Hierarchically porous biochar derived from aerobic granular sludge for high-performance membrane capacitive deionization.
Membrane capacitive deionization (MCDI) is a cost-effective desalination technique known for its low energy consumption. The performance of MCDI cells relies on the properties of electrode materials. Activated carbon is the most widely used electrode material. However, the capacitive carbon available on the market is often expensive. Here, we developed hierarchically porous biochar by combining carbonization and activation processes, using easily acquired aerobic granular sludge (AGS) from biological sewage treatment plants as a precursor. The biochar had a specific surface area of 1822.07 m2 g-1, with a micropore area ratio of 58.65% and a micropore volume of 0.576 cm3 g-1. The MCDI cell employing the biochar as electrodes demonstrated a specific adsorption capacity of 34.35 mg g-1, comparable to commercially available activated carbon electrodes. Our study presents a green and sustainable approach for preparing highly efficient, hierarchically porous biochar from AGS, offering great potential for enhanced performance in MCDI applications.


Identification of HLA-A2 restricted epitopes of glypican-3 and induction of CTL responses in HLA-A2 transgenic mice.
Hepatocellular carcinoma (HCC) is a malignant tumor with high mortality, but lacks effective treatments. Carcinoembryonic antigen glypican-3 (GPC3) is a tumor-associated antigen overexpressed in HCC but rarely expressed in healthy individuals and thus is one of the most promising therapeutic targets. T cell epitope-based vaccines may bring light to HCC patients, especially to the patients at a late stage. However, few epitopes from GPC3 were identified to date, which limited the application of GPC3-derived epitopes in immunotherapy and T cell function detection. In this study, a total of 25 HLA-A0201 restricted GPC3 epitopes were in silico predicted and selected as candidate epitopes. Then, HLA-A0201+/GPC3+ HCC patients' PBMCs were collected and co-stimulated with the candidate epitope peptides in ex vivo IFN-γ Elispot assay, by which five epitopes were identified as real-world epitopes. Their capacity to elicit specific CD8+ T cells activation and proliferation was further confirmed by in vitro co-cultures of patients' PBMCs with peptide, in vitro co-cultures of healthy donors' PBLs with DCs and peptide, T2 cell binding assay as well as HLA-A2 molecule stability assay. Moreover, the in vivo immunogenicity of the five validated epitopes was confirmed by peptides cocktail/poly(I:C) vaccination in HLA-A0201/DR1 transgenic mice. Robust epitope-specific CD8+ T cell responses and cytotoxicity targeting HepG2 cells were observed as detected by IFN-γ Elispot, intracellular IFN-γ staining and cytolysis assay. This study provided novel GPC3 CTL epitopes for the development of T cell epitope vaccines and evaluation of GPC3 specific T cell responses.


MERISTEM ACTIVITYLESS (MAL) is involved in root development through maintenance of meristem size in rice.
Rice MERISTEM ACTIVITYLESS (MAL), a RING-H2 finger domain (RFD)-containing gene, regulates meristem cell viability after the initiation of root primordia mediated by cytokinin signaling. Genes in the RING-H2 finger domain (RFD) family play various roles during plant development and in biotic/abiotic stress responses. Rice gene MERISTEM ACTIVITYLESS (MAL), being contained in the RING-H2 finger domain (RFD), is characterized by a transmembrane domain at the N-terminal and a C3H2C3 zinc finger domain at the C-terminal. To elucidate the physiological and molecular functions of MAL, we generated MAL knockdown transgenic plants by RNA interference. MAL RNA-interfered (MRi) transgenic plants exhibited a phenotype with shorter crown root length and lower crown root number, accompanied by a lower cell division rate. The low division rate was observed in the root meristem exactly where MAL was expressed. Furthermore, transcriptome data revealed that cell wall macromolecule metabolism-related genes and redox-related genes were enriched in MAL RNAi lines. Most of these differentially expressed genes (DEGs) were induced by exogenous cytokinin. Hence, we conclude that MAL, as a novel regulatory factor, plays a major role in maintaining cell viability in the meristem after the initiation of root primordial formation, mediated by cytokinin signaling and reactive oxygen species (ROS).


Atypical presentation of West Nile virus in a newly diagnosed human immunodeficiency virus patient in New York City.
Central nervous system manifestations of West Nile virus (WNV) infection include meningitis, encephalitis, and poliomyelitis-like syndrome. We describe a 44-year-old man with no past medical history who presented with a meningoencephalitic syndrome and spastic paralysis bilaterally in the upper and lower extremities, hyperreflexia, and myoclonus and was ultimately diagnosed as being infected with HIV and WNV.


Targeting oncogene expression to endothelial cells induces proliferation of the myelo-erythroid lineage by repressing the Notch pathway.
Human oncogenes involved in the development of hematological malignancies have been widely used to model experimental leukemia. However, models of myeloid leukemia rarely reproduce the human disease in full, due to genetic complexity or to difficulties in targeting leukemia initiating cells. Here, we used a zebrafish genetic model to induce the expression of oncogenic RAS in endothelial cells, including the hemogenic endothelium of the dorsal aorta that generates hematopoietic cells, and observed the development of a myelo-erythroid proliferative disorder. In larvae, the phenotype is characterized by disruption of the vascular system and prominent expansion of the caudal hematopoietic tissue. In few surviving juveniles, increased number of immature hematopoietic cells and arrest of myeloid maturation was found in kidney marrow. Peripheral blood showed increased erythroblasts and myeloid progenitors. We found that the abnormal phenotype is associated with a downregulation of the Notch pathway, whereas overexpressing an activated form of Notch together with the oncogene prevents the expansion of the myelo-erythroid compartment. This study identifies the downregulation of the Notch pathway following an oncogenic event in the hemogenic endothelium as an important step in the pathogenesis of myelo-erythroid disorders and describes a number of potential effectors of this transformation.


The DEER database: a bridge connecting drugs, environmental effects, and regulations.
Variability in patient drug responses is observed with increasing frequency, necessitating the establishment of causal associations between factors and drug response phenotypes. This individual variability can be caused by genetic factors and environmental factors (ENFs). Although pharmacogenetics has been instrumental in describing genetic variations, frameworks for understanding the association between ENFs (particularly chemical ENFs) and drug responses are lacking. In this study, we constructed a novel database, DEER, for interpretations of chemical ENF effects on drug responses. DEER includes computational predictions of the associations between chemical ENFs and drug responses. Putative regulatory intermediates such as transcription factors, cytochrome P450s (CYPs), drug targets, and transporters as well as chemical similarities are provided to support our predictions. DEER currently encompasses 579 drugs, 401 chemical ENFs, and 9247 predicted drug-ENF associations. The entire dataset can be easily queried through a search page. The results can be downloaded, and each drug-ENF association with intermediary factors can be displayed via a graphical viewer. DEER is available at http://bsb.kiz.ac.cn:90/DEER/. We expect this approach and resource to be valuable for personalized medicine and drug development.


Diverse responses between human pancreatic cancer cell lines to native alpha 1-antitrypsin and its C-terminal fragment.
Previous studies imply that human pancreatic cancer cells have a wide heterogeneity and their exposure to various agents may give unpredictable results in clinical situations. The cell lines LPC-3, -5 and -10, established from primary cultures of pancreatic adenocarcinoma, were exposed to 5 microM of AAT or its C-terminal peptide C-36 for 24 hours and analysed for cytokines by an enzyme-linked immunosorbent assay and for NF kappa B by the electrophoretic mobility shift assay. Native AAT lowers TGF-beta 1 levels and increases NF-kappa B activity in LPC-3 cells, while C-36 increases TGF-beta 1 levels and up-regulates NF-kappa B in LPC-5 cells. In LPC-10 cells AAT lowers TGF-beta 1. However, both AAT and C-36 fail to cause a change in NF-kappa B expression. For LPC-10 cells treated with C-36 IL-6 and TNF-alpha levels also increase. Our findings provide evidence that human cancer cell lines originating from primary pancreatic tumors do not have a uniform response to the same stimulus which shows a great heterogenicity among pancreatic cancer cells. Serine proteinase inhibitor, AAT, dependent on its molecular form, is also found to exert diverse effects on the properties of tumour cells confirming the complexity of cell-protein interaction.


Fermentative metabolism in grape berries: isolation and characterization of pyruvate decarboxylase cDNA and analysis of its expression throughout berry development.
The involvement of pyruvate decarboxylase (PDC) in the control of alcohol production during ripening of fruit tissues under aerobic conditions has been only partially studied. Enzymological studies showed a significant increase in PDC activity during the ripening of oranges and pears, concurrently with the induction of ethanol production. In tomato, on the other hand, the induction of ethanol production and ADH gene expression after the onset of ripening was not accompanied by induction of PDC activity. The isolation of PDC cDNA from fruits has not yet been reported, nor has its expression pattern during fruit development. We report here the isolation of a cDNA clone encoding for a grape PDC and the characterization of its expression throughout berry development. The pattern of PDC gene expression throughout berry development, combined with earlier findings on constitutive PDC activity in the berry, may suggest that PDC is not the limiting factor for the production of ethanol in the berry, which is induced only after the onset of berry ripening. Alternatively, the induction of ADH gene expression, which occurs only after the onset of ripening in both tomatoes and grape berries, may serve as a regulator of ethanol production in response to a ripening-related cue.


Cytokine medicines in clinical practice: current issues.
Cytokine medicines have been licensed for the treatment of rheumatoid arthritis since 2000. The rheumatology community has accrued a large amount of experience in the use of these medications. This experience has led to the development of guidelines for their use that include ongoing vigilance for long term adverse events and efficacy using the Biologics Register. Delivery of these expensive therapies has prompted extensive system developments within rheumatology. The cytokine medicines have provided important tools to probe the pathogenesis of rheumatoid and other inflammatory diseases. Further cytokine medicines, in various stages of development, are on the horizon and continue to stimulate excitement within this fast expanding field.


Involvement of the small protein tyrosine phosphatases TC-PTP and PTP1B in signal transduction and diseases: from diabetes, obesity to cell cycle, and cancer.
As in other fields of biomedical research, the use of gene-targeted mice by homologous recombination in embryonic stem cells has provided important findings on the function of several members of the protein tyrosine phosphatase (PTP) family. For instance, the phenotypic characterization of knockout mice has been critical in understanding the sites of action of the related PTPs protein tyrosine phosphatase 1B (PTP1B) and T-cell-PTP (TC-PTP). By their increased insulin sensitivity and insulin receptor hyperphosphorylation, PTP1B null mice demonstrated a clear function for this enzyme as a negative regulator of insulin signaling. As well, TC-PTP has also been recently involved in insulin signaling in vitro. Importantly, the high identity in their amino acid sequences suggests that they must be examined simultaneously as targets of drug development. Indeed, they possess different as well as overlapping substrates, which suggest complementary and overlapping roles of both TC-PTP and PTP1B. Here, we review the function of PTP1B and TC-PTP in diabetes, obesity, and processes related to cancer.


Feasibility of a single-stage tracheostomy decannulation protocol with endoscopy in adult patients.
Gradual decrease in tube size and tube capping are considered the standard of care for tracheostomy decannulation. Both of these actions result in increased airway resistance. Immediate decannulation may offer a more tolerable approach. To assess the feasibility of immediate tracheostomy decannulation compared with the traditional decannulation methods. This study is a single institute, case-control retrospective study of patients between the years 2009 to 2014. The study group included all patients who underwent immediate decannulation, whereas the control group comprised patients who underwent traditional staged decannulation. An immediate decannulation protocol included admission to the intensive care unit, a comprehensive evaluation, decannulation, 24 hours of monitoring, and observation until discharge. Twenty-nine patients were included in the study group and 20 in the control group. No significant statistical difference was found between the two groups in the patients' medical history and tracheostomy data, except for the Acute Physiology and Chronic Health Evaluation II score and duration of the deflated cuff, which were significantly higher in the control group. A significant difference was found in the complication rate between the groups. In the staged decannulation group, four patients failed decannulation and required reinsertion of the tracheostomy cannula, whereas there were no such failures in the immediate decannulation group. Hospitalization duration after decannulation of the study group patients was significantly shorter than that of the control group. Immediate decannulation may offer a safe alternative for weaning from tracheostomy. It may also reduce the duration of the weaning process and hospitalization. 3b Laryngoscope, 126:2057-2062, 2016.


Congenital dislocation of the knee at birth - Part I: Clinical signs and classification.
Congenital dislocation of the knee (CDK) is rare, and clinical semiology at birth is not always suitably analyzed. Existing classifications fail to guide treatment. The aim of the present study was to develop a CDK classification for the neonatal period. A classification based on neonatal severity of clinical signs is easy to implement on simple criteria. Fifty-one CDKs (40 patients) seen neonatally were included. Three types could be distinguished in terms of reduction and stability: type I, easily reducible CDK, with reduction snap when the femoral condyles pass in flexion, remaining stable in flexion; type II, "recalcitrant" dislocation, reducible by posteroanterior "piston" but unstable, with iterative dislocation once posteroanterior pressure on the condyles is relaxed; and type III, irreducible. The number of anterior skin grooves, global range of motion, flexion deficit and reduction stability were noted for each type. Mean age at first consultation was 5.6 days (range: 0-30). CDK was type I, II and III in respectively 28, 16 and 7 cases. Number of skin grooves, flexion and baseline range of motion were greater in type I than types II and III. The present neonatal clinical classification is original, logical and simple. It may be useful for prognosis and guiding treatment. IV, single-center retrospective series.


Impact of surgical resection on the survival in Mexican patients with gastric cancer: A meta-analysis and systematic review.
Gastric cancer (GC) is one of the most frequent cancer types in Mexico. The primary method used as a treatment is surgical resection. The role of surgery in increasing survival is controversial. This study aimed to determine whether surgical resection increases the survival of patients with GC in a Mexican population. A systematic review of literature searches (Evidence-based MEDLINE/PubMed, Web of Science, Cochrane Library, and SciELO) and meta-analysis were performed based on the Preferred Reporting Items for Systematic Reviews and Meta-Analysis criteria. The published articles from 2000 to the current time were divided into cross-sectional and randomized studies. The inclusion criteria were survival, surgical resections, patients treated in Mexico, and primary GC. The effect estimation was calculated using the risk ratio (RR). The random-effects model and a confidence interval (CI) of 95% were used. The RR of the pooled studies was 1.09 (95% CI, 0.71-1.67). RR of 0.82 (95% CI, 0.63-1.07) was obtained in cross-sectional studies, and randomized studies showed a RR of 2.08 (95% CI, 0.25-17.07). This work is the first systematic study that assesses the role of surgery on the survival of patients with GC in the Mexican population, the results showed that surgical resection did not improve survival in patients with GC.


Early Hepatic Artery Thrombosis After Liver Transplantation: What is the Impact of the Arterial Reconstruction Type?
Hepatic artery thrombosis (HAT) is the most severe vascular complication occurring after liver transplantation, with an incidence ranging from 2 to 9% in adults. Although the ideal arterial reconstruction is often described as a short and non-redundant anastomosis fashioned between the recipient and donor hepatic arteries, there is no strong evidence about this ideal reconstruction in the literature. The aim of this study was to assess the impact of the type of arterial reconstruction on early HAT after primary liver transplantation. We retrospectively reviewed a contemporary MELD era cohort of 282 patients who underwent deceased donor primary liver transplantation from 2007 to 2012. Graft artery was classified as "short" when the section was located at the proper/common hepatic artery or "long" when the celiac trunk was used for anastomosis. Recipient arterial sites for arterial anastomosis were classified in three sites: (1) "distal" (proper hepatic artery or common hepatic artery/gastro-duodenal bifurcation), (2) "intermediate" (common hepatic artery) and (3) "proximal" (celiac trunk-splenic artery-aorta). We used univariate and multivariate analyses to assess the impact of different types of arterial reconstruction on early HAT. Of 282 primary liver transplantations, 17 patients (6%) developed early HAT. Patients with and without early HAT had comparable demographic and operative data. The main anastomotic combination was short graft artery on the recipient-common hepatic artery (n = 111, 39%). A long graft artery was used in 91 patients (32%) and was associated with hepatic artery variations (56%; n = 51; p = 0.001). Arterial reconstructions using a long graft artery (p = 0.003), a recipient proximal site as celiac trunk-splenic artery-aorta (p = 0.02) and the combination of a long graft artery on the recipient distal hepatic artery (p = 0.02) were significantly associated with early HAT. The early HAT rate in patients with a long graft artery was not significantly different between patients with or without donor arterial variation (respectively, 12% (n = 6/51) vs. 12% (n = 5/40); p = 1). In multivariate analysis, the use of a long graft artery, whatever the recipient anastomosis site, was an independent risk factor of early HAT (OR 3.2; 95% CI 1.2-9; p = 0.02). The type of arterial reconstruction used for arterial anastomosis during primary liver transplantation has an impact on the occurrence of early HAT. The use of a long graft artery is an independent risk factor of early HAT. Thereby, we recommend the use of a short graft artery with a direct path when feasible to reduce the occurrence of early HAT after primary liver transplantation.


An alternate method of descenting skunks.
Striped skunk (Mephitis mephitis) scent glands were ligated closed with waxed dental floss to allow them to be handled during toxicological studies without fear of scenting. This descenting technique was more rapid and less traumatic than scent gland removal. Thirty-four skunks were kept for less than or equal to 127 days and did not display behavioral or physical abnormalities due to this procedure.


Impaired Sensitivity to Thyroid Hormones is Associated with Mild Cognitive Impairment in Euthyroid Patients with Type 2 Diabetes.
The prevalence of mild cognitive impairment (MCI) in patients with type 2 diabetes (T2D) is rapidly increasing. Thyroid hormones are key regulators of cognitive function in adults. The purpose of this study was to investigate the relationship between thyroid hormone sensitivity and MCI in euthyroid T2D patients. A total of 400 euthyroid T2D patients were enrolled in this cross-sectional study, including 218 patients with normal cognition and 182 MCI patients. The Montreal Cognitive Assessment (MoCA) was used to evaluate cognitive function. The free triiodothyronine to free thyroxine (FT3/FT4) ratio was calculated as a measure of peripheral sensitivity to thyroid hormones; the thyroid-stimulating hormone index (TSHI), thyrotrophic thyroxine resistance index (TT4RI) and thyroid feedback quantile-based index (TFQI) were calculated as measures of central sensitivity to thyroid hormones. Linear regression analysis and logistic regression analysis were performed to explore the relationships between these indices of thyroid hormone sensitivity and the MoCA score and MCI, respectively. Compared with the normal cognitive function group, patients in the MCI group had higher TSHI, TT4RI and TFQI but a lower FT3/FT4 ratio (P<0.05). The MoCA score was positively correlated with the FT3/FT4 ratio but negatively correlated with TSHI, TT4RI and TFQI (P< 0.05). Multivariate logistic regression analysis showed that a low FT3/FT4 ratio and high TSHI, TT4RI and TFQI were independently associated with MCI (P<0.05). After adjustment for confounding factors, the odds ratio (OR) for the association between MCI and the highest tertile of the FT3/FT4 was 0.455 (95% CI: 0.264-0.785), for the highest tertile of TSHI, the OR was 2.380 (95% CI: 1.376-4.119), for the highest tertile of TT4RI, the OR was 2.342 (95% CI:1.353-4.054), and for the highest tertile of TFQI, the OR was 2.536 (95% CI: 1.466-4.387) (P< 0.05). Impaired sensitivity to thyroid hormones is associated with MCI in euthyroid T2D patients.


The value of right heart contrast echocardiography combined with migraine rating scale in evaluating the efficacy of patent foramen ovale closure.
To investigate the clinical values of right heart contrast transthoracic echocardiography (cTTE) combined with migraine rating scale in evaluating the efficacy of patent foramen ovale (PFO) closure. From January 2018 to December 2021, a total of 68 hospitalized patients, 21 males and 47 females, who were treated with transcatheter closure of PFO-induced migraine in the Heart Center of the First Affiliated Hospital of Tsinghua University were selected, with the age of 38.4 ± 11.9 years old. The changes of right heart contrast transthoracic echocardiography (cTTE), visual analogue pain score(VAS), headache impact test-6(HIT-6) and migraine disability assessment questionnaire(MIDAS) before and 6 months after PFO occlusion were compared. Pre-operative cTTE data show that 36 patients (52.9%) had moderate right-to-left shunt (RLS), and 32 patients (47.1%) had massive RLS. cTTE was reexamined 6 months after operation and 1 case in the moderate RLS group had minimal RLS, 2 cases in the large RLS group had minimal RLS, and no shunts were seen for the rest. The VAS, HIT-6 and MIDAS scores before and 6 months after the operation were 7.65 ± 1.39 vs. 1.28 ± 1.53, 70.78 ± 6.82 vs. 41.53 ± 6.07, and 30.60 ± 13.24 vs. 1.93 ± 3.87, respectively. All the indexes 6 months after the operation significantly improved compared with the preoperative baseline (P < 0.05). cTTE combined with migraine evaluation scale could be used as an objective index to evaluate the clinical effect of PFO occlusion.


Construction, identification and application of HeLa cells stably transfected with human PEPT1 and PEPT2.
The purpose of this study was to construct stably transfected HeLa cells with human peptide transporters (hPEPT1/hPEPT2) and to identify the function of the transfected cells using the substrate JBP485 (a dipeptide) and a typical substrate for PEPTs, glycylsarcosine (Gly-Sar). An efficient and rapid method was established for the preparation and transformation of competent cells of Escherichia coli. After extraction and purification, hPEPT1/hPEPT2-pcDNA3 was transfected into HeLa cells by the liposome transfection method, respectively. HeLa-hPEPT1/hPEPT2 cells were selected by measuring the protein expression and the uptake activities of JBP485 and Gly-Sar. A simple and rapid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of JBP485 and Gly-Sar in biological samples. The Michaelis-Menten constant (K(m)) values of Gly-Sar uptake by the hPEPT1 and hPEPT2-expressing transfectants were 1.03 mM and 0.0965 mM, respectively, and the K(m) values of JBP485 uptake were 1.33 mM for PEPT1 and 0.144 mM for PEPT2. The uptake of Gly-Sar was significantly inhibited by JBP485 with a K(i) value of 8.11 mM (for PEPT1) and 1.05 mM (for PEPT2). Maximal uptake of Gly-Sar were detected at pH 5.8 (for PEPT1) and pH 6.5 (for PEPT2), suggesting that both HeLa-hPEPT1 and HeLa-hPEPT2 were H(+) dependent transporters. Stably transfected HeLa-hPEPT1/HeLa-hPEPT2 cells were constructed successfully, and the functions of hPEPT1/hPEPT2 were identified using their substrates, JBP485 and Gly-Sar. The transfected cells with transporters were used to investigate drug-drug interactions (DDIs) between JBP485 and other substrates (cephalexin or lisinopril) of PEPT1 and PEPT2.


The Effect of Botulinum Toxin A on NADPH oxidase system and Ischemia Reperfusion Injury.
While studies aimed at overcoming ischemia-reperfusion (IR) injury using various materials are becoming popular, studies using botulinum toxin type A (BoNTA) are still limited. This study tested the hypotheses that BoNTA can protect flaps from IR injury by inhibiting the NADPH oxidase system and suppressing ROS (reactive oxygen species) production. The subjects were Sprague-Dawley rats (n = 76). In 4 rats, the effects of different dose of BoNTA on superoxide production was evaluated through lucigenin enhanced chemiluminescence assay (LECL) using SD rats' thoracic aorta ring. In 60 SD rats, The BoNTA and normal saline-pretreated superficial inferior epigastric artery (SIEA) flaps were clamped for 0, 1, and 4 hours, and reperfused. On the 5th day after the opeartion, well-maintained flaps were grossly inspected, survival rates were analyzed, and histological analysis was also performed. In 12 rats, after making IR injury through the same model, SIEA flap segments and femoral vessels were obtained, and ROS production was evaluated through LECL and dihydroethidium (DHE) staining. In LECL, the experimental group produced a smaller amount of superoxide than the control group through NADPH oxidase inhibition (p < .05). There was no significant difference between the experimental and control group in the 0, and 1 hour IR groups, but the experimental group (90%) showed a higher survival rate than the control group (60%) in the 4 hours IR group (p = .028). In the measurement of ROS production through LECL and DHE staining, there was no significant difference in the 0, and 1 hour IR groups, but a significant difference was shown in the 4 hours IR group in both the SIEA flaps and femoral vessels (p < .05). This study verified hypothesis that BoNTA can protect flaps from IR injury by inhibiting the NADPH oxidase system and suppressing ROS production. Based on this research model, future research should be expanded into studies on subtypes or subunits of NADPH oxidase, and the findings from the present study are expected to contribute and lead to clinical studies on BoNTA, which has already been proven to be clinically safe.


Lysis of fresh human tumour cells by autologous tumour-associated lymphocytes: two distinct types of autologous tumour killer cells induced by co-culture with autologous tumour.
The specific and natural killer (NK)-restricted nature of auto-tumour cytotoxicity of tumour-associated lymphocytes was studied in cancer patients with malignant pleural effusions. Large granular lymphocytes (LGL) and small T lymphocytes were isolated from carcinomatous pleural effusions by centrifugation on discontinuous Percoll gradients. Tumour cells freshly isolated from pleural effusions were classified according to their susceptibility to lysis by Percoll-purified LGL from the blood of normal donors in a 4-h 51Cr release assay. Of 12 NK-sensitive tumour samples, 11 were killed by autologous fresh effusion LGL, whereas only 2 were lysed by autologous T cells. Neither LGL nor T cells were cytotoxic to NK-resistant autologous tumour cells. T cells and LGL were each cultured in vitro with autologous tumour cells for 6 days. Effusion LGL maintained their auto-tumour killing activity in 10 of 12 autologous mixed lymphocyte-tumour cultures (MLTC) with NK-sensitive tumour, while LGL lost the activity when cultured alone. Removal of high-affinity sheep erythrocyte-rosetting cells from Percoll-purified LGL enriched effector cells. Autologous MLTC-derived LGL could also kill NK-sensitive allogeneic effusion tumour cells and K562 cells, as did fresh LGL. In autologous MLTC LGL failed to acquire lytic function to NK-resistant autologous tumour cells. In contrast, in vitro activation of effusion T cells with autologous tumour cells induced auto-tumour killer cells in 9 of 12 NK-sensitive tumour samples and 3 of 6 NK-resistant tumour cases. However, cultured T cells were incapable of killing allogeneic tumour cells and K562 cells. In the autologous MLTC effusion T cells proliferated vigorously in response to autologous tumour cells, whereas LGL showed no proliferation. The enrichment of blasts from cultured T cells on discontinuous Percoll gradients resulted in an enhancement of auto-tumour cytotoxicity, with no reactions recorded in blast-depleted, small, resting T cells. These results indicate that two distinct types of auto-tumour-recognising lymphocytes, LGL and T cells, are present in carcinomatous pleural effusions of cancer patients and that each effector type recognises different membrane moieties of autologous effusion tumour cells.


Polypeptide composition of purified QH2:cytochrome c oxidoreductase from beef-heart mitochondria.
1. The polypeptide composition of purified QH2: cytochrome c oxidoreductase prepared by three different methods from beef-heart mitochondria has been determined. Polyacrylamide gel electrophoresis in the presence of dodecyl sulphate resolves eight intrinsic polypeptide bands; when, in addition, 8 M urea is present and a more highly cross-linked gel is used, the smallest polypeptide band is resolved into three different bands. 2. The identity of several polypeptide bands has been established by fractionation. The two heaviest polypeptides (bands 1 and 2) represent the so-called core proteins, band 3 the hemoprotein of cytochrome b, band 4 the hemoprotein of cytochrome c1, band 5 and Rieske Fe-S protein, band 6 a polypeptide associated with cytochrome c1 and identified with the so-called oxidation factor, and band 7 a polypeptide peptide associated with cytochrome b. 3. The validity of molecular weight estimate for the polypeptides of the enzyme based on their mobility on dodecyl sulphate gels has been examined. The polypeptides of bands 1, 2 and 3 showed anomalous migration rates. The molecular weights of the other polypeptides have been estimated from their relative mobilities on either dodecyl sulphate gels or 8 M urea-dodecyl sulphate gels as 29 000, 24 000, 12 000, 8000, 6000, 5000 and 4000, respectively. 4. The stoicheiometry of the different polypeptides in the intact complex was determined using separate staining factors for the individual polypeptide band.


Discovery of pyrido[4,3-d]pyrimidinone derivatives as novel Wee1 inhibitors.
Wee1 has emerged as a potential target in cancer therapy due to its critical role in the regulation of the cell cycle. Here, we describe a series of Wee1 inhibitors with a novel scaffold that are potent inhibitors of this kinase (IC50 = 19-1485 nM). These inhibitors demonstrated robust cytotoxicity in MV-4-11 and T47D cell lines (MV-4-11 IC50 = 660-2690 nM, T47D IC50 = 2670-20,000 nM) and displayed good stability in mouse liver microsomes in vitro. Additionally, compound 34 showed remarkable selectivity (more than 500-fold) over the other 9 kinases. Further mechanistic studies demonstrated that compound 34 displayed measurable effects on downstream biomarkers and induced cancer cell apoptosis and cell cycle arrest in the G0/G1 phase. Taken together, these results show that compound 34, potentially a leading Wee1 inhibitor, warrants further investigation.


Asian medical staff attitudes towards witnessed resuscitation.
To assess and compare local Emergency Department medical and nursing staff attitudes towards witnessed resuscitation. Over a period from October 2002 to March 2003, we conducted a self-administered survey of doctors and nurses working in the Emergency Department of the Singapore General Hospital (SGH). We issued 160 forms and received 132 responses, giving a response rate of 82.5%. Should relatives be present during resuscitation? Eighty percent of doctors and 78% of nurses said no. However 32.1% of doctors and 24.1% of nurses had received requests from relatives of patients to be present during resuscitation in the last 6 months. The most frequent reasons for not wanting relatives to be present were: concern that watching the resuscitation process will be a traumatic experience for relatives, relatives might ask too many questions and interfere with resuscitation, relatives might cause stress for staff performing resuscitation, and medico-legal issues might arise. A total of 78.4% of doctors felt that the decision to allow family presence during resuscitation should be made by the senior doctor and not the nursing officer (P=0.001). However, nurses were more likely to feel that it should be a team decision (P<0.001). Locally, we found that medical staff are generally not in favour of witnessed resuscitation. More research is needed to assess attitudes of the general public, and whether this diverges greatly from medical staff attitudes.


Age-adjusted CSF β2-microglobulin and lactate are increased and ACE is decreased in patients with multiple sclerosis, but only lactate correlates with clinical disease duration and severity.
We compared cerebrospinal fluid levels of lactate, β2-microglobulin and angiotensin-converting enzyme (ACE) between 438 untreated patients with multiple sclerosis and 276 patients with non-inflammatory neurological disorders. Age-adjusted β2-microglobulin and lactate were significantly higher and ACE was significantly lower in MS patients than in controls. β2-microglobulin and ACE positively correlated with high significance both in MS patients and controls. While disease duration negatively correlated and progression index, defined as EDSS score divided by disease duration in years, positively correlated with age-adjusted lactate levels, both did neither correlate with β2-microglobulin nor with ACE. Both CSF β2-microglobulin and ACE deserve further investigation as biomarkers of multiple sclerosis pathophysiology.


Ginsenoside Rf2, a new dammarane glycoside from Korean red ginseng (Panax ginseng).
A new dammarane glycoside named ginsenoside Rf2 has been isolated from Korean red ginseng (Panax ginseng) and its chemical structure has been elucidated as 6-O-[alpha-L-rhamnopyranosyl (1-->2) beta-D-glucopyranosyl]dammarane-3 beta, 6 alpha, 12 beta, 20(R), 25-pentol by chemical and spectral methods.


Kinetics of the glycation of bovine serum albumin by mannose and fucose in vitro.
Glycation of bovine serum albumin was measured for mannose and fucose at 37 degrees C. Mannose as well as fucose demonstrated an initial rapid increase in rate of formation of total adducts followed by a slower secondary reaction. The equilibrium constant for Schiff base formation was almost two times larger for mannose than fucose, although the Schiff base formed by fucose rearranged 1.5 times faster than that for mannose. Both sugars showed parallel lines for the formation of total and acid stable products after three hours. Discussion integrates new mechanistic data with previously suggested mechanisms.


Potential participation of calpain in platelet activation studied by use of a cell penetrating calpain inhibitor (calpeptin).
Employing a cell penetrating calpain inhibitor (calpeptin), the role of calpain in platelet activation was examined. In washed platelets (WPs) both thrombin and collagen-induced platelet aggregation were dose-dependently inhibited by calpeptin. The addition of plasma to WPs interfered with the action of calpeptin, however more than 3 min preincubation of calpeptin with WPs completely abolished the influence of plasma. In thrombin-activated WPs with calcium, the increase of intracellular calcium concentration, [Ca2+]i, and the production of inositol triphosphate (IP3) were dose-dependently inhibited by calpeptin. The generation of thromboxane B2 (TxB2) was inhibited by calpeptin in collagen and thrombin-activated WPs. In [3H]-arachidonic acid (AA)-labelled platelets, calpeptin increased the amount of [3H]-AA liberated by inhibiting [3H]-AA degradation after collagen or thrombin stimulation. When [14C]-AA degradation by the platelet suspension was observed, calpeptin inhibited TxB2 and hydroxyheptadecatrienoic acid (HHT) generation but increased prostaglandin (PG) E1, E2, 12-hydroxyeicosatetraenoic acid (12HETE) and AA. Based on these findings, calpain may be involved in the activation phospholipase C and thromboxane synthetase.


Intraoperative assessment of human spinal cord perfusion using near infrared spectroscopy with indocyanine green tracer technique.
Despite the significant interest in the assessment of human cerebral perfusion, investigations into human spinal cord perfusion (SCP) are scarce. Current intraoperative monitoring of spinal cord relies on the assessment of neural conduction as a surrogate for SCP. However, there are various inherent limitations associated with the use of these techniques. Near infrared spectroscopy (NIRS) has been successfully used for monitoring and assessment of human cerebral perfusion and has shown promising results in intraoperative assessment of SCP in animal models. The aim of this study was to investigate whether it is possible to monitor physiological changes in human SCP intraoperatively using NIRS with indocyanine green (ICG) tracer technique. We used this technique to calculate the human spinal cord carbon dioxide (CO₂) reactivity index. In addition, we investigated whether the lamina causes significant attenuation of NIRS signals. Intraoperative human experimental study. Eighteen patients undergoing elective posterior cervical spine surgery. Carbon dioxide reactivity of human SCP. Nine patients underwent transdural assessment of SCP, with an additional nine patients undergoing translaminar measurements. Patients' SCP was continuously monitored using an NIRO-500 NIRS monitor via a set of purpose built optodes. Their arterial ICG concentration was simultaneously assessed using a pulse dye densitometer. Patients' end-tidal CO₂ was gradually increased by 7.5 mm Hg and then returned back to baseline. Three sets of measurements were taken: baseline, hypercapnic, and return to baseline. After hypercapnia, SCP increased by a mean of 57.2 ± 23.3% in the transdural group and 46.6 ± 36.3% in the translaminar group. Carbon dioxide reactivity index was 7.6 ± 3.2%ΔSCP/mm Hg in the transdural group and 6.4 ± 5.3 %ΔSCP/mm Hg in the translaminar group. There was no significant difference in the increase in SCP (p=.475) or the CO₂ reactivity index (p=.581) observed between the transdural and the translaminar groups. Intraoperative NIRS with ICG tracer technique can identify an increase in the SCP in response to hypercapnia. It is possible to use this technique for monitoring SCP over the dura and the lamina. This technique could potentially be used to provide insight in to the pathophysiology and autoregulation of commonly acquired spinal cord conditions. Further research assessing the use of NIRS for monitoring of SCP is required.


Fasciocutaneous Flaps for Refractory Intermediate Stage Osteoradionecrosis of the Mandible-Is It Time for a Shift in Management?
Historically, free tissue transfer has been reserved for advanced stage osteoradionecrosis (ORN) of the jaw with pathologic fracture or cutaneous fistula. The purpose of this study was to evaluate if in patients with recalcitrant intermediate stage ORN, would mandibular debridement in combination with vascularized soft tissue coverage result in durable symptom and disease resolution. A retrospective cohort study of patients with mandibular ORN was performed comparing a mandibular preserving approach with a fasciocutaneous forearm flap for progressive treatment-resistant intermediate stage ORN to mandibulectomy and vascularized bone flap (VBF) reconstruction for advanced stage ORN. The primary outcome was ORN resolution. The sample was composed of 35 patients. One-hundred percent of patients undergoing a mandibular preserving approach experienced ORN resolution compared with 83.3% in the segmental mandibulectomy and VBF reconstruction group (P = .28). Patients in the mandibular preservation group experienced a shorter hospitalization (6 vs 9 days; P = .07), decreased length of surgery (384.9 vs 406.3 minutes; P = .01), and less delayed healing requiring local wound care (9.1 vs 45.8% of patients; P = .06). A mandibular preserving approach is successful at arresting intermediate stage ORN and is associated with a decreased operative time, a shorter hospitalization, and less need for prolonged wound care when compared with VBF reconstruction for advanced stage ORN.


Survey on immunotherapy practice patterns: dose, dose adjustments, and duration.
Practical issues dealing with the administration of allergen immunotherapy (AIT) by European and US allergists are not well known. Several concerns are only partially covered by guidelines. To survey AIT practice patterns among worldwide members of the American Academy of Allergy, Asthma and Immunology (AAAAI). A web-based survey was conducted among AAAAI members on dosing, dose adjustment after missed doses, and duration of AIT. A total of 1,201 replies (24.7% response rate of which 10% of responses were from non-US and non-Canada members). A total of 57% to 65% of the US-Canadian dosing falls within the recommended Practice Parameter ranges (9.4%-19% too low). Dose adjustment after missed doses is based on time elapsed since the last administered dose by 77% of US-Canadian and 58% of non-US-Canadian allergists. Doses are reduced when a patient comes in more than 14 days for 5 weeks after the last administration and initial dosing restarted after more than 30 days for 12 weeks since last administration during the build-up or maintenance stage. After missing 1 to 3 doses, the dosing schedules were mostly followed (build-up phase: repeat last dose, reduce by 1 dose, reduce by 2doses; maintenance phase: reduce by 1 dose, reduce by 2 doses, reduce by 3 doses). AIT is prescribed for a median of 3 years by non-US-Canadian allergists but for a median of 5 years by 75% of US-Canadian allergists. Main reasons for continuing beyond 5 years were "after stopping, symptoms reappeared" or "patient afraid to relapse." Many patients receive less than recommended doses. Two areas in which to plan further research are establishment of an optimal dose-adjustment plan for missed applications and exploration of the maximum appropriate duration of immunotherapy.


Tapping basement membrane motifs: Oral junctional epithelium for surface-mediated soft tissue attachment to prevent failure of percutaneous devices.
Teeth, long-lasting percutaneous organs, feature soft tissue attachment through adhesive structures, hemidesmosomes, in the junctional epithelium basement membrane adjacent to teeth. This soft tissue attachment prevents bacterial infection of the tooth despite the rich - and harsh - microbial composition of the oral cavity. Conversely, millions of percutaneous devices (catheters, dental, and orthopedic implants) fail from infection yearly. Standard of care antibiotic usage fuels antimicrobial resistance and is frequently ineffective. Infection prevention strategies, like for dental implants, have failed in generating durable soft tissue adhesion - like that seen with the tooth - to prevent bacterial colonization at the tissue-device interface. Here, inspired by the impervious natural attachment of the junctional epithelium to teeth, we synthesized four cell adhesion peptide (CAPs) nanocoatings, derived from basement membranes, to promote percutaneous device soft tissue attachment. The two leading nanocoatings upregulated integrin-mediated hemidesmosomes, selectively increased keratinocyte proliferation compared to fibroblasts, which cannot form hemidesmosomes, and expression of junctional epithelium adhesive markers. CAP nanocoatings displayed marked durability under simulated clinical conditions and the top performer CAP nanocoating was validated in a percutaneous implant murine model. Basement membrane CAP nanocoatings, inspired by the tooth and junctional epithelium, may provide an alternative anti-infective strategy for percutaneous devices to mitigate the worldwide threat of antimicrobial resistance. STATEMENT OF SIGNIFICANCE: Prevention and management of medical device infection is a significant healthcare challenge. Overzealous antibiotic use has motivated alternative material innovations to prevent infection. Here, we report implant cell adhesion peptide nanocoatings that mimic a long-lasting, natural "medical device," the tooth, through formation of cell adhesive structures called hemidesmosomes. Such nanocoatings sidestep the use of antimicrobial or antibiotic elements to form a soft-tissue seal around implants. The top performing nanocoatings prompted expression of hemidesmosomes and defensive factors to mimic the tooth and was validated in an animal model. Application of cell adhesion peptide nanocoatings may provide an alternative to preventing, rather that necessarily treating, medical device infection across a range of device indications, like dental implants.


Gambling Harm Experienced by Children Exposed to Parental Gambling: An Online Survey of Australians.
Although child wellbeing is known to be negatively affected by gambling, relatively little is known about the specific harms experienced by children exposed to parental gambling problems. The current study aimed to better understand gambling harm directly attributed to regular parental gambling in key areas of child wellbeing: financial, psychological, interpersonal wellbeing and intergenerational transmission of problem gambling. Using data from a national survey of Australian adults exposed to parental gambling under the age of 18 (n = 211), the results show that parental gambling was related significant levels of financial harm, abuse, neglect as well as relational and psychological problems as a direct result of parental gambling. The likelihood of experiencing gambling harms was positively associated with parental problem gambling severity. Harmful impacts of parental gambling as a child were also associated with a range of psychological problems in adulthood including depression, anxiety, Post-Traumatic Stress Disorder and intimate partner violence victimisation. Parental problem gambling severity was negatively associated with own lifetime gambling problems, suggesting a specific pattern of intergenerational transmission of problem gambling in children of regular, or heavy, gamblers. This research highlights the need for more supports for families with children in which at least one parent gambles regularly.


Cranial and cervical nerve injuries after carotid endarterectomy: a prospective study.
The purpose of this study was to review the outcome of patients who had cranial and cervical nerve injuries after carotid endarterectomy (CEA). This prospective study reviewed 200 consecutive CEAs. Preoperative and postoperative cranial nerve assessment was completed on all patients. Neurologic evaluation included routine direct fiberoptic laryngoscopy. Patients found to have no neurologic injury had no further follow-up. Patients with postoperative peripheral neurologic dysfunction were enrolled for regular long-term follow-up to assess delayed recovery. Overall, 25 (12.5%) nerve injuries were identified in 24 patients. There were 11 (5.5%) hypoglossal, 8 (4%) recurrent laryngeal, 2 (1%) superior laryngeal, 2 (1%) marginal mandibular, and 2 (1%) greater auricular nerve injuries. None of the patients were lost to follow-up. All nerve dysfunctions were transient, with all but 4 nerves recovering completely within 6 months. The recovery took from 1 week to 37 months, with a mean recovery time of 5.8 months. Two patients with recurrent laryngeal nerve dysfunction were found to have prolonged full recovery time (i.e., 31 and 37 months, respectively). Two patients successfully underwent contralateral CEA, although movement of the opposite vocal cord was not fully restored. Cranial nerve injury after CEA is a common occurrence and can be classified as a "major" or "minor" complication, depending on the severity of the clinical consequences. Extended follow-up will identify the specific subset of patients with a late complete nerve recovery.


[Surgical therapy of rectal carcinoma. Portrayal of the current health care status within the scope of a district quality assurance study].
In an era of economic pressure, patient's participation in decision-making and evidence-based medicine, the need for quality assurance of medical processes in daily routine becomes increasingly important. The aim of this quality assessment study is to demonstrate the real state of surgical care of patients with rectal carcinoma in the context of a field study on quality management in oncology. Prolective cohort-study, small-area-analysis. Between 01.01.1997 and 31.12.1998, all patients with newly diagnosed rectal carcinoma and treatment in the county of Marburg-Biedenkopf (252.975 inhabitants, 3 hospitals) were included in the study. Documentation included all variables (n = 82) relevant for primary surgical therapy according to guidelines and standards. 146 consecutive patients fulfilled the inclusion criteria. 93% underwent resection of the tumor. In 6.6% of these cases rigid rectoscopy for determination of the tumor's distance from the anal verge was not performed. In 16.9% the colon was not examined completely. In 98.5% the state of distant metastases was documented preoperatively. 93% of the patients with tumors in the upper/middle third of the rectum and curative-intent surgery (n = 68) received a sphincter-saving operation. For all curative-intent procedures negative resection margins were confirmed histologically. Complication rates were 31.6% for anterior resection and 46.9% for abdominoperineal extirpation. Comparative analysis of quality indicators revealed a predominantly good quality of care. Improving efforts can now be concentrated on few areas of suboptimal quality in the setting of internal quality management. Critical results require detailed analysis of the single cases. Thus, conditions requiring deviation from guidelines become apparent.


Immobilization of α-transglucosidase on silica-coated magnetic nanoparticles and its application for production of isomaltooligosaccharide from the potato peel.
In this study, the production of isomaltooligosaccharide from potato peel starch was carried out in three steps: liquefaction, saccharification, and transglucosylation. Further, cloning α-transglucosidase gene from Aspergillus niger (GH31 family), transforming into E. coli BL21 (DE3), overexpressing and purifying the resulting protein for the production of α-transglucosidase. The generated α-transglucosidase was then bound with magnetic nanoparticles, which improved reusability up to 5 cycles with more than 60% activity. All the modifications were characterized using the following methods: Fourier transform infra-red analysis, Transmission Electron Microscopy, Field Emission Scanning Electron Microscopy, Energy Dispersive X-ray spectroscopy, X-Ray Diffraction Spectroscopy, Thermogravimetric Analysis, and Dynamic Light Scattering (DLS) analysis. Further, the optimum conditions for transglucosylation were determined by RSM as follows: enzyme-to-substrate ratio 6.9 U g-1, reaction time 9 h, temperature 45 °C, and pH 5.5 with a yield of 70 g l-1 (± 2.1). MALDI-TOF-MS analysis showed DP of the IMOs in ranges of 2-10. The detailed structural characterization of isomaltooligosaccharide by GC-MS and NMR suggested the α-(1 → 4) and α-(1 → 6)-D-Glcp residues as major constituents along with minor α-(1 → 2) and α-(1 → 3) -D-Glcp residues.


Mechanism for microbial population collapse in a fluctuating resource environment.
Managing trade-offs through gene regulation is believed to confer resilience to a microbial community in a fluctuating resource environment. To investigate this hypothesis, we imposed a fluctuating environment that required the sulfate-reducer Desulfovibrio vulgaris to undergo repeated ecologically relevant shifts between retaining metabolic independence (active capacity for sulfate respiration) and becoming metabolically specialized to a mutualistic association with the hydrogen-consuming Methanococcus maripaludis Strikingly, the microbial community became progressively less proficient at restoring the environmentally relevant physiological state after each perturbation and most cultures collapsed within 3-7 shifts. Counterintuitively, the collapse phenomenon was prevented by a single regulatory mutation. We have characterized the mechanism for collapse by conducting RNA-seq analysis, proteomics, microcalorimetry, and single-cell transcriptome analysis. We demonstrate that the collapse was caused by conditional gene regulation, which drove precipitous decline in intracellular abundance of essential transcripts and proteins, imposing greater energetic burden of regulation to restore function in a fluctuating environment.


Prescription patterns and opioid usage in sinonasal surgery.
Excess opioid use after surgery contributes to opiate misuse and diversion. Understanding opioid prescribing and utilization patterns after sinonasal surgery is critical in designing effective practice protocols. In this study we aim to identify factors associated with variable opioid usage and further delineate optimal prescription patterns for sinonasal surgery. All patients undergoing sinonasal surgery within a single health-care system from March 2017 to August 2018 were sent electronic postoperative surveys. Data were collected on the amount of opioid required, pain control, presurgical opiate use, and narcotic disposal. Additional data collected from the electronic medical record included demographics, type of surgery performed, and total amount of opioid prescribed, including refills. Three-hundred sixty four patients were included. A mean number of 25.3 tablets were prescribed per patient, yet the mean taken was just 11.8 tablets. Excess opioids were prescribed 84.9% of the time with a mean excess narcotic in oral morphine equivalents of 152.5. Among patients, 11.8% reported using no opioids, whereas 52.1% used <50% and 36.1% used >50% of their narcotic prescription. Patients used 9.3% of their full prescription and only 2.6% required a refill. The amount used was not associated with complexity of endoscopic sinus surgery, type of opiate prescribed, gender, distance living from hospital, or current opioid usage before surgery (p > 0.05). The addition of septoplasty and/or turbinoplasty was associated with variation in opioid usage (p < 0.001). A total of 76.1% of patients incorrectly discarded/stored excess opiates. Opioids are overprescribed after sinonasal surgery. The amount of postoperative opiate prescribed should be greatly reduced and may be based on the specific procedures performed. Improved patient education regarding disposal of excess narcotics may help to curtail future opioid diversion.


Isochronal visualization of transcription and proteasomal proteolysis in cell culture or in the model organism, Caenorhabditis elegans.
Investigation of differential gene regulation by protein degradation requires analysis of the spatial and temporal association between proteolysis and transcription. Here, we describe the isochronal visualization of proteasomal proteolysis and transcription in cell culture or in vivo in the model organism Caenorhabditis elegans. This includes localization of proteasome-dependent proteolysis by fluorescent degradation products of model and endogenous substrates of the proteasome in combination with immunolabelling of RNA polymerase II and transcription in situ run-on assays.


On mouse and man: neutrophil gelatinase associated lipocalin is not involved in apoptosis or acute response.
Neutrophil gelatinase-associated lipocalin (NGAL) is a siderphore binding molecule present in the specific granules of neutrophils and induced in a variety of epithelial cells during inflammation. Its mouse orthologue, 24p3, is also an acute phase protein synthesized in the liver and adipose tissue during inflammation. 24p3 has recently been implicated in apoptosis of myeloid cells. We investigated whether similar features are characteristics of NGAL. First, isolated normal myeloid bone marrow cells were incubated with NGAL for 6 and 24 hr and analyzed for apoptosis by annexin V binding and by propidium iodide labeling. We found no indication that NGAL induces significant apoptosis in myeloid cells. Second, a human sepsis model where normal volunteers were given endotoxin 2 ng/kg intravenously, showed no evidence that NGAL is an acute phase protein. The plasma level of NGAL reflected the number of circulating neutrophils and was completely different from the kinetics of C-reactive protein. We thus conclude that major differences exist between mouse and man with regards to the role of this lipocalin in myelopoiesis and inflammation.


Exploration of Caregiver Experiences of Conservatively Managed End-Stage Kidney Disease to Inform Development of a Psychosocial Intervention: The ACORN Study Protocol.
End-stage kidney disease (ESKD) is an overwhelming illness that impacts not just patients, but also their informal carers. Patients who opt for conservative management rather than dialysis experience difficult symptoms and the psychosocial consequences of their condition. Informal carers of patients who choose conservative management can also experience high levels of psychosocial burden, yet there is little guidance on how best to support informal carers, and no evidence on psychosocial interventions to address unmet needs. The aim of this study is to explore the experiences and unmet needs of informal carers of patients with ESKD receiving conservative management in order to inform the development of a psychosocial intervention. This qualitative study will consist of three stages: (I) semi-structured interviews with informal carers in England and Northern Ireland, (II) focus groups with healthcare professionals and informal carers, and (III) national workshops to refine the components of a psychosocial intervention. Informal carers of patients with ESKD who are receiving conservative management experience a high psychosocial burden, but there is limited evidence on how best to provide support, particularly as the patient nears the end of life. To our knowledge this study will be the first to address this gap by exploring the experiences and unmet needs of informal carers, with the aim of informing the development of a psychosocial intervention to support the health and wellbeing of informal carers.


DichroIDP: a method for analyses of intrinsically disordered proteins using circular dichroism spectroscopy.
Intrinsically disordered proteins (IDPs) are comprised of significant numbers of residues that form neither helix, sheet, nor any other canonical type of secondary structure. They play important roles in a broad range of biological processes, such as molecular recognition and signalling, largely due to their chameleon-like ability to change structure from unordered when free in solution to ordered when bound to partner molecules. Circular dichroism (CD) spectroscopy is a widely-used method for characterising protein secondary structures, but analyses of IDPs using CD spectroscopy have suffered because the methods and reference datasets used for the empirical determination of secondary structures do not contain adequate representations of unordered structures. This work describes the creation, validation and testing of a standalone Windows-based application, DichroIDP, and a new reference dataset, IDP175, which is suitable for analyses of proteins containing significant amounts of disordered structure. DichroIDP enables secondary structure determinations of IDPs and proteins containing intrinsically disordered regions.


Antiferromagnetic spintronics.
Antiferromagnetic materials are internally magnetic, but the direction of their ordered microscopic moments alternates between individual atomic sites. The resulting zero net magnetic moment makes magnetism in antiferromagnets externally invisible. This implies that information stored in antiferromagnetic moments would be invisible to common magnetic probes, insensitive to disturbing magnetic fields, and the antiferromagnetic element would not magnetically affect its neighbours, regardless of how densely the elements are arranged in the device. The intrinsic high frequencies of antiferromagnetic dynamics represent another property that makes antiferromagnets distinct from ferromagnets. Among the outstanding questions is how to manipulate and detect the magnetic state of an antiferromagnet efficiently. In this Review we focus on recent works that have addressed this question. The field of antiferromagnetic spintronics can also be viewed from the general perspectives of spin transport, magnetic textures and dynamics, and materials research. We briefly mention this broader context, together with an outlook of future research and applications of antiferromagnetic spintronics.


Comparative analysis of the immunostimulatory properties of different adjuvants on the immunogenicity of a prototype parainfluenza virus type 3 subunit vaccine.
The immunogenicity of a parainfluenza virus type 3 (PIV-3) subunit vaccine consisting of affinity-purified haemagglutinin-neuraminidase (HN) and fusion (F) surface glycoproteins was tested in guinea-pigs and hamsters. The ability of several different immunopotentiating agents to enhance the antibody response of animals to the PIV-3 surface glycoproteins was evaluated. The immunity induced by HN and F alone was compared with the response elicited by purified proteins combined with Freund's complete adjuvant, aluminium phosphate, Syntex's threonyl-muramyl dipeptide (MDP) SAF-MF formulation, or Ribi's adjuvant formulation containing BCG cell wall skeleton (CWS), trehalose dimycolate (TDM) and monophosphoryl lipid A (MPL) in a 2% squalene-in-water emulsion. Purified proteins were also incorporated into three different liposome formulations prepared by the detergent dialysis procedure. Immunization of guinea-pigs and hamsters with two 15 micrograms doses of the PIV-3 surface glycoproteins administered in the absence of adjuvant elicited high haemagglutination inhibition, neutralization and anti-fusion titres. The liposome preparations failed to enhance the antibody titres. Ribi's adjuvant formulation was effective at inducing a good secondary response to the purified proteins while the immunostimulatory effects of aluminium phosphate, Syntex and Freund's adjuvants were clearly demonstrated in both primary and secondary responses. When administered without adjuvant, a 15 microgram dose of the HN and F mixture was capable of protecting hamsters against live virus challenge. The immunoprotective dose of the purified proteins could be reduced to at least 0.1 microgram by the addition of aluminium phosphate, Syntex or Freund's adjuvants.


46,XX aromatase deficiency: A single-center experience with the varied spectrum and recurrent variants, and a systematic review of hormonal parameters.
Aromatase deficiency is a rare disorder, with only a few cases reported in India. We describe a single-center experience in western India, with a systematic review of genetically proven 46,XX aromatase deficiency patients to evaluate hormonal parameters. Retrospective review of case records, collating phenotypic and genotypic data and molecular modeling. Systematic review of 46,XX aromatase deficiency, analyzing data on gonadotropins, estrogen and androgens. In the seven patients from our center, presentation was frequent in childhood or adolescence (4/7: delayed puberty or hyperandrogenism), with maternal virilization (4/7), predominance of Prader III/IV (5/7), and initial rearing as females (6/7). Three patients had hypoplastic ovaries. One patient had spontaneous regular menses. We report three novel (p.Arg115Pro, p.Arg192Pro, and c.145+1_145+4delins) and two recurrent variants (p.Val370Met, and c.145+1_145+4delins) in western and northern India, respectively. On systematic review (n=43), gonadotropins were elevated (FSH>LH) across ages (except preterm infants), androgens were elevated in about one-third of cases during childhood and puberty, and estradiol was lower than in controls in mini-puberty and puberty. Spontaneous thelarche and streak ovaries were significantly more frequent in patients with non-truncating and truncating variants, respectively. We report uncommon presentations with possible founder variants, and highlight hormonal parameters across ages. Serum FSH levels were elevated except in preterms, and can be used as a diagnostic marker.


Proteins secreted by the parasitic nematode Nippostrongylus brasiliensis act as adjuvants for Th2 responses.
Infections with parasitic helminths such as Nippostronglyus brasiliensis induce dominant type 2 responses from antigen-specific T helper cells. The potency of the Th2 bias can also drive Th2 responses to bystander antigens introduced at the same time as infection. We now report that the Th2-promoting effect of infection can be reproduced with soluble N. brasiliensis excretory-secretory proteins (NES) released by adult parasites in vitro. Immunization of BALB/c mice with NES results in the production of IL-4 with elevated total serum IgE and specific IgG1 antibodies. NES is also able to stimulate IL-4 and polyclonal IgE production in other mouse strains (C57BL/6, B10.D2, CBA). These features are seen whether NES is administered without adjuvant as soluble protein in phosphate-buffered saline or with complete Freund's adjuvant which normally favors Th1 responses. Thus, NES possesses intrinsic adjuvanticity. Moreover, co-administration of hen egg lysozyme (HEL) with NES in the absence of other adjuvants results in generation of HEL-specific lymphocyte proliferation, IL-4 release and IgG1 antibody responses, documenting that NES can act as an adjuvant for third-party antigens. Proteinase K digestion or heat treatment of NES before immunization abolished the IL-4-stimulating activity, indicating that the factors acting to promote Th2 induction are proteins secreted by the adult parasite.


Impact of postoperative compression dressing using polyethylene foam pad on the multimodal protocol for swelling control following total knee arthroplasty: a randomized controlled trial.
The multimodal approach is commonly applied to improve knee swelling after total knee arthroplasty. Although the use of a knee compression pad may improve knee swelling, its impact in the multimodal protocol remains unclear. A total of 57 knees were randomly assigned to one of 2 groups receiving treatment with either a compression dressing using a polyethylene foam pad designed to effectively compress the postoperative knee combined with an elastic bandage or a compression dressing with use of an elastic bandage only. All patients were treated with the multimodal approach, including an external cooling system, intravenous tranexamic acid, no pneumonic tourniquet, and periarticular multidrug injection. In intention-to-treat analysis, the circumferences of the thigh measured on postoperative days 1 and 7 were not significantly different between the 2 groups. In addition, there were no differences between the 2 groups in knee flexion angle, perioperative total blood loss, or complication rate. The polyethylene foam pad had no significant beneficial effect on knee swelling in patients undergoing total knee arthroplasty using the multimodal approach.


Fructose-1,6-diphosphatase and glucose-6-phosphatase in newborn rats with intrauterine growth retardation.
The activities of two gluconeogenic enzymes, glucose-6-phosphatase and fructose-1,6-diphosphatase were examined in the normal and intrauterine growth retarded (IUGR) rat during the first 5 days of life. The fructose-1,6-diphosphatase activity, 1.54 +/- 0.10 mumol/min/g liver (means +/- SEM) in control and 1.47 +/- 0.20 in the IUGR rats, increased in both groups on days 2--4 but remained significantly lower in the IUGR rats through day 4 (4.53 +/- 0.6 mumol/min/g liver in control and 3.09 +/- 0.22 mumul/min/g liver in the IUGR rats, P less than 0.01). The glucose-6-phosphatase activity increased similarly in both groups. The weight of the IUGR rats remained lower through the third postnatal day (6.47 +/- 0.42 compared to 8.64 +/- 0.27 g in control rats). Blood glucose concentrations at birth were 117 +/- 11 mg/dl in control rats and 73 +/- 11 mg/dl in the IUGR rats (P less than 0.01). Although the glucose concentrations increased in both groups on days 2--4, the IUGR rats maintained relatively lower levels (P less than 0.01). The results indicate that IUGR fetal rats do not have augmented gluconeogenesis in spite of hypoglycemia. In addition, effective gluconeogenesis in the neonatal period appears to be delayed.


A monoclonal antibody based solid-phase enzyme-binding assay to measure levels of placental alkaline phosphatase in serum of women during pregnancy.
Monoclonal antibodies raised against highly purified placental alkaline phosphatase (PALP) reacted specifically with PALP and were found not to cross-react with the liver, kidney and intestinal enzymes. One of these antibodies (AP-3) was selected to develop an enzyme-binding assay to measure levels of PALP in the sera of 11 women throughout pregnancy. PALP activity rose sharply after 28 weeks of gestation reaching a peak at delivery followed by an immediate drop within 4 days to values approaching non-pregnant levels. The potential of using this assay as an index of placental function to monitor progress of a pregnancy is discussed.


"Crack smoke" is a respirable aerosol of cocaine base.
The smoking of cocaine base [corrected] ("crack") has emerged as a significant substance abuse problem. A detailed characterization of cocaine smoke is a prerequisite for studies of its pharmacokinetics, abuse potential and toxicity. Model pipes were used to generate cocaine smoke analogous to that inhaled by human "crack" abusers. Using procedures to minimize pyrolysis, cocaine base smoke was determined to be 93.5% cocaine particles with the remainder being cocaine vapor. The average particle size generated from all model pipes was 2.3 mu which is small enough to ensure deposition into the alveolar region of the human lung. Although this particle size is eminently respirable [corrected] by primates, a much smaller fraction will reach the alveolar region of rodents. Special generating procedures would therefore be required to expose rodents to meaningful doses of airborne cocaine that mimic the rapid absorption achieved by "crack" smokers.


NK Cells and ILC1s in Cancer Biology.
Natural killer (NK) cells are cytotoxic innate lymphocytes that can kill tumor cells. While a majority of the early studies on the role of NK cells in cancer focused on hematopoietic tumors, there has been a growing interest in the role of NK cells in solid tumors. NK cells are grouped with innate lymphoid cells (ILCs) that include ILC1, a closely related but distinct cell whose role in antitumor immunity is incompletely understood. In this review we focus primarily on the role of NK cells in solid tumors and review the limited data available on the role of ILC1s in cancer.


A flow-injection ultrafiltration sampling chemiluminescence system for on-line determination of drug-protein interaction.
A flow-injection ultrafiltration sampling chemiluminescence system for on-line determination of cimetidine-bovine serum albumin (BSA) interaction is proposed in this paper. Cimetidine can be oxidized by N-bromosuccinimide (NBS) and sensitized by fluorescein to produce high chemiluminescence emission in basic media. The concentration of cimetidine is linear with the CL intensity in the range 3 x 10(-7) - 1 x 10(-4) mol L(-1) with a detection limit of 1 x 10(-7) mol L(-1) (3 sigma). The drug and protein were mixed in different molar ratios in 0.067 mol L(-1) phosphate buffer, pH 7.4, and incubated at 37 degrees C in a water bath. The ultrafiltration probe was utilized to sample the mixed solution at a flow rate of 5 micro L min(-1). The data obtained by the proposed ultrafiltration flow-injection chemiluminescence method was analyzed with Scrathard analysis and a Klotz plot. The estimated association constant ( K) and the number of the binding site ( n) on one molecule of BSA by Scrathard analysis and Klotz plot were 3.15 x 10(4) L mol(-1) and 0.95, 3.25 x 10(4) L mol(-1) and 0.92, respectively. The proposed system proved that flow-injection chemiluminescence analysis coupled with on-line ultrafiltration sampling is a simple and reliable technique for the study of drug-protein interaction.


Inhibition effect of secondary metabolites accumulated in a pervaporation membrane bioreactor on ethanol fermentation of Saccharomyces cerevisiae.
The secondary metabolites accumulated in a pervaporation membrane bioreactor during ethanol fermentation were mostly composed of acetic acid, lactic acid, propionic acid, citric acid, succinic acid and glycerol. The inhibition effect of these compounds at a broad concentration range was studied through ethanol fermentation by Saccharomyces cerevisiae. An increasing concentration of the secondary metabolites led to longer lag time and a reduction of cell growth. The specific cell growth rate, cell yield, ethanol productivity were only 0.061 h(-1), 0.024, 0.47 g L(-1) h(-1) respectively, when the medium contained 3.12 g of acetic acid, 10.23 g of lactic acid, 2.72 g of propionic acid, 1.35 g of citric acid, 2.26 g of succinic acid and 49.25 g of glycerol per liter (a concentration level in pervaporation membrane bioreactor at later fermentation period). By increasing pH of the medium to 6.0-8.0, the inhibition of these secondary metabolites could be greatly relieved.


Maturation of postural sway in young children.
The postural sway of 76 healthy young children aged from two to 14 years was investigated to identify age-related changes in the extent of sway, the effects of eye-closure, and the spectral composition of sway. Postural sway was measured from the excursions of the centre of pressure of ground reaction forces, and was analysed for both time and frequency. Within the age-range of the children tested, postural sway decreased linearly with age. Boys tended to become more stable at a faster rate than girls, but started from an initially greater level of instability. The children's Romberg quotients, which provide an index of the influence of eye-closure on sway magnitude, were remarkably low in comparison with adult values. Power spectral analysis of the postural sway showed that the principal energy was contained below 0.7 Hz for both lateral and antero-posterior sway, but that very young children had some power in the 0.8 to 2.0 Hz range. Taken together, these results confirm and extend the previously established view that children use visual information to control balance in a manner different from that of adults, and that it is not until after the age of seven years that adult-like balance-control strategies begin to appear. Also, in contrast to adults, the appearance of sway at high frequencies in children is not necessarily associated with any pathology.


Neotricitrinols A-C, unprecedented citrinin trimers with anti-osteoporosis activity from the deep-sea-derived Penicillium citrinum W23.
Marine fungi are prolific source for the discovery of structurally diverse and bioactive molecules. In our search for new anti-osteoporosis compounds from deep-sea-derived fungi, we prioritized a fungus whose extract exhibited moderate activity and rich chemical diversity. The investigation of this strain afforded a class of citrinins, including three new citrinin trimers, neotricitrinols A-C (1-3), and three known dimeric/monomeric precursors (4-6). Neotricitrinols A-C (1-3) feature a unique octacyclic carbon scaffold among the few reported citrinin trimers with their absolute configurations established by spectroscopic analysis, theoretical-statistical approaches (GIAO-NMR, TDDFT-ECD/ORD calculations), DP4+ probability analysis as well as biogenetic consideration. A plausible biosynthetic pathway linking 1-3 from the common intermediate metabolite penicitrinol A (4) was proposed. Biologically, neotricitrinol B (2) showed potential anti-osteoporosis activity by promoting osteoblastogenesis and inhibiting adipogenic differentiation on primary bone mesenchymal stem cells, while displaying no cytotoxicity.


Sympathetic nervous system plays a major role in acute cold/restraint stress inhibition of host resistance to Listeria monocytogenes.
BALB/c mice exposed to acute cold/restraint stress (ACRS) had significantly lower host resistance to Listeria monocytogenes (LM) than controls. The stress hormones corticosterone (CORT) and norepinephrine (NE), which are known to modulate immune responses, were evaluated as the cause of the decline in immune defense. The involvement of CORT and NE was investigated by pretreating mice with the CORT synthesis inhibitor metyrapone and the chemical sympathectomy drug 6-hydroxydopamine (6-OHDA), respectively. LM burdens in spleen and liver were determined three days post-infection. 6-OHDA significantly decreased the LM burden in both control and stressed animals. 6-OHDA also completely blocked the stress effects observed in spleens while only partially affecting the liver. The 6-OHDA-uptake inhibitor desipramine aided confirmation that peripheral sympathetic adrenergic nerves and NE depletion, rather than the direct action of 6-OHDA, were responsible for the decreased susceptibility to LM. The results suggest that the peripheral sympathetic nervous system (SNS) postganglionic neurotransmitter NE plays a major role in LM host resistance and has significant tissue-dependent effects after ACRS. In contrast, metyrapone-treated animals had further decreased host resistance to LM, suggesting a potential protective effect of CORT after ACRS. Altogether, the results suggest that stress hormones play an important role in stress-modulated host resistance and that NE is the major hormone involved in ACRS-induced suppression of host resistance.


Bivalent SMAC Mimetics for Treating Cancer by Antagonizing Inhibitor of Apoptosis Proteins.
Inhibitors of apoptosis proteins (IAPs) inhibit caspase activity, allowing various cancers to reduce programmed cell death (apoptosis) and resist drug treatment. The second mitochondrial-derived activator of caspases (SMAC) protein is an endogenous IAP antagonist, which can be considered as a potential anticancer therapy. Small-molecule SMAC mimetics based on the Ala-Val-Pro-Ile motif have been validated as potent IAP antagonists. In particular, most bivalent SMAC mimetics, which target both the baculovirus IAP repeat 2 (BIR2) and BIR3 domains in X-linked IAP (XIAP), antagonize IAPs better than the corresponding monovalent mimetics. Here we focus on strategies for designing bivalent small-molecule SMAC mimetics and progress in using them to antagonize IAPs. We also consider their clinical potential. Our discussion will hopefully help guide further study of these interesting mimetics.


The role of computed tomography in the diagnosis of brain tumors in infants and children.
CT has considerably facilitated the diagnosis of intracranial tumors, especially in infants and children. The authors review their experience with a series of 260 children with proven brain tumors. In most cases CT is so conclusive that other diagnostic techniques are no longer often needed. Pneumoencephalography in particular has largely been replaced by CT in recent years in the authors' departments, and the frequency of central ventriculography with contrast material has decreased substantially. Particular diagnostic difficulties with regard to tumors near to or within the caudal brain stem, some posterior fossa tumors, small vermian tumors, and tumors of the cerebral hemispheres are discussed.


Candidate genes for late-onset Alzheimer's disease: focus on chromosome 12.
In recent years, there was an increasing interest on candidate genes may play an important role in the development of Alzheimer's disease (AD). Several genome wide screens have undertaken so far or expanded recently, and suggested a number of genomic areas that may contain novel susceptibility genes for AD, in particular most compelling have been the findings on chromosome 12. Polymorphisms in different susceptibility genes on chromosome 12 (A2M, LRP1, CP2 and OLR1) are now being suggested as possible genetic markers for increased risk of developing AD. However, many of these studies are controversial and have shown conflicting results. Thus far, the search for the chromosome 12 Alzheimer's gene must continue and there are several other genes in this region that we are looking at. In this article, we focused on the current knowledge of the genetics of familial late-onset and sporadic AD linked to the chromosome 12, and the future search for other candidate genes.


Can income-based co-payment rates improve disparity? The case of the choice between brand-name and generic drugs.
Higher income population tend to prefer brand-name to generic drugs, which may cause disparity in access to brand-name drugs among income groups. A potential policy that can resolve such disparity is imposing a greater co-payment rate on high-income enrollees. However, the effects of such policy are unknown. We examined how patients' choice between brand-name and generic drugs are affected by the unique income-based co-payment rates in Japan; 10% for general enrollees and 30% for those with high income among the elderly aged 75 and over. We drew on cross-sectional price variation among commonly prescribed 311 drugs using health insurance claims data from a large prefecture in Japan between October 2013 and September 2014 to identify between-income-group differences in responses to differentiated payments. Running 311 multivariate logistic regression models controlling individual demographics, the median estimate indicated that high-income group was 3% (odds ratio = 0.97) less likely to choose a generic drug than the general-income group and the interquartile estimates ranged 0.92-1.02. The multivariate feasible generalized least squares model indicated high-income group's higher likelihood to choose brand-name drugs than the general-income group without co-payment rate differentiation (p < 0.001). Such gap in the likelihood was attenuated by 0.4% (p = 0.027) with an US$1 increase in the difference in additional payment/month for brand-name drugs between income groups - no gap with US$10 additional payment/month. This attenuation was observed in drugs for chronic diseases only, not for acute diseases. Income-based co-payment rates appeared to reduce disparity in access to brand-name drugs across income groups, in addition to reducing total medical expenditure among high-income group who shifted from brand-name drugs to generic ones due to larger drug price differences.


Effects on cell cycle progression and cytoskeleton organization of five Bothrops spp. venoms in cell culture-based assays.
Snake envenomation is a neglected tropical disease. In Brazil, the Bothrops genus is responsible for about 86% of snakebite accidents. Despite extensive evidence of the cytotoxicity of snake venoms, the cellular and molecular mechanisms involved are not fully understood, especially regarding the effects on cell cycle progression and cytoskeleton organization. Traditionally, the effectiveness and quality control tests of venoms and antivenoms are assessed by in vivo assays. Despite this, there is a rising effort to develop surrogate in vitro models according to the 3R principle (Replacement, Reduction, and Refinement). In this study, we treated rat liver cells (BRL-3A) with venoms from five Bothrops species (B. jararaca, B. jararacussu, B. moojeni, B. alternatus, and B. neuwiedi) and analyzed cell viability and IC50 by MTT assay, cell cycle phases distribution by flow cytometry, and morphology and cytoskeleton alterations by immunofluorescence. In addition, we evaluated the correlation between IC50 and the enzymatic and biological activities of each venom. Our results indicated that Bothrops spp. venoms decreased the cell viability of rat liver BRL-3A cells. The rank order of potency was B. jararacussu > B. moojeni > B. alternatus > B. jararaca > B. neuwiedi. The mechanisms of cytotoxicity were related to microtubules and actin network disruption, but not to cell cycle arrest. No clear correlation was found between the IC50 and retrieved literature data of in vitro enzymatic and in vivo biological activities. This work contributed to understanding cellular and molecular mechanisms underlying the Bothrops spp. venom cytotoxicity, which can help to improve envenomation treatment, as well as disclose potential therapeutic properties of snake venoms.


Morphological studies of the spleen in splenomegalic liver cirrhosis comparing with the spleen in idiopathic portal hypertension (so-called Banti's syndrome without liver cirrhosis).
Morphological changes of the spleen in splenomegalic liver cirrhosis (SLC) were studied. Comparisons with the normal spleen and the spleen of idiopathic portal hypertension (IPH) were made by (1) light microscopy with histometry, (2) scanning electron microscopy (SEM) with histometry, and (3) SEM of the spleen vascular replica. Histometrical studies by light microscopy showed that in both SLC and IPH, the white pulp volume was decreased in random units of splenic tissue but increased in the whole spleen, whereas the red pulp volume was increased both in random units of splenic tissue and in the whole spleen. The increase in the total volume of the white pulp was less marked in SLC than in IPH. SEM histometry demonstrated in the red pulp an increase in small venous sinuses and narrowing of the Billroth cord in SLC and IPH. Narrowing of the Billroth cord was more marked in SLC than in IPH. SEM of the white pulp showed channels formed by reticulum cells around the central artery in SLC and IPH. The channels are thought to correspond to so-called "follikuläre Fibroadenie" or periarterial fibrosis. The spleen vascular replica demonstrated open arterial termination into the tissue spaces in the Billroth cord in SLC, IPH and the normal. Venous sinuses were arranged in bundles in SLC and IPH.


Choroidal mass as an initial presentation of lung cancer.
Choroidal mass may be a presenting sign of systemic malignancy. The diagnosis of metastatic ocular tumor is important because it can indicate poor prognosis. We report a case of a 45-year-old man presenting to our clinic with the complaint of decreased vision in the left eye, which is the initial finding of non-small cell lung-carcinoma.


Modulatory effect of fibre-enriched cake on alloxan-induced diabetic toxicity in rat brain tissues.
Diabetes is a metabolic disorder characterized by hyperglycaemia and it is fast becoming a scourge in sub-Saharan Africa. The nutritional properties of developed fibre-enriched cake and its protective potential against diabetic induced neurotoxicity in rats were investigated. Fibre-enriched cake was developed from selected fruits and analysed for its nutritional and sensory attributes. Rats were induced with diabetes by a single intraperitoneal injection of alloxan and treated with the formulated cake. After 14 days treatment, the rats were sacrificed by cervical dislocation. Their brain tissues were accessed for reduced glutathione (GSH), catalase, superoxide dismutase (SOD) activities, protein content and lipid peroxidation as well as lipid profiles which cover for total cholesterol, triglycerides, HDL and LDL. Induction of diabetes led to significant reduction (p < 0.05) of GSH, catalase, SOD activities and protein content. Feeding on the formulated cake led to their significant increase. Decreased lipid peroxidation, total cholesterol, LDL and triglycerides, and increased concentration of HDL were also observed on feeding with the cake. These results indicate an antioxidant protective potential of the fibre-enriched cake against diabetic-induced brain toxicity. Thus, it can serve as an adjunct to dietary therapy for diabetes.


The AO tubular external fixator in the treatment of open fractures and infected non-unions of the shaft of the femur.
A series of 14 cases of open fractures of the shaft of the femur and another group of 13 cases of infected non-union of the shaft of the femur are reported. They were treated with the AO tubular external fixator. The complications and results are assessed and it is concluded that the AO external fixator system has a valuable part to play in these difficult cases.


Sticking like sticky tape: tree frogs use friction forces to enhance attachment on overhanging surfaces.
To live and clamber about in an arboreal habitat, tree frogs have evolved adhesive pads on their toes. In addition, they often have long and slender legs to facilitate not only long jumps, but also to bridge gaps between leaves when climbing. Both adhesive pads and long limbs are used in conjunction, as we will show in this study. Previous research has shown that tree frogs change from a crouched posture (where the limbs are close to the body) to a sprawled posture with extended limbs when clinging on to steeper inclines such as vertical or overhanging slopes. We investigated this change in posture in White's tree frogs (Litoria caerulea) by challenging the frogs to cling onto a tiltable platform. The platform consisted of an array of 24 three-dimensional force transducers, which allowed us to measure the ground reaction forces of the frogs during a tilt. Starting from a crouched resting position, the normal forces on the forelimbs changed sign and became increasingly negative with increasing slope angle of the platform. At about 106° ± 12°, tilt of the platform the frogs reacted by extending one or two of their limbs outwards. At a steeper angle (131° ± 11°), the frogs spread out all their limbs sideways, with the hindlimbs stretched out to their maximum reach. Although the extension was strongest in the lateral direction, limbs were significantly extended in the fore-aft direction as well. With the extension of the limbs, the lateral forces increased relative to the normal forces. The large contribution of the in-plane forces helped to keep the angle between the force vector and the platform small. The Kendall theory for the peeling of adhesive tape predicts that smaller peel angles lead to higher attachment forces. We compare our data with the predictions of the Kendall model and discuss possible implications of the sliding of the pads on the surface. The forces were indeed much larger for smaller angles and thus can be explained by peeling theory.


Mesenchymal stem cells shift the pro-inflammatory phenotype of neutrophils to ameliorate acute lung injury.
Mesenchymal stem cell (MSC) treatment plays a major role in the management of acute lung injury (ALI), and neutrophils are the initial line of defense against ALI. However, the effect of MSCs on neutrophils in ALI remains mostly unknown. We investigated the characteristics of neutrophils in lung tissue of ALI mice induced by lipopolysaccharide after treatment with MSCs using single-cell RNA sequencing. Neutrophils separated from lung tissue in ALI were co-cultured with MSCs, and then samples were collected for reverse transcription-polymerase chain reaction and flow cytometry. During inflammation, six clusters of neutrophils were identified, annotated as activated, aged, and circulatory neutrophils. Activated neutrophils had higher chemotaxis, reactive oxygen species (ROS) production, and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase scores than aged neutrophils. Circulatory neutrophils occurred mainly in healthy tissue and were characterized by higher expression of Cxcr2 and Sell. Activated neutrophils tended to exhibit higher expression of Cxcl10 and Cd47, and lower expression of Cd24a, while aged neutrophils expressed a lower level of Cd47 and higher level of Cd24a. MSC treatment shifted activated neutrophils toward an aged neutrophil phenotype by upregulating the expression of CD24, thereby inhibiting inflammation by reducing chemotaxis, ROS production, and NADPH oxidase. We identified the immunosuppressive effects of MSCs on the subtype distribution of neutrophils and provided new insight into the therapeutic mechanism of MSC treatment in ALI.


Recent advances in point spread function engineering and related computational microscopy approaches: from one viewpoint.
This personal hybrid review piece, written in light of my recipience of the UIPAB 2020 young investigator award, contains a mixture of my scientific biography and work so far. This paper is not intended to be a comprehensive review, but only to highlight my contributions to computation-related aspects of super-resolution microscopy, as well as their origins and future directions.


In vivo evaluation of a transdermal codrug of 6-beta-naltrexol linked to hydroxybupropion in hairless guinea pigs.
6-Beta-naltrexol is the major active metabolite of naltrexone, NTX, a potent mu-opioid receptor antagonist used in the treatment of alcohol dependence and opioid abuse. Compared to naloxone, NTX has a longer duration of action largely attributed to 6-beta-naltrexol. This study was carried out in order to determine percutaneous absorption of a transdermal codrug of naltrexol, 6-beta-naltrexol-hydroxybupropion codrug (CB-NTXOL-BUPOH), in hairless guinea pigs as well as to evaluate the safety of 6-beta-naltrexol for development as a transdermal dosage form. This codrug may be useful in the simultaneous treatment of alcohol dependence and tobacco addiction. The carbonate codrug traversed the skin at a faster rate than 6-beta-naltrexol. 6-Beta-naltrexol equivalent steady state plasma concentrations of 6.4 ng/ml were obtained after application of the codrug as compared to 1.2 ng/ml from 6-beta-naltrexol base. The steady state plasma concentration of hydroxybupropion after codrug application was 6.9 ng/ml. Skin sensitization and irritation tested in the hairless guinea pigs using the Buehler method revealed that 6-beta-naltrexol had no skin sensitizing potential. The method was validated with a known sensitizer, p-phenylenediamine, which induced sensitization in 90% of the animals. 6-beta-Naltrexol caused only mild transient skin irritation after the initial application of the patch. During subsequent applications, erythema was slightly increased but no skin damage was observed. In conclusion, a transdermal codrug of 6-beta-naltrexol could be a viable alternative treatment for alcohol and opiate abuse.


NER-factor DDB2 regulates HIF1α and hypoxia-response genes in HNSCC.
Cancers in the oral/head & neck region (HNSCC) are aggressive due to high incidence of recurrence and distant metastasis. One prominent feature of aggressive HNSCC is the presence of severely hypoxic regions in tumors and activation of hypoxia-inducible factors (HIFs). In this study, we report that the XPE gene product DDB2 (damaged DNA binding protein 2), a nucleotide excision repair protein, is upregulated by hypoxia. Moreover, DDB2 inhibits HIF1α in HNSCC cells. It inhibits HIF1α in both normoxia and hypoxia by reducing mRNA expression. Knockdown of DDB2 enhances the expression of angiogenic markers and promotes tumor growth in a xenograft model. We show that DDB2 binds to an upstream promoter element in the HIF1Α gene and promotes histone H3K9 trimethylation around the binding site by recruiting Suv39h1. Also, we provide evidence that DDB2 has a significant suppressive effect on expression of the endogenous markers of hypoxia that are also prognostic indicators in HNSCC. Together, these results describe a new mechanism of hypoxia regulation that opposes expression of HIF1Α mRNA and the hypoxia-response genes.