute coronary syndrome) and TRA 2P-TIMI 50 (stable atherosclerosis) reducing ischemic events with vorapaxar came at the cost of bleeding. TRACER compared vorapaxar to placebo in 12,944 patients who had non-ST-segment elevation acute coronary syndromes on top of contemporary treatment including dual antiplatelet therapy (aspirin and clopidogrel). Vorapaxar reduced ischemic events non-significantly, but increased bleeding significantly, therefore not justifying triple antiplatelet therapy in this setting. Follow-up was stopped early because of bleeding. TRA 2P-TIMI 50 examined 26,449 patients who had a history of myocardial infarction, ischemic stroke, or peripheral arterial disease. Vorapaxar reduced ischemic events and increased bleeding both significantly. Recruitment of patients with prior stroke was stopped early. Net clinical outcome and subgroup analyses suggested that vorapaxar could be beneficial for patients with prior myocardial infarction - but no history of stroke.


Thoracic endovascular aortic repair - a Brazilian experience in 255 patients over a period of 112 months.
The aim of this study is to analyze the immediate and late evolution for death and reintervention in a thoracic endovascular aortic repair (TEVAR) group, over a follow-up period of 112 months. Retrospective data of 255 patients, from 1998 to 2007, were obtained. The most prevalent diseases were thoracic aortic aneurysms (89), thoracic and abdominal aneurysms (85) and thoracic aortic dissections (61). The mean age was 63.2 years and 67.1% were male. Three hundred and three endoprostheses were used. Causes of morbidity, in the immediate postoperative period, were hyperthermia (45.9%), endoleaks (9.8% - being 7.1% type I), vascular complications (5.2%), renal insufficiency (3.1%) and neurological complications (3.1%). There were two (0.8%) hospital deaths and 17 (6.7%) late deaths. Time of follow-up was up to 112 months (mean of 60 months). The Kaplan-Meier curve analysis showed an increase of reintervention, compared with death, after a follow-up period of 42 months. Freedom from death at 36, 60 and 112 months was 96%, 89.1%, 85.1% and for reintervention, for the same periods was 93.6%, 82.7%, 57.2%, respectively. This study showed low incidence of prostheses related morbidity and immediate mortality. After a period of 42 months there was an increase on the percentual tax of reintervention.


Vascular endothelial growth factor (VEGF) as a target of bevacizumab in cancer: from the biology to the clinic.
Angiogenesis is important in the growth and progression of solid tumours. The main pro-angiogenic factor, namely vascular endothelial growth factor (VEGF), also known as vascular permeability factor, is a potent angiogenic cytokine that induces mitosis and also regulates the permeability of endothelial cells. The soluble isoform of VEGF is a dimeric glycoprotein of 36-46 kDa, induced by hypoxia and oncogenic mutation and it binds to two specific tyrosine-kinase receptors: VEGF-1 (flt-1) and VEGF-2 (KDR/flk1). An increase in VEGF expression in tumour tissue or some blood compartments (i.e. serum or plasma) has been found in solid and haematological malignancies of various origins and is associated with metastasis formation and poor prognosis. Bevacizumab, a recombinant humanised monoclonal antibody developed against VEGF, binds to soluble VEGF, preventing receptor binding and inhibiting endothelial cell proliferation and vessel formation. Pre-clinical and clinical studies have shown that bevacizumab alone or in combination with a cytotoxic agent decreases tumour growth and increases median survival time and time to tumour progression. Bevacizumab is the first anti-angiogenetic treatment approved by the American Food and Drug Administration in the first-line treatment of metastatic colorectal cancer. It has shown preliminary evidence of efficacy for breast, non-small-cell lung, pancreatic, prostate, head and neck and renal cancer as well as haematological malignancies. Common toxicities associated with bevacizumab include hypertension, proteinuria, bleeding episodes and thrombotic events. This review summarises the critical role of VEGF and discusses the data available on bevacizumab, from the humanisation of its parent murine monoclonal antibody (mAb) A.4.6.1 to its use in cancer clinical trials.


Cardiac arrest and therapeutic hypothermia decrease isoform-specific cytochrome P450 drug metabolism.
Mild therapeutic hypothermia is emerging clinically as a neuroprotection therapy for individuals experiencing cardiac arrest (CA); however, its effects combined with disease pathogenesis on drug disposition and response have not been fully elucidated. We determined the activities of four major hepatic-metabolizing enzymes (CYP3A, CYP2C, CYP2D, and CYP2E) during hypothermia after experimental CA in rats by evaluating the pharmacokinetics of their probe drugs as a function of altered body temperature. Animals were randomized into sham normothermia (37.5-38°C), CA normothermia, sham hypothermia (32.5-33°C), and CA hypothermia groups. Probe drugs (midazolam, diclofenac, dextromethorphan, and chlorzoxazone) were given simultaneously by intravenous bolus after temperature stabilization. Multiple blood samples were collected between 0 and 8 h after drug administration. Pharmacokinetic (PK) analysis was conducted using a noncompartmental approach and population PK modeling. Noncompartmental analysis showed that the clearance of midazolam (CYP3A) in CA hypothermia was reduced from sham normothermia rats (681.6 ± 190.0 versus 1268.8 ± 348.9 ml · h(-1) · kg(-1), p < 0.05). The clearance of chlorzoxazone (CYP2E) in CA hypothermia was also reduced from sham normothermia rats (229.6 ± 75.6 versus 561.89 ± 215.9 ml · h(-1) · kg(-1), p < 0.05). Population PK analysis further demonstrated the decreased clearance of midazolam (CYP3A) was associated with CA injury (p < 0.05). The decreased clearance of chlorzoxazone (CYP2E1) was also associated with CA injury (p < 0.01). Hypothermia was found to be associated with the decreased volume of distribution of midazolam (V(1)), dextromethorphan (V(1)), and peripheral compartment for chlorzoxazone (V(2)) (p < 0.05, p < 0.05, and p < 0.01, respectively). Our data indicate that hypothermia, CA, and their interaction alter cytochrome P450-isoform specific activities in an isoform-specific manner.


Regulation of Arabidopsis COPINE 1 gene expression in response to pathogens and abiotic stimuli.
The copines are a widely distributed class of calcium-dependent, phospholipid-binding proteins of undetermined biological function. Mutation of the Arabidopsis CPN1 (COPINE 1) gene causes a humidity-sensitive lesion mimic phenotype with increased resistance to a bacterial and an oomyceteous pathogen, constitutive pathogenesis-related gene expression, and an accelerated hypersensitive cell death defense response. Here, we show that the disease resistance phenotype of the cpn1-1 mutant was also temperature sensitive, demonstrate increased CPN1 gene transcript accumulation in wild-type plants under low-humidity conditions, and present a detailed analysis of CPN1 gene transcript accumulation in response to bacterial pathogens. In wild-type plants, CPN1 transcript accumulation was rapidly, locally, and transiently induced by both avirulent and virulent strains of Pseudomonas syringae pv tomato bacteria. However, induction of CPN1 transcript accumulation by avirulent bacteria was much faster and stronger than that induced by virulent bacteria. Bacterial induction of CPN1 transcript accumulation was dependent on a functional type III bacterial protein secretion system. In planta expression of the avrRpt2 avirulence gene was sufficient to trigger rapid CPN1 transcript accumulation. CPN1 transcript accumulation was induced by salicylic acid treatment but was not observed during lesion formation in the lesion mimic mutants lsd1 and lsd5. These results are consistent with CPN1 playing a role in plant disease resistance responses, possibly as a suppressor of defense responses including the hypersensitive cell death defense response. The results also suggest that CPN1 may represent a link between plant disease resistance and plant acclimation to low-humidity and low-temperature conditions.


Implementation of an intensified antibiotic stewardship programme targeting third-generation cephalosporin and fluoroquinolone use in an emergency medicine department.
Early initiation of antimicrobial treatment for acute infection is an important task in the emergency department (ED) with a likely impact on the hospital-wide antibiotic use pattern. We implemented an antibiotic stewardship (ABS) programme focused on non-trauma emergency patients at a large university hospital centre targeting broad-spectrum cephalosporin and fluoroquinolone use. Guidelines and focused discussion groups emphasised reduced prescription of third-generation cephalosporins and fluoroquinolones and encouraged penicillins. Antibiotic consumption expressed as monthly drug density in WHO-Anatomical Therapeutic Chemical defined and locally recommended daily doses (DDD and RDD) per 100 patient days was analysed before (January 2008 to October 2011) and after starting the intervention (January 2012 to October 2013). We performed a before-and-after uncontrolled interventional study using interrupted time-series (ITS) analysis in one ED to investigate ABS intervention-related effects in a quasiexperimental research setting. The mean monthly total antibiotic use density declined from 111 RDD (138 DDD) per 100 patient days before the intervention to 86 RDD (128 DDD) per 100 patient days after starting the intervention. Among the different antibacterial drug classes, the consumption of third-generation cephalosporins showed the largest reduction and dropped significantly by -68% between preintervention and postintervention periods. Using the RDD dataset, ITS confirmed a highly significant postintervention change in level of third-generation cephalosporins (-15.2, 95% CI (-24.08 to -6.311)) and a corresponding increase in the use of aminopenicillin/betalactamase inhibitor formulations (+6.6, 95% CI (4.169 to 9.069)). The drug use densities for fluoroquinolones and for overall antibiotics declined, however, the postinterventional level changes missed statistical significance--overall (95% CI (-39.99 to 0.466), fluoroquinolones 95% CI (-11.72 to 4.333)). An intensified ABS programme using non-restrictive tools targeting third-generation cephalosporin and fluoroquinolone use in the setting of a large academic hospital emergency medicine department is feasible and effective. The intervention may serve as a model for other emergency medicine departments at hospitals with a similar structure and baseline situation.


Impact of vitamin A on clinical outcomes in haemodialysis patients.
Patients on maintenance haemodialysis treatment experience an excessive risk of cardiovascular disease and mortality. The vitamin A concentration is known to be higher in these patients compared to the general population where elevated vitamin A concentrations are associated with adverse outcome. The impact of vitamin A on morbidity and mortality in end-stage renal disease patients is controversial and is the topic of this study. We analysed plasma retinol and retinol-binding protein 4 (RBP4) in 1177 diabetic haemodialysis patients, who participated in the German Diabetes and Dialysis Study (median follow-up 4 years). By Cox regression analyses hazard ratios (HRs) were determined for pre-specified, adjudicated end points according to baseline concentrations. Patients had a mean age of 66 ± 8 years, mean retinol and RBP4 concentrations of 3.28 (0.71-7.44) and 4.02 (1.28-10.1) μmol/L, respectively. Patients with retinol concentrations in the first quartile (<2.6 μmol/L) had an almost 2-fold increased risk of all-cause mortality compared to patients of the fourth quartile [>3.9 μmol/L; HR 1.81, 95% confidence interval (CI) 1.43-2.30]. There was a strong association between low retinol and the risk of sudden cardiac death (SCD, HR 2.22, 95% CI 1.41-3.50) and fatal infection (HR 2.19, 95% CI 1.26-3.82). Patients with RBP4 concentrations in the lowest quartile (<3.0 μmol/L) were more likely to die of any cause (HR 1.43, 95% CI 1.14-1.80), experience SCD (HR 1.97, 95% CI 1.28-3.03) and cardiovascular events (HR 1.43, 95% CI 1.10-1.85). This large cohort study shows a strong association of low retinol and RBP4 concentrations with SCD and all-cause mortality in diabetic haemodialysis patients.


Removal of trace arsenic(V) and phosphate from water by a highly selective ligand exchange adsorbent.
A highly selective ligand exchange type adsorbent was developed for the removal of trace arsenic(V) (As(V)) and phosphate from water. This adsorbent was prepared by loading zirconium(IV) on monophosphonic acid resin. This adsorbent was able to remove toxic anions efficiently at wide pH ranges. However, low pH was preferable for maximum breakthrough capacity in an adsorption operation. The effect of a large amount of competing anions such as chloride, bicarbonate, and sulfate on the adsorption systems of As(V) and phosphate anions was investigated. The experimental findings revealed that the As(V) and phosphate uptakes were not affected by these competing anions despite the enhancement of the breakthrough points and total adsorption. Phosphate anion was slightly preferable than As(V) in their competitive adsorption by the adsorbent. The adsorbed As(V) and phosphate on the Zr(IV)-loaded resin were quantitatively eluted with 0.1 mol/L sodium hydroxide solution, and the adsorbent was regenerated by 0.5 mol/L sulfuric acid. During several cycles of adsorption-elution-regeneration operations, no Zr(IV) was detected in the column effluents. Therefore, the Zr(IV)-loaded monophosphonic acid resin is an effective ligand exchange adsorbent for removing trace concentrations of As(V) and phosphate from water.


Different roles for the stress-activated protein kinase pathway in the regulation of trehalose metabolism in Schizosaccharomyces pombe.
The Wis1p-Sty1p mitogen-activated protein kinase cascade is a major signalling system in the fission yeast Schizosaccharomyces pombe for a wide range of stress responses. It is known that trehalose functions as a protective metabolite to counteract deleterious effects of environmental stresses. Herein it is reported that the expression of genes related to trehalose metabolism in S. pombe, ntp1(+) (neutral trehalase) and tps1(+) [trehalose-6-phosphate (T6P) synthase], is partially regulated by the Sty1p kinase under salt-induced osmotic stress and conditions of slight oxidative stress and is fully dependent on this kinase under severe oxidative stress. This control is carried out through transcription factors Atf1p/Pcr1p during osmotic stress and through Pap1p during exposure to low levels of oxidative stress. However, all three transcription factors are needed for gene expression under conditions of extreme oxidative stress. In addition, a role for Sty1p in the modulation of post-transcriptional activation of trehalase mediated by Pka1p/Sck1p kinases, as well as in the activity of T6P synthase under such stressful conditions has been demonstrated. These results reveal a novel dual action of the Wis1p-Sty1p pathway in the regulation of trehalose metabolism in fission yeast.


Cases of human immunodeficiency virus infection and tuberculosis--early experiences of different aspects.
An outpatients department based survey conducted in Calcutta amongst 1349 established cases of tuberculosis (TB) revealed 0.67% human immunodeficiency virus (HIV) infected cases. Those affected by HIV and TB did not show any deviation from epidemiological pattern of HIV infection in India. All contracted HIV infection by heterosexual route, mostly from Bombay (47.8%) followed by West Bengal (30.4%). In follow-up study of a cohort of 36 HIV seropositives over 3 years, 10(27.7%) developed TB. Of the 23 HIV infected cases with TB, lesions were mostly pulmonary (n = 18, 78.3%) followed by pleural effusion (n = 3;13%). Low incidence of Mycobacterium avium (intracelluarae) complex and tuberculous lymphadenopathy one case each and 52.2% positivity with 14.5 mm mean induration diameter in intradermal test with one TU PPD-RT23 are deviations from previous reports. Low incidence of cough (43.5%), marked weight loss (100%) and fever (100%) were the cardinal clinical features. TB infection on pattern suggestive of reactivation of dormant pulmonary lesions lower rate (11%) of treatment failure and infection caused by organisms other than Mycobacterium tuberculosis were other findings of the study. Importance of serosurveillance to unearth more TB cases amongst HIV infected cases for early treatment and isoniazid prophylaxis is stressed upon.


Blimp-1 homolog Hobit identifies effector-type lymphocytes in humans.
Human cytomegalovirus (CMV) induces the formation of effector CD8(+) T cells that are maintained for decades during the latent stage of infection. Effector CD8(+) T cells appear quiescent, but maintain constitutive cytolytic capacity and can immediately produce inflammatory cytokines such as IFN-γ after stimulation. It is unclear how effector CD8(+) T cells can be constitutively maintained in a terminal stage of effector differentiation in the absence of overt viral replication. We have recently described the zinc finger protein Homolog of Blimp-1 in T cells (Hobit) in murine NKT cells. Here, we show that human Hobit was uniformly expressed in effector-type CD8(+) T cells, but not in naive or in most memory CD8(+) T cells. Human CMV-specific but not influenza-specific CD8(+) T cells expressed high levels of Hobit. Consistent with the high homology between the DNA-binding Zinc Finger domains of Hobit and Blimp-1, Hobit displayed transcriptional activity at Blimp-1 target sites. Expression of Hobit strongly correlated with T-bet and IFN-γ expression within the CD8(+) T-cell population. Furthermore, Hobit was both necessary and sufficient for the production of IFN-γ. These data implicate Hobit as a novel transcriptional regulator in quiescent human effector-type CD8(+) T cells that regulates their immediate effector functions.


Contemporary innovations in two-dimensional transition metal dichalcogenide-based P-N junctions for optoelectronics.
Two-dimensional transition metal dichalcogenides (2D-TMDCs) with various physical characteristics have attracted significant interest from the scientific and industrial worlds in the years following Moore's law. The p-n junction is one of the earliest electrical components to be utilized in electronics and optoelectronics, and modern research on 2D materials has renewed interest in it. In this regard, device preparation and application have evolved substantially in this decade. 2D TMDCs provide unprecedented flexibility in the construction of innovative p-n junction device designs, which is not achievable with traditional bulk semiconductors. It has been investigated using 2D TMDCs for various junctions, including homojunctions, heterojunctions, P-I-N junctions, and broken gap junctions. To achieve high-performance p-n junctions, several issues still need to be resolved, such as developing 2D TMDCs of superior quality, raising the rectification ratio and quantum efficiency, and successfully separating the photogenerated electron-hole pairs, among other things. This review comprehensively details the various 2D-based p-n junction geometries investigated with an emphasis on 2D junctions. We investigated the 2D p-n junctions utilized in current rectifiers and photodetectors. To make a comparison of various devices easier, important optoelectronic and electronic features are presented. We thoroughly assessed the review's prospects and challenges for this emerging field of study. This study will serve as a roadmap for more real-world photodetection technology applications.


A Rare Case of Cecal Bascule in a Child With Normal Neurodevelopment.
Cecal volvulus is a rare cause of bowel obstruction in adults and an extremely rare presentation in children. One form known as a cecal bascule has only previously been reported in children with neurodevelopmental issues or with severe chronic constipation. We present the case of a 10-year-old boy who presented with an acute history of left lower quadrant abdominal pain, who upon investigation was found to have a cecal bascule.


Inter-alpha-trypsin inhibitor and its related proteins in Syrian hamster urine and plasma.
Urinary excretion of trypsin inhibitor increased after injection of a carcinogen, N-nitrosobis(2-oxopropyl)amine, into Syrian hamsters. Two inhibitors were purified to apparent homogeneity from urine collected during the course of the carcinogenesis experiment. Their complete amino acid sequences were determined by Edman degradation of the intact proteins and partially degraded fragments. One corresponded to a hamster liver cDNA clone that hybridized with human bikunin probe [Ide et al, (1994) Biochim, Biophys. Acta 1209, 286-292], except that the protein sequence lacked C-terminal serine and the other was trypstatin, the C-terminal half of the bikunin molecule. Three proteins containing covalently linked bikunin were also identified in pooled blood plasma. They were all dissociated into heavy and light chains by treatment with chondroitinase ABC or 50 mM NaOH, but not by heating at 100 degrees C in the presence of sodium dodecyl sulfate and dithiothreitol, N-terminal amino acid sequence analyses of the native chains and partially degraded fragments thereof revealed that these proteins are (i) human-type inter-alpha-trypsin inhibitor, consisting of heavy chains 1 and 2 and bikunin, (ii) bovine-type inter-alpha-trypsin inhibitor, consisting of heavy chains 2 and 3 and bikunin, and (iii) pre-alpha-trypsin inhibitor, consisting of heavy chain 3 and bikunin. Heterodimer of bikunin/heavy chain 1 or bikunin/heavy chain 2 was not detected. These results suggest that the composition, and hence function, of the inter-alpha-trypsin inhibitor family differs considerably from species to species.


Evaluation of Using Behavioural Changes to Assess Post-Operative Pain in the Guinea Pig (Cavia porcellus).
To manage pain effectively in people and animals, it is essential to recognise when pain is present and to assess its intensity. Currently there is very little information regarding the signs of post-surgical pain or its management in guinea pigs. Studies from other rodent species indicate that behaviour-based scoring systems can be used successfully to detect pain and evaluate analgesic efficacy. This preliminary study aimed to establish whether behaviour-based scoring systems could be developed to assess post-surgical pain in guinea pigs. This prospective, randomised, placebo-controlled study used 16 guinea pigs, and evaluated changes in behaviour following either anaesthesia alone or anaesthesia and orchiectomy. Behaviour was assessed using a combination of manual and automated scoring of remotely obtained video footage. A small number of behaviours were identified that appeared to have high specificity for pain caused by orchiectomy. However, the behaviours were displayed infrequently. The most common was a change in posture from standing to recumbency, sometimes with one hind leg extended either to the side or behind the body. A composite behaviour score incorporating these abnormal behaviours differentiated between the effects of surgery and anaesthesia alone (p<0.0001), and between animals that received analgesia post-operatively compared to an untreated group (p<0.0001). Although behavioural changes occurred in these guinea pigs after orchiectomy, the changes were relatively subtle and the individual specific pain-related behaviours occurred infrequently. However, it may prove possible to develop a behaviour-based scoring system for routine use in this species using a combination of pain-related behaviours.


Hapten-specific T cell responses to 4-hydroxy-3-nitrophenyl acetyl. VI. Evidence for different T cell receptors in cells that mediate H-21-restricted and H-2D-restricted cutaneous sensitivity responses.
We have previously shown that cross-reactive sensitivity (CS) responses induced by 4-hydroxy-3-nitrophenyl acetyl-O-succinimide (NP-O-Su) and elicited by its 5-iodo analogue, 4-hydroxy-5-iodo-3-nitrophenyl acetyl-O-succinimide were observed in strains of mice possessing the Igh-1b allotype, but not in strains bearing allotypes Igh-1c or Igh-1j. These CS responses are mediated by T cells and can be transferred to naive recipients that are homologous at either the H-2K, H-2I, or H-2D regions of the major histocompatibility complex. We now extend our analysis of cross-reactive 4-hydroxy-3-nitrophenyl-acetyl (NP)-induced CS responses to inbred strains of mice expressing additional Igh-1 allotypes. In contrast to NP-induced delayed-type hypersensitivity responses, which only display 4-hydroxy-5-iodo-3-nitrophenyl acetyl (NIP) cross-reactivity in Igh-1b-bearing mice, cross-reactive CS responses can also be elicited in NP-primed mice carrying the Igh-1d, Igh-1e, or Igh-1f allotypes. Moreover, cross-reactive NP-induced CS responses could be transferred by NP-O-Su-primed lymph node cells from the AKR (Igh-1d) strain, into naive recipients homologous at the H-2D region, but only non-cross-reactive NP responses could be transferred into strains homologous at the H-2I region. Furthermore, the lack of cross-reactivity in the Igh-1j-bearing C3H strain was not the result of an inability of these mice to recognize NP in association with H-2K/D products, because NP-O-Su-primed cells from C3H donors transferred NP-specific CS responses into both H-2D and H02I homologous recipients. The results are discussed with respect to the nature of the T cell receptors that control NP responses.


Effectiveness and safety analysis of titanium mesh grafting versus bone grafting in the treatment of spinal Tuberculosis: a systematic review and meta-analysis.
To systematically assess the safety and effectiveness of titanium mesh grafting compared with bone grafting in the treatment of spinal tuberculosis. Electronic databases, including PubMed, Embase, Web of Science, and Cochrane Library, were searched from their inception until April 2023. The outcome indicators for patients treated with titanium mesh grafting or bone grafting for spinal tuberculosis include surgical duration, intraoperative blood loss, graft fusion time, American Spinal Injury Association (ASIA) Spinal Cord Injury Grade E assessment, VAS score, lumbar pain score, post-graft kyphotic angle, and postoperative complications. The Newcastle-Ottawa Scale (NOS) and the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) approach were used for quality assessment and evidence grading of clinical studies. Funnel plots and Begg's test were employed for bias assessment. A total of 8 studies were finally included, comprising 523 patients, with 267 cases of titanium mesh fixation and 256 cases of bone grafting. The meta-analysis showed no significant statistical differences in surgical duration (Weighted Mean Difference (WMD) = -7.20, 95% Confidence Interval (CI): -28.06 to 13.67, P = 0.499), intraoperative blood loss (WMD = 16.22, 95% CI: -40.62 to 73.06, P = 0.576), graft fusion time (WMD = 0.97, 95% CI: -0.88 to 2.81, P = 0.304), ASIA Spinal Cord Injury Grade E assessment (Relative Risk (RR) = 1.03, 95% CI: 0.97 to 1.09, P = 0.346), and overall complications (RR = 0.87, 95% CI: 0.49 to 1.55, P = 0.643). Differences in VAS score, ODI lumbar pain score, and post-graft kyphotic angle between the titanium mesh grafting group and the bone grafting group were not significant within the 95% CI range. The rate of postoperative implant subsidence was slightly lower in bone grafting than in titanium mesh grafting (RR = 9.30, 95% CI: 1.05 to 82.22, P = 0.045). Both bone grafting and titanium mesh grafting are effective and safe for the surgery, with no significant statistical differences in the results. Considering the limitations of the present study, large-scale randomized controlled trials are warranted to further verify the reliability of this finding.


Identification of miRNAs in a Model of Retinal Degenerations.
We investigated the expression profile of and identify all microRNAs (miRNAs) that potentially regulate inflammation in a light-induced model of focal retinal degeneration. Sprague Dawley (SD) rats aged 90 to 140 postnatal days were exposed to 1000 lux white fluorescent light for 24 hours. At 24 hours, and 3 and 7 days after exposure, the animals were euthanized and retinas processed for RNA. Expression of 750 miRNAs at 24 hours of exposure was assessed using low density array analysis. Significantly modulated miRNAs and their target mRNAs were used to assess the potential biological effects. Expression of seven miRNAs, potentially modulating inflammation, was investigated across a protracted time course after light exposure using quantitative PCR. Photoreceptor cell death was analyzed using TUNEL. Intense light exposure for 24 hours led to differential expression of a number of miRNAs, 37 of which were significantly modulated by 2-fold or more. Of those, 19 may potentially regulate the inflammatory immune response observed in the model. MicroRNAs -125-3p, -155, -207, -347, -449a, -351, and -542-3p are all upregulated at 24 hours of exposure along with peak photoreceptor cell death. The MiRNAs -542-3p and -351 reached maximum expression at 7 days after exposure, while -125-3p, -155, -207, -347, and -449 reached a peak expression at 3 days. The results of the study show that miRNAs are modulated in response to light damage (LD). These miRNAs potentially regulate the inflammatory immune response, triggered as a result of the acute retinal damage, which is a key mediator of retinal degeneration in this model and age-related macular degeneration.


The Construction of the Self-Induced Sal System and Its Application in Salicylic Acid Production.
The design and construction of more complex and delicate genetic control circuits suffer from poor orthogonality in quorum sensing (QS) systems. The Sal system, which relies on salicylic acid as a signaling molecule, is an artificially engineered regulatory system with a structure that differs significantly from that of natural QS signaling molecules. Salicylic acid is an important drug precursor, mainly used in the production of drugs such as aspirin and anti-HIV drugs. However, there have been no reports on the construction of a self-induced Sal system in single cells. In this study, a high-copy plasmid backbone was used to construct the regulatory proteins and a self-induced promoter of salicylic acid in E. coli by adjusting the precise regulation of key gene expression; the sensitivity and induction range of this system were improved. Subsequently, the exogenous gene pchBA was introduced in E. coli to extend the shikimate pathway and synthesize salicylic acid, resulting in the construction of the first complete self-induced Sal system. Finally, the self-induced Sal System was combined with artificial trans-encoded sRNAs (atsRNAs) to repress the growth-essential gene ppc and accumulate the precursor substance PEP, thereby increasing the titer of salicylic acid by 151%. This construction of a self-induced artificial system introduces a new tool for selecting communication tools and induction systems in synthetic biology and metabolic engineering, but also demonstrates a self-inducible pathway design strategy for salicylic acid biosynthesis.