Monocarboxylate transporter 1 mediates DL-2-Hydroxy-(4-methylthio)butanoic acid transport across the apical membrane of Caco-2 cell monolayers.
The methionine hydroxy analogue DL-2-hydroxy-(4-methylthio)butanoic acid (DL-HMB) is a supplementary source of methionine commonly added to commercial animal diets to satisfy the total sulfur amino acid requirement. In this study, we characterized DL-HMB transport across the apical membrane of Caco-2 cells to identify the transport mechanism involved in the intestinal absorption of this methionine source. DL-HMB transport induced a significant decrease in intracellular pH (pH(i)) and was inhibited in the presence of the protonophore carbonyl cyanide 4-(trifluoromethoxy)-phenylhydrazone. Moreover, both Na(+) removal and 5-(N-ethyl-N-isopropyl)amiloride, an inhibitor of apical Na(+)/H(+) exchanger (NHE3), significantly reduced substrate uptake and pH(i) recovery, suggesting cooperation between H(+)-dependent DL-HMB transport and NHE3 activity. cis-Inhibition experiments with L-Ala, beta-Ala, D-Pro, betaine, or glycyl-sarcosine excluded the participation of systems proton amino acid transporter 1 and peptide transporter 1. In contrast, alpha-cyano-4-hydroxycinnamate, phloretin, L-lactate, beta-hydroxybutyrate, butyrate, and pyruvate, inhibitors and substrates of monocarboxylate transporter 1 (MCT1), significantly reduced DL-HMB uptake. Dixon plot analysis of L-lactate transport in the presence of DL-HMB revealed a competitive interaction (inhibition constant, 17.5 +/- 0.11 mmol/L), confirming the participation of system MCT1. The kinetics of DL-HMB uptake was described by a model involving passive diffusion and a single low-affinity, high-capacity transport mechanism (K(D), 1.9 nL/microg protein; K(m), 13.1 +/- 0.04 mmol/L; and V(max), 43.6 +/- 0.14 pmol/microg protein) compatible with MCT1 kinetic characteristics. In conclusion, the methionine hydroxy analogue is transported in Caco-2 cell apical membrane by a transport mechanism with functional characteristics similar to those of MCT1.


A novel, cell-type-specific mechanism for estrogen receptor-mediated gene activation in the absence of an estrogen-responsive element.
The estrogen receptor (ER) typically activates gene transcription by binding to estrogen-responsive elements (EREs). The brain creatine kinase (BCK) promoter is responsive to estrogen but contains no ERE-related sequence. To investigate the mechanism of estrogen induction, we have introduced the estrogen receptor into HeLa cells and primary rat cardiomyocytes and fibroblasts along with 195 bp of BCK promoter linked to a chloramphenicol acetyltransferase (CAT) reporter gene. A 10-fold stimulation of CAT activity was observed in the presence of beta-estradiol in both HeLa and rat primary fibroblasts, but no induction was observed in primary rat cardiomyocytes. In contrast, a control vitellogenin gene construct which contains a typical ERE was induced in an ER-dependent manner in all cell types studied. Estrogen induction in HeLa was not sensitive to cycloheximide and was blocked by the ER antagonists tamoxifen and ICI 164,384. Analysis of 5' deletion and linker-scanning mutations indicates sequences between bp -45 and -75 including a TA-rich sequence and a CCAAT sequence to be crucial for stimulation of the BCK promoter by the ER. BCK estrogen induction is dependent on the DNA-binding domain and transactivation domain TAF2 of the ER. However, direct DNA binding is probably not required. Taken together, these results suggest a novel mechanism for ER-mediated gene activation. This mechanism is consensus ERE independent and cell type specific and requires interactions between the ER and molecules capable of interacting with the BCK promoter TA-rich region.


[Depression during pregnancy and preterm delivery. Epistemological aspects in research].
This paper examines recent research on the association between depression during pregnancy and preterm delivery. Dissemination of scientific information on this disease entity (depression) means that it is often present in common-sense discourse, although frequently used rather imprecisely. While various areas from the mental health field approach depression in very different ways, they suggest the influence of emotional states on organic outcomes. Still, this relationship remains difficult to prove by conventional scientific research methods. One problem in establishing associations between depression and preterm delivery appears to be due to epistemological aspects related to the theoretical demarcation of depression.


Biologically enhanced healing of the rotator cuff.
Failure of rotator cuff repair is a well-documented problem. Successful repair is impeded by muscle atrophy, fat infiltration, devascularization, and scar tissue formation throughout the fibrocartilagenous transition zone. This case study exemplifies a technique to biologically augment rotator cuff healing. Clinically, pain and function improved. Postoperative magnetic resonance imaging evaluation confirmed construct integrity. Biological enhancement of the healing process and physiologically based alterations in rehabilitation protocols can successfully treat complicated rotator cuff tears. Prospective studies with larger sample sizes and continued follow-up are necessary to assess the definitive efficacy of this treatment modality.


Effect of the CO-2-bicarbonate buffer system on the water and ion contents of rat renal cortical slices.
(1) Partial replacement of medium chloride by bicarbonate, with pHmaintained constant by addition of CO2, lead to a greater rate of swelling when rat renal corti cal slices were leached in nitrogenated medium at 0--2 degree C. This greater rate of swelling was still present when 1 mM iodoacetate was added to the leaching media. (2) Rat renal cortical slices incubated in oxygenated bicarbonate medium at 25 degree C maintained a slightly greater tissue water content than did slices incubated in phosphate-buffered media. Inclusion of malate in the incubation medium also led to a greater steady-state water content when slices were incubated at 25 degree C in oxygenated media. These results suggest that that particular cell volume which is maintained by metabolism and the double Donnan system is determined at least in part by the nature of the extracellular anions. (3) The rates of net sodium extrusion and of net potassium uptake were not significantly altered by presence of CI2 and bicarbonate in the incubation medium.


Very echogenic amniotic fluid: ultrasonography-amniocentesis correlation.
Very echogenic amniotic fluid has been variably attributed to meconium, blood, or vernix caseosa. However, most previous reports have been case reports, and most cases have not had proof by amniocentesis. In a larger series of patients with proof by amniocentesis, we sought to determine the relative frequency of these substances as causes of very echogenic amniotic fluid. We retrospectively identified obstetric sonograms in which the amniotic fluid was homogeneously filled with innumerable echogenic particles. The cause of the increased echogenicity was determined by fluid appearance at amniocentesis. Of 86 cases identified, immediate proof by amniocentesis was available in 19 patients for whom the gestational age ranged from 32.8 to 39.4 weeks. Vernix was present in 18 (95%) patients and meconium in one (5%) patient. Very echogenic amniotic fluid in the third trimester is most often due to vernix and infrequently due to meconium. This sonographic finding is not a reliable indicator of meconium or blood in amniotic fluid and should not typically alter antenatal management.


A Prospective Study of the Age at Menarche in North Indian Girls, Its Association With the Tanner Stage, and the Secular Trend.
Background Menarche is an important event in a female's reproductive health. However, the age at menarche is variable and has implications on the future health of the girl. The purpose of this study was to estimate the mean age at menarche of North Indian adolescent girls and its association with the Tanner stage and to study the trend of menarcheal age in India in the 21st century. Materials & methods A longitudinal descriptive study was conducted on 470 healthy girls aged nine to 16 years in expectant menarche (Tanner stage II). They were followed for six months to check for the attainment of menarche. Those who achieved menarche were grouped in Group I and the rest in Group II. The data were analyzed using the independent t-test. Results Menarche was achieved by 263 girls in the six-month follow-up period. The mean (SD) age of menarche was 13.13 (1.23) years. Group I girls were mostly in Tanner stage IV and above. Group II girls had a mean (SD) age of 11.53 (1.1) years and were mostly in Tanner stage III or below. The estimated decline in the age of menarche in the 21st century was about 0.41 years per decade. Conclusion The girls who achieved menarche had a significantly higher age and Tanner stage of sexual maturity than girls who did not achieve menarche in the study period. Tanner stage is a better measure to estimate the pubertal onset than age. The studies on the age of menarche in the current century reveal that the declining trend in the menarcheal age is continuing in India.


Appropriate care for people with learning disabilities.
How can nurses meet the healthcare needs of people with learning disabilities in the community? Solutions include addressing communication difficulties and issues of consent, and developing professionals' education.


Antithrombotic activity of losartan in two kidney, one clip hypertensive rats. A study on the mechanism of action.
The potential antithrombotic action of losartan, an AT1 receptor antagonist, administered to two-kidney, one-clip rats (2K1C) in an experimental model of venous thrombosis was evaluated. The involvement of nitric oxide (NO) in this effect was also studied. Venous stasis was induced by ligation of the vena cava. Losartan after single dose (10 mg/kg, p.o.) significantly reduced the venous thrombus growth. The antithrombotic action of losartan in 2K1C rats was abolished by N(G)-nitro-L-arginine methyl ester (L-NAME, 30 mg/kg s.c.) and restored by L-arginine (1000 mg/kg s.c.). Platelet adhesion to fibrillar collagen significantly decreased after administration of losartan. No changes in primary hemostasis and platelet aggregation were observed. Moreover, coagulation parameters such as activated partial thromboplastin time, prothrombin time and euglobulin clot lysis time were found unchanged after losartan administration either in systemic circulation or at the place of thrombus formation. Our results indicate that antithrombotic activity of losartan in 2K1C rats is NO--dependent; observed inhibition of platelet adhesion could also play a role in this phenomenon.


Are Physical Activities Associated With Perceived Stress? The Evidence From the China Health and Nutrition Survey.
Psychological stress is a negative affective state. The association between physical activity and psychological stress was commonly reported in previous researches. Few published studies with large sample sizes have explored such an association in Chinese population. The current research aims to assess the association between perceived stress and physical activity preferences, as well as the association between risk of high perceived stress and physical activity behaviors (e.g., sports-, transport-, occupational-related physical activity, sedentary activities and time in bed). The data were collected from the China Health and Nutrition Survey (CHNS) in 2015. Generalized linear models and logistic regression models were used to explore the association between perceived stress and physical activity. In total, 11,066 participants were included in the current analysis. For physical activity preferences, we found that "like" preference of all six mentioned domains of activities contributed to lower perceived stress (i.e., lower perceived stress scale -14 score, all β < 0 and p < 0.05). For physical activity behaviors, the results indicated that none/low intensity physical activity behaviors (e.g., sports-, occupational- related, sedentary) were associated with risk of high perceived stress, except that low intensity of transport-related activities seemed to be protective from high stress. However, the association between intensity of physical activity behaviors and perceived stress was not simply the higher the better. High intensity of all these physical activity behaviors was also associated with high perceived stress. Our findings suggest that positive preferences and moderate physical activity behaviors were associated with low perceived stress. The findings herein highlight the effect of regulating physical activity on perceived stress, as well as inform potential strategies to reduce psychological stress.


[RANK INDICES METHOD AND ITS USE FOR THE COMPARATIVE ANALYSIS OF POPULATION HEALTH].
There is presented a calculation method aimed to elevate the informative value of the integral indices of the social and hygienic monitoringfor purposes of comparative analysis. The method of rank indices is based on the ranking of monitoring objects on the values of primary indices on the base of which there are calculated the integral such indices as, for example, life expectancy. There are presented results of the use of this method for the comparative analysis of mortality rate in WHO Member States for the period of 1990-2011. There were revealed specialfeatures of mortality trends which cannot be detected when using only mortality rates or the life expectancy. In particular, for Russia there was shown that, in spite of the downward trend in child and adolescent mortality rate observed in the last decade, the country's world rankings for these indices fail to achieve the level of 1990. This means that the competitiveness of the country, sharply declined in the 90's, was not restored until now. There are described some features of the use of the method of rank indices for the analysis of indices of the environment state, public health and its socio-economic determinants.


"I Gotta Go With Modern Technology, So I'm Gonna Give 'em the Narcan": The Diffusion of Innovations and an Opioid Overdose Prevention Program.
Fatal opioid overdoses can be prevented by opioid overdose prevention programs (OOPPs). The present study qualitatively examined the diffusion process of an OOPP among 30 persons who inject drugs (PWIDs) in an opioid-saturated community. Purposive sampling was used to recruit participants into three groups based on familiarity with the OOPP. Findings revealed that participants often adopted the OOPP, which was offered by a local harm reduction organization, if first exposed by staff hosting and implementing it. Barriers to adoption included belief that OOPP training was lengthy or unnecessary, lack of perceived relative advantage, nonengagement with the host organization, and trepidation of administering withdrawal-causing medication to fellow PWIDs. Participants outside of networks diffusing the OOPP were isolated from other PWIDs. Staff from the host organization were influential in encouraging OOPP adoption, which underscores their importance in the effort to reduce fatal overdoses.


Synthesis of hexagonal BaTa2O6 nanorods and influence of defects on the photocatalytic activity.
Hexagonal barium tantalate (BaTa2O6) nanorods were synthesized by a hydrothermal method based on the reaction of concentrated Ba(OH)2 solution and Ta2O5. BaTa2O6 samples show a uniform cylindrical structure with diameters of 5-30 nm and the lengths of 50-200 nm. The formation of BaTa2O6 nanorods follows a dissolution-recrystallization mechanism and is governed by hydrothermal temperature and time. BaTa2O6 nanorod samples prepared at 270 degrees C for 72 h have exhibited the highest photocatalytic activity in the degradation of Rhodamine B (RhB) in aqueous solution under UV radiation. Hydrogen-related defects were detected in BaTa2O6 nanorods, which originate from the oxygen octahedron. The number of defects was dependent on the hydrothermal temperature, and the photocatalytic activities of BaTa2O6 nanorods increase with the decrease of defect amounts. On the basis of the experiment results, the difference in photocatalytic activities for samples is mainly caused by lattice defects, which can act as inactivation centers.


Genomic profiling of long non-coding RNA and mRNA expression associated with acquired temozolomide resistance in glioblastoma cells.
Temozolomide (TMZ) is an alkylating chemotherapeutic agent widely used in anti-glioma treatment. However, acquired TMZ resistance represents a major clinical challenge that leads to tumor relapse or progress. This study investigated the genomic profiles including long non-coding RNA (lncRNA) and mRNA expression associated with acquired TMZ resistance in glioblastoma (GBM) cells in vitro. The TMZ-resistant (TR) of GBM sub-cell lines were established through repetitive exposure to increasing TMZ concentrations in vitro. The differentially expressed lncRNAs and mRNAs between the parental U87 and U87TR cells were detected by human lncRNA microarray method. In this study, we identified 2,692 distinct lncRNAs demonstrating >2-fold differential expression with 1,383 lncRNAs upregulated and 1,309 lncRNAs downregulated. Moreover, 4,886 differential mRNAs displayed 2,933 mRNAs upregulated and 1,953 mRNAs downregulated. Further lncRNA classification and subgroup analysis revealed the potential functions of the lncRNA-mRNA relationship associated with the acquired TMZ resistance. Gene ontology and pathway analysis on mRNAs showed significant biological regulatory genes and pathways involved in acquired TMZ resistance. Moreover, we found the ECM‑receptor interaction pathway was significantly downregulated and ECM related collagen Ι, fibronectin, laminin and CD44 were closely associated with the TR phenotype in vitro. Our findings indicate that the dysregulated lncRNAs and mRNAs identified in this work may provide novel targets for overcoming acquired TMZ resistance in GBM chemotherapy.


Structure and activity of insulin, XIII. Specificity of the arginine-guanidino group in biologically active tetrapeptides of the insulin sequence B 22-25 (Arg-Gly-Phe-Phe).
The biologically active partial sequence Arg-Gly-Phe-Phe (position B 22-25 of the insulin B chain) in the form of the synthetic tetrapeptidamide, was compared in several bioassays with the following analogous synthetic peptides: homoarginyl-, ornithyl-, lysyl-, citrullyl-, alanyl- and NG-nitroarginyl-Gly-Phe-Phe-NH2. The syntheses of the lysyl- and alanyl-tetrapeptidamides are described. After intraperitoneal injection of the peptides in doses of 3-100 mumol per 100 g rat, together with [U-14C]glucose, the natural sequence Arg-Gly-Phe-Phe showed the highest insulin like activity (incorporation of labeled carbon into the diaphragm). The activity of the homoarginyl peptide was a little weaker. The ornithyl- and the lysyl-peptide, however, showed a remarkably diminished activity. The activity of the citrullyl-peptide was even lower and the alanyl-peptide was inactive. In vitro assays with rat diaphragm showed the same range of effects for the elevation of glucose uptake and glycogen content of the diaphragm. The activity decreased in the following order: Arg- greater than Har- greater than Orn- greater than Cit-Gly-Phe-Phe-NH2. Alanyl- and Nitroarginyl-Gly-Phe-Phe-NH2 were without effect. In isolated fat cells the glucose oxidation was enhanced significantly only by the arginyl-peptide. The results show that among the structures examined the guanidino group carried by the C5 chain of arginine is the most effective. The results are in accordance with our preceding work [1] using semisynthetic insulins obtained from natural A-chain and synthetic B-chain variants. In these products the replacement of Arg B 22 by ornithine or lysine also led to drastically diminished activity and after replacement of Arg B 22 by alanine the activity also disappeared.


Lipofuscin inhibits the proteasome by binding to surface motifs.
Lipofuscin, a highly oxidized aggregate, consists of covalently cross-linked proteins, lipids, and sugar residues and is one of the major life-span-limiting factors in postmitotic aging cells. An artificial model of this material, showing characteristics and effects comparable to those of the natural form, has turned out to be very useful for in vitro studies. Artificial lipofuscin was used to investigate its effects on the viability of human fibroblasts, its rate of uptake, and its ability to inhibit the proteasomal system. The inhibition of the proteasomal system is one of the major aspects of the cytotoxic effects of lipofuscin. We present here that this proteasomal inhibition is due to proteasomal binding to the lipofuscin surface motifs, degradable by protease K. Furthermore, removal of the surface peptide structures by protease K strongly reduces the cytotoxic effects of lipofuscin and binding of cellular proteins and proteasomes to intracellular protein aggregates.


A polyoxyethylene-substituted bimetallic europium helicate for luminescent staining of living cells.
The homoditopic ligand H2LC3 has been designed to form neutral triple-stranded bimetallic helicates of overall composition [Ln2(LC3)3]. The grafting of the polyoxyethylene fragments ensures water solubility and also favors cell penetration while being amenable to further derivatization. The ligand pKa values have been determined by spectrophotometric titration and range from 3.5 (sum of the first two) to 10.3. The thermodynamic stability of the helicates is large at physiological pH (logbeta23 in the range 22-23). The ligand triplet state has an adequate energy (0-phonon transition at approximately 20,800 cm(-1)) for sensitizing the luminescence of EuIII (Q=11%). Analysis of the EuIII emission spectrum points to an overall pseudo D3 symmetry for the metal environment. No significant effect of [Eu2(LC3)3] is observed on the viability of several cancerous cell lines (MCF-7, HeLa, Jurkat, and 5D10). The cell imaging properties of the EuIII helicate are demonstrated for the HeLa cell line by luminescence microscopy. Bright EuIII emission is seen for helicate concentration>50 microM and after 20-30 min loading time. The helicate stains the cytoplasm and the permeation mechanism is likely to be endocytosis.


Scientific data from precipitation driver response model intercomparison project.
This data descriptor reports the main scientific values from General Circulation Models (GCMs) in the Precipitation Driver and Response Model Intercomparison Project (PDRMIP). The purpose of the GCM simulations has been to enhance the scientific understanding of how changes in greenhouse gases, aerosols, and incoming solar radiation perturb the Earth's radiation balance and its climate response in terms of changes in temperature and precipitation. Here we provide global and annual mean results for a large set of coupled atmospheric-ocean GCM simulations and a description of how to easily extract files from the dataset. The simulations consist of single idealized perturbations to the climate system and have been shown to achieve important insight in complex climate simulations. We therefore expect this data set to be valuable and highly used to understand simulations from complex GCMs and Earth System Models for various phases of the Coupled Model Intercomparison Project.


Childbirth education in 1980.
A questionnaire was developed to elicit information from childbirth educators regarding their preparation; their clients; their business arrangements; and their opinions on class content, national guidelines, and certification. From 776 questionnaires mailed, 238 usable responses were obtained. Persons of varing backgrounds enter childbirth education through a variety of routes, practice in a variety of settings, and use varied content. A majority agree that there is a need for national guidelines for preparation and content.


RNase L Suppresses Androgen Receptor Signaling, Cell Migration and Matrix Metalloproteinase Activity in Prostate Cancer Cells.
The interferon antiviral pathways and prostate cancer genetics converge on a regulated endoribonuclease, RNase L. Positional cloning and linkage studies mapped Hereditary Prostate Cancer 1 (HPC1) to RNASEL. To date, there is no correlation of viral infections with prostate cancer, suggesting that RNase L may play additional roles in tumor suppression. Here, we demonstrate a role of RNase L as a suppressor of androgen receptor (AR) signaling, cell migration and matrix metalloproteinase activity. Using RNase L mutants, we show that its nucleolytic activity is dispensable for both AR signaling and migration. The most prevalent HPC1-associated mutations in RNase L, R462Q and E265X, enhance AR signaling and cell migration. RNase L negatively regulates cell migration and attachment on various extracellular matrices. We demonstrate that RNase L knockdown cells promote increased cell surface expression of integrin β1 which activates Focal Adhesion Kinase-Sarcoma (FAK-Src) pathway and Ras-related C3 botulinum toxin substrate 1-guanosine triphosphatase (Rac1-GTPase) activity to increase cell migration. Activity of matrix metalloproteinase (MMP)-2 and -9 is significantly increased in cells where RNase L levels are ablated. We show that mutations in RNase L found in HPC patients may promote prostate cancer by increasing expression of AR-responsive genes and cell motility and identify novel roles of RNase L as a prostate cancer susceptibility gene.


[Problems of epidural droperidol administration].
We describe three cases of extrapyramidal reactions apparently caused by epidural administration of droperidol. These patients suffered from chronic pain and was treated with epidural lidocaine and droperidol. Two patients received continuous administration of droperidol, and experienced acute dystonia and another after a single dose, developed akathisia. Adverse reactions occurred at 15, 20 and 24 hours after the administration of droperidol. The first patient received droperidol 6 mg, the second 8.5 mg and the third 5 mg. We consider that extrapyramidal reactions are due to overdoses because the patients who had been given less than 2.5 mg of droperidol a day, showed no adverse reaction. Although we use higher doses with NLA or for management of fever than with epidural administration of droperidol, we seldom encounter cases of side effects with droperidol. Epidurally administrated droperidol spreads rostral within the neuraxis and causes delayed extrapyramidal reactions as epidural morphine develops delayed respiratory depression. We must be careful in caring patients suffering from chronic pain with continuous epidural administration of droperidol.


Chest Wall Keloids Depicted by 18 F-Piflufolastat PET/CT Imaging.
Keloids are pathological scars from exuberant fibroproliferative collagen response and excessive extracellular matrix production usually extending beyond the original wound margins. Although keloids are mostly of dermatological concern, they could be incidentally depicted on scintigraphic planar and PET/CT imaging and could mimic other types of skin diseases. The authors present a case of chest wall keloids documented on 18 F-piflufolastat PET/CT during the evaluation of prostate cancer recurrence.


Staphylococcal enterotoxin D is a promiscuous superantigen offering multiple modes of interactions with the MHC class II receptors.
Dimerization of MHC class II molecules on the cell surface of human THP-1 monocytic cell line is a requirement for staphylococcal superantigen (SAG)-induced cytokine gene expression. The capacities of various SAG to induce this response are governed by their modes of interaction with MHC class II molecules. Staphylococcal enterotoxin A (SEA), with its two binding sites, dimerizes MHC class II molecules and subsequently induces cytokine gene expression in THP-1 cells. Here, we demonstrate that staphylococcal enterotoxin D (SED) and staphylococcal enterotoxin E (SEE) induce, similarly, IL-1beta and TNF-alpha gene expression in these cells. Using mutated toxins that lost their binding site with the MHC class II alpha- or beta-chain, we demonstrate that this response is also mediated by the dimerization of MHC class II molecules through two binding sites. Furthermore, SED forms Zn2+-dependent homodimers that allow multiple modes of MHC class II clustering, including ligation of alpha-chains (alpha/alpha), beta-chains (beta/beta), or the alpha- and beta-chains of two different class II molecules. The beta/beta interaction following Zn2+-dependent SED/SED homodimer formation seems to be mediated by the appearance of a novel binding site on SED that interacts with histidine 81 of the MHC class II beta-chain. The different modes of SED interactions also influence SED-induced T cell activation where simultaneous ligation of the alpha- and beta-chains is essential for optimal response. These various modes of SED binding may be used to preserve bivalency regardless of variability in the MHC class II alpha/beta/peptide complexes.


A comparative, two-dimensional 14N ESEEM characterization of reduced [2Fe-2S] clusters in hyperthermophilic archaeal high- and low-potential Rieske-type proteins.
Proteins of the Rieske and Rieske-type family contain a [2Fe-2S] cluster with mixed ligation by two histidines and two cysteines, and play important roles in various biological electron transfer reactions. We report here the comparative orientation-selected ESEEM and HYSCORE studies of the reduced clusters from two hyperthermophilic Rieske-type proteins; a high-potential, archaeal Rieske protein called sulredoxin (SDX) from Sulfolobus tokodaii with weak homology to the cytochrome bc-associated Rieske proteins, and a low-potential, archaeal homolog of an oxygenase-associated Rieske-type ferredoxin (ARF) from Sulfolobus solfataricus. (14)N ESEEM and HYSCORE spectra of SDX and ARF show well-defined variations, which are primarily determined by changes of quadrupole couplings (up to 50% depending on the selected orientation) of the two coordinated nitrogens. These are due to variations in coordination geometry of the histidine imidazole ligands rather than to variations of hyperfine couplings of these nitrogens, which do not exceed 8-10%. The measured quadrupole couplings and their differences in the two proteins are consistent with those calculated using the reported crystal structures of high- and low-potential Rieske proteins. These results suggest that exploration of quadrupole tensors might provide a more accurate method for characterization of the histidine coordination in different proteins and mutants than hyperfine tensors, and might have potential applications in a wider range of biological systems.


Global Tuberculosis.
Tuberculosis (TB) infections remain a common but deadly disease. TB control in health care and correctional settings presents numerous public health challenges for occupational and correctional health nurses.


Characterization and distribution of protein kinase C isoforms in human skin fibroblasts.
In the present study we have examined the distribution of several isoforms of protein kinase C, a lipid-regulated serine/threonine kinase essential for signal transduction and cell regulation, in cultured human skin fibroblasts. By Western blot analysis we have detected the presence of at least three of the known protein kinase C isoforms. The calcium-dependent protein kinase C alpha was primarily associated with the cytosolic fraction. Three non-calcium-dependent isoforms, protein kinases C epsilon, C delta, and C zeta, were also detected. Protein kinases C zeta and C delta were present primarily in the cytosol, while protein kinase C epsilon was associated primarily with the membrane fraction. Binding and activity studies were consistent with the pattern of expression and distribution defined by Western blot analysis. These results provide a useful frame of reference for the study of isoform-specific effects of protein kinase C in the regulation of cell growth and metabolism.


Anion-π interactions suppress phase impurities in FAPbI3 solar cells.
Achieving both high efficiency and long-term stability is the key to the commercialization of perovskite solar cells (PSCs)1,2. However, the diversity of perovskite (ABX3) compositions and phases makes it challenging to fabricate high-quality films3-5. Perovskite formation relies on the reaction between AX and BX2, whereas most conventional methods for film-growth regulation are based solely on the interaction with the BX2 component. Herein, we demonstrate an alternative approach to modulate reaction kinetics by anion-π interaction between AX and hexafluorobenzene (HFB). Notably, these two approaches are independent but work together to establish 'dual-site regulation', which achieves a delicate control over the reaction between AX and BX2 without unwanted intermediates. The resultant formamidinium lead halides (FAPbI3) films exhibit fewer defects, redshifted absorption and high phase purity without detectable nanoscale δ phase. Consequently, we achieved PSCs with power conversion efficiency (PCE) up to 26.07% for a 0.08-cm2 device (25.8% certified) and 24.63% for a 1-cm2 device. The device also kept 94% of its initial PCE after maximum power point (MPP) tracking for 1,258 h under full-spectrum AM 1.5 G sunlight at 50 ± 5 °C. This method expands the range of chemical interactions that occur in perovskite precursors by exploring anion-π interactions and highlights the importance of the AX component as a new and effective working site to improved photovoltaic devices with high quality and phase purity.


Endonucleolytic inhibition assay of DNA/Fok I transducer as a sensitive platform for homogeneous fluorescence detection of small molecule-protein interactions.
This paper reported a novel homogeneous fluorescence assay strategy for probing small molecule-protein interactions based on endonucleolytic inhibition of a DNA/Fok I transducer. The transducer could cyclically cleave fluorescence-quenched probes to yield activated fluorescence signal, while protein binding to the small molecule label would prevent Fok I from approaching and cleaving the fluorescence-quenched probes. Because of the efficient signal amplification from the cyclic cleavage operation, the developed strategy could offer high sensitivity for detecting small molecule-protein interactions. This strategy was demonstrated using folate and its high-affinity or low-affinity binding proteins. The results revealed that the developed strategy was highly sensitive for detecting either high- or low-affinity small molecule-protein interactions with improved selectivity against nonspecific protein adsorption. This strategy could also be extended for assays of candidate small-molecule ligands using a competitive assay format. Moreover, this strategy only required labeling the small molecule on a DNA heteroduplex, circumventing protein modifications that might be harmful for activity. In view of these advantages, this new method could have potential to become a universal, sensitive, and selective platform for quantitative assays of small molecule-protein interactions.


Isoflurane binds and stabilizes a closed conformation of the leukocyte function-associated antigen-1.
We previously demonstrated that isoflurane targets lymphocyte function-associated antigen-1 (LFA-1), a critical adhesion molecule for leukocyte arrest. However, it remains to be determined how isoflurane interacts with the full ectodomain LFA-1 and modulates its conformation and function. Isoflurane binding sites on the full ectodomain LFA-1 were probed by photolabeling using photoactivatable isoflurane (azi-isoflurane). The adducted residues were determined by liquid chromatography/mass spectrometry analysis. Separately, docking simulations were performed to predict binding sites. Point mutations were introduced around isoflurane binding sites. The significance of isoflurane's effect was assessed in both intracellular adhesion molecule-1 (ICAM-1) binding assays and epitope mapping of activation-sensitive antibodies using flow cytometry. Two isoflurane binding sites were identified using photolabeling and were further validated by the docking simulation: one at the hydrophobic pocket in the ICAM-1 binding domain (the αI domain); the other at the βI domain. Mutagenesis of the α'1 helix showed that isoflurane binding sites at the βI domain were significantly important in modulating LFA-1 function and conformation. Epitope mapping using activation-sensitive antibodies suggested that isoflurane stabilized LFA-1 in the closed conformation. This study suggested that isoflurane binds to both the αI and βI domains allosteric to the ICAM-1 binding site, and that isoflurane binding stabilizes LFA-1 in the closed conformation.


Relaxing Wrinkles in Jammed Interfacial Assemblies.
Dynamic covalent bonding has emerged as a mean by which stresses in a network can be relaxed. Here, the strength of the bonding of ligands to nanoparticles at the interface between two immiscible liquids affect the same results in jammed assemblies of nanoparticle surfactants. Beyond a critical degree of overcrowding induced by the compression of jammed interfacial assemblies, the bonding of ligands to nanoparticles (NPs) can be broken, resulting in a desorption of the NPs from the interface. This reduces the areal density of nanoparticle surfactants at the interface, allowing the assemblies to relax, not to a fluid state but rather another jammed state. The relaxation of the wrinkles caused by the compression reflects the tendency of these assemblies to eliminate areas of high curvature, favoring a more planar geometry. This enabled the generation of giant vesicular and multivesicular structures from these assemblies.


Effects of the promoter open complex formation on gene expression dynamics.
Little is known about the biological mechanisms that shape the distribution of intervals between the completion of RNA molecules (T(p)RNA) , and thus transcriptional noise. We characterize numerically and analytically how the promoter open complex delay (tau(P)) and the transcription initiation rate (k(t)) shape T(p)RNA. From this, we assess the noise and mean of transcript levels and show that these can be tuned both independently and simultaneously by tau(P) and k(t). Finally, we characterize how tau(P) affects bursting in RNA production and show that the tau(P) measured for a lac promoter best fits independent measurements of the burst distribution of the same promoter. Since tau(P) affects noise in gene expression, and given that it is sequence dependent, it is likely to be evolvable.


Hemangiomatosis of the colon and peritoneum: case report and management discussion.
A 3-month-old infant presented with ascites, anemia, minimal rectal bleeding, and thrombocytopenia. He was found to have several hundred small cavernous hemangiomas of the colon and peritoneal surfaces. Treatment consisted of laser surgery, subtotal colectomy, and steroids. A brief review of the literature on intestinal hemangiomatosis is included as well as the complications of the disease and current therapy.


Changes of serum leptin and endocrine and metabolic parameters after 7 days of energy restriction in men and women.
Circulating leptin decreases during fasting in rodents and humans; however, the mechanism of the decrease is unknown. The aim of this study was to examine the relationship between decrements of serum leptin concentrations and changes of hormonal (insulin and cortisol) and metabolic (glucose, ketones, and fatty acids) parameters involved in the metabolic adaptation to energy restriction in normal-weight humans. Because there are marked gender differences in circulating leptin, both men and women were studied. The body mass index (BMI), percent body fat (% body fat), and serum leptin, insulin, cortisol, glucose, beta-hydroxybutyrate,(BOHB), and nonesterified fatty acids (NEFA) were determined in 11 men and 13 women (age, 20 to 41 years; BMI, 21.2 to 26.8 kg/m2) before and during 7 days of energy restriction (-68% +/- 1% of daily energy requirements). Weight loss averaged about 4% in both men and women. Leptin in men was 3.7 +/- 0.5 and decreased to 2.1 +/- 0.4 ng/mL (percent change [%delta], -36% +/- 6.0%, P < .0005) during restriction. Concurrently, insulin decreased from 7.2 +/- 0.6 to 1.8 +/- 0.3 microU/mL (%delta, -74% +/- 4%, P < .0001). In contrast, leptin was higher in women before (16.2 +/- 1.9 ng/mL) and after (6.0 +/- 0.8 ng/mL) restriction and decreased more than in men (%delta, -61% +/- 4%, P < .02 v men), whereas the decrease of insulin in women was less than in men: 10.1 +/- 1.9 to 6.1 +/- 1.0 microU/mL (%delta, -31% +/- 9%, P < .0025; P < .0005 v men), perhaps because glucose decreased less in women than in men. Overall, the changes of leptin during fasting were independently correlated with the changes of glucose (r = .53, P < .007), NEFA (r = .53, P < .01), and BOHB (r = .65, P < .001). In addition, the change of leptin correlated with a combined index of the parameters that reflect decreased glucose availability and increased lipolysis ([deltaglucose + deltainsulin + deltaNEFA]/3, r = .73, P < .0001) or a combined index of parameters that would be expected to limit glucose uptake by adipocytes ([deltaglucose + deltainsulin + deltacortisol]/3, r = .48, P < .02). We conclude that there are significant differences between men and women in the responses of leptin and insulin to energy restriction. Furthermore, decreases of circulating leptin during negative energy balance are related to changes of endocrine and metabolic parameters, suggesting that leptin secretion may be regulated by alterations of adipocyte glucose and lipid metabolism, ie, decreased glucose uptake and metabolism and increased lipolysis.


[Extended salvage pelvic and retroperitoneal lymph node dissection due to prostate cancer relapse].
Treatment of a biochemical prostate cancer relapse represents a difficult clinical dilemma, which has remained without a definitive solution so far. Based on clinical studies, we combine radical prostatectomy with extended pelvic lymph node dissection in intermediate and high risk patients as a routine procedure at our clinic. In this paper, we report on a case of extended salvage lymphadenectomy performed due to biochemical prostate cancer recurrence. The 56-year-old patient came to our clinic in April 2012 with a finding of lymph node metastasis according to PET-CT imaging. Laparoscopic radical retropubic prostatectomy with lymphadenectomy had been performed in 2008 [pT3a, N0 (0/4), M0, R0, GS 5+4=9, iPSA 26.67 ng/mL], and followed by radiotherapy as of September 2009. The extended salvage lymphadenectomy was performed in April 2012 due to a PSA-level rise up to 24 ng/mL and the aforementioned PET-CT findings. A total of 22 lymph nodes were removed, among them 3 lymph nodes with metastases. In the fossa obturatoria on the right we identified a walnut-size lymph node relapse with tumour necrosis, which fully corresponded to the PET-CT scan. The PSA level subsequently dropped to 0.4 ng/mL postoperatively, and further to the current value of 0.02 ng/mL (August 2012).


Centrosomal CAP350 protein stabilises microtubules associated with the Golgi complex.
A comprehensive model of how the centrosome organises the microtubule network in animal cells has not yet been elucidated. Here we show that the centrosomal large CAP-Gly protein CAP350 is not only present at the centrosome, but is also present as numerous dots in the pericentrosomal area. Using in vitro and in vivo expression of partial constructs, we demonstrated that CAP350 binds microtubules through an N-terminal basic region rather than through its CAP-Gly domain. CAP-Gly-containing domains of CAP350 are targeted not only to the centrosome but also to a Golgi-like network. Interestingly, full-length GFP-tagged CAP350 bound preferentially to microtubules in the pericentrosomal area. These results indicate that the large CAP350 protein has a dual binding ability. Overexpression of CAP350 promoted an increase in the stability of the whole microtubule network, as judged by a significant decrease in the number of EB1 comets and by an enhanced microtubule resistance to Nocodazole treatment. In support of this, CAP350 depletion decreased microtubule stability. Moreover, both depletion and overexpression of CAP350 induced specific fragmentation of the Golgi complex while maintaining a juxtanuclear localisation. We propose that CAP350 specifically stabilises Golgi-associated microtubules and in this way participates in the maintenance of a continuous pericentrosomal Golgi ribbon.


[Spontaneous pneumomediastinum mimicking Boerhaave's syndrome].
Spontaneous pneumomediastinum is an uncommon benign disorder that usually occurs in young men. Because of its clinical features and physiopathologic mechanism, other, more serious disorders that could require urgent treatment, such as Boerhaave's syndrome or spontaneous esophageal rupture, must be ruled out. We report the case of a 19-year-old man with no relevant history, who presented mediastinal emphysema after an episode of repeated vomiting in the context of alcohol abuse. Imaging techniques help to assess esophageal involvement.


Induction of fetal hemoglobin by cell-cycle-specific drugs and recombinant erythropoietin.
During the last several years, studies in humans and experimental animals have identified several compounds that induce fetal hemoglobin in the adult. These include: cell-cycle-specific drugs, other cytotoxic drugs, butyric acid analogs, and erythropoietin. Several of these compounds induce fetal hemoglobin indirectly by triggering kinetics of rapid erythroid regeneration. High doses of erythropoietin increase the frequency of erythroid progenitors programmed to hemoglobin F. This results in transient increases of hemoglobin F-containing cells (F cells) in the peripheral blood. Erythropoietin and hydroxyurea increase F cells in a cooperative fashion. Although high doses of erythropoietin can induce F cell production in humans, the practical relevance of such observations is unclear


Locoweed (Oxytropis sericea)-induced lesions in mule deer (Odocoileius hemionus).
Locoweed poisoning has been reported in wildlife, but it is unknown whether mule deer (Odocoileius hemionus) are susceptible. In areas that are heavily infested with locoweed, deer and elk (Cervus elaphus nelsoni) have developed a spongiform encephalopathy, chronic wasting disease (CWD). Although these are distinct diseases, no good comparisons are available. The purpose of this study was to induce and describe chronic locoweed poisoning in deer and compare it with the lesions of CWD. Two groups of four mule deer were fed either a complete pelleted ration or a similar ration containing 15% locoweed (Oxytropis sericea). Poisoned deer lost weight and developed a scruffy, dull coat. They developed reluctance to move, and movement produced subtle intention tremors. Poisoned deer had extensive vacuolation of visceral tissues, which was most severe in the exocrine pancreas. Thyroid follicular epithelium, renal tubular epithelium, and macrophages in many tissues were mildly vacuolated. The exposed deer also had mild neuronal swelling and cytoplasmic vacuolation that was most obvious in Purkinje cells. Axonal swelling and dystrophy was found in many white tracts, but it was most severe in the cerebellar peduncles and the gracilis and cuneate fasciculi. These findings indicate that deer are susceptible to locoweed poisoning, but the lesions differ in severity and distribution from those of other species. The histologic changes of locoweed poisoning are distinct from those of CWD in deer; however, the clinical presentation of locoweed poisoning in deer is similar. Histologic and immunohistochemical studies are required for a definitive diagnosis.


Multiparametric homogeneous method for identification of ligand binding to G protein-coupled receptors: receptor-ligand binding and β-arrestin assay.
Two homogeneous assay systems have been combined to provide a new cell-based functional assay. The assay can be used to identify ligand binding to β(2)-adrenergic receptors, but also the downstream response can be determined in the same assay. Both the quenching resonance energy transfer (QRET) and the DiscoveRx PathHunter assay formats allow the use of intact cells. The homogeneous QRET technique is a single-label approach based on nonspecific quenching of the time-resolved luminescence, enabling agonist and antagonist receptor binding measurements. The commercial PathHunter assay is in turn based on enzyme fragment complementation, which can be detected on the basis of chemiluminescence signal. In the PathHunter technology the enzyme complementation is recorded immediately downstream of agonist-induced receptor activation. The new multiparametric detection technology combines these two assay methods enabling the identification of agonist, and antagonist binding to the receptor, and the agonist-induced response. Using the QRET and the PathHunter methods a panel of β(2)-adrenergic receptor ligands (epinephrine, terbutaline, metaproterenol, salmeterol, propranolol, alprenolol, bisoprolol, ICI 118,551, and bucindolol) was tested to prove the assay performance. The signal-to-background ratio for tested ligands ranged from 5 to 11 and from 6 to 18 with QRET and PathHunter, respectively. Combined homogeneous assay technique can provide an informative method for screening purposes and an efficient way to monitor receptor-ligand interaction, thus separating agonist from antagonist.


The feasibility and usability of DNA-dot bioconjugation to antibody for targeted in vitro cancer cell fluorescence imaging.
DNA-protein bioconjugation is an appealing target-tracking strategy. The new capability of DNA molecule as a biological nanomaterial endows unique fluorescence and physicochemical properties to be applied in bioimaging. Progression in targeted imaging is contingent on the conjugation of diagnostic nanoparticles to biomolecular signatures, particularly antibody-based ligands. Here, we have reported our recent experience, DNA-dot synthesis and characterization, the covalent conjugation of DNA-dot to goat F(ab')2 IgG and Epidermal Growth Factor Receptor (EGFR) antibodies, DNA-dot@antibody coupling confirmation, and fluorescent targeted imaging of lung cancer cell line. As a result, the average size of DNA-dot was 4.5-5 nm which was conjugated to amine-rich antibodies with returned PO4-1 groups on the DNA-dot surface via PN bond. The synthetic DNA-dots were conjugated to the goat F(ab')2 IgG and tested for fluorescent detection usability by indirect Dot-blot assay. Also, DNA-dot@EGFR conjugates identified lung cancer cells with 84-92% specificity and 100% sensitivity in five concentrations, associated with 0.0025 to 0.04 g 100 μL-1 DNA-dot. The results demonstrated that bioconjugated DNA-dot can do the diagnosis profiling of molecular biomarkers. Generally, DNA-dot bioconjugation with antibody is implemented within two days and biomarker detection takes one day. Consequently, DNA-dot@antibody is potentially a toxic-free, swift, and efficient method of antibody labeling that opens up new horizons in fluorescent nanoimaging in the field of cancer cell detection.


Numerical simulations of stochastic circulatory models.
Properties of two of the stochastic circulatory models theoretically introduced by Smith et al., 1997, Bull. Math. Biol. 59, 1-22 were investigated. The models assumed the gamma distribution of the cycle time under either the geometric or Poisson elimination scheme. The reason for selecting these models was the fact that the probability density functions of the residence time of these models are formally similar to those of the Bateman and gamma-like function models, i.e., the two common deterministic models. Using published data, the analytical forms of the probability density functions of the residence time and the distributions of the simulated values of the residence time were determined on the basis of the deterministic models and the stochastic circulatory models, respectively. The Kolmogorov-Smirnov test revealed that even for 1000 xenobiotic particles, i.e., a relatively small number if the particles imply drug molecules, the probability density functions of the residence time based on the deterministic models closely matched the distributions of the simulated values of the residence time obtained on the basis of the stochastic circulatory models, provided that parameters of the latter models fulfilled selected conditions.


There's always a villain to punish: group processes contributing to violence and its remediation.
This paper considers the widespread use of violent metaphors, such as "combat" and "war," to represent the current social, psychological, and political problems within the United States. I apply Lakoff and Johnson's (1980) thesis that metaphor shapes thought, policy, and behavior. I examine how use of such metaphors inclines the national consciousness toward violence and punishment for it. In addition, I discuss shame and humiliation as psychological precursors of violence, particularly as these play out in the exclusion and extrusion via group scapegoating of individuals and whole groups from active participation in an esteemed or powerful other group. Included within the concept of "violence" are those harmful social policies that invalidate the experiences of disempowered people within the United States. I consider the role of group processes in resolving the injuries wrought by violence, particularly as these operate within such restorative justice projects as the Glencree Ex-Combatants Programme in Northern Ireland. Lessons emerge from restorative justice projects installed internationally for ameliorating conflict within and between "victim" groups in the United States.


Interstitial brachytherapy as the sole treatment for tongue metastasis from breast carcinoma.
Breast carcinoma metastasize more frequently to the bone, liver lung and brain as compared to other anatomical sites. As such, advanced breast tumours with secondaries involving the rarer sites often pose when a therapeutic challenge for the clinician to find a good balance between offering the most effective therapy and its potential toxicity. We report the first case of tongue metastasis from breast cancer that was successfully treated with interstitial brachytherapy, resulting in good local control and no severe side effects.


Conservation of trans-acting circuitry during mammalian regulatory evolution.
The basic body plan and major physiological axes have been highly conserved during mammalian evolution, yet only a small fraction of the human genome sequence appears to be subject to evolutionary constraint. To quantify cis- versus trans-acting contributions to mammalian regulatory evolution, we performed genomic DNase I footprinting of the mouse genome across 25 cell and tissue types, collectively defining ∼8.6 million transcription factor (TF) occupancy sites at nucleotide resolution. Here we show that mouse TF footprints conjointly encode a regulatory lexicon that is ∼95% similar with that derived from human TF footprints. However, only ∼20% of mouse TF footprints have human orthologues. Despite substantial turnover of the cis-regulatory landscape, nearly half of all pairwise regulatory interactions connecting mouse TF genes have been maintained in orthologous human cell types through evolutionary innovation of TF recognition sequences. Furthermore, the higher-level organization of mouse TF-to-TF connections into cellular network architectures is nearly identical with human. Our results indicate that evolutionary selection on mammalian gene regulation is targeted chiefly at the level of trans-regulatory circuitry, enabling and potentiating cis-regulatory plasticity.


First optical hyperfine structure measurement in an atomic anion.
We have investigated the hyperfine structure of the transition between the 5d{7}6s{2} {4}F{9/2}{e} ground state and the 5d{6}6s{2}6p {6}D{J}{o} excited state in the negative osmium ion by high-resolution collinear laser spectroscopy. This transition is unique because it is the only known electric-dipole transition in atomic anions and might be amenable to laser cooling. From the observed hyperfine structure in 187Os- and 189Os- the yet unknown total angular momentum of the bound excited state was found to be J=9/2. The hyperfine structure constants of the {4}F{9/2}{e} ground state and the {6}D{9/2}{o} excited state were determined experimentally and compared to multiconfiguration Dirac-Fock calculations. Using the knowledge of the ground and excited state angular momenta, the full energy level diagram of 192Os- in an external magnetic field was calculated, revealing possible laser cooling transitions.


Control of hyperbilirubinemia in glucose-6-phosphate dehydrogenase-deficient newborns using an inhibitor of bilirubin production, Sn-mesoporphyrin.
Hyperbilirubinemia in new-borns with glucose-6-phosphate dehydrogenase (G6PD) deficiency is a serious clinical problem because of the severity and unpredictability of its course. An innovative approach to this problem is suggested by previous experience with Sn-mesoporphyrin (SnMP), a potent inhibitor of bilirubin production, in moderating neonatal hyperbilirubinemia caused by ABO incompatibility, immaturity, and unspecified mechanisms. To compare the effectiveness of the preventive and therapeutic uses of SnMP in ameliorating the course of bilirubinemia of G6PD-deficient neonates. Neonates born at the Metera Maternity Hospital, Athens, Greece, and found to be G6PD-deficient by cord blood testing were stratified by sex and gestational age (210-265 days and >265 days) and randomized in pairs to receive SnMP (6 micromol/kg birth weight, intramuscularly) either on the first day of life (preventive use) or if and when the plasma bilirubin concentration (PBC) level reached an age-specific threshold level for intervention (therapeutic use). In the case of failure of SnMP to control the rise of PBC levels, the protocol defined precisely the threshold PBC levels for switchover to phototherapy (PT) and, if necessary, exchange transfusion. PBC was measured daily until a declining value was obtained and the case was closed. A total of 86 G6PD-deficient neonates were randomized: 42 in the preventive arm and 44 in the therapeutic arm. Of the latter, 20 (45%) reached PBC levels requiring therapeutic intervention and thus received SnMP. Regardless of the trial arm, none of the 86 neonates required PT, whereas in a previous study in the same population, 33% of G6PD-deficient neonates required PT. In the intrapair sequential analysis, the favored arm was decided on the criterion of the age at closure of the case being shorter by at least 1 day. After plotting 30 untied pairs in the sequential analysis graph, the preventive use of SnMP proved to be the favored arm, and the trial was stopped. At this point, there were 2 unpaired neonates, 12 tied pairs, 22 pairs in which the preventive use of SnMP was favored and 8 pairs in which the therapeutic use of SnMP was favored. In the group analysis, infants in the preventive group, compared with those in the therapeutic group, had a lower maximum PBC level (8.2 +/- 3.1 and 10.9 +/- 2.8 mg/dL, respectively), which was reached at an earlier age (63.5 +/- 34.8 and 82.2 +/- 24.7 hours, respectively) as well as a lower closing PBC level (7.2 +/- 2.9 and 9.6 +/- 2.5 mg/dL, respectively) and an earlier age at closing (89.1 +/- 35.6 and 110.8 +/- 23.6 hours, respectively). Moreover, a PBC level of >/=8.0 mg/dL, a level at which jaundice is clearly visible, was not reached by 52% of the neonates in the preventive arm and 16% of the neonates in the therapeutic arm. In G6PD-deficient neonates, a single dose of SnMP administered preventively or therapeutically entirely supplanted the need for PT to control hyperbilirubinemia. The preventive use of SnMP offers practical advantages in populations with a high enough prevalence of G6PD deficiency to justify cord blood screening.


Systemic vasculitis complicating hairy cell leukaemia treatment with cladribine.
A single course of cladribine has been used commonly over the past decade to treat hairy cell leukaemia, with an impressive rate of complete remission and few serious adverse effects. Although vasculitis has been reported in the course of hairy cell leukaemia, it has only rarely been reported as the consequence of cladribine treatment. We describe a 73-year-old woman who developed serious systemic vasculitis with associated thrombocytopenia in the course of treatment with cladribine.


Nitroprusside after open-heart surgery.
The effects of intravenous infusion of sodium nitroprusside were studied in 11 children immediately after open-heart surgery for congenital heart disease. The patients were selected because, following bypass, their cardiac index was below 2,0 L/min/m2 and their systemic vascular resistance exceeded 30 units. In order to eliminate the effects of preload, mean left atrial pressure was maintained at a constant level by blood transfusion. During infusion of nitroprusside the mean decrease of mean arterial pressure was 18.6%, of systemic vascular resistance was 53.7%, and the increase in cardiac index was 76.9%. All children recovered.


Effective suppression of the Kirsten rat sarcoma viral oncogene in pancreatic tumor cells via targeted small interfering RNA delivery using nanoparticles.
The objective of this study was to establish an efficient carrier for small interfering RNA (siRNA) delivery targeting pancreatic tumor cells. A copolymer consisting of a single-chain variable fragment targeted to human CD44 variant 6 (scFv(CD44v6)) functional group conjugated to polyethylene glycol-poly-L-lysine was synthesized and assembled into micelles encapsulating the siRNAs. Flow cytometry and Western blot assays were performed to evaluate the transfection efficiency and gene-silencing effect of the siRNAs. Afterward, (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, Transwell, soft agar colony formation, and enzyme-linked immunosorbent assays were performed to evaluate the biological functions of PANC-1 cells after Kirsten rat sarcoma viral oncogene knockdown. In vivo assays were performed using a BALB/c (nu/nu) mouse model subcutaneously injected with PANC-1 xenografts. Real-time in vivo fluorescence imaging was used to monitor the tumor homing of the nanoparticles. The scFv(CD44v6) enabled more efficient delivery of siRNAs and exhibited enhanced gene silencing compared with nontargeted nanoparticles. Furthermore, targeted delivery of the siRNAs induced a potent inhibitory effect on cell proliferation, colony formation, invasion, and vascular endothelial growth factor production. The animal assays revealed that single-chain variable fragment nanoparticles accumulated in the tumor tissue and enhanced the inhibition of tumor growth in vivo. The scFv(CD44v6)-conjugated nanocarriers provide a highly efficient and safe platform for systemic gene therapy for pancreatic cancer.


Long non-coding RNA H19 enhances the pro-apoptotic activity of ITF2357 (a histone deacetylase inhibitor) in colorectal cancer cells.
Introduction: Long non-coding RNA H19 (lncH19) is highly expressed in colorectal cancer (CRC) and plays critical roles in tumor development, proliferation, metastasis, and drug resistance. Indeed, the expression of lncH19 usually affects the outcomes of chemo-, endocrine, and targeted therapies. ITF2357 (givinostat) is a histone deacetylase inhibitor (HDACi) that revealed a significant anti-tumor action by inducing apoptosis in different tumor models, including leukemia, melanoma, and glioblastoma. However, no data are present in the literature regarding the use of this compound for CRC treatment. Here, we investigate the role of lncH19 in ITF2357-induced apoptosis in CRC cells. Methods: The HCT-116 CRC cell line was stably silenced for H19 to investigate the role of this lncRNA in ITF2357-induced cell death. Cell viability assays and flow cytometric analyses were performed to assess the anti-proliferative and pro-apoptotic effects of ITF2357 in CRC cell lines that are silenced or not for lncH19. RT-PCR and Western blot were used to study the effects of ITF2357 on autophagy and apoptosis markers. Finally, bioinformatics analyses were used to identify miRNAs targeting pro-apoptotic factors that can be sponged by lncH19. Results: ITF2357 increased the expression levels of H19 and reduced HCT-116 cell viability, inducing apoptosis, as demonstrated by the increase in annexin-V positivity, caspase 3 cleavage, and poly (ADP-ribose) polymerase (PARP-1) degradation. Interestingly, the apoptotic effect of ITF2357 was much less evident in lncH19-silenced cells. We showed that lncH19 plays a functional role in the pro-apoptotic activity of the drug by stabilizing TP53 and its transcriptional targets, NOXA and PUMA. ITF2357 also induced autophagy in CRC cells, which was interpreted as a pro-survival response not correlated with lncH19 expression. Furthermore, ITF2357 induced apoptosis in 5-fluorouracil-resistant HCT-116 cells that express high levels of lncH19. Conclusion: This study shows that lncH19 expression contributes to ITF2357-induced apoptosis by stabilizing TP53. Overall, we suggest that lncH19 expression may be exploited to favor HDACi-induced cell death and overcome 5-fluorouracil chemoresistance.


Abdominal trauma: never underestimate it.
Introduction. We present a case of a sports injury. The initial presentation and clinical examination belied serious intra-abdominal injuries. Case Presentation. A 16-year-old male patient came to emergency department after a sports-related blunt abdominal injury. Though on clinical examination the injury did not seem to be serious, FAST revealed an obscured splenorenal window. The CT scan revealed a large left renal laceration and a splenic laceration that were managed with Cook coil embolization. Patient remained tachycardic though and had to undergo splenectomy, left nephrectomy, and a repair of left diaphragmatic rent. Patient had no complication and had normal renal function at 6-month followup. Conclusion. The case report indicates that management of blunt intra-abdominal injury is complicated and there is a role for minimally invasive procedures in management of certain patients. A great deal of caution is required in monitoring these patients, and surgical intervention is inevitable in deteriorating patients.


Novel SYNGAP1 variant in a patient with intellectual disability and distinctive dysmorphisms.
We describe a novel de novo heterozygous variant in SYNGAP1 (c.1741C>T, p.R581W), identified through targeted resequencing in an 8-year-old boy with intellectual disability, autism spectrum disorder, distinctive dysmorphic features, and no seizures. Our data strongly suggest that the SYNGAP1 variant is causative of intellectual disability in this patient.


6-(7-Nitro-2,1,3-benzoxadiazol-4-ylthio) Hexanol Inhibits Proliferation and Induces Apoptosis of Endometriosis by Regulating Glutathione S-Transferase Mu Class 4.
Endometriosis is a chronic disease associated with a disrupted oxidative balance and chronic inflammation. In this study, we investigated the role of glutathione S-transferase Mu class 4 (GSTM4) in endometriosis and determined whether 6-(7-nitro-2,1,3-benzoxadiazol-4-ylthio) hexanol (NBDHEX) regulates GSTM4 expression to affect cellular functions and oxidative stress. GSTM4 expression was detected by immunohistochemistry in endometrium from 15 endometriosis patients and 15 healthy controls. Western blotting was used to detect the expression of GSTM4, proliferating cell nuclear antigen (PCNA), matrix metalloproteinase-9 (MMP-9), Survivin, B-cell lymphoma-extra-large (Bcl-XL), Bax, kelch-like ECH-associated protein 1 (Keap1), and nuclear factor-erythroid 2-related factor 2 (Nrf2) in primary endometrial stromal cells with endometriosis (EESC) and normal endometrial stromal cells (NESC). The effects of NBDHEX on cell proliferation, migration, and invasion were evaluated using Cell Counting Kit-8 (CCK8) and Transwell assays. Apoptosis was detected by flow cytometry. The expression of GSTM4 was significantly increased in endometrium from endometriosis patients. Upon NBDHEX treatment, ESC exhibited reduced proliferation, migration and invasion abilities, and increased apoptosis. NBDHEX decreased the expression of endometriosis prognostic markers (PCNA and MMP-9) and anti-apoptotic proteins (Survivin and Bcl-xl), while it increased the expression of the apoptotic protein Bax. It had no effect on Keap1 expression, and it decreased the expression of Nrf2. The effect of siRNA-mediated knockdown of GSTM4 was similar to that of suppressing GSTM4 expression with NBDHEX treatment. These results indicate that GSTM4 is highly expressed in endometriosis and its expression is inhibited by NBDHEX. Decreased expression of GSTM4 inhibits cell growth, migration, and invasion, and negatively regulates Nrf2 to affect oxidative stress-induced apoptosis. Our results suggest that GSTM4 may play a role in ameliorating the progression of endometriosis. NBDHEX may have therapeutic potential in the treatment of endometriosis.


Impairments of cells and genomic DNA by environmentally transformed engineered nanomaterials.
Enormous increase in the production of nanomaterials and their growing applications in the device technology, biotechnology and biomedical areas suggest the need for developing models for predicting the environmental health and safety (EHS) risks posed by such nanomaterials. We hypothesize that CdSe quantum dots (QDs) and ZnO nanoparticles (NPs) encompassed in liposomes or not and transformed by simulated solar UV light can be model systems for studying the environmental toxicity of engineered nanomaterials. In this study, human lung epithelial adenocarcinoma cells (H1650) are exposed to photoirradiated CdSe QDs or ZnO nanopowder included or not in liposomes. The release of cadmium and zinc ions from the nanomaterials exposed to solar simulated UV radiation is detected and quantified by measuring the steady-state and time resolved fluorescence of the metal ion sensor tetracarboxyphenylporphyrin (TCPP) or the commercial Measure iT Pd/Cd sensor. Viability of cells treated with nanomaterials exposed to solar simulated UV radiation for different durations is measured by MTT assay. Enhanced etching of the nanoparticles exposed to solar simulated UV radiation results in the release of toxic levels of heavy metal ions, which considerably lower the viability of H1650 cells is due to the deactivation of DNA repair enzymes as evidenced by the pinching off of nuclear DNA in comet assays and DNA samples in electrophoresis. Results from this study highlight the need to obtain not only quantitative information about the environmental risks posed by engineered nanomaterials but also environment friendly nanomaterials for practical applications.


Longevity, growth rate and related traits among strains of Tribolium castaneum.
Longevity of eight laboratory strains of the flour beetle Tribolium castaneum, with various geographic backgrounds, was studied under constant laboratory conditions of 33 degrees C and 70% relative humidity in standard medium (95% whole wheat flour and 5% dried yeast) during a period of 227 days starting from the egg stage. The eggs were collected from the same parents, first a few days after emergence and afterwards at intervals of 13, 9, 10 and 11 days. Mean survival time (MST) was found to be strain-specified. It ranges from 128.6 days for KJ (Kyoto, Japan) to 174.2 days for ES (Edinburgh, Scotland). MST was highly correlated with the percentage of adults alive after 227 days, which did not change the ranking order of strain longevity. Parental age had no effect on longevity. The mean adult longevity of the strains was correlated with the available data on adult weight, growth rate, viability and productivity. There was no relationship between adult weight and longevity. LIfe span was found to depend on growth rate (measured as 13-day larval weight), percent viability (from 13-day larvae to adulthood) and productivity. Developmental time was also found to influence adult life span within certain limits (two extreme strains deviated). The data suggest that ageing and death in T. castaneum is under genetic control and support the idea that ageing, allied to development, is genetically controlled.


[A survey on exposure to rabies among the primary school student in the area with a high incidence of rabies in China].
To understand the current situation and characteristics of potential rabies exposure to primary school students in the area with a high incidence of rabies of China, and to provide evidence for developing the intervention measures. A population-based investigation was conducted in Guizhou and Guangdong provinces which had been the severe rabies epidemic areas in southern China. Two urban primary schools, two county schools, four township schools and four village schools were selected in the 2 provinces. Samples of the students were from each grade of the 12 schools for their potential rabies exposure information. Among 2408 primary school students being interviewed, 290 person-times exposure were found during January of 2007 to May of 2008. The total exposure rate in the observed students in 2007 was 9.22%, but higher in Guizhou province (16.92%) The rate of exposure was higher in male students than in female, and was also higher in the students from rural than from the urban areas. More exposures were seen in summer season, from June to August. The kinds of animals that causing the exposures would include dogs (74.14%) and cats (25.86%). 32.07% of the total exposures were related to playing with animals. Arms and legs were the most affected areas of the body including, 37.59% of the wounds appeared below the knees. The rate of potential rabies exposure was high in primary school students, especially in the rural areas. Playing with dogs and cats appeared to be the major behavior causing the exposures among the students.


Th1 cell adjuvant therapy combined with tumor vaccination: a novel strategy for promoting CTL responses while avoiding the accumulation of Tregs.
We have previously described a method for adoptive immunotherapy of cancer based on antigen-specific T(h)1 cells. However, efficient induction of anti-tumor responses using T(h)1 cells remains a formidable challenge, especially for MHC class II-negative tumors. In the present study, we sought to develop a novel strategy to eradicate established tumors of the MHC class II-negative, ovalbumin (OVA)-expressing EG-7 cells. Tumor-bearing mice were intradermally treated with OVA-specific T(h)1 cells, combined with the model tumor antigen (OVA), near the tumor-draining lymph node (DLN). We found that tumor growth was significantly inhibited by this strategy and approximately 50-60% of tumor-bearing mice were completely cured. Tumor eradication was crucially dependent on the generation of OVA/H-2K(b)-specific CTLs in the tumor DLNs and tumor site. The injected T(h)1 cells were mainly distributed in tumor DLNs, where they vigorously proliferated and enhanced the activation of dendritic cells. Strikingly, we also found that the accumulation of CD4(+)CD25(+) regulatory T cells (Tregs) was significantly inhibited in tumor DLNs by T(h)1 cell adjuvant therapy and this abrogation was associated with IFNgamma secreted by T(h)1 cells. These results identify T(h)1 cell adjuvant therapy combined with tumor vaccination as a novel approach to the treatment of human cancer.


Extended swimming exercise reduces inflammatory and peripheral neuropathic pain in rodents.
Physical exercise is often recommended to patients who have chronic pain. However, only a small number of studies report exercise-induced analgesia in the setting of inflammatory pain, and even fewer relate long-term exercise to reductions in neuropathic pain. To address these questions, we evaluated the effect of extended swimming exercise in animal models of inflammatory (intraplantar injection of dilute formalin) and neuropathic (partial peripheral nerve injury) pain. We found that 9 days of swimming exercise in 37 degrees C water for 90 min/d decreased licking and flinching responses to formalin, as compared with nonexercised control animals. In addition, 18 to 25 days of swimming decreased nerve injury-induced cold allodynia and thermal hyperalgesia in rats, and 7 days of swimming decreased nerve injury-induced thermal hyperalgesia in mice. Our data indicate that swimming exercise reduces behavioral hypersensitivity in formalin- and nerve injury-induced animal models of persistent pain. Surprisingly, few animal studies have investigated the effects of extended exercise on chronic pain. Our results support the use of exercise as a nonpharmacological approach for the management of peripheral neuropathic pain.


Techniques for Robotic-Assisted Surgical Myocardial Revascularization.
The two current strategies for robotic-assisted, surgical myocardial revascularization are minimally invasive direct coronary artery bypass (MIDCAB) and totally endoscopic coronary artery bypass grafting (TECAB). We discuss the rationale underlying the benefits of robotic assistance in surgical myocardial revascularization, and detail the technical steps to safely and effectively perform these two procedures.


A Comparative Study of Three Types of Rapid Maxillary Expansion Devices in Surgically Assisted Maxillary Expansion: A Finite Element Study.
The aim of this study was to analyze the displacement pattern and stress distribution during surgically assisted rapid maxillary expansion (RME) with three different types of RME devices by constructing a finite element model. A finite element model is constructed from the computerized tomography scans. According to the type of RME device, 3 groups were simulated on this mesh model. The experimental groups were as follows; Group I (tooth borne appliance), Group II (bone borne appliance), and Group III (hybrid appliance). A Le fort I osteotomy with bilateral pterygomaxillary disjunction and midpalatal split osteotomy cuts were incorporated in all the groups. The displacement pattern and stress distribution for each 1 mm of activation of appliance up to 10 mm is noted and analyzed. The data were analyzed using Student's t-test, Analysis of Variance and Duncan new multiple range test. Tooth borne appliance has more rotational tendencies. The bone borne and the hybrid appliance exhibited similar stress patterns for the dissipation of the forces produced by RME appliances. The pivoting effect decreased with the hybrid and the bone borne appliance and can be utilized in patient with hyper divergent growth. The pivoting effect is least with the hybrid appliances.


Mapping the association between back pain and type 2 diabetes: A cross-sectional and longitudinal study of adult Spanish twins.
Back pain and type 2 diabetes often co-occur, resulting in greater impact on people's health and complexity in their care. Plausible causal mechanisms for this association have been proposed, yet the nature of the link remains unclear. We therefore explored the direction of the association between type 2 diabetes and chronic back pain in twins, controlling for genetics and early environmental confounding. 2,096 and 1,098 twins were included in the cross-sectional and longitudinal analyses, respectively. Any or severe (≥ 9) low back pain (LBP), neck pain (NP), and spinal pain (concurrent LBP and NP) and type 2 diabetes were investigated. Sequential analyses were performed using logistic regression. Firstly, twins were analysed unpaired (adjusted age and gender): total sample analyses. Then, to control for genetic and shared environmental factors, a co-twin case-control analysis was performed including monozygotic and dizygotic twin pairs discordant for back pain (cross-sectional only). In the cross-sectional total sample analyses, type 2 diabetes was associated with chronic spinal pain (OR 1.61; 95%CI 1.12 to 2.31), severe chronic spinal pain (OR 3.33; 95%CI 1.47 to 7.53), chronic NP (OR 1.37; 95%CI 1.01 to 1.85), severe chronic NP (OR 2.28; 95%CI 1.24 to 4.21), and severe chronic LBP (OR 1.63; 95%CI 1.00 to 2.64). After further adjustment for genetic and shared environmental factors, none of the associations remained significant. The longitudinal analyses indicated that the presence of type 2 diabetes did not increase the risk of future back pain, or vice-versa, after two to four years. Chronic back pain (spinal pain, NP, or LBP) was associated with the prevalence of type 2 diabetes. Associations are stronger for severe cases of pain. Future research should investigate the temporal relationships between these conditions with longer follow up in twins.


Administration of lower doses of radium-224 to ankylosing spondylitis patients results in no evidence of significant overall detriment.
The use of low doses of radium-224 (224Ra) chloride for the treatment of ankylosing spondylitis was stopped following the discovery that patients treated with it had a higher than control incidence of leukaemia and other cancers. This was so even though the treatment resulted in decreased pain and increased mobility-both of which are associated with decreased mortality. It was decided to re-analyze the epidemiological data looking at all causes of death. The risk of leukaemia, solid cancer, death from non-cancer causes and from all causes in a study populations of men that received either the typical dose of 5.6 to 11.1 MBq of 224Ra, any dose of 224Ra or no radium were compared using the Cox proportional hazard model. For patients that received the typical dose of 224Ra agreed with the excess cancer was similar to that reported in previous studies. In contrast, these patients were less likely to die from non-cancer diseases and from all causes of death than the control patients. No excess mortality was also found in the population of all males that received the radionuclide. It is concluded that 224Ra treatment administered at low doses to patients with ankylosing spondylitis did not impact mortality from all causes. The study demonstrates the need to consider all causes of death and longevity when assessing health impacts following irradiation.


SNP-SNP interactions as risk factors for aggressive prostate cancer.
Prostate cancer (PCa) is one of the most significant male health concerns worldwide. Single nucleotide polymorphisms (SNPs) are becoming increasingly strong candidate biomarkers for identifying susceptibility to PCa. We identified a number of SNPs reported in genome-wide association analyses (GWAS) as risk factors for aggressive PCa in various European populations, and then defined SNP-SNP interactions, using PLINK software, with nucleic acid samples from a New Zealand cohort. We used this approach to find a gene x environment marker for aggressive PCa, as although statistically gene x environment interactions can be adjusted for, it is highly impossible in practicality, and thus must be incorporated in the search for a reliable biomarker for PCa. We found two intronic SNPs statistically significantly interacting with each other as a risk for aggressive prostate cancer on being compared to healthy controls in a New Zealand population.


Sleep and breathing disorders in myotonic dystrophy type 2.
In patients who exhibit myotonic dystrophy type 1 (DM1), sleep disorders and breathing impairments are common; however, in those with DM type 2 (DM2), limited studies on polysomnography (PSG) and none on phrenic compound motor action potential (CMAP) have been performed, which is the aim of this study. Sixteen patients with DM2 were questioned about respiratory symptoms. They underwent PSG with morning arterial gas analyses (AGA). Respiratory functions and phrenic CMAPs were studied. The data were compared to those of 16 healthy controls and 25 patients with DM1. Daytime tiredness is the most common symptom, but orthopnea was reported in 13% of patients with DM2. A detailed sleep architecture analysis revealed a significantly greater proportion of time in stage 3 and REM sleep, and a shorter time in stage 2 in the DM2 than in controls. Lower respiratory volumes and pressures, abnormalities in AGA, night oxygen desaturation and higher EtCO2 are present in DM2, but are less pronounced than in the DM1 population. Small CMAP amplitudes were presented in 12% of patients with DM2, correlating with smaller respiratory functions and poorer sleep quality. AHI was abnormal in 38% of DM2, mainly due to obstructive apneas. PSG did not reveal hypoventilation. Diaphragm weakness and sleep apneas might be present in patients with DM2; therefore, we suggest regular questioning about symptoms of respiratory insufficiency and monitoring of phrenic CMAP. PSG should be recorded, when patients have suggestive symptoms, abnormalities in AGA or higher BMI.


Antiviral activity of diallyl trisulfide against H9N2 avian influenza virus infection in vitro and in vivo.
Diallyl trisulfide (DATS) is a garlic-derived organosulfur compound. As it has been shown to have anti-viral activity, we hypothesized that it may alleviate infections caused by H9N2 avian influenza virus (AIV), which is prevalent in poultry with pandemic potential. Human lung A549 epithelial cells were treated with three different concentrations of DATS 24 h before (pre-treatment) or one hour after (post-treatment) H9N2 AIV infection. Culture supernatants were collected 24 h and 48 h post-infection and analyzed for viral titers and levels of inflammatory and anti-viral immune responses. For in vivo experiments, BABL/c mice were administered daily by intraperitoneal injection with DATS (30 mg/kg) for 2 weeks starting 1 day after H9N2 AIV infection. Clinical signs, lung pathology, and inflammatory and anti-viral immune responses were assessed 2, 4, and 6 days after infection. Both pre-treatment and post-treatment of A549 cells with DATS resulted in reduced viral loads, increased expression of anti-viral genes (RIG-I, IRF-3, and interferon-β), and decreased expression of inflammatory cytokines (TNF-α and IL-6). These effects were also observed in H9N2 AIV-infected mice treated with DATS. Such treatment also reduced lung edema and inflammation in mice. Results suggest that DATS has anti-viral activity against H9N2 AIV and may be used as an alternative treatment for influenza virus infection.


Landmark article Sept 17, 1938: Sodium diphenyl hydantoinate in the treatment of convulsive disorders. By H. Houston Merritt and Tracy J. Putnam.
In a previous study it has been shown that sodium diphenyl hydantoinate is effective in preventing electrically induced convulsive seizures in cats. The drug is relatively nontoxic and well tolerated by the usual laboratory animals. A clinical trial of sodium diphenyl hydantoinate was made in 200 patients with frequent convulsive seizures which had not been relieved by the previous modes of therapy. In 142 such patients who have received the treatment for periods varying from two to eleven months, grand mal attacks were relieved in 58 per cent and greatly decreased in frequency in an additional 27 per cent; petit mal attacks were relieved in 35 per cent and greatly decreased in frequency in an additional 49 per cent, and psychic equivalent attacks were relieved in 67 per cent and greatly decreased in frequency in 33 per cent. There were no fatalities. A toxic dermatitis occurred in ten patients (5 per cent), nonthrombocytopenic purpura in one patient and minor (in many instances, transient) toxic reactions, tremors, ataxia, dizziness and the like in approximately 15 per cent.


Demonstration of the heterogeneity of epitopes of the platelet-specific alloantigen, Baka.
It is well known that the platelet-specific alloantigen, Baka is carried on glycoprotein (GP) IIb, but little is known about the biochemical characteristics of its epitopes. To clarify the characteristics of the epitopes, we examined the interaction of four anti-Baka sera (Yam, Lin, Kl and MO) with their epitopes, either with or without modifications by sodium dodecyl sulphate (SDS) and/or neuraminidase. By immunoprecipitation, all four antisera bound to the intact GP IIb/IIIa complex from a Baka-positive subject. In contrast, immunoblotting demonstrated that Yam, Lin and Kl bound to SDS-denatured GP IIb, while MO did not. When blotted GP IIb was treated with neuraminidase, Yam and Lin did not bind to desialylated GP IIb, while Kl still did. When the purified GP IIb/IIIa complex or washed platelets were treated first with neuraminidase followed by immunoblotting, the molecular weight of GP IIb decreased from 145 kD to 138 kD; Yam did not bind to desialylated GP IIb, but Kl did. Furthermore, to eliminate the effect of SDS, we examined the interaction of Yam and Lin with neuraminidase-treated platelets using flow cytometry. The results were the same as those obtained using immunoblotting. Our results thus demonstrate that the expression of the Baka epitopes is not uniform and that sialic acid contributes to the expression of some actual allogenic epitopes.


Targeted Next-Generation Sequencing Revealed Novel Mutations in Chinese Ataxia Telangiectasia Patients: A Precision Medicine Perspective.
Ataxia telangiectasia (AT) is an autosomal recessive disease characterized by progressive cerebellar ataxia, oculocutaneous telangiectasia and immunodeficiency due to mutations in the ATM gene. We performed targeted next-generation sequencing (NGS) on three unrelated patients and identified five disease-causing variants in three probands, including two pairs of heterozygous variants (FAT-1:c.4396C>T/p.R1466X, c.1608-2A>G; FAT-2:c.4412_4413insT/p.L1472Ffs*19, c.8824C>T/p.Q2942X) and one pair of homozygous variants (FAT-3: c.8110T>G/p.C2704G, Hom). With regard to precision medicine for rare genetic diseases, targeted NGS currently enables the rapid and cost-effective identification of causative mutations and is an updated molecular diagnostic tool that merits further optimization. This high-throughput data-based strategy would propel the development of precision diagnostic methods and establish a foundation for precision medicine.


Sacroiliitis as an initial manifestation of acute myelogenous leukemia.
Sacroiliitis is the most pathognomonic and earliest manifestation of ankylosing spondylitis. We herein report a 28-year-old female patient who presented with sacroiliitis as an initial manifestation of acute myelogenous leukemia (AML). She had a 3-month history of anemia and walking difficulty. Bone marrow findings revealed an increase of blasts with trilineage dysplasia. Although she was initially diagnosed with myelodysplastic syndrome (MDS), blasts rapidly increased and AML developed 1 month after the diagnosis of MDS with Sacroiliitis. Induction chemotherapy failed to induce a complete remission of AML, but it did effectively treat the sacroiliitis. However, the sacroiliitis relapsed when the leukemia cells progressed thereafter. Oral corticosteroids helped ameliorate the sacroiliitis. She underwent bone marrow transplantation (BMT) from an HLA-identical sister during a nonremission period; however, the leukemic cells began to rapidly increase from day 30 after BMT. The close relationship between the occurrence of sacroiliitis and AML suggested that autoimmune sacroiliitis was a paraneoplastic phenomenon of AML in this patient. Although autoimmune disorders develop in a substantial number of MDS patients, they are rarely observed in de novo AML. No previous report has described sacroiliitis as the initial manifestation of de novo AML.


Antibiotic Therapy for Chorioamnionitis to Reduce the Global Burden of Associated Disease.
Chorioamnionitis is associated with significant maternal and neonatal morbidity and mortality throughout the world. In developed countries, great progress has been made to minimize the impact of chorioamnionitis, through timely diagnosis and appropriate treatment. In the global setting, where many women deliver outside the healthcare facilities, this diagnosis is frequently overlooked and not properly treated. In addition to its impact on maternal health, a significant proportion of neonatal morbidity and mortality can be prevented by both recognition and access to readily available treatment. With the increasing focus on saving the most vulnerable members of society, we echo the need for providing parturient women with suspected chorioamnionitis universal access to appropriate therapy. We describe known effective antibiotic therapies for chorioamnionitis and provide an overview of additional potential antimicrobial treatments that might be effectively implemented in areas with limited access to care.


Events of Molecular Changes in Epithelial-Mesenchymal Transition.
EMT is the process by which epithelial cells, characterized by well-developed intercellular contacts, transdifferentiate into motile and invasive mesenchymal cells. This process is associated with the loss of transmembrane intercellular adhesion molecule E-cadherin and disruption of cell-cell junctions along with acquisition of migratory properties. EMT is integral in embryonic development, wound healing, and stem cell behavior; however, its aberrant activation by micro-environmental alterations and abnormal stimuli can lead to cancer progression. Here, we review the different molecular changes associated with EMT that are responsible for downregulation of epithelial genes. Increased knowledge of the EMT process is essential for therapeutic targeting of cancer cells.


Infections associated with clozapine: a pharmacovigilance study using VigiBase®.
Introduction: Clozapine is primarily reserved for treatment-resistant schizophrenia due to safety concerns associated with its use. Infections have been reported with clozapine, which may lead to elevated serum levels of the drug. However, the existing literature on this topic is limited. Therefore, we conducted a study using VigiBase® to investigate the potential over-reporting of infections associated with clozapine, to explore the presence of dose-dependency, and to investigate the underlying mechanism. Methods: Disproportionality analyses were performed using VigiBase to assess the association between clozapine and all types of infections, the association between clozapine-associated infections and neutropenia, the association between clozapine-associated infections and agranulocytosis, the dose-effect relationship between clozapine and infections, and the interaction between clozapine and the main strong CYP450 inhibitors using reports carried out until 11 April 2023. Results: A statistically significant signal of infections was observed with clozapine, as indicated by an information component of 0.43 [95% CI: (0.41-0.45)]. The most commonly reported infections were respiratory and gastrointestinal in nature. Neutropenia showed weaker association with clozapine-associated reports of infections compared to other clozapine-associated reports [X2 (1, N = 204,073) = 454; p < 0.005], while agranulocytosis demonstrated a stronger association with clozapine-associated reports of infections [X2 (1, N = 204,073) = 56; p < 0.005]. No evidence of dose-dependency was observed. Among the 17 tested CYP inhibitors, significant drug-drug interactions were found with clarithromycin, metronidazole, valproic acid, lansoprazole, omeprazole, amiodarone, and esomeprazole. Discussion: Our study revealed a significant safety signal between clozapine use and infections, predominantly respiratory and gastrointestinal infections. The co-administration of clozapine with valproic acid or proton pump inhibitors may potentially contribute to an increased risk of infection. Further vigilance is warranted in clinical practice, and consideration of therapeutic drug monitoring of clozapine in cases involving concomitant use of these drugs or in the presence of infections may be beneficial.


Genetic association between 1425G/A SNP in PRKCH and hypertrophic cardiomyopathy in a Chinese population.
Hypertrophic cardiomyopathy is a heterogeneous myocardial disorder with a broad spectrum of clinical presentation and morphologic features. Previous reports indicated that protein kinase C pathway as a major determinant of cardiac hypertrophy and heart failure. Population-based analyses of the association between PRKCH gene (encoded PKCη) and HCM has not been performed yet. The purpose of this study is to investigate the association of the nonsynonymous SNP (1425G/A) in PRKCH gene and hypertrophic cardiomyopathy in a Chinese population. 323 patients with HCM and 326 controls were examined using a case-control methodology. The 1425G/A SNP in PRKCH was genotyped by allele-specific real-time PCR assay. The 1425G/A SNP in PRKCH increased the risk of HOCM (hypertrophic obstructive cardiomyopathy) (OR=1.427, 95% confidence interval, 1.013 to 2.012, P=0.046) under a dominant model. After age- and sex-adjustment, the significant associations remained in HOCM (for GG +AG versus AA, OR= 2.497, 95% confidence interval, 1.01 to 6.17; P=0.047). The 1425G/A SNP in PRKCH increases the risk of hypertrophic obstructive cardiomyopathy in the Chinese population.


Ultrastructural features of collagen in thyroid carcinoma tissue observed by polarization second harmonic generation microscopy.
Changes in collagen ultrastructure between malignant and normal human thyroid tissue were investigated ex vivo using polarization second harmonic generation (SHG) microscopy. The second-order nonlinear optical susceptibility tensor component ratio and the degree of linear polarization (DOLP) of the SHG signal were measured. The ratio values are related to the collagen ultrastructure, while DOLP indicates the relative amount of coherent signal and incoherent scattering of SHG. Increase in ratio values and decrease in DOLP were observed for tumor tissue compared to normal thyroid, indicating higher ultrastructural disorder in tumor collagen.