Integrative transcriptomic and network pharmacology analysis reveals the neuroprotective role of BYHWD through enhancing autophagy by inhibiting Ctsb in intracerebral hemorrhage mice.
In this study, we aimed to combine transcriptomic and network pharmacology to explore the crucial mRNAs and specific regulatory molecules of Buyang Huanwu Decoction (BYHWD) in intracerebral hemorrhage (ICH) treatment. C57BL/6 mice were randomly divided into three groups: sham, ICH, and BYHWD. BYHWD (43.29 g/kg) was administered once a day for 7 days. An equal volume of double-distilled water was used as a control. Behavioural and histopathological experiments were conducted to confirm the neuroprotective effects of BYHWD. Brain tissues were collected for transcriptomic detection. Bioinformatics analysis were performed to illustrate the target gene functions. Network pharmacology was used to predict potential targets for BYHWD. Next, transcriptomic assays were combined with network pharmacology to identify the potential differentially expressed mRNAs. Immunofluorescence staining, real-time polymerase chain reaction, western blotting, and transmission electron microscopy were performed to elucidate the underlying mechanisms. BYHWD intervention in ICH reduced neurological deficits. Network pharmacology analysis identified 203 potential therapeutic targets for ICH, whereas transcriptomic assay revealed 109 differentially expressed mRNAs post-ICH. Among these, cathepsin B, ATP binding cassette subfamily B member 1, toll-like receptor 4, chemokine (C-C motif) ligand 12, and baculoviral IAP repeat-containing 5 were identified as potential target mRNAs through the integration of transcriptomics and network pharmacology approaches. Bioinformatics analysis suggested that the beneficial effects of BYHWD in ICH may be associated with apoptosis, animal autophagy signal pathways, and PI3K-Akt and mTOR biological processes. Furthermore, BYHWD intervention decreased Ctsb expression levels and increased autophagy levels in ICH. Animal experiments in combination with bioinformatics analysis confirmed that BYHWD plays a neuroprotective role in ICH by regulating Ctsb to enhance autophagy.


Overexpressed fibroblast growth factor receptor 2 in the invasive front of colorectal cancer: a potential therapeutic target in colorectal cancer.
Fibroblast growth factor receptor 2 (FGFR2) is considered a novel therapeutic target for various cancer. We used a silencing strategy to clarify the effect of reduced FGFR2 expression in human colorectal cancer (CRC) cells. The invasive front of cancer cells exhibited stronger FGFR2 expression than the surface area of the cancers. FGFR2 shRNA-transfected LoVo cells inhibited cell migration, invasion and tumor growth in vitro and in vivo. Thus, FGFR2 plays important roles in CRC progression in association with tumor cell migration, invasion and growth, and FGFR2 might be a novel therapeutic target for CRC.


Expression of RSUME is Associated With Poor Prognosis in Clear Cell Renal Carcinoma: Involvement of ROS Related Metabolism.
RSUME (RWD domain-containing protein SUMO Enhancer), RWD domain containing 3 (RWDD3) gene product, is upregulated by hypoxia and expressed in organs prone to develop von Hippel-Lindau (VHL) syndrome tumors. We evaluated RSUME prognostic value in clear cell renal cell carcinoma (ccRCC) based mainly on the dataset (KIRC) from patients in The Cancer Genome Atlas (TCGA). Wilcoxon signed-rank test and one-way analysis of variance (ANOVA) followed by Tukey's test were used to evaluate relationships between clinicopathological features and RSUME expression and univariate and multivariate Cox regression analysis methods were used to evaluate prognostic factors. The biological function of RSUME was assessed by gene set enrichment analysis (GSEA). For validation, total amount of ROS was detected in ccRCC cell lines using dichlorofluorescin diacetate. RSUME is highly expressed in tumor tissues compared with normal tissues (P = .006, P = .039, P = .002, P = .036, P < .001) and associates with tumor T (P = .018) and tumor M (P = .036) advanced stages and higher extent cysts (P = .005). RSUME expression appears to be an independent risk factor for overall survival (OS) (P = .002) and disease-specific survival (DSS) (P = .026) in ccRCC patients. GSEA showed enrichment of relevant glycerophospholipid- and ROS-related pathways in RSUME high-expression phenotype. ROS diminished levels in RSUME-silenced ccRCC cell lines validated RSUME relevance in ROS-related pathways. RSUME high expression may predict poor prognosis in ccRCC and impact through its action on metabolism and ROS related pathways.


The dose makes the medicine.
Dose and time considerations in the development and use of a drug are important for assessing actions and side effects, as well as predictions of safety and toxicity. This article deals with epistemological aspects of dose selection by probing into the linguistic and cultural roots for the measure of medicine mediated by the medical doctor. Because toxicity is related to dose, historic and recent views suggest that less can be more. At low, medium and high dose levels, effects can differ not only quantitatively but also qualitatively. Dose-related target activation and recognition of enantiodromic thresholds between beneficial and toxic effects require elucidation of underlying events. Such studies, including hormesis and microdosing, call for extended ADME procedures with high-resolution methods in addition to the current low-resolution approaches. Improved information of drug logistics and target pharmacokinetics enables effective drug selection, dose determination and prediction. It also allows considerations of systems biology [i.e. integral (gestalt) pharmacology] exemplified by the drug homunculus, as in the case of vitamin D, that might lead to new paradigms and drug design.


The Portuguese validation of the short form of the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12).
Sexual well-being is an important aspect of women's life. The objective of this study was to validate the Portuguese-translated version of the short form of the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire--PISQ-12. Sixty-four women were enrolled to participate in the process of validating the PISQ-12 (study group). A further 68 subjects were allocated to a control group for comparison between the group of women with pelvic organ prolapse/urinary incontinence and the asymptomatic group. The PISQ-12 presented good internal consistency (Cronbach's alpha of 0.79 for the study group and 0.80 for the control group). The test-retest reliability using the intraclass correlation coefficient was 0.77. Mean score on the PISQ-12 was significantly higher in the control group than in the study group (42.7 ± 3.9 vs 27.8 ± 9.3) The Portuguese version of the PISQ-12 was reliable for the assessment of sexual function in women with pelvic organ prolapse/urinary incontinence.


Detecting transcriptionally engaged RNA polymerase in eukaryotic cells with permanganate genomic footprinting.
Analysis of the distribution of RNA polymerase II on the genomes of Drosophila and human cells using in vivo protein-DNA crosslinking reveals that RNA polymerase II (Pol II) is concentrated at the 5'-ends of thousands of genes. This appears to be irrespective of transcription levels. Hence, a potential regulatory step in the transcription of many genes occurs after Pol II has associated with the promoter. The protein-DNA crosslinking technique widely used to monitor Pol II and other proteins on chromosomes in vivo, however, does not reveal if Pol II is transcriptionally engaged on DNA. Genomic footprinting with potassium permanganate provides one method for detecting transcriptionally engaged Pol II. Using this approach, we have determined that the Pol II associated with the promoters of many genes has initiated transcription but paused in the region 20-50 nucleotides from the start. Here we describe the application of this method in Drosophila and human cells. The method should prove useful in assessing if promoter bound Pol II has engaged in transcription and for investigating the establishment and regulation of transcriptionally engaged Pol II.


Accelerated Resolution Therapy (ART): a Review and Research to Date.
To describe and summarize published research on accelerated resolution therapy (ART), a promising and relatively new psychotherapy with the potential to offer rapid and effective resolution of a wide range of psychiatric symptoms. Unlike most evidence-based psychotherapies, ART is a predominately imaginative therapy that relies upon the rescripting of distressing events and metaphors as one of its key therapeutic elements. The number of studies conducted on ART is limited, primarily consisting of one randomized, controlled trial (RCT) with 57 subjects and two large cohort studies involving 80 and 117 subjects, respectively. However, a growing body of research in the neuroscience field involving the initial creation (consolidation), activation, and reconsolidation of memories may also be relevant and is summarized herein. ART appears to be an effective, efficient, and versatile form of psychotherapy. Future studies, particularly high-quality RCTs, are needed to more fully understand the potential reach of this promising therapeutic modality.


The insertion of D-beta-hydroxybutyrate apodehydrogenase into phospholipid monolayers and phospholipid vesicles.
The strong interaction of D-beta-hydroxybutyrate dehydrogenase with phospholipid monomolecular films is demonstrated by the surface pressure increase of a film compressed up to 33 mN/m. Although the D-beta-hydroxybutyrate apodehydrogenase was able to penetrate many phospholipid monolayers, it interacted preferentially with negatively charged monolayers such as those made from diphosphatidylglycerol. The weakest interaction was found with phosphatidylcholine, which is the reactivating phospholipid for the enzyme. These interactions were dependent on the phospholipid chain length, ionic strength, and pH. At basic pH the apoenzyme lost its specificity for negatively charged phospholipids, suggesting the deprotonation of a cationic amino acid residue of the enzyme polypeptide chain. The charge effects are in agreement with results obtained using phospholipid vesicles. Beside the electrostatic interactions, the influence of phospholipid chain length and the ionic strength indicate that D-beta-hydroxybutyrate apodehydrogenase penetrates into the hydrophobic part of the lipid interface.


Adverse pregnancy outcomes are associated with multiple maternal thrombophilic factors.
To determine to what extent adverse pregnancy outcomes are associated with thrombophilia. We studied 31 women who had HELLP syndrome, placental abruption, fetal growth restriction or unexplained stillbirth (study group), matched with 12 controls. All women were tested for: Factor V, Prothrombin, methylenetetrahydrofolate reductase gene (MTHFR) mutations; for Protein C, S and Antithrombin III deficiency; for lupus anticoagulant. Correlation with 24h BP monitoring and uterine Doppler velocimetry indexes at 22-24 weeks' gestation was performed. Women with multiple thrombophilic factors had a significant lower birth weight (1568.33+/-146.8 g versus 2546.45+/-438 g), higher 24 h mean diastolic blood pressure at second trimester (76.3+/-12.5 mmHg versus 65.2+/-7.8 mmHg) and higher RI of uterine arteries (0.69+/-0.05 versus 0.50+/-0.15) than women with single thrombophilic factor. Multiple thrombophilic factors carry a major additional risk for adverse maternal and fetal outcomes and correlate well with placental maladaptation as indicated by uterine Doppler velocimetry and 24h BP monitoring.


Idiopathic hypogonadotropic hypogonadism in a male runner is reversed by clomiphene citrate.
To assess the efficacy of estrogen antagonist therapy on the function of the hypothalamic-pituitary-testicular axis in a young male runner with significant morbidity attributable to idiopathic hypogonadotropic hypogonadism. An uncontrolled case study. The outpatient endocrinology clinic of a university tertiary referral center. A 29-year-old male who has run 50 to 90 miles per week since 15 years of age and who presented with a pelvic stress fracture, markedly decreased bone mineral density, and symptomatic hypogonadotropic hypogonadism. Clomiphene citrate (CC) at doses up to 50 mg two times per day over a 5-month period. Serum concentrations of LH, FSH, and T before and after CC therapy, as well as clinical indicators of gonadal function. Barely detectable levels of LH and FSH associated with hypogonadal levels of T were restored to the normal range with CC therapy. The patient experienced improved erectile function, increased testicular size and sexual hair growth, and an improved sense of well being. Exercise-induced hypogonadotropic hypogonadism exists as a clinical entity among male endurance athletes, and CC may provide a safe and effective treatment option for males with debilitating hypogonadism related to endurance exercise.


Predictive value of neutrophil-to-lymphocyte-ratio in neoadjuvant-treated patients with breast cancer.
Breast cancer (BC) is the most common malignancy among women and prognosis is strongly influenced by tumor subtype. Neoadjuvant chemotherapy (NAC) is the standard treatment for both locally advanced- and early-stage triple-negative and Her2-positive BC. Pathologic complete response (pCR) to NAC is an important predictor of patient outcomes. Neutrophil-to-lymphocyte-ratio (NLR) in peripheral blood is associated with prognosis in various malignancies. Here, we investigated the value of the pretreatment NLR as a response predictor in neoadjuvant-treated patients with BC. A retrospective chart analysis of 862 patients with invasive BC treated with NAC at the Heidelberg University Hospital during 2003-2015 was conducted. NLR was calculated as the ratio of the absolute neutrophil and lymphocyte counts in peripheral blood, and pCR was defined as absence of invasive or in situ carcinoma in breast and axillary lymph nodes. A total of 151 patients with invasive BC who underwent NAC were included in this study. NLR tended to be higher in the pCR group than the non-pCR group (p < 0.1). Analyses of BC subtypes demonstrated that NLR was significantly higher in the pCR- compared with the non-pCR group (3.304 vs. 2.379, respectively; p = 0.048) in patients with luminal B/Her2-negative tumors. Further, we found a significant difference in NLR according to remission status in postmenopausal patients (2.861 vs. 2.313, respectively; p = 0.043). NLR was significantly higher only for patients achieving pCR in the Luminal B/Her2-negative and postmenopausal subgroups. Hence, NLR is a candidate additional predictive factor in patients with Luminal B/Her2-negative BC.


Infection of the urinary tract after anti-reflux surgery.
Two hundred twenty-three children who underwent anti-reflux surgery were evaluated. Pyelonephritic scarring was more apt to occur with greater degrees of reflux. Postoperative infection was principally confined to the bladder and occurred primarily in females. The beneficial effect of anti-reflux surgery is discussed.


Changes in pain perception and pain-related somatosensory evoked potentials in humans produced by exposure to oscillating magnetic fields.
Nociception has been reported to be influenced by exposure to magnetic fields (MFs). The aim of this study was to investigate the effects of 2 h exposure to weak, oscillating MFs on pain perception thresholds and on pain-related somatosensory evoked potentials (SEPs). In 11 healthy volunteers, pain perception thresholds and pain-related SEPs were assessed by intracutaneous electrical stimulation. After sham treatment, pain thresholds significantly increased, whereas after MFs a slight non-significant decrease in thresholds was found. After both treatments pain-related SEP amplitude was reduced, but this decrease was more evident and statistically significant only after MF exposure. The increase found in thresholds after sham exposure may be due to stress-induced analgesia (SIA) and the contrasting behaviour recorded after MF exposure might indicate a suppression of SIA. The significant reduction in pain-related SEP amplitude observed after MF exposure provides the first evidence that human SEPs are influenced by MFs.


A micro method for enumerating T and B cells in mouse peripheral blood.
A micro method has been devised to assess T and B cells in mouse peripheral blood. This technique permits simultaneous determination of B lymphocytes and monocytes. T lymphocytes are determined by a cytotoxicity assay. Tests require only 0.2 ml of whole blood and avoid the loss of certain cell populations secondary to flotation separation techniques. Percentages of IgM and IgG bearing B lymphocytes, Thy-1 bearing T lymphocytes as well as monocytes in mouse peripheral blood were determined in a total of 50 mice of 11 different strains.


Early efficacy of teprotumumab for the treatment of dysthyroid optic neuropathy: A multicenter study.
To study post-interventional findings in patients with dysthyroid optic neuropathy (DON) treated with teprotumumab. In this multicenter observational Case series, patients with DON were treated with teprotumumab, an insulin-like growth factor I receptor inhibitor (10 mg/kg for the first infusion then 20 mg/kg for subsequent infusions, every three weeks for a total 8 infusions). This study included patients with acute and chronic thyroid eye disease (TED) with DON who had failed conventional therapies and were not candidates for surgical decompression. Data collected included best corrected visual acuity (BCVA), color vision, RAPD when present, and orbital CT or MRI. Proptosis, clinical activity score (CAS), Gorman diplopia score (GDS), and Humphrey visual fields (HVF) were also evaluated.Ten patients (6 women, 4 men) with an average age 64 years old were included in this study. Mean follow up after completion of infusions was 15 weeks. Baseline visual acuity (VA) impairment ranged from hand motion (HM) to 20/25 in affected eyes. All patients had pre-treatment orbital CT or MRI that confirmed orbital apex compression. Seventy percent of patients had objective improvement in DON after 2 infusions of teprotumumab measured as significant improvement in visual acuity, resolution of RAPD, or both. After completion of treatment, affected eyes had a mean BCVA improvement of 0.87 logMAR (p=0.0207), proptosis reduction of 4.7 mm (p<0.00001), CAS improvement of 5.25 points (p<0.00001), and GDS improvement of 0.75 points (p=0.160). All 6 patients who presented with an RAPD had resolution or improvement of RAPD. All 7 patients who presented with color vision deficits had normalization or improvement of color vision. Teprotumumab infusions resulted in medical decompression and objective resolution or improvement of dysthyroid optic neuropathy. Most patients had rapid improvement of visual acuity and reversal of RAPD. Post-infusion imaging demonstrated reduction in extraocular muscle size that correlated with improvement in visual dysfunction. However, patients who presented with longstanding severe visual loss had limited improvement. There was no recurrence of DON after completion of teprotumumab in our cohort.


Phthalimide-tethered isatins as novel poly(ADP-ribose) polymerase inhibitors: Design, synthesis, biological evaluations, and molecular modeling investigations.
Humanity is currently facing various diseases with significant mortality rates, particularly those associated with malignancies. Numerous enzymes and proteins have been identified as highly promising targets for the treatment of cancer. The poly(ADP-ribose) polymerases (PARPs) family comprises 17 members which are essential in DNA damage repair, allowing the survival of cancer cells. Unlike other PARP family members, PARP-1 and, to a lesser extent, PARP-2 show more than 90% activity in response to DNA damage. PARP-1 levels were shown to be elevated in various tumor cells, including breast, lung, ovarian, and prostate cancer and melanomas. Accordingly, novel series of phthalimide-tethered isatins (6a-n, 10a-e, and 11a-e) were synthesized as potential PARP-1 inhibitors endowed with anticancer activity. All the synthesized molecules were assessed against PARP-1, where compounds 6f and 10d showed nanomolar activities with IC50 = 15.56 ± 2.85 and 13.65 ± 1.42 nM, respectively. Also, the assessment of the antiproliferative effects of the synthesized isatins was conducted on four cancer cell lines: leukemia (K-562), liver (HepG2), and breast (MCF-7 and HCC1937) cancers. Superiorly, compounds 6f and 10d demonstrated submicromolar IC50 values against breast cancer MCF-7 (IC50 = 0.92 ± 0.18 and 0.67 ± 0.12 µM, respectively) and HCC1937 (IC50 = 0.88 ± 0.52 and 0.53 ± 0.11 µM, respectively) cell lines. In addition, compounds 6f and 10d induced arrest in the G2/M phase of the cell cycle as compared to untreated cells. Finally, in silico studies, including docking and molecular dynamic simulations, were performed to justify the biological results.


Undegraded human immunoglobulin for intravenous use.
A report is presented on the development and the properties of a nondegraded human immunoglobulin preparation suitable for intravenous administration. Intravenous tolerance is achieved by elimination of anticomplementary activity through modification with beta-propiolactone. The natural antibody characteristics, such as antibody activity, molecular weight, and half-life, are retained. The findings presented open up new perspectives for an extension of the range of indication for intravenous immunoglobulin.


The fluorescence induction kinetics as a non-destructive tool for investigating spruce treated with ozone.
The scattering coefficient of yellow spruce needles exceeds that of green needles by a factor of 2, whereas the fluorescence efficiency is approximately equal for both needle colours. As shown by the angular distribution the fluorescence light is diffusely emitted. However, the scattered light consists of a diffuse and a reflecting portion below 20 degrees with a ratio of the intensities of 1 : 2 at perpendicular observation (0 degrees). Control measurements show that in the rejection region the effective transmission of cut-off-filters commonly used to separate fluorescence light and excitation light exceeds the value calculated from the filter specifications by a factor of 100. Therefore, the portion of the scattered light in the measuring signal must be controlled if the fluorescence induction kinetics is measured from specimen of different colour. A device for the determination of the fluorescence induction kinetics is described which employs a He--Ne laser, a mechanically working shutter with an opening time of 4 ms for the excitation, and a computer for data storage and device control. Two filters select the fluorescence components at 685 nm and 730 nm and they reduce the portion of the scattered light in the measuring signal to 0.18% and 0.55%, respectively. In order to consider the temporal development of the fluorescence kinetics the sampling rate is reduced from 2 kHz to 1 Hz. From the data stored in the computer maximum value Fp, and steady-state-value Fs are determined for both fluorescence components. Measurements on 4-year-old spruce exposed to ozone-concentrations of 0, 300 ppb, 600 ppb, and 1000 ppb were repeated every week. With increasing concentration and duration of treatment Rfd = (Fp-Fs)/Fs was decreased for both fluorescence components. With the highest ozone concentration a reduction of Rfd of 23% and 24%, respectively, was obtained for the two fluorescence components after three weeks.


Immunohistochemical characterization of glial fibrillary acidic protein (GFAP)-expressing cells in a rat liver cirrhosis model induced by repeated injections of thioacetamide (TAA).
Hepatic stellate cells, the principal fibrogenic cell type in the liver, are known to express the astrocyte marker glial fibrillary acidic protein (GFAP). However, the exact role of GFAP-expressing cells in liver fibrosis remains to be elucidated. In this study, cellular properties of GFAP-expressing cells were investigated in a rat model of liver cirrhosis. Six-week-old male F344 rats were injected intraperitoneally with thioacetamide (100 mg/kg BW, twice a week) and examined at post first injection weeks 5, 10, 15, 20 and 25. Appearance of GFAP-expressing myofibroblasts peaked at week 15, associated with fibrosis progression. The majority of GFAP-expressing myofibroblasts co-expressed vimentin, desmin and alpha-smooth muscle actin. Some GFAP-positive myofibroblasts co-expressed nestin (neural stem cell marker), while a few co-expressed A3 (mesenchymal stem cell marker) and Thy-1 (immature mesenchymal cell marker). A few GFAP expressing cells underwent both mitosis and apoptosis. These results indicate that there is a dynamic participation of GFAP-expressing myofibroblasts in rat liver cirrhosis, and that they are mainly derived from hepatic stellate cells, and partly from cells in the stem cell lineage. These findings, which were shown for the first time in detail, would be useful to understand the role of GFAP-expressing myofibroblasts in the pathogenesis of chemically induced liver cirrhosis.


About camel breeds: A reevaluation of current classification systems.
This article revaluates the current dromedary breed classifications based on Leese's (1927) division into hills, riverine and plains camel according to camel breeding zones, and the even earlier distinction by Cross (1917) and Leonard (1894) between baggage and riding camels. It is suggested that these classifications are inappropriate to portray the wide variety of camel breeds actually existing, since they do not accurately reflect the ecological context of camel breeding and also depict incompletely the actual uses to which camels are put in camel pastoral societies. The breeding practices of traditional camel pastoral societies are described and discussed, and it is concluded that these have been conducive to the development of a large number of specialized camel breeds. It is pointed out that many camel breeds are endangered, and that it is imperative to take steps for their documentation. ZUSAMMENFASSUNG: Kamelrassen: Revision des gegenwärtigen Klassifikationssystems In diesem Artikel werden die heute gebräuchlichen Klassifikationssysteme für Dromedarrassen einer Prüfung unterzogen: Diese beruhen auf der Einteilung von Leese (1927) nach Kamelzuchtgebieten in Berg-, Fluß- und Flachlandkamele und der noch älteren Unterscheidung von Cross (1917) und Leonard (1894) zwischen Reit- und Lastkamelen. Es wird vorgeschlagen, daß diese Einteilungen der tatsächlichen Variabilität zwischen Kamelrassen nicht gerecht werden, weil sie die ökologischen Bedingungen, unter denen Kamelzucht betrieben wird, nicht richtig reflektieren und die tatsächlichen Nutzungsweisen, denen das Kamel in Kamelhirtenkulturen unterzogen wird, nur unvollständig wiedergeben. Die Zuchtmethoden traditioneller Kamelhirten werden beschrieben und diskutiert, und es wird die Schlußfolgerung gezogen, daß diese für die Herausbildung vieler spezialisierter Rassen ideale Vorbedingungen darstellten. Auf die große Gefahr des Verlustes vieler dieser Rassen und die Notwendigkeit diese zu katalogisieren wird hingewiesen.


Optimal Thyroid Hormone Replacement Dose in Immune Checkpoint Inhibitor-Associated Hypothyroidism Is Distinct from Hashimoto's Thyroiditis.
Background: Immune checkpoint inhibitors (ICI) have revolutionized the treatment of many advanced cancers but are recognized to cause treatment-limiting immune-related adverse events (IrAE). ICI-associated thyroiditis is the most common endocrine IrAE and usually resolves to permanent hypothyroidism. Optimal thyroid hormone replacement in these patients remains unclear. We report the levothyroxine (LT4) dose needed to achieve stable euthyroid state in patients with hypothyroidism from ICI-associated thyroiditis, with comparison to patients with Hashimoto's thyroiditis (HT) and athyreotic state. Methods: We conducted a retrospective study of adults with ICI-associated hypothyroidism treated with LT4 at an academic medical center. Patient data were collected from the electronic medical record. Cases had ICI exposure followed first by hyperthyroidism and then subsequent hypothyroidism. Controls were HT (positive thyroid autoantibodies, requiring LT4) and athyreotic (total thyroidectomy or radioiodine ablation, requiring LT4) patients. Patients with central hypothyroidism, thyroid cancer, pregnancy, gastrointestinal stromal tumors, and use of L-triiodothyronine were excluded. Our primary outcome compared LT4 dose needed to achieve euthyroid state (thyrotropin 0.3-4.7 mIU/L over >6 consecutive weeks) for ICI-associated hypothyroidism, HT, and athyreotic patients, considering the impact of age and possible interfering medications by linear regression modeling. Secondary analysis considered the impact of endocrine specialty care on the time to euthyroid state. Results: One hundred three patients with ICI-associated thyroiditis were identified. Sixty-six of the 103 patients achieved euthyroid state; 2 with intrinsic thyroid gland function recovery and 64 on LT4. The mean LT4 dose achieving stable euthyroid state was 1.45 ± standard deviation (SD) 0.47 mcg/[kg·day] in ICI-associated hypothyroidism, 1.25 ± SD 0.49 mcg/[kg·day] in HT, and 1.54 ± SD 0.38 mcg/[kg·day] in athyreotic patients, using actual body weight. The difference in dose between ICI-associated hypothyroidism and HT was statistically significant (p = 0.0093). Dosing differences were not explained by age or use of interfering medications. Conclusions: ICI-associated thyroiditis represents an increasingly recognized cause of hypothyroidism. Our study demonstrates that patients with ICI-associated hypothyroidism have different thyroid hormone dosing requirements than patients with HT. Based on our findings and prior reports, we recommend that in patients with ICI-associated thyroiditis LT4 therapy be started at an initial weight-based dose of 1.45 mcg/[kg·day] once serum free thyroxine levels fall below the reference range.


Synthesis of milk specific fatty acids and proteins by dispersed goat mammary-gland epithelial cells.
The method now described for preparation of dispersed lactating goat mammary-gland cells gives a high yield of morphologically and functionally normal mammary cells. The cells synthesize specific goat milk fatty acids in the right proportions, and they respond to hormones by increased protein synthesis. The cells can be frozen and thawed without losing the above properties, which makes them an excellent tool for metabolic and hormonal studies.


Blindness as an ictal phenomenon: investigations with EEG and SPECT in two patients suffering from epilepsy.
Blindness is a rare ictal phenomenon in epileptic seizures. It can occur as an aura, as the seizure itself, or postictally. We investigated two such patients, in one of whom blindness manifested as an aura prior to tonic clonic seizures; the interictal EEG exhibited a spike-wave focus bioccipitally. In the second patient blindness occurred postictally. An ictal SPECT, carried out at the onset of the seizure demonstrated marked hyperperfusion in both occipital regions.


In vitro and in vivo anti-tumor activity of L-glutamic acid gamma-monohydroxamate against L1210 leukemia and B16 melanoma.
A glutamine analogue, L-glutamic acid gamma-monohydroxamate (GAH) demonstrated complete cytotoxicity against L1210 cells in culture and marked anti-tumoral activity in vivo against L1210 leukemia and B16 melanoma. In vitro, GAH caused concentration-dependent inhibition of L1210 cell growth, with complete cell death being reached at 72 hr and at a 500 microM concentration. A minimal incubation time of 38 hr with 500 microM GAH was necessary to obtain complete cell death at 72 hr. During incubation, GAH is metabolized to hydroxylamine. Hydroxylamine acts as the active form of GAH, since the concentration-dependent inhibition of cell growth caused by hydroxylamine is the same as that observed with GAH. The cytotoxic effects of GAH and hydroxylamine on L1210 cells were not reversed or prevented by L-glutamine or L-glutamic acid and purine nucleosides but were prevented or reversed by pyruvate, 2-oxaloacetate and 2-oxoglutarate. In vivo, GAH considerably increased survival of mice bearing L1210 leukemia or a solid tumor, the B16 melanoma. Antitumor activity of GAH against L1210 leukemia and B16 melanoma was schedule-dependent. The administration of GAH 3 times daily was more effective than a twice daily treatment and the maximum ILS was observed using split-dose schedules on days 1 through 3 and 7 through 9 without noticeable toxicity. Under these conditions hydroxylamine is highly toxic, suggesting that in vivo GAH might act as an hydroxylamine releaser in the tumor cells and is not significantly metabolized in the body.


Trpac1, a pH response transcription regulator, is involved in cellulase gene expression in Trichoderma reesei.
Fungi grow over a relatively wide pH range and adapt to extracellular pH through a genetic regulatory system mediated by a key component PacC, which is a pH transcription regulator. The cellulase production of the filamentous fungi Trichoderma reesei is sensitive to ambient pH. To investigate the connection between cellulase expression regulation and ambient pH, an ortholog of Aspergillus nidulans pacC, Trpac1, was identified and functionally characterized using a target gene deletion strategy. Deleting Trpac1 dramatically increased the cellulase production and the transcription levels of the major cellulase genes at neutral pH, which suggested Trpac1 is involved in the regulation of cellulase production. It was further observed that the expression levels of transcription factors xyr1 and ace2 also increased in the ΔTrpac1 mutant at neutral pH. In addition, the ΔTrpac1 mutant exhibited conidiation defects under neutral and alkaline pH. These results implied that Trpac1 in involved in growth and development process and cellulase gene expression in T. reesei.


Digital gene expression analysis of reproductive toxicity of benzo[a]pyrene in male scallop chlamys farreri.
Benzo[a]pyrene (BaP) is a representative polycyclic aromatic hydrocarbon (PAH) and is studied widely for its strong toxicity and wide distribution. Although BaP pollution in marine environment is increasing, molecular mechanisms underlying reproductive toxicity of BaP in marine mollusks have been seldom systematically studied, especially in males. In this study, genes that regulated reproductive responses of Chlamys farreri under BaP stress were analyzed through digital gene expression (DGE) sequencing with testis tissues. A total of 12,485,055 and 14,454,127 clean reads were generated from control and BaP exposure DGE libraries, respectively. After comparing two libraries, 1051 differentially expressed genes were detected, with 223 up-regulated and 828 down-regulated genes. Gene ontology (GO) annotation and kyoto encyclopedia of genes and genomes (KEGG) pathway analyses were performed on all genes to understand their biological functions and processes. The results showed that numerous enriched, differentially expressed genes related to aromatic compound catabolic processes, spermatid development, microtubule-based movement, energy production and immune response. Quantitative real-time PCR was performed to verify the expressed genes of DGE. The study generated data to show the overall reproductive transcription responses of male C. farreri under BaP stress, and it also can serve as the reference for future study of organic pollutions in aquatic mollusks.


Single Particle Analysis for High-Resolution 2D Electron Crystallography.
Electron crystallography has been used for decades to determine three-dimensional structures of membrane proteins embedded in a lipid bilayer. However, high-resolution information could only be retrieved from samples where the 2D crystals were well ordered and perfectly flat. This is rarely the case in practice. We implemented in the FOCUS package a module to export transmission electron microscopy images of 2D crystals for 3D reconstruction by single particle algorithms. This approach allows for correcting local distortions of the 2D crystals, yielding much higher resolution reconstructions than otherwise expected from the observable diffraction spots. In addition, the single particle framework enables classification of heterogeneous structures coexisting within the 2D crystals. We provide here a detailed guide on single particle analysis of 2D crystal data based on the FOCUS and FREALIGN packages.


Is colonic electrical activity a similar phenomena to small-bowel electrical activity?
This study was designed to investigate colonic spike bursts regarding 1) their migration behavior, 2) their pressure correlates, and 3) comparing colonic short spike bursts with spike bursts from migrating myoelectric complex from the small bowel. Rectosigmoid electromyography and manometry were recorded simultaneously in seven normal volunteers and electromyography alone in five others during two hours of fasting and for two hours after one 2,100-kJ meal. One patient with an ileostomy was also studied by the same method to record the migrating myoelectric complex from the terminal ileum during fasting. Three kinds of spike bursts were observed in the pelvic colon: rhythmic short spike bursts, migrating long spike bursts, and nonmigrating long spike bursts. The meal significantly increased the number of migrating and nonmigrating long spike bursts (from 25 to 38.7 percent of the recording time; P < 0.01). These bursts of potentials showed a peak 15 minutes after the meal, which may be caused by the gastrocolic reflex. Migrating long spike bursts started anywhere along the rectosigmoid and migrated from there aborad 82 percent of the time and orad or in both directions in 10 or 7 percent of the time, respectively. They originated pressure waves 99 percent of the time. Short spike bursts were more frequent before the meal (15.1 percent before and 9.6 percent after the meal), but the difference was not significant; they neither propagated nor initiated pressure waves detected by the mini-balloon. Migrating long spike bursts were the only potentials that migrated, sometimes for short distances. Short spike bursts are a different phenomenon from the small-bowel migrating myoelectric complex because they do not migrate; they can occur during the postprandial period and never originated intraluminal pressure waves.


Monopolar electromyographic signals recorded by a current amplifier in air and under water without insulation.
It was recently proposed that one could use signal current instead of voltage to collect surface electromyography (EMG). With EMG-current, the electrodes remain at the ground potential, thereby eliminating lateral currents. The purpose of this study was to determine whether EMG-currents can be recorded in Tap and Salt water, as well as in air, without electrically shielding the electrodes. It was hypothesized that signals would display consistent information between experimental conditions regarding muscle responses to changes in contraction effort. EMG-currents were recorded from the flexor digitorum muscles as participant's squeezed a pre-inflated blood pressure cuff bladder in each experimental condition at standardized efforts. EMG-current measurements performed underwater showed no loss of signal amplitude when compared to measurements made in air, although some differences in amplitude and spectral components were observed between conditions. However, signal amplitudes and frequencies displayed consistent behavior across contraction effort levels, irrespective of the experimental condition. This new method demonstrates that information regarding muscle activity is comparable between wet and dry conditions when using EMG-current. Considering the difficulties imposed by the need to waterproof traditional bipolar EMG electrodes when underwater, this new methodology is tremendously promising for assessments of muscular function in aquatic environments.


Dependent personality traits and information processing: assessing the interpretation of ambiguous information using the Thematic Apperception Test.
This study was designed to investigate interpretation bias in people with dependent personality traits. Eight Thematic Apperception Test (TAT) cards were administered to participants (N=56) who scored high or low on DSM-III-R dependent personality pathology. Two independent judges rated the TAT stories using a rating list based on the cognitive model of the dependent and paranoid personality disorder. Controlling for self-esteem, the dependent interpretation bias appeared to be specific for dependent personality pathology. SEM analysis supported a mediation model in which beliefs mediate the relationship between DSM-III-R traits and interpretation bias. The findings in this study support the hypothesis that people with dependent traits are characterized by a schema-related interpretation bias and that this bias is mediated by dependent beliefs.


[Duplex sonographic studies on the pathogenesis of splenic hemodynamics in liver cirrhosis].
The haemodynamic role of the spleen in portal hypertension remains unclear. Duplex sonography was undertaken prospectively to discover the relationship of splenomegaly and splenic vein flow to type and severity of liver cirrhosis and portal vein flow, as well as to the degree of oesophageal varices. 89 patients (54 men, 35 women; mean age 52 [26-81] years), diagnosed in the second half of 1993 as having liver cirrhosis were consecutively included in the study. In 40 patients the cirrhosis was in stage A (according to Child-Pugh classification), in 31 in stage B and in 18 in stage C. Portal vein flow fell significantly with increasing degree of cirrhosis (stage A: 8.29 cm/s; stage B 7.03 cm/s; stage C 5.26 cm/s; P < 0.05). Splenic vein flow differed significantly only between stages B and C (stage A: 9.22 cm/s; stage B: 9.46 cm/s; stage C: 7.87 cm/s; B vs C, P < 0.05). There was no correlation between portal vein flow, splenic vein flow, degree of splenomegaly and extent of oesophageal varices. The results may be explained by the presence of collateral circulations and by the differing pathophysiological part played by the spleen in liver cirrhosis of different aetiologies.


Interactions between selenium and group Va-metalloids (arsenic, antimony and bismuth) in the biliary excretion.
The interrelationship between the biliary excretion of exogenous group Va-metalloids (arsenic, antimony and bismuth) and selenium, as well as endogenous glutathione has been studied in rats injected intravenously with sodium selenite and one of the group Va-metalloids. Arsenic, antimony and bismuth appeared in the bile of rats together with large amounts of non-protein thiols (NPSH, representing glutathione and its SH-containing degradation products) and, with the exception of bismuth, they caused choleresis. Significant interactions were observed in the hepatobiliary disposition between selenium and each of the group Va-metalloids, however, their outcomes were not uniform. When coadministered with sodium arsenite or arsenate, selenite enhanced the initial biliary excretion of arsenic 2- and 8-fold, respectively, without further increasing the concomitant excretion of NPSH or the choleretic effect of arsenicals. However, selenite augmented neither the excretion of antimony or bismuth, nor the simultaneous biliary release of NPSH. In turn, arsenite, arsenate and antimony potassium tartrate increased the initial biliary excretion of selenium more than 10-fold and enhanced the accumulation of selenium in blood (exclusively in the erythrocytes). In contrast, administration of bismuth ammonium citrate diminished both the biliary excretion and the erythrocytic accumulation of selenium, while causing retention of selenium in the blood plasma. In rats receiving arsenic or antimony with selenite, the time courses of the biliary excretion of these group Va-metalloids, selenium and NPSH were similar. It is hypothesised that incorporation of selenol metabolites of selenite into the glutathione complexes of arsenic and antimony, resulting in cholephilic ternary complexes, accounts for the arsenic- and antimony-induced augmentation of the hepatobiliary transport of selenium. However, additional chemical and/or dispositional mechanisms are thought to be responsible for the selenite-induced increase in biliary excretion of arsenic.


Rapid purification of topoisomerase I from human breast cancer cells by high-performance liquid chromatography.
The DNA regulatory enzyme topoisomerase I (TpI) from human breast cancer cells has been analyzed by high-performance liquid chromatography (HPLC) for the first time. Cells were homogenized in Tris buffer and TpI activity was extracted with 0.5 M sodium chloride. Negatively supercoiled plasmid pBR322 was used as the substrate to monitor TpI activity, as judged by relaxed products, analyzed on 1% agarose gels. HPLC in the anion-exchange mode (HPIEC) provided an approximately 6-fold purification of the enzyme. Enhanced purification was subsequently obtained by chromatography of a HPIEC eluate on size-exclusion columns (30- to 60-fold). Recovery of TpI from size-exclusion columns, whether used in multistep analysis or as the first step, was dependent on inclusion of organic solvent, 1-propanol (0.5%, v/v), in the mobile phase. Marked resolution of TpI activity was observed with HPIEC on a SynChrom CM-300 column. Enzyme activity was noted in the void volume, at 150-200 mM phosphate and at 250-350 mM phosphate. TpI purification was 10- and 120-fold in the latter two peaks, respectively. Silver-stained polyacrylamide gels of TpI-containing activity, eluted from a CM-300 column, showed considerable purification of all but the void volume fraction. A distinct protein band at approximately 88-90 kD was seen in the peak eluted from the CM-300 column with 250-350 mM phosphate. These results indicate that HPLC is useful for rapid purification of the labile enzyme, TpI, in the analysis of its structure-function relationship.


Role of the Nrf2-ARE pathway in acrylamide neurotoxicity.
Acrylamide (ACR) intoxication is associated with selective nerve terminal damage in the central and peripheral nervous systems. As a soft electrophile, ACR could form adducts with nucleophilic sulfhydryl groups on cysteine residues of kelch-like erythroid cell-derived protein with CNS homology-associated protein 1 (Keap1) leading to dissociation of the transcription factor, nuclear factor erythroid 2-related factor 2 (Nrf2). Nrf2 activation of the antioxidant-responsive element (ARE) and subsequent upregulated gene expression of phase II detoxification enzymes and anitoxidant proteins might provide protection in neuronal regions with transcriptional capabilities (e.g., cell body). In contrast, non-transcriptional cell regions (axons, nerve terminals) might be vulnerable to electrophile-induced damage. To test this possibility, immunoblot analysis was used to measure protein products of Nrf2-activated ARE genes in nerve terminals and in cytosolic/nuclear factions of neuronal cell bodies isolated from rats intoxicated at two different ACR dose-rates; i.e., 50mg/kg/d×10 days, 21mg/kg/d×38 days. To detect possible differences in cell-specific induction, the cytoprotective response to ACR intoxication was determined in hepatic cells. Results show that control brain and hepatic cell fractions exhibited distinct subcellular distributions of Nrf2, Keap1 and several ARE protein products. ACR intoxication, however, did not alter the levels of these proteins in synaptosomal, brain cytoplasm or liver cell fractions. These data indicate that ACR was an insufficient electrophilic signal for ARE induction in all subcellular fractions tested. Because a cytoprotective response was not induced in any fraction, nerve terminal vulnerability to ACR cannot be ascribed to the absence of transcription-based defense mechanisms in this neuronal region.


Comparison of intravenous ibuprofen and paracetamol in the treatment of fever: A randomized double-blind study.
Fever is one of the frequent reasons for admission to the emergency department. Studies comparing oral forms of non-steroidal anti-inflammatory drugs (NSAIDs) and paracetamol with intravenous (IV) forms for fever are common in the literature. Our study is the first emergency department study comparing IV forms of ibuprofen and paracetamol in the treatment of febrile patients. A randomized, double-blind study was conducted in a tertiary university emergency department for a six-month period. Patients aged 18-65 years who had a fever of ≥38.0 °C were included. Patients were administered 400 mg of IV ibuprofen and 1000 mg of IV paracetamol. The primary aim of the study was to determine whether there was a difference in the effect of the two drugs on fever. The secondary aim was to investigate whether there was a difference in terms of numeric rating scale (NRS) measurements and the need for additional antipyretic therapy. A total of 200 people, 100 of whom were female, were included in the study. The mean age was 30.77 ± 10.61 years. The mean initial temperature for ibuprofen and paracetamol was 38.79 ± 0.470 °C and 38.70 ± 0.520 °C, respectively, with no difference noted between the groups (p = 0.380). It was found that both drugs significantly provided fever control in the first 30 min (p < 0.001), with no difference between them in terms of fever reduction (p = 0.980). Both drugs significantly improved in accompanying symptoms, although both drugs did not show superiority to each other (p = 0.0226). When evaluated in terms of a need for rescue medication, no significant difference was found between the two drugs (p = 0.404). No side effects were encountered during the study. In adult age group patients admitted to the emergency department with high fever, the IV forms of 1000 mg paracetamol and 400 mg ibuprofen effectively and equally reduce complaints, such as fever and accompanying pain. They can be effectively used as each other's rescue medicine and as an alternative to each other in patients with comorbid diseases.


Advances in Diagnosis and Treatment for Leptomeningeal Disease in Melanoma.
Melanoma has one of the highest incidences of causing leptomeningeal disease (LMD) among solid tumors. LMD patients have very poor prognosis with a dismal survival despite aggressive management. In this article, we review the current approaches in the management of patients with LMD secondary to melanoma, including updates in diagnosis, treatment, up-to-date clinical studies, and future directions. Cerebrospinal fluid (CSF) cytology remains the gold standard for diagnosis, and alternatively, MRI based on clinical presentation can be used. Other approaches such as "liquid biopsies" that detect circulating tumor cells and cell-free DNA have the potential to considerably enhance the diagnosis of LMD from melanoma. As for treatment options, several systemic therapies, involving systemic targeted and immunotherapies have evolved that showed to have possible benefit in LMD patients. Intrathecal chemotherapy, cellular therapy, and immunotherapy are currently under evaluation in Phase I/II clinical trials. In addition, new radiation therapy approaches such as proton cranial-spinal irradiation (CSI) are currently under investigation. LMD management still remains challenging. Future studies are critical to elucidate the pathophysiology of LMD in order to develop new urgently needed diagnostic tools and therapies. Clinical trials ought to be expanded to include patients with LMD. Future clinical studies should also integrate tissue interrogation, scientifically designed therapies, and aggressive, early intervention in patients with suspected LMD.


Improvement in the visualization of superficial arm veins being evaluated for access and bypass.
Duplex ultrasound mapping of arm veins is being performed with increasing frequency. Unlike ultrasound testing in other areas, this has never been subjected to a gold standard invasive test to determine accuracy. Duplex mapping appears to have a good predictive value whenever large veins are demonstrated preoperatively, but its ability to accurately measure minimum-sized veins is unproven. In this study, we compared diameter measurements obtained under six different conditions and used the maximum diameter as the comparison gold standard. A 12-MHz linear probe was used to measure the cephalic and basilic vein cross-sectional diameters at the wrist level in 24 normal volunteers under the following conditions: (1) resting supine with a room temperature of 23 degrees to 24 degrees C, (2) supine with a tourniquet inflated to 65 mm Hg, (3) sitting with the arm dangling, (4) sitting with a tourniquet, (5) sitting after a 2-minute immersion in warm water (44 degrees C), and (6) same with tourniquet. Half the subjects underwent the protocol in a different order. Vein diameters were significantly larger after submersion in warm water compared with supine (P < .05, pair-wise multiple comparison procedure, Student-Newman-Keuls method). Assuming the sitting position (from supine) resulted in a decreased arm vein diameter 58% of the time. In 25% of the normal subjects, the cephalic vein size was <2 mm, which increased to >2 mm after warming. All subjects had either a cephalic or a basilic vein at the wrist that was >3.1 mm after warming. Use of warm water immersion before vein diameter measurement in a sitting position, without a tourniquet, will result in significantly larger diameter findings in normal arm veins. These diameters are likely to more closely resemble the venous diameter after distension with arterial pressure. Further studies are needed to see if warming in patients could result in increased utilization of autogenous arm vein for dialysis access and bypass.


Age-Specific Prevalence of Binge and High-Intensity Drinking Among U.S. Young Adults: Changes from 2005 to 2015.
This study examined changes during the past decade, from 2005 to 2015, in binge and high-intensity drinking in 7 separate age groups of U.S. 12th graders and young adults. National longitudinal data (N = 6,711) from Monitoring the Future were used to examine trends in consuming 5+, 10+, and 15+ drinks on the same occasion in the past 2 weeks from ages 18 to 29/30 overall and by gender. Results were compared with trends in past 12-month and 30-day alcohol use for the same age groups. Between 2005 and 2015, binge (5+) and high-intensity drinking (10+, 15+) generally decreased for individuals in their early 20s, remained somewhat stable for individuals in their mid-20s, and increased for individuals at the end of young adulthood (age 29/30). The observed historical trends in binge and high-intensity drinking were similar to those for past 12-month and past 30-day alcohol use for those aged 18 to 20, but diverged for most other age groups in young adulthood. Trends were generally similar for men and women, except that the increase in prevalence began earlier in young adulthood for women than for men. Binge and high-intensity drinking among U.S. 12th graders and young adults are dynamic phenomena. Prevention and intervention efforts aimed at reducing the harms resulting from 5+, 10+, and 15+ drinking should acknowledge and focus on differences in trends in these behaviors by age and gender.


Effects of Fengycins and Iturins on Fusarium oxysporum f. sp. physali and Root Colonization by Bacillus velezensis Bs006 Protect Golden Berry Against Vascular Wilt.
Bacillus velezensis Bs006 has shown antagonistic activity on Fusarium oxysporum f. sp. physali and biocontrol activity against Fusarium wilt (FW) in golden berry (Physalis peruviana). We hypothesized that strain Bs006 has the ability to synthesize antimicrobial cyclic lipopeptides (CLPs) like other members of the same species. However, if so, the real effects of CLPs on F. oxysporum f. sp. physali and their potential as a biocontrol tool against Physalis-FW have not been elucidated. In this study the CLPs profile of Bs006 in liquid culture and antagonist-plant-pathogen interactions were characterized. Also, the potential effects of supernatant free of bacteria against F. oxysporum f. sp. physali and FW were explored and compared with the effects of pure CLPs. Ultraperformance liquid chromatography-electrospray ionization-mass spectrometry analysis revealed the capacity of Bs006 to synthesize homologous compounds of iturins, surfactins, and fengycins in liquid culture and on the inhibition zone against F. oxysporum f. sp. physali in dual confrontation tests. Bs006 supernatant reduced the germination and growth of F. oxysporum f. sp. physali and caused vacuolization, swelling, and lysis of F. oxysporum f. sp. physali cells in a concentration-dependent manner. Pure fengycins affected the development of F. oxysporum f. sp. physali from 11 mg/liter and iturins from 21 mg/liter. In a gnotobiotic system, Bs006 colonized the root surface of golden berry, inhibited the growth of F. oxysporum f. sp. physali, and produced CLPs. Individual application of Bs006 and supernatant protected the plants from F. oxysporum f. sp. physali infections by 37 to 53%, respectively. Meanwhile, fengycins reduced the disease progress by 39%. These results suggest further studies to select an optimum combination of Bs006 and supernatant or CLPs, which might be a good option as biofungicide against F. oxysporum f. sp. physali.


Tissue magnesium status in diabetes mellitus.
Leucocyte and erythrocyte magnesium was assayed in 17 healthy subjects and 17 insulin dependent diabetic patients. Plasma magnesium concentration (mean +/- standard error of mean) was significantly lower in the diabetic patients (0.80 +/- 0.02 mmol/l), compared with the healthy subjects (0.90 +/- 0.02 mmol/l, p less than 0.001), but the leucocyte and erythrocyte magnesium content was not significantly different in the diabetic patients (34.5 +/- 0.8 and 6.2 +/- 0.2 mmol/kg dry solids) compared with the healthy subjects (35.5 +/- 0.8 and 6.5 +/- 0.11 mmol/kg dry solids). In a separate study skeletal muscle obtained by needle biopsy was also assayed. Plasma magnesium in 10 diabetic patients (0.74 +/- 0.01 mmol/l) was significantly lower than in 16 healthy subjects (0.85 +/- 0.02 mmol/l, p less than 0.001), but there was no significant difference in the mean muscle magnesium content (43.0 +/- 0.7 compared with 40.7 +/- 0.9 mmol/kg dry solids in the diabetic patients).


Noncoordinated transcription in the absence of protein synthesis in yeast.
In the yeast Saccharomyces cerevisiae we have carried out a detailed study of the response of transcription to the inhibition of translation. Measurement of the incorporation of labeled bases and nucleosides into the nucleoside triphosphate pools revealed that amino acid deprivation brings about a 10- to 50-fold inhibition of such labeling. Therefore accurate comparisons of RNA synthesis using such precursors are difficult to obtain. To overcome this problem we have turned to the use of L-[methyl-3H] methionine as a precursor, because the labeling of the S-adenosylmethionine pool is relatively unaffected by the rate of protein synthesis. Using this precursor, we have observed that in the absence of protein synthesis the transcription of ribosomal RNA is reduced by 80%, the transcription of messenger RNA is reduced by about 25%, and the transcription of transfer RNA is reduced by less than 20%. These results are obtained when protein synthesis is inhibited either by deprivation of an amino acid or by the addition of cycloheximide. Ribosomal precursor RNA synthesized in the absence of protein synthesis is fully methylated. We conclude that the transcription of rRNA is the primary target of stringent control. Furthermore the inhibition of protein synthesis, itself, may be the trigger for this response.


The prevalence of diabetes among tuberculosis patients in Denmark.
It is estimated that 25% of the world's population are infected with Mycobacterium tuberculosis and that 463 million people are living with diabetes mellitus (DM), a number that is increasing. Patients with DM have three times the risk of developing tuberculosis (TB) and there is significant interaction between DM and TB, suggesting that DM affects not only risk of TB but also TB presentation, treatment response and outcome. The aim was determining the prevalence of DM among TB patients in Denmark and to assess risk factors. Patient files from all notified TB cases in Denmark from 2009 to 2014 were retrospectively assessed. In total, 1912 patients were included and 5.0% had DM. Patients with DM were older, had more comorbidities, came from outside Denmark, and had a higher mortality compared to non-DM-patients. None of the patients from Greenland had DM. Patients with low socio-economic status had a low prevalence of DM. We found a higher prevalence of DM among Danish-born < 54 year and migrant ≥ 75 year compared to a Danish background population. We found a higher prevalence of DM among TB patients with known risk factors, and a surprisingly low prevalence among patients with low socioeconomic status and patients from Greenland.


Zinc inhibition of potassium efflux in depolarized frog muscle and its modification by external hydrogen ions and diethylpyrocarbonate treatment.
Efflux of 42K+ was measured in frog sartorius muscles equilibrated in hyperosmotic depolarizing solutions. At the internal potentials obtained, K+ passes mainly through the inward rectifier potassium channels. Inhibition of K+ efflux by external Zn2+ (0.25 to 15 mM) differs in three significant ways from inhibition by Ba2+. (1) The dose-response relation does not correspond to action at a single site. (2) The Zn2+-sensitivity of K+ efflux does not depend on [K+]0 at constant internal potential. (3) Zn2+ inhibition is reduced by hydrogen ions, while Ba2+ inhibition is unaffected. Further, the Ba2+-sensitivity of K+ efflux is not altered by a half-inhibiting Zn2+ concentration, suggesting that the two ions do not interact at a common site. The histidine-modifying reagent diethylpyrocarbonate (DEPC) reduces Zn2+ inhibition. After DEPC treatment Zn2+ inhibition is further reduced by low pH. DEPC has little effect on Ba2+ inhibition. Zn2+ inhibition is not altered by treatment with the sulfhydryl reagents 5,5'-dithio-bis(2-nitrobenzoic acid) or dithiothreitol. The results can be described by either of two models in which two sites can bind Zn2+ and one or both of the sites may also bind H+. When both sites bind Zn2+, K+ efflux is inhibited, and a third site may then bind H+. The effects of DEPC can be accounted for by a decrease in H+ affinity of the first two sites by a factor of 50, and a decrease in Zn2+ affinity of these sites and of the H+ affinity of the third site by about one order of magnitude.


Pregnancy resumption following contraceptive discontinuation: Hazard survival analysis of the Indonesia Demographic and Health Survey Data 2007, 2012 and 2017.
The objective of this research is to estimate the probability of pregnancy resumption after discontinuing reversible contraceptives-pills, injectables, implants and IUDs, and to examine the factors associated with the resumption of fertility. The study uses pregnancy calendar data from Indonesia Demographic and Health Surveys (IDHS) of 2007, 2012 and 2017. A hazard model survival method is used for estimating the time needed to resume pregnancy since discontinuing reversible contraceptives. Retrospective data on 4,573; 5,183 and 5,989 episodes of reversible contraceptive discontinuation at the three surveys respectively have been analysed. This study shows that women regained fecundity within one year of discontinuing IUD, pill, injectables or implants. Women using IUD could resume their pregnancy faster than those using implants, pills and injectables. Over the three IDHS 2007, 2012 and 2017 the age-specific percentages of women becoming pregnant after one year of contraceptive discontinuation vary between 72 and 85 for IUD, 75 and 81 for pills, 72 and 76 for implants and 64 and 67 for injectables, with the percentages being higher among younger women. The analysis further shows that length of contraceptive use, parity, prior sexually transmitted infections, knowledge of fertile period, household wealth status and place of residence have no impact on occurrence of pregnancy after contraceptive discontinuation. The analysis disproves a myth that reversible contraceptives make women infertile. Depending on the type of reversible contraceptive used, 65% to 85% of the women were able to conceive after one year of discontinuation.


Recurrent bacteraemia in sheep infected persistently with Anaplasma phagocytophilum.
Following experimental or natural infection with Anaplasma phagocytophilum, the causative agent of tick-borne fever (TBF), sheep may be infected persistently for several months or years. In the present study, quantitative real-time polymerase chain reaction was used to investigate the duration and magnitude of primary bacteraemia and to establish whether the organism is present continuously in the peripheral blood after the period of primary bacteraemia and the cessation of clinical signs. Persistent infection was characterized by a clearly defined period of primary bacteraemia followed by recurrent cycles of bacteraemia, usually lasting a few days and of lower magnitude, interspersed by negative periods of variable duration in which bacterial DNA could not be detected. During a 150-day period of consecutive sampling of four sheep, A. phagocytophilum was detected on 64.25 ± 4.9 occasions, which means that on average bacterial DNA was detected in 42.8 ± 3.3 percent of all samples, with the positive days falling into 15-20 distinct cycles. Primary bacteraemia lasted for 15.5 ± 2.33 days, but secondary and subsequent cycles of bacteraemia were short-lived, with 61% of the cycles lasting only 1-2 days and 39% lasting for 3 or more days. Secondary and subsequent cycles of bacteraemia were not accompanied by febrile responses or other clinical features of TBF. For three animals, bacterial DNA was detected at 311, 318 and 358 days post infection, indicating the long-term persistence of A. phagocytophilum within peripheral blood.


A Severe Case of Recalcitrant Pompholyx.
Pompholyx is an inflammatory vesiculobullous skin disease of the hands and feet belonging to the spectrum of eczema. To report a severe case of pompholyx, its clinical presentation, and its management. A medical chart review was conducted on a patient with this condition. The patient was patch tested according to the North American Contact Dermatitis Group (NACDG) Standard Screening Series, and results were negative. In the past, she was treated with corticosteroid creams as well as narrow band ultraviolet B (UVB), and Psoralen ultraviolet A (PUVA) therapies, and none were beneficial. When the frequency of the episodes increased, methotrexate was introduced but failed to control the condition. Next, mycophenolate mofetil was started and gradually increased to 3.5 g daily along with a separate trial of radiotherapy with marked success. Pompholyx can be challenging to manage, and treatment can involve an assortment of therapies over a prolonged period of time.


Nanoscale Substrate Roughness Hinders Domain Formation in Supported Lipid Bilayers.
Supported lipid bilayers are model membranes formed at solid substrate surfaces. This architecture renders the membrane experimentally accessible to surface-sensitive techniques used to study their properties, including atomic force microscopy, optical fluorescence microscopy, quartz crystal microbalance, and X-ray/neutron reflectometry, and allows integration with technology for potential biotechnological applications such as drug screening devices. The experimental technique often dictates substrate choice or treatment, and it is anecdotally recognized that certain substrates are suitable for a particular experiment, but the exact influence of the substrate has not been comprehensively investigated. Here, we study the behavior of a simple model bilayer, phase-separating on a variety of commonly used substrates, including glass, mica, silicon, and quartz, with drastically different results. The distinct micron-scale domains observed on mica, identical to those seen in free-floating giant unilamellar vesicles, are reduced to nanometer-scale domains on glass and quartz. The mechanism for the arrest of domain formation is investigated, and the most likely candidate is nanoscale surface roughness, acting as a drag on the hydrodynamic motion of small domains during phase separation. Evidence was found that the physicochemical properties of the surface have a mediating effect, most likely because of the changes in the lubricating interstitial water layer between the surface and bilayer.


Impact of polysorbate 65 on tripalmitin crystal growth and release stability of hot melt coated multiparticulate systems.
Hydrochlorothiazide (HCT) multiparticulate systems (MPS) were hot melt coated with the binary mixture of tripalmitin (PPP) and polysorbate 65 (PS 65) to gain an immediate release profile. Once, HCT MPS were produced with a constant ratio of PPP/PS 65 (90:10) at three different coating amounts (15, 25, and 60%w/w) and once the PPP/PS 65 ratio was varied on 98:2 and 80:20, by keeping the coating amount at 60%w/w. PS 65 induced the polymorphic transformation of PPP from the α-form to its most stable β-form right after the hot melt coating (HMC). A release alteration of HCT, either accelerated or decelerated, occurred after the storage under accelerated conditions. The effect of the API core on the lipid lamellar configuration, the thermal behavior of lipid coating, and the effect of PS 65 concentration on the crystal growth of PPP were investigated via X-ray diffraction and DSC. While a low amount of PS 65 was sufficient to promote crystal growth of PPP and resulted in a decelerated release of HCT from the coating, a higher PS 65 concentration favored phase separation of PPP and PS 65 and led to an accelerated release. The increase in PS 65 reinforced the molecular interaction with the lipophilic HCT, reflected in less crystal growth and decelerated release. The knowledge presented in this study supports understanding the instability of binary emulsifier-lipid coating systems, paving the way for developing robust HMC formulations.


The Advantages, Challenges, and Future of Human-Induced Pluripotent Stem Cell Lines in Type 2 Long QT Syndrome.
Type 2 long QT syndrome (LQT2) is the second most common subtype of long QT syndrome and is caused by mutations in KCHN2 encoding the rapidly activating delayed rectifier potassium channel vital for ventricular repolarization. Sudden cardiac death is a sentinel event of LQT2. Preclinical diagnosis by genetic testing is potentially life-saving.Traditional LQT2 models cannot wholly recapitulate genetic and phenotypic features; therefore, there is a demand for a reliable experimental model. Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) meet this challenge. This review introduces the advantages of the hiPSC-CM model over the traditional model and discusses how hiPSC-CM and gene editing are used to decipher mechanisms of LQT2, screen for cardiotoxicity, and identify therapeutic strategies, thus promoting the realization of precision medicine for LQT2 patients.


Identifying communicative functions in discourse with content types.
Texts are not monolithic entities but rather coherent collections of micro illocutionary acts which help to convey a unitary message of content and purpose. Identifying such text segments is challenging because they require a fine-grained level of analysis even within a single sentence. At the same time, accessing them facilitates the analysis of the communicative functions of a text as well as the identification of relevant information. We propose an empirical framework for modelling micro illocutionary acts at clause level, that we call content types, grounded on linguistic theories of text types, in particular on the framework proposed by Werlich in 1976. We make available a newly annotated corpus of 279 documents (for a total of more than 180,000 tokens) belonging to different genres and temporal periods, based on a dedicated annotation scheme. We obtain an average Cohen's kappa of 0.89 at token level. We achieve an average F1 score of 74.99% on the automatic classification of content types using a bi-LSTM model. Similar results are obtained on contemporary and historical documents, while performances on genres are more varied. This work promotes a discourse-oriented approach to information extraction and cross-fertilisation across disciplines through a computationally-aided linguistic analysis.


Developmental plasticity and evolutionary explanations.
Developmental plasticity looks like a promising bridge between ecological and developmental perspectives on evolution. Yet, there is no consensus on whether plasticity is part of the explanation for adaptive evolution or an optional "add-on" to genes and natural selection. Here, we suggest that these differences in opinion are caused by differences in the simplifying assumptions, and particular idealizations, that enable evolutionary explanation. We outline why idealizations designed to explain evolution through natural selection prevent an understanding of the role of development, and vice versa. We show that representing plasticity as a reaction norm conforms with the idealizations of selective explanations, which can give the false impression that plasticity has no explanatory power for adaptive evolution. Finally, we use examples to illustrate why evolutionary explanations that include developmental plasticity may in fact be more satisfactory than explanations that solely refer to genes and natural selection.


Usefulness of convalescent plasma transfusion for the treatment of severely ill COVID-19 patients in Pakistan.
Convalescent plasma(CP) was utilized as potential therapy during COVID-19 pandemic in Pakistan. The study aimed at appraisal of CP transfusion safety and usefulness in COVID pneumonia. Single arm, MEURI study design of non-randomized open label trial was conducted in five centers. Patients werecategorized as moderately severe, severe, and critical. The primary endpoint was a) improvement in clinical status and change in category of disease severity; secondary endpoint was b) CP ability to halt disease progression to invasive ventilation. CP transfused to hospitalized patients. Statistical tests including median (interquartile ranges), Mann-Whitney U test, Fisher's exact test using SPSS ver. 23, ANOVA and Chi-square test were applied for the analysis of results parameters before and after CP treatment. SOFA score was applied for multiorgan failure in severe and critical cases. A total of 50 adult patients; median age 58.5 years (range: 29-92 years) received CP with infusion titers; median 1:320 U/mL (Interquartile range 1:80-1:320) between April 4 to May 5, 2020. The median time from onset of symptoms to enrollment in trial was 3 to 7 days with shortness of breath and lung infiltration as severity criterion. In 35 (70%) recipients, oxygen saturation improved from 80 to 95% within 72h, with resolution of lung infiltrates. Primary endpoint was achieved in 44 (88%) recipients whereas secondary endpoint was achieved in 42 (84%). No patient experienced severe adverse events. A high SOFA score (> 7) correlated with deaths in severe and critical patients. Eight (16%) patients expired due to comorbidities; cardiac arrest in 2 (4%), multiorgan failure secondary to cytokine storm in 5 (10%) and ventilator associated complications in 1 (2%). CP transfusion can be used as a safe and useful treatment in moderately severe and severe patients. The trial registration number is NCT04352751 ( https://www.irct.ir/search/result?query=IRCT20200414047072N1 ). Trial Registration date is 28th April 2020.


β-Propiolactone (BPL)-inactivation of SARS-Co-V-2: In vitro validation with focus on saliva from COVID-19 patients for scent dog training.
β-Propiolactone (BPL) is an organic compound widely used as an inactivating agent in vaccine development and production, for example for SARS-CoV, SARS-CoV-2 and Influenza viruses. Inactivation of pathogens by BPL is based on an irreversible alkylation of nucleic acids but also on acetylation and cross-linking between proteins, DNA or RNA. However, the protocols for BPL inactivation of viruses vary widely. Handling of infectious, enriched SARS-CoV-2 specimens and diagnostic samples from COVID-19 patients is recommended in biosafety level (BSL)- 3 or BSL-2 laboratories, respectively. We validated BPL inactivation of SARS-CoV-2 in saliva samples with the objective to use saliva from COVID-19 patients for training of scent dogs for the detection of SARS-CoV-2 positive individuals. Therefore, saliva samples and cell culture medium buffered with NaHCO3 (pH 8.3) were comparatively spiked with SARS-CoV-2 and inactivated with 0.1 % BPL for 1 h (h) or 71 h ( ± 1 h) at 2-8 °C, followed by hydrolysis of BPL at 37 °C for 1 or 2 h, converting BPL into non-toxic beta-hydroxy-propionic acid. SARS-CoV-2 inactivation was demonstrated by a titre reduction of up to 10^4 TCID50/ml in the spiked samples for both inactivation periods using virus titration and virus isolation, respectively. The validated method was confirmed by successful inactivation of pathogens in saliva samples from COVID-19 patients. Furthermore, we reviewed the currently available literature on SARS-CoV-2 inactivation by BPL. Accordingly, BPL-inactivated, hydrolysed samples can be handled in a non-laboratory setting. Furthermore, our BPL inactivation protocols can be adapted to validation experiments with other pathogens.


Construction of accurate acrylic resin provisional restorations.
A technique has been described for making accurate fitting acrylic resin provisional restorations with attention to marginal adaptation and ease of fabrication.


Effective quantum Brownian dynamics in presence of a rapidly oscillating space-dependent time-periodic field.
We explore the Brownian dynamics in the quantum regime (by investigating the quantum Langevin and Smoluchowski equations) in terms of an effective time-independent Hamiltonian in the presence of a rapidly oscillating field. We achieve this by systematically expanding the time-dependent system-reservoir Hamiltonian in the inverse of driving frequency with a systematic time-scale separation and invoking a quantum gauge transformation within the framework of Floquet theorem.


Individual and community-level determinants of childhood vaccination in Ethiopia.
Vaccines are one of our most important tools for preventing outbreaks and keeping the world safe. Most unvaccinated children live in the poorest countries including Ethiopia. Therefore, this study aimed to identify the determinants of vaccination coverage among children aged12-23 months in Ethiopia. A cross-sectional secondary data were obtained from the 2016 Ethiopian Demographic and Health Survey data (EDHS). A total of 1929 children were included. A Multilevel Proportional Odds Model was used to identify the individual and community-level factors associated with child vaccination. Among 1, 929 children, only 48.6% (95% CI: 46.3 to 50.8%) were fully vaccinated while 37.8% (95% CI: 35.7 to 40.1%) were partially vaccinated.. The multilevel ordinal logistic regression model reveled that housewife mother (AOR =1.522, 95%CI: 1.139, 2.034), institutional delivery (AOR =2.345, 95%CI: 1.766, 3.114),four or above antenatal care visits (AOR = 2.657; 95% CI: 1.906, 3.704), children of mothers with secondary or higher education (AOR = 2.008; 95% CI: 1.209, 3.334),Children whose fathers primary education (AOR = 1.596; 95% CI: 1.215, 2.096), from the rich households (AOR = 1.679; 95% CI: 1.233, 2.287) were significantly associated with childhood vaccination. Child vaccination coverage in Ethiopia remains low. Therefore, there is a need to increase child vaccination coverage by promoting institutional delivery and prenatal care visits, as well as maternal tetanus immunization. Besides, public initiatives needed to improve child vaccination coverage, women's and husband's education, poor women, and further advancement of health care services for poor women, housewife women, women living in remote areas should be made to maintain further improvements in child vaccination. Furthermore, policies and programs aimed at addressing cluster variations in child vaccination need to be formulated and their implementation must be strongly pursued.


JARID1B modulates lung cancer cell proliferation and invasion by regulating p53 expression.
Although three therapeutic modalities (surgical resection, chemotherapy, and radiotherapy) have been established, long-term survival for lung cancer patients is still generally poor. Until now, the mechanisms of lung cancer genesis remain elusive. The JARID1B is a histone demethylase that has been proposed as oncogene in several types of human cancer, but its clinical significance and functional role in human non-small cell lung cancer (NSCLC) remain unclear. In present study, we found that JARID1B was overexpressed in lung cancer cell lines and lung cancer tissues but not in normal lung tissues. The proliferation and invasive potential of lung cancer cells was significantly increased by ectopic expression of JARID1B. Contrarily, RNA interference targeting JARID1B in lung cancer cells significantly decreased the proliferation and invasive potential of cells. Moreover, we also found that the expression of p53 was modulated by JARID1B. Overexpressed JARID1B cell exhibited greatly decreased p53 expression, whereas silencing of JARID1B expression dramatically increased p53 expression at both the messenger RNA (mRNA) and protein levels. Inhibition of p53 by small interfering RNA (siRNA) reversed the shJARID1B-induced suppression of proliferation and invasion. Our results collectively suggested that JARID1B expressed in lung cancer played a role in lung cancer cells proliferation and invasion, which may be partly associated with the p53 expression.


Sequential EMT-MET induces neuronal conversion through Sox2.
Direct neuronal conversion can be achieved with combinations of small-molecule compounds and growth factors. Here, by studying the first or induction phase of the neuronal conversion induced by defined 5C medium, we show that the Sox2-mediated switch from early epithelial-mesenchymal transition (EMT) to late mesenchymal-epithelial transition (MET) within a high proliferation context is essential and sufficient for the conversion from mouse embryonic fibroblasts (MEFs) to TuJ+ cells. At the early stage, insulin and basic fibroblast growth factor (bFGF)-induced cell proliferation, early EMT, the up-regulation of Stat3 and Sox2, and the subsequent activation of neuron projection. Up-regulated Sox2 then induced MET and directed cells towards a neuronal fate at the late stage. Inhibiting either stage of this sequential EMT-MET impaired the conversion. In addition, Sox2 could replace sequential EMT-MET to induce a similar conversion within a high proliferation context, and its functions were confirmed with other neuronal conversion protocols and MEFs reprogramming. Therefore, the critical roles of the sequential EMT-MET were implicated in direct cell fate conversion in addition to reprogramming, embryonic development and cancer progression.


Inducible nitric oxide synthase and bcl-2 expression in nasopharyngeal cancer: correlation with outcome of patients after radiotherapy.
The expression of inducible nitric oxide synthase (iNOS) and bcl-2 proteins was evaluated and the prognostic significance determined in nasopharyngeal cancer (NPC) patients treated by radiotherapy. Tissue sections from 55 patients with NPC were assessed for iNOS and bcl-2 protein expression by immunohistochemistry, immunoelectron microscopy, and in situ hybridization before treatment. The markers were correlated with apoptosis (detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay) and clinicopathologic parameters. All NPC sections exhibited positive iNOS and bcl-2 immunoreactivity, with a mean percentage of 6.24% +/- 0.58% and 17.09% +/- 2.48%, respectively. A significant positive correlation was observed between iNOS expression and the apoptotic index (p < 0.0001, Pearson's r = 0.8518), and bcl-2 expression correlated inversely with apoptosis (p = 0.0001; Pearson's r = -0.6170). A significant inverse correlation was found between iNOS and bcl-2 immunoreactivity (p < 0.0001, Pearson's r = -0.7144). Bcl-2 but not iNOS expression was associated with the stage of the tumor according to the criteria of the American Joint Committee on Cancer (1997) (p < 0.0001). Patients who had recurrence of the tumor and metastasis after radiotherapy had a lower expression of iNOS (p = 0.014 and p = 0.035, respectively), although overall survival was not significantly different statistically. Higher bcl-2 expression was also associated with local tumor recurrence (p = 0.005) but not with metastasis or overall survival. It appears that iNOS and bcl-2 expression may be potentially useful biomarkers for predicting the outcome of radiotherapy in NPC patients.


Bronchodilating effects of natriuretic and vasorelaxant peptides compared to salbutamol in asthmatics.
In animal studies, the bronchial effects of urodilatin (URO, CDD/ANP-95-126, INN: ularitide) were superior to those of cardiodilatin/atrial natriuretic peptide (CDD, CDD/ANP-99-126). We compared the bronchodilating properties of intravenous URO and CDD in 36 clinically stable asthmatics showing a beta 2-agonist-induced increase of the FEV1 by > or = 15%. Any aerosol medication was discontinued for at least 8 h prior to the study. After baseline measurements of lung function parameters (FEV1, VC, PEF, MEF75, MEF50, MEF25) an intravenous infusion of 5.7, 11.4 or 17.1 pmol/kg/min URO or CDD was administered for 40 min in the morning. All measurements were repeated every 10 min during the infusion, for 30 min thereafter, and after the inhalation of 1.25 mg salbutamol (SALB). Both peptides had significant effects. While 11.4 pmol/kg/min URO dilated the central airways (FEV1, PEF, MEF75) slightly more potently than the peripheral bronchioles (MEF50, MEF25), 17.1 pmol/kg/min URO was as effective as SALB at all levels of the tracheobronchial tree. CDD reached only 50% of the SALB effect without a predominant localization of its action. The cardiovascular parameters revealed a significantly stronger vasorelaxant activity of CDD. In conclusion, the dose-dependent bronchodilating properties of intravenous URO were significantly superior to those of CDD.


Temperature exposure during pregnancy and birth outcomes: An updated systematic review of epidemiological evidence.
Seasonal patterns of birth outcomes have been observed worldwide, and there was increasing evidence that ambient temperature played as a trigger of adverse birth outcomes, such as preterm birth (PTB), low birth weight (LBW), and stillbirth. To systematically review updated epidemiological evidence about the relationship between temperature exposure during pregnancy and PTB, LBW, and stillbirth, we searched for related studies published in English from electronic databases and references of identified papers. We only included original articles that directly reported the effects of prenatal temperature exposure on birth outcomes. The characteristics and main findings of included studies were examined. A total of 36 epidemiological studies were finally included in this review. Most of these studies focused on PTB and LBW, while less attention has been paid to stillbirth that was relatively rare in the occurrence. Several designs including ecological (e.g., descriptive and time-series) and retrospective cohort studies (e.g., case-crossover and time-to-event) were applied to assess temperature effects on birth outcomes. Temperature metrics and exposure windows varied greatly in these investigations. Exposure to high temperature was generally found to be associated with PTB, LBW, and stillbirth, while several studies also reported the adverse impact of low temperature on birth outcomes of PTB and LBW. Despite no conclusive causality demonstrated, the current evidence for adverse effect on birth outcomes was stronger for heat than for cold. In summary, the evidence linking birth outcomes with ambient temperature was still very limited. Consequently, more related studies are needed worldwide and should be conducted in diversified climate zones, so as to further ascertain the association between temperature and birth outcomes. Future studies should focus on more sophisticated study designs, more accurate estimation of temperature exposure during pregnancy, and more efficient methods to find out the exposure windows, as well as cold-related effects on birth outcomes.


Mobility of schizophrenic patients, non-psychotic patients and the general population in a case register area.
This preliminary study attempts to evaluate the effect of a community psychiatric service on residential mobility by comparing a cohort of schizophrenic patients over a 5-year period with that of non-psychotic and general population control groups in South Verona, a case register area in Northern Italy. No significant differences were found between the groups for different types of movement (within South Verona/outside catchment area of South Verona/outside City of Verona), except for movements within South Verona between the schizophrenic group and neurotic group. The former tended to move less than the latter. The sociodemographic factors associated with movements of the general population in Verona were being male and single, whereas place of birth was less important. The results showed that in the two patient groups as well as in the general population in South Verona, the percentages of those who moved outside the case-register area ranged between 1.5%-4.4% over a one-year period, and between 7.3%-13.8% over a five-year period, indicating a relatively consistent denominator for the Case Register.


The helicase core accessory regions of the phage BFK20 DnaB-like helicase gp43 significantly affect its activity, oligomeric state and DNA binding properties.
The multifunctional phage replication protein gp43 is composed of an N-terminal prim-pol domain and a C-terminal domain similar to the SF4-type replicative helicases. We prepared four mutants all missing the prim-pol domain with the helicase core flanked by accessory N- and C-terminal regions truncated to varying extents. The shortest fragment still possessing strong ssDNA-dependent ATPase activity and helicase activity was gp43HEL519-983. The other proteins tested were gp43HEL557-983, gp43HEL519-855 and gp43HEL519-896. Removal of the 38 N-terminal residues in gp43HEL557-983, or the 128 and 87 C-terminal residues in gp43HEL519-855 and gp43HEL519-896, resulted in a significant decrease in the ATPase activities. The 38-amino acid N-terminal region has probably a function in modulating DNA binding and protein oligomerization. Deletion of the 87 C-terminal residues resulted in a twofold increase in the unwinding rate. This region is likely indispensable for binding to DNA substrates.


Single dietary amino acids control resting egg production and affect population growth of a key freshwater herbivore.
The enormous success of the genus Daphnia in freshwater ecosystems is at least partially due to their cyclical parthenogenetic life cycle, in which asexual and sexual reproduction alternate periodically. This temporal change between reproductive strategies allows for (1) rapid population growth via subitaneously developing eggs when environmental conditions are appropriate and (2) the maintenance of genetic diversity via sexual reproduction and the production of resting eggs when environmental conditions deteriorate. We show here that dietary amino acids are involved in triggering the switch between reproductive modes in Daphnia pulex. Supplementation experiments demonstrate that specific dietary amino acids, in particular arginine and histidine, avert crowding-induced resting egg production, enhance subitaneous reproduction by increasing algal food quality and, as a combined effect of both processes, increase population growth rates. These findings suggest that the availability of single dietary amino acids potentially affects the seasonal dynamics and long-term persistence of Daphnia populations in the field, which may have consequences for the efficiency of carbon transfer and thus the trophic structure of freshwater food webs.


Downregulation of microRNA-27b-3p via aberrant DNA methylation contributes to malignant behavior of gastric cancer cells by targeting GSPT1.
Epigenetics play a vital role in the initiation and development of cancers, including gastric cancer (GC). In the present study, we aimed to explore potential up- and downstream mechanisms of miR-27b-3p in GC. The expression level of miR-27b-3p in GC cells and tissues (n = 80) was measured by quantitative RT-PCR. The mimics, inhibitors, and negative controls of miR-27b-3p were transfected into cell lines to perform the gain and loss of function study. Cell proliferation, migration, and invasion assays were utilized to assess biological behaviors caused by miR-27b-3p in vitro. Common target genes were predicted using four biological software programs and used for gene functional enrichment analysis. GSPT1 was selected for target gene verification using dual luciferase assays and its expression level was detected by western blot. The MKN-45 cell line was treated with 5-aza-2'-deoxycytidine (5-Aza-dC) and the methylation level was measured by methylation-specific PCR (MSP). miR-27b-3p was significantly downregulated in the GC cell lines and tissues compared with the normal group. The expression of miR-27b-3p was determined to be negatively associated with TNM stage and tumor size using statistical analysis. Overexpression of miR-27b-3p inhibited MKN-45 and SGC-7901 cell proliferation, invasion, and migration. Gene functional enrichment analysis indicated that the target genes were involved in several signaling pathways. Dual luciferase assays showed that miR-27b-3p combined with the 3'-untranslated region of GSPT1 mRNA. MSP demonstrated that miR-27b-3p promoter CpG island was hyper-methylated and 5-Aza-dC was able to partially reverse the methylation. Our study data indicated that miR-27b-3p is downregulated by aberrant DNA methylation in GC. In addition, miR-27b-3p suppresses GC cell proliferation, invasion, and migration via negative expression regulation of GSPT1, which could be a potential therapeutic target.


Whole-bowel irrigation as a treatment for acute lithium overdose.
To determine if gastrointestinal decontamination using whole-bowel irrigation (WBI) was an effective treatment for acute ingestion of sustained-release lithium. In a two-phase, crossover protocol, ten normal volunteers ingested in each phase 0.80 mEq/kg sustained-release lithium carbonate. In the second phase, WBI was begun one hour after lithium ingestion, and 10 L of polyethylene glycol solution were administered over five hours. Serum samples were collected every half hour for six hours, every hour for an additional six hours, and then every 24 hours for as long as 72 hours after ingestion. These samples were analyzed for lithium concentration. The area under the lithium serum concentration-versus-time curve was calculated for each phase. The average area under the lithium serum concentration-versus-time curve in the WBI phase was 67% +/- 11% less than that in the control phase (P less than .0005 using a two-tailed Student's t test). The mean serum lithium concentration was significantly decreased (P = .03) within one hour of beginning WBI. WBI is an effective treatment for acute ingestion of sustained-release lithium. We recommend it as the decontaminant procedure of choice in this situation.


Fractional contribution of lung, nasal and gastrointestinal absorption to the systemic level following nose-only aerosol exposure in rats: a case study of 3.7- micro m fluorescein aerosols.
Because absorption takes place from multiple sites of aerosol deposition, it is generally difficult to interpret systemic levels following nose-only inhalation in laboratory rodents. Therefore, this study attempted to determine the fractional contribution of lung, nasal and gastrointestinal (GI) absorption to the observed systemic level following nose-only aerosol exposure in rats using fluorescein as a model powder solute. Rats were treated orally with vehicle or activated charcoal, the latter diminishing GI absorption of fluorescein, and were subsequently nose-only exposed to 3.7- micro m fluorescein aerosols at 25.2 micro g/l(air) for 10 min. While fluorescein similarly disappeared from the lung at a half-life of 0.23 hr, its plasma concentrations in the charcoal-treated group were significantly lower than those in the charcoal-untreated (vehicle) group. This suggests that significant portions of fluorescein were transported by nasopharyngeal and tracheobronchial mucociliary clearances following aerosol exposure and were absorbed from the GI tract. Despite the lack of GI absorption in the charcoal-treated animals, it was estimated that this nose-only exposure of fluorescein allowed 25.7 and 82.5 micro g/kg of simultaneous lung and nasal deposition, respectively, followed by their absorption composing the observed systemic level in this group (AUC(0- infinity ) 137.49 ng/ml h). Thus, assuming linear pharmacokinetics of fluorescein, the extent of absorption (AUC(0- infinity )) due to such nasal deposit (82.5 micro g/kg) was estimated to be 47.00 ng/ml h using the AUC(0- infinity ) obtained in an independent study of intranasal powder insufflation at 34.5 micro g/kg in the charcoal-treated rats (AUC(0- infinity ) 19.66 ng/ml h). As a result, the AUC(0- infinity ) due to 25.7 micro g/kg of the lung deposit was deconvoluted to be 90.49 ng/ml h and finally, the absolute bioavailability (F%) of the "lung-region-specific" deposition and absorption of fluorescein was estimated to be 55.0%. It is observed therefore, that lung, nasal and GI absorption accounted for 24.2, 12.5 and 63.3% of the total fluorescein absorption, respectively, following nose-only exposure of 3.7- micro m aerosols. This study addresses the common methodological insufficiency of nose-only inhalation studies in rodents, which have been neglected in most cases, and provides the appropriate kinetic interpretation for their observed systemic level.