The smoke-free university: anticipating health education needs of faculty and staff.
A survey instrument was developed to assess the anticipated impact of smoke-free university policy on faculty and staff and to recommend health education interventions based on stages of change theory and employee interest. Four months before the policy was implemented, the 25-item anonymous questionnaire was sent to a random sample of 1,000 faculty/staff. A majority of university workers (85.5%) are not smokers and approve of the no-smoking policy. Smokers (14.5%) are older than nonsmokers, are more likely to have a high school versus higher education, and are more likely to be maintenance or clerical workers. Most smokers smoke less than one pack per day and smoke outdoors during work hours. More smokers were in later stages of change (planning to stop soon) than expected. Interest in availability of smoking cessation activities differed by smoking status and change stage.


Wnt Signaling Pathways: A Role in Pain Processing.
The wingless-related integration site (Wnt) signaling pathway plays an essential role in embryonic development and nervous system regulation. It is critically involved in multiple types of neuropathic pain (NP), such as HIV-related NP, cancer pain, diabetic neuralgia, multiple sclerosis-related NP, endometriosis pain, and other painful diseases. Wnt signaling is also implicated in the pain induced by sciatic nerve compression injury and selective spinal nerve ligation. Thus, the Wnt signaling pathway may be a potential therapeutic target for NP.


Bioavailability of cadmium from shellfish and mixed diet in women.
Dietary intake and uptake of cadmium (Cd) were studied in nonsmoking women, 20-50 years of age, consuming a mixed diet low in shellfish (N = 34) or with shellfish once a week or more (N = 17). Duplicate diets were collected during 4 consecutive days for the determination of Cd content. The women kept detailed dietary records, and the intake of energy and various nutrients was calculated. The shellfish diets (median 22.3 micrograms Cd/day) contained twice as much Cd as the mixed diets (median 10.5 micrograms Cd/day; p < 0.0001). Cadmium in feces corresponded to 100 and 99% of that in duplicates of shellfish diets and mixed diets, respectively, indicating a low average absorption of the dietary Cd. In spite of the differences in the daily intake of Cd, there was no statistically significant difference in the concentrations of Cd in blood (B-Cd, shellfish group 0.25 micrograms/liter, mixed diet group 0.23 micrograms/liter) or urine (U-Cd, 0.10 micrograms Cd/liter in both groups). This indicates a lower absorption of Cd in the shellfish group than in the mixed diet group or a difference in the kinetics. A higher gastrointestinal absorption of Cd in the mixed diet group could partly be explained by lower body iron stores as measured by the concentrations of serum ferritin (S-fer, median 18 micrograms/liter, compared to 31 micrograms/liter in the shellfish group). In the mixed diet group, S-fer was negatively correlated with B-Cd and the main determining for B-Cd besides U-Cd in the multiple regression analysis, indicating an increased absorption of Cd at low body iron stores. When women with S-fer exceeding 20 micrograms/liter were compared, the higher dietary intake of Cd in the shellfish group compared to the mixed diet group (24 versus 10 micrograms/day) resulted in higher B-Cd (0.26 versus 0.16 micrograms/liter), although not in proportion to the difference in Cd intake. Thus, there seems to be differences in the bioavailability and/or kinetics of dietary Cd related to the type of diet. This is, to our knowledge, the first study where the influence of various types of diets and nutritional factors on the intake and uptake of cadmium in human subjects has been studied.


Photodynamic therapy inhibits the injury-induced fibrotic response of vascular smooth muscle cells.
excessive deposition of extracellular matrix (ECM) proteins plays a key role in the intervention-related vascular fibroproliferative response, resulting in intimal hyperplasia (IH). Cytokines, such as platelet-derived growth factor (PDGF), released after vascular injury and deposited in the ECM, are known to stimulate production of matrix proteins. Photodynamic therapy (PDT), the combination of light and a photosensitive dye to produce free radicals, is a novel approach to inhibit experimental IH by the local eradication of smooth-muscle cells (SMC) and alteration of ECM. This in vitro study examined whether PDT can inhibit the fibrotic response of vascular SMC. the effect of PDT on important pro-fibrotic factors was determined by performing PDT of isolated ECM, injured SMC and pure PDGF. SMC production of collagen was monitored by cellular [3H]-proline incorporation. untreated SMC seeded on ECM demonstrated an increase of 50% in collagen production ( p <0.0001) as compared to SMC on an empty plate. This increase was also seen when SMC was incubated with the conditioned media of mechanically injured SMC, or pure PDGF. However, after PDT of ECM, injured SMC or PDGF, there was an inhibition of 40% ( p <0.05) in SMC-collagen production. these findings indicate that PDT can interfere with factors that lead to the vascular fibrotic response. In this way, PDT, with its cytotoxic and extracellular effects, can promote healing of the vessel wall without the stimulus of fibrosis that can lead to restenosis.


Effect of in vivo fetal infusion of dexamethasone at 0.75 GA on fetal ovine resistance artery responses to ET-1.
At 110-111 days gestation, instrumented fetal sheep were administered saline or dexamethasone (2.2 microgram. kg(-1). h(-1) iv) for 48 h. Measurement of fetal blood pressure showed a greater increase in dexamethasone-treated (n = 6) compared with control (n = 5) fetuses (7.3 +/- 2.3 vs. 0.6 +/- 2.3 mmHg, P < 0.05). Fetuses were delivered by cesarean section, and the femoral muscle and brain were obtained under halothane anesthesia. Femoral and middle cerebral arteries (approximately 320-micrometer internal diameter) were evaluated using wire myography. Sensitivity to KCl (2.5-125 mM) and the magnitude of the maximal vasoconstriction to 125 mM K(+) were similar in femoral and middle cerebral arteries from dexamethasone-treated vs. control fetuses. Acetylcholine-induced vasorelaxation was similar in femoral arteries from control and dexamethasone-treated fetuses. Middle cerebral arteries did not relax to acetylcholine. Sensitivity to endothelin-1 (ET-1; 0.1 pM-0.1 microM) and magnitude of the ET-1-induced vasoconstriction were greater in femoral arteries from dexamethasone-treated vs. control fetuses (P < 0.05). Autoradiographical studies with receptor-specific ligands demonstrated increased ET(A)-receptor binding, the principal receptor subtype, in femoral muscle vessels (P < 0.001) but decreased ET(A)-receptor binding in middle cerebral arteries (P < 0.01) from dexamethasone-treated compared with control fetuses. Relatively little ET(B)-receptor binding was evident in all tissues examined. We conclude that hyperreactivity to ET-1, due to increased ET(A)-receptor binding, may be involved in the dexamethasone-induced increase in peripheral vascular resistance in fetal sheep in vivo.


Control of Upper Airway Motoneurons During REM Sleep.
The loss of tone in upper airway muscles contributes to disorders of breathing during sleep. In an animal model of rapid eye movement sleep atonia, decrements in the activity of upper airway motoneurons are caused by withdrawal of excitation mediated by serotonin and other transmitters, rather than by state-dependent inhibition.


Direct-imaging-based quantification of Bacillus cereus ATCC 14579 population heterogeneity at a low incubation temperature.
Bacillus cereus ATCC 14579 was cultured in microcolonies on Anopore strips near its minimum growth temperature to directly image and quantify its population heterogeneity at an abusive refrigeration temperature. Eleven percent of the microcolonies failed to grow during low-temperature incubation, and this cold-induced population heterogeneity could be partly attributed to the loss of membrane integrity of individual cells.


DNA binding by cut homeodomain proteins is down-modulated by protein kinase C.
The Drosophila and mammalian Cut homeodomain proteins contain, in addition to the homeodomain, three other DNA binding regions called Cut repeats. Cut-related proteins thus belong to a distinct class of homeodomain proteins with multiple DNA binding domains. Using nuclear extracts from mammalian cells, Cut-specific DNA binding was increased following phosphatase treatment, suggesting that endogenous Cut proteins are phosphorylated in vivo. Sequence analysis of Cut repeats revealed the presence of sequences that match the consensus phosphorylation site for protein kinase C (PKC). Therefore, we investigated whether PKC can modulate the activity of mammalian Cut proteins. In vitro, a purified preparation of PKC efficiently phosphorylated Cut repeats, which inhibited DNA binding. In vivo, a brief treatment of cells with calphostin C, a specific inhibitor of PKC, led to an increase in Cut-specific DNA binding, whereas phorbol 12-myristate 13-acetate, a specific activator of PKC, caused a decrease in DNA binding. The PKC phosphorylation sites within the murine Cut (mCut) protein were identified by in vitro mutagenesis as residues Thr415, Thr804, and Ser987 within Cut repeats 1-3, respectively. Cut homeodomain proteins were previously shown to function as transcriptional repressors. Activation of PKC by phorbol 12-myristate 13-acetate reduced transcriptional repression by mCut, whereas a mutant mCut protein containing alanine substitutions at these sites was not affected. Altogether, our results indicate that the transcriptional activity of Cut proteins is modulated by PKC.


Left ventricular function during graded exercise in patients with coronary artery disease and in control subjects.
The left ventricular function of 30 patients with coronary artery disease and 11 control subjects was studied by electrocardiography gated cardiac blood pool scintigraphy as the participants lay on their backs and either rested or exercised on a cycle ergometer at graded levels on intensity. The control subject showed a progressive increase in ejection fraction from rest (51% +/- 7%) to intermediate (56% +/- 10%, P less than 0.05) and maximum levels of exercise (64% +/- 10%, P less than 0.001). All the patients showed a decrease in ejection fraction from rest (42% +/- 16%) to their maximal level of exercise (36% +/- 11%, P less than 0.001). However, the response of some of the patients to intermediate exercise ranged from a decrease or no change to an increase in ejection fraction. Thus, exercise at maximal intensity is necessary to induce the left ventricular dysfunction that is diagnostic of coronary artery disease.


Mayer-Rokitansky-Küster-Hauser Syndrome: MR Manifestations Of Typical And Atypical Cases.
The Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome is a group of congenital malformations arising due to agenesis or arrest of the growth of paired Mullerian ducts leading to absence of uterus, fallopian tubes and upper part of vagina. MRI has an unmatched role in diagnosing MRKH syndrome. The aim of the present study is to elaborate MRI manifestations of typical and atypical MRKH syndrome and its common and rare associations. It was a cross sectional analysis of 19 cases in the Radiology Department of a public sector tertiary health care hospital of Rawalpindi over a period of one year. All patients clinically diagnosed as MRKH syndrome were included in the study. MRI pelvis of the study patients was performed and evaluated by two experienced radiologists. The mean age of study patients was 22.2 years. Out of 19 study patients, 16 (84%) had type I while 3 (16%) had type II MRKH syndrome. None of the patients had normally cited uterus of normal morphology. Bilateral ovaries with follicular activity were identified in all patients. A well-defined leiomyoma was identified arising from the left uterine bud in one of these patients. All three patients with MRKH II syndrome had ectopic pelvic kidneys. One of the patients showed a neoplastic mass lesion arising from ectopic left kidney. MRI is an excellent imaging modality for accurate diagnosis and evaluation of other system anomalies in MRKH syndrome. Rudimentary uterine buds are commonly seen in these patients and may have functioning endometrium. Ovaries are of normal appearance but frequently ectopic.


Postselection-Free Entanglement Dynamics via Spacetime Duality.
The dynamics of entanglement in "hybrid" nonunitary circuits (for example, involving both unitary gates and quantum measurements) has recently become an object of intense study. A major hurdle toward experimentally realizing this physics is the need to apply postselection on random measurement outcomes in order to repeatedly prepare a given output state, resulting in an exponential overhead. We propose a method to sidestep this issue in a wide class of nonunitary circuits by taking advantage of spacetime duality. This method maps the purification dynamics of a mixed state under nonunitary evolution onto a particular correlation function in an associated unitary circuit. This translates to an operational protocol which could be straightforwardly implemented on a digital quantum simulator. We discuss the signatures of different entanglement phases, and demonstrate examples via numerical simulations. With minor modifications, the proposed protocol allows measurement of the purity of arbitrary subsystems, which could shed light on the properties of the quantum error correcting code formed by the mixed phase in this class of hybrid dynamics.


Differences in workers' compensation claim rates for within-province and out-of-province workers in British Columbia, Canada, 2010-2017.
Little is known about the work-related injury and illness risk of out-of-province workers. This study examines whether there are differences in work-related injury and illness claim rates between within-province and out-of-province workers in British Columbia (BC), Canada. Workers' compensation claim data for injuries and illnesses in BC from 2010 to 2017 were linked with denominator data from Statistics Canada. Multivariable negative binomial regression estimated the claim rate ratio (RR) and 95% confidence intervals (CI) for out-of-province workers with all, health care-only (HCO), short-term disability, long-term disability, and fatality (SLF), and serious injury (SI) claims, compared to within-province workers. Compared to within-province workers, out-of-province workers had a lower total claim rate (RR: 0.54, 95% CI: 0.52-0.57), adjusting for sex, age, industry sector, and year. Differences in rates differed by claim type, with the largest differences for HCO claims (RR: 0.49, 95% CI: 0.47-0.52) and smallest differences for SI claims (RR: 0.85, 95% CI: 0.78-0.92). Sex-stratified models showed larger differences for males than females, with older female out-of-province workers having elevated SI claim rates. Industry-specific models showed that, even in sectors with high proportions of out-of-province workers' claims, these workers have lower claim rates than within-province workers. Out-of-province workers generally have lower claim rates than within-province workers. The overall duration of work exposure, and underreporting or underclaiming, are factors that may explain these lower claim rates. Understanding the determinants and differences of these claim rates may improve the administration and adjudication of claims while also identifying where further prevention measures may be merited.


CHH DNA methylation increases at 24-PHAS loci depend on 24-nt phased small interfering RNAs in maize meiotic anthers.
Plant phased small interfering RNAs (phasiRNAs) contribute to robust male fertility; however, specific functions remain undefined. In maize (Zea mays), male sterile23 (ms23), necessary for both 24-nt phasiRNA precursor (24-PHAS) loci and Dicer-like5 (Dcl5) expression, and dcl5-1 mutants unable to slice PHAS transcripts lack nearly all 24-nt phasiRNAs. Based on sequence capture bisulfite-sequencing, we find that CHH DNA methylation of most 24-PHAS loci is increased in meiotic anthers of control plants but not in the ms23 and dcl5 mutants. Because dcl5-1 anthers express PHAS precursors, we conclude that the 24-nt phasiRNAs, rather than just activation of PHAS transcription, are required for targeting increased CHH methylation at these loci. Although PHAS precursors are processed into multiple 24-nt phasiRNA products, there is substantial differential product accumulation. Abundant 24-nt phasiRNA positions corresponded to high CHH methylation within individual loci, reinforcing the conclusion that 24-nt phasiRNAs contribute to increased CHH methylation in cis.


Microbial signalling in colonic motility.
Sensory nerve endings within the wall of the gastrointestinal (GI) tract may respond to bacterial signalling, providing the basis for key biological processes that underlie intestinal motility and microbial homeostasis. Enteric neurons and smooth muscle cells are well known to express an array of receptors, including G-protein coupled receptors and ligand-gated ion channels, that can sense chemical ligands and other bacterially-derived substances. These include short chain fatty acids, secondary bile acids and lipopolysaccharide. For neural detection of microbial activators to occur, luminal substances must first interact with enterocytes for direct signalling or cross paracellularly. Recent studies indicate that bacterial-derived microvesicles can cross the gut epithelial barrier and affect motility. This suggests a possible intercellular communication pathway between the GI tract and the ENS. We explore the idea that bacterial microvesicles can behave as a delivery package for communication between microbe and host.


A Proposed Methodology to Assess the Accuracy of 3D Scanners and Casts and Monitor Tooth Wear Progression in Patients.
The aim of this study was to detail and assess the capability of a novel methodology to 3D-quantify tooth wear progression in a patient over a period of 12 months. A calibrated stainless steel model was used to identify the accuracy of the scanning system by assessing the accuracy and precision of the contact scanner and the dimensional accuracy and stability of casts fabricated from three different types of impression materials. Thereafter, the overall accuracy of the 3D scanning system (scanner and casts) was ascertained. Clinically, polyether impressions were made of the patient's dentition at the initial examination and at the 12-month review, then poured in type IV dental stone to assess the tooth wear. The anterior teeth on the resultant casts were scanned, and images were analyzed using 3D matching software to detect dimensional variations between the patient's impressions. The accuracy of the 3D scanning system was established to be 33 μm. 3D clinical analysis demonstrated localized wear on the incisal and palatal surfaces of the patient's maxillary central incisors. The identified wear extended to a depth of 500 μm with a distribution of 4% to 7% of affected tooth surfaces. The newly developed 3D scanning methodology was found to be capable of assessing and accounting for the various factors affecting tooth wear scanning. Initial clinical evaluation of the methodology demonstrates successful monitoring of tooth wear progression. However, further clinical assessment is needed.


Transradial access for arteriovenous fistuloplasty in Singapore.
Endovascular balloon angioplasty is standard therapy for dysfunctional arteriovenous fistula in end-stage renal failure patients on hemodialysis. Venous antegrade or retrograde puncture of the fistula is typically performed to gain access for fistuloplasty. Transradial approach for brachiocephalic or brachiobasilic arteriovenous fistulas offers an alternative method of access with the advantage of addressing multi-focal juxta-anastomotic and venous stenosis from the same approach. We aim to review the efficacy, outcomes and complication rates of transradial access for arteriovenous fistuloplasty among patients in Singapore. A retrospective review of 195 endovascular fistuloplasties from September 2017 to August 2019, at a tertiary university hospital Vascular Surgery unit. Of 195 fistuloplasties, 43 (22%) were transradial approach (23 brachiocephalic arteriovenous fistulas, 20 brachiobasilic arteriovenous fistulas) in 33 patients (67% male and mean age = 65 years). Of these 43 procedures, 11 (26%) were performed as balloon-assisted maturation fistuloplasties while 32 (74%) were performed for mature arteriovenous fistulas with multi-focal juxta-anastomosis and venous stenosis. Technical success rate was 95% with mean procedure duration at 43.5 ± 14.6 min. Mean pre- and post-fistuloplasty dialysis access flow rates increased from 502 to 952 ml/min (p < 0.001). Post-intervention primary patency was 100%, 66% and 20% at 1, 6 and 12 months, respectively. There were four patients with non-limb-threatening radial artery thrombosis (9.3%) while there was no radial artery pseudoaneurysm or post-procedural bleeding. Transradial approach for arteriovenous fistuloplasty is a safe and feasible option in patients requiring balloon-assisted maturation or with multi-focal juxta-anastomotic and venous stenosis.


[Kampo Medicine for Intractable Brain and Neurological Diseases].
In Japan, 147 types of oral Kampo extract products are covered by the public insurance system; however, to the best of our knowledge, there are no large-scale randomized clinical trials. Therefore, the level of evidence or these products is low compared with that for medicines that have been introduced as insurance reimbursement drugs through randomized clinical trials. However, when conventional western medical treatments have not been effective or when patients desire a more effective therapy, Kampo extract products are one of the treatment options. Kampo extract products involve the adding of crude drugs and comprise a multi-component system. Therefore, it may be effective for various symptoms. I recommend that specialists use these products as complementary alternative medicines. I also introduce the "Huaier extract granule," which is an orally bioavailable traditional Chinese medicine composed of granules containing an aqueous extract of Trametes robiniophila Murr (Huaier), a mushroom found on hardwood tree trunks, with potential immune-regulating activity. Huaier has immunity enhancement activity, such as antineoplastic property, as well as immunity regulating activity against diseases, such as asthma, psoriasis, and immunoglobulin A nephritis. Therefore, for some intractable diseases, especially when the disease is thought to involve an immune disorder, Huaier can be an additional treatment option.


Structure of Plant Cell Walls : XXVI. The Walls of Suspension-Cultured Sycamore Cells Contain a Family of Rhamnogalacturonan-I-Like Pectic Polysaccharides.
Considerable information has been obtained about the primary structures of suspension-cultured sycamore (Acer pseudoplatanus) cell-wall pectic polysaccharides, i.e. rhamnogalacturonan I, rhamnogalacturonan II, and homogalacturonan. However, these polysaccharides, which are solubilized from the walls by endo-alpha-1,4-polygalacturonase, account for only about half of the pectic polysaccharides known to be present in sycamore cell walls. We now report that, after exhaustive treatment with endo-alpha-1,4-polygalacturonase, additional pectic polysaccharides were extracted from sycamore cell walls by treatment with Na(2)CO(3) at 1 and 22 degrees C. These previously uncharacterized polysaccharides accounted for approximately 4% of the cell wall. Based on the glycosyl and glycosyl-linkage compositions and the nature of the products obtained by treating the quantitatively predominant NaCO(3)-extracted polysaccharides with lithium metal dissolved in ethylenediamine, the polysaccharides were found to strongly resemble rhamnogalacturonan I. However, unlike rhamnogalacturonan I that characteristically had equal amounts of 2- and 2,4-linked rhamnosyl residues in its backbone, the polysaccharides extracted in Na(2)CO(3) at 1 degrees C had markedly disparate ratios of 2- to 2,4-linked rhamnosyl residues. We concluded that polysaccharides similar to rhamnogalacturonan I but with different degrees of branching are present in the walls of suspension-cultured sycamore cells.


Tocotrienol levels in various tissues of Sprague-Dawley rats after intragastric administration of tocotrienols.
A tocotrienol (T3) mixture was intragastricaly administered to Sprague-Dawley rats, and the T3 levels in various tissues were measured 0, 4, 8 and 24 hr after the administration. In blood clots, brain, thymus, testes, vice-testes and muscles, T3 homologues were not detected at all. In epididymal adipose, renal adipose, subcutaneous adipose and brown adipose tissues and in the heart, the T3 levels were maintained or increased for 24 hr after the administration. In the serum, liver, mesenteric lymph node, spleen and lungs, the T3 levels were highest 8 hr after the T3 administration. These results suggest that the distribution and metabolism of T3 in the rat vary considerably among different tissues.


Species Comparison of the Role of p38 MAP Kinase in the Female Reproductive System.
The p38 mitogen-activated protein kinases (MAPKs) are members of discrete signal transduction pathways that have significant regulatory roles in a variety of biological processes, depending on the cell, tissue and organ type. p38 MAPKs are involved in inflammation, cell growth and differentiation and cell cycle. In the female reproductive system, p38 MAPKs are known to regulate various aspects of the reproductive process such as mammalian estrous and menstrual cycles as well as early pregnancy and parturition. p38 MAPKs have also been implicated in alterations and pathologies observed in the female reproductive system. Therefore, pharmacologic modulation of p38 MAPKs, and inter-connected signaling pathways (e.g., estrogen receptor signaling, c-fos, c-jun), may influence reproductive physiology and function. This article provides a critical, comparative review of available data on the roles of p38 MAPKs in the mammalian female reproductive system and in reproductive pathophysiology in humans and preclinical species. We first introduce fundamental differences and similarities of the mammalian female reproductive system that should be considered by toxicologists and toxicologic pathologists when assessing the effects of new pharmacologic agents on the female reproductive system. We then explore in detail the known roles for p38 MAPKs and related molecules in female reproduction. This foundation is then extended to pathological conditions in which p38 MAPKs are thought to play an integral role.


Uptake of exogenous serine is important to maintain sphingolipid homeostasis in Saccharomyces cerevisiae.
Sphingolipids are abundant and essential molecules in eukaryotes that have crucial functions as signaling molecules and as membrane components. Sphingolipid biosynthesis starts in the endoplasmic reticulum with the condensation of serine and palmitoyl-CoA. Sphingolipid biosynthesis is highly regulated to maintain sphingolipid homeostasis. Even though, serine is an essential component of the sphingolipid biosynthesis pathway, its role in maintaining sphingolipid homeostasis has not been precisely studied. Here we show that serine uptake is an important factor for the regulation of sphingolipid biosynthesis in Saccharomyces cerevisiae. Using genetic experiments, we find the broad-specificity amino acid permease Gnp1 to be important for serine uptake. We confirm these results with serine uptake assays in gnp1Δ cells. We further show that uptake of exogenous serine by Gnp1 is important to maintain cellular serine levels and observe a specific connection between serine uptake and the first step of sphingolipid biosynthesis. Using mass spectrometry-based flux analysis, we further observed imported serine as the main source for de novo sphingolipid biosynthesis. Our results demonstrate that yeast cells preferentially use the uptake of exogenous serine to regulate sphingolipid biosynthesis. Our study can also be a starting point to analyze the role of serine uptake in mammalian sphingolipid metabolism.


Gene expression analysis of peripheral blood cells reveals Toll-like receptor pathway deregulation in colorectal cancer.
Colorectal cancer is the leading cause of cancer-related deaths worldwide. The disease is curable when detected at an early stage. However, the compliance rate with current screening recommendations remains poor. An accurate, minimally invasive blood test that has the potential for greater patient compliance would be a welcome addition to the current methods. Recent data have shown that gene expression profile of peripheral blood cells can reflect disease states and thus have diagnostic value. In this study, genome-wide gene expression profiling of peripheral blood cells from 20 healthy controls and 20 colorectal cancer patients were performed using PAXgene™ technology and Affymetrix GeneChip® microarrays. We identified a list of 1,469 genes that were differentially expressed between the healthy controls and cancer patients. Gene annotation and functional enrichment analysis revealed that those genes are mainly related to immune functions. Particularly, a set of genes belonging to the Toll-Like Receptor pathways were up-regulated in the colorectal cancer patients. These findings provide a new understanding of blood gene expression profile in colorectal cancer. Our result may serve as the basis for further development of blood biomarkers for the diagnosis and treatment of colorectal cancer.


Barriers to Alzheimer disease drug discovery and development in the biotechnology industry.
The major barrier to Alzheimer disease (AD) drug discovery and development in the biotechnology industry is scale. Most biotechnology companies do not have the personnel or expertise to carry a drug from the bench to the market. Much effort in the industry has been directed toward the elucidation of molecular mechanisms of AD and the identification of new targets. Advances in biotechnology have generated new insights into disease mechanisms, increased the number of lead compounds, and accelerated biologic screening. The majority of costs associated with drug development are in clinical testing and development activities, many of which are driven by regulatory issues. For most biotechnology companies, the costs of such trials and the infrastructure necessary to support them are prohibitive. Another significant barrier is the definition of therapeutic benefit for AD drugs; Food and Drug Administration (FDA) precedent has established that a drug must show superiority to placebo on a performance-based test of cognition and a measure of global clinical function. This restrictive definition is biased toward drugs that enhance performance on memory-based tests. Newer AD drugs are targeted toward slowing disease progression; however, there is currently no accepted definition of what constitutes efficacy in disease progression. Despite these obstacles, the biotechnology industry has much to offer AD drug discovery and development. Biotechnology firms have already developed essential technology for AD drug development and will continue to do so. Biotechnology companies can move more quickly; of course, the trick is to move quickly in the right direction. Speed may offset some of the problems associated with lack of scale. Additionally, biotechnology companies can afford to address markets that may be too restricted for larger pharmaceutical companies. This advantage will have increasing importance, as therapies are developed to address subtypes of AD.


Transplantation of allogeneic CD34+ blood cells.
Pluripotent stem cells of hematopoiesis and lymphopoiesis are among the CD34+ cells in blood or bone marrow. After granulocyte-colony stimulating factor (G-CSF) treatment, 1% to 2% of the mononuclear cells in blood are CD34+ cells, which can be procured by leukapheresis. We investigated the potential of CD34+ blood cells for reconstituting hematopoiesis and lymphopoiesis after allogeneic transplantation. HLA-identical sibling donors of 10 patients with hematologic malignancies were treated with G-CSF (filgrastim), 5 microgram/kg subcutaneously twice daily for 5 to 7 days. CD34+ cells were selected from the apheresis concentrates by immunoadsorption, concomitantly the number of T cells was reduced 100- to 1,000-fold. After transplantation, five patients received cyclosporine A for graft-versus-host disease (GvHD) prophylaxis (group I); five patients additionally received methotrexate (group II). G-CSF and erythropoietin were given to all patients. Mean numbers of 7.45 x 10(6) CD34+ and 1.2 x 10(6) CD3+ cells per kilogram were transplanted. In group I, the median times of neutrophil recovery to 100, 500, and 1,000 per mm3 were 10, 10, and 11 days, respectively. Group II patients reached these neutrophil levels after 10, 14, and 15 days, respectively. Platelet transfusions were administered for a median of 18 days in group I and 30 days in group II, and red blood cells for 9 and 12 days, respectively. Between day 30 and 60, lymphocytes reached levels of 353 +/- 269 cells per mm3. The median grades of acute GvHD were III in group I and I in group II. Two patients in group I died from acute GvHD. Two leukemic relapses occurred in group II. Complete and stable donor hematopoiesis was shown in all patients with a median follow up of 370 (45 to 481) days. Allogeneic blood CD34+ cells can successfully reconstitute hematopoiesis and lymphopoiesis. Reduction of T cells by CD34+ blood cell enrichment and cyclosporine A alone might not be sufficient for prophylaxis of severe acute GvHD.


Endoscopic Ultrasound Guided Rendezvous Drainage of Biliary Obstruction Using a New Flexible 19-Gauge Fine Needle Aspiration Needle.
Background and Aim. A successful endoscopic ultrasound guided rendezvous (EUS-RV) biliary drainage is dependent on accurate puncture of the bile duct and precise guide wire manipulation across the ampulla of Vater. We aim to study the feasibility of using a flexible 19-gauge fine aspiration needle in the performance of EUS-RV biliary drainage. Method. This is a retrospective case series of EUS-RV biliary drainage procedures at a single center. Patients who failed ERCP during the same session for benign or malignant biliary obstruction underwent EUS-RV using a flexible, nitinol covered, 19-gauge needle for biliary access and guide wire manipulation. Result. 24 patients underwent EUS-RV biliary drainage via extrahepatic access while 1 attempt was via intrahepatic access. The technical success rate was 80%, including 83.3% of cases via extrahepatic access. There was no significant difference in success rate of inpatient and outpatient procedures, benign or malignant indications, or type of guide wire used. Adverse events included mild pancreatitis (3 patients) and cholangitis (1 patient). Conclusion. A flexible 19-gauge needle for biliary access can be safe and effective when used to perform EUS-RV biliary drainage. Direct comparison between the nitinol needle and conventional metal needles in the performance of EUS guided biliary drainage is needed.


Acrylonitrile induces autolysis Bacillus subtilis.
Acrylonitrile (AN) is an industrial chemical used in the manufacture of plastics and other polymers. AN has been reported to be an acute toxin and is a known carcinogen in rodents. When AN was mixed with suspensions of Bacillus subtilis, the bacteria began autolysis. It was determined that AN is partially converted to cyanide, a strong protonophore in B. subtilis. Autolytic enzymes in B. subtilis become active when the protonmotive force is dissipated. The amount of cyanide produced from AN, however, was not enough to promote autolysis in exponential B. subtilis. This is the first report showing that AN may induce autolytic reactions in bacteria. It is suggested the autolysis of B. subtilis may be useful in the environmental monitoring of AN. In addition, the metabolism of AN by bacilli may be useful in bioremediation.


Regulation of m6A Methylation as a New Therapeutic Option against COVID-19.
The rapid spread of SARS-CoV-2 and the resulting pandemic has led to a spasmodic search for approaches able to limit the diffusion of the disease. The epigenetic machinery has aroused considerable interest in the last decades, and much evidence has demonstrated that this type of modification could regulate the early stages of viral infection. Recently it was reported that N6-methyladenosine (m6A) influences SARS-CoV-2 replication, although its role remains to be further investigated. The knockdown of enzymes involved in the m6A pathway could represent an optimal strategy to deepen the epigenetic mechanism. In the present study, we blocked the catalytic activity of the fat mass and obesity-associated protein (FTO) by using the selective inhibitor rhein. We observed a strong broad-spectrum reduction of infectivity caused by various coronaviruses, including SARS-CoV-2. This effect could be due to the modulation of m6A levels and could allow identification of this modification as a new therapeutic target to treat SARS-CoV-2 infection.


Influence of pneumoperitoneum on tumor growth and pattern of intra-abdominal tumor spreading: in vivo study of a murine model.
The employment of laparoscopy in the treatment of locally advanced colorectal cancer is still questioned by many surgeons, mainly due to the fear of spreading tumors by the laparoscopic procedure. However, it is still unknown whether laparoscopy is actually associated with increased tumor dissemination, especially for those with tumor invasion through the serosa. The main purpose of this study is to investigate the effect of pneumoperitoneum on the intra-abdominal tumor growth and spreading of colon cancer cells by an animal study of murine model. Under anesthesia, 10(6) tumor cells (CT26) were inoculated into the lower abdominal cavity of BALB/c mice by a mini-laparotomy. The mice were randomized to undergo pneumoperitoneum with CO2 (n=10), helium (n=10), or to act as controls (n=10). Pneumoperitoneum was established over 20 min at a pressure of 10cm H2O. The distribution pattern and the weight of peritoneal tumor growth of each mouse were recorded and analyzed at 15 days after surgery. The mean ratios of the tumor mass over the total body weight of the mice were 0.77+/-1.13% (control group), 4.30+/-0.86% (CO2 pneumoperitoneum), and 2.17+/-0.88% (helium pneumoperi-toneum). The mean tumor weight ratio (3.23+/-1.38%) of the pneumoperitoneal group was 4 times larger than that of the control group (p<0.001). Regarding the use of different insufflation gases over tumor growth, CO2 accelerates tumor growth more significantly than helium (p<0.001). Intraabdominal distribution (p=0.047) and diaphragm spreading (p<0.001) were significantly greater in the pneumoperitoneum group than the control group. The results of this animal study imply that presence of a pneumoperitoneum enhances the implantation and growth of free intraperitoneal malignant colon cancer cells in this in vivo mouse model. The results of the current study suggest that insufflation during pneumoperitoneum may play an important role in the development of peritoneal dissemination when there are free tumor cells in the intra-abdominal cavity, and the effect of using CO2 might be greater than that of using helium.


Leprosy: clinical and epidemiological study in patients above 60 years in Espírito Santo State - Brazil.
Leprosy is a chronic infectious disease that is endemic in Brazil and little studied in patients over 60 years old. The aim of this study was the epidemiological and clinical description of cases of leprosy in individuals older than 60 years, notified in the State of Espírito Santo - Brazil. It was an observational, descriptive and retrospective study of leprosy patients notified between 2001 and 2011. Out of 16,025 notifications, 2,510 (15.6%) were of patients over 60 years of age; the distribution among the period was egalitarian except in the last 2 years, when there was a mild reduction of cases; the average was 70 years of age (±7 years); 46% were women, being 62% of those paucibacillary cases; 1,145 (50.5%) were dark-skined people in particular among paucibacillary cases; 1,638 (72.9%) were illiterates or poorly instructed n particular among multibacillary cases (P=0.022); 59.9% patients were clinically multibacillary cases and 37.4% had positive bacilloscopy; 37.9% patients had clinical alterations on peripheral nerves, 36.7% of all the multibacillary cases were classified as grade I and 15.3% as grade in assessment of disability. This was a retrospective study, which used secondary data generated by physicians and notified by other professionals, whar could have enabled possible errors on original data. Leprosy in this age group suggests a long incubation period with reactivation of latent focus or late infection. Men were more affected, as well as the afro descendent race was statistically significant in the paucibacillary cases (P=0.000) and illiterate/poor education in multibacillary cases (P=0.022). Nearly 40% of patients had positive bacilloscopy and grade I/II disability, demonstrating a late diagnosis.


Some aspects of DNA polymerase functioning.
The group of DNA polymerases was studied using some new nucleoside 5'-triphosphate analogs with termination substrate properties. Among DNA polymerases tested the least specific appeared reverse transcriptases of retroviruses and the most specific were DNA polymerases alpha type from high eucaryotes including mammalians.


Short communication: Influence of shortening the dry period of Swedish dairy cows on plasmin activity in milk.
The aim of this study was to evaluate the influence of shortening the dry period of Swedish dairy cows on plasmin activity and casein composition in milk. Swedish Holstein and Swedish Red cows, 45 in total, were assigned to a dry period of either 4 or 8wk. Milk samples were taken 10 and 5wk prepartum, and 6 and 12wk postpartum. Plasmin activity and plasminogen activity were measured with a spectrophotometric assay. Casein composition was measured by capillary zone electrophoresis. Prepartum plasminogen activity increased by 22% between 10 and 5wk prepartum, whereas no change in plasmin activity was observed during the same period. Cows with a 4-wk dry period had 61% higher plasmin activity in postpartum milk than cows with an 8-wk dry period. Cows of third or greater parity tended to have a stronger increase in plasmin activity as a result of applying a short dry period than cows of second parity. Although the αS1- and β-casein fractions declined with increasing plasmin activity, no dry period effects were found. Based on postpartum differences in plasmin activity, it was concluded that particularly multiparous cows require more than 4wk between lactations for recovery of the mammary epithelium. Changes in casein composition as an effect of plasmin activity are not expected to have a great effect on processing quality of milk, although future work is needed to verify this.


Mechanistic crystallography. Mechanism of inactivation of gamma-aminobutyric acid aminotransferase by (1R,3S,4S)-3-amino-4-fluorocyclopentane-1-carboxylic acid as elucidated by crystallography.
(1R,3S,4S)-3-Amino-4-fluorocyclopentane-1-carboxylic acid (7) was previously shown to be a mechanism-based inactivator of gamma-aminobutyric acid aminotransferase (GABA-AT) [Qiu, J. and Silverman, R. B. (2000) J. Med. Chem. 43, 706-720]. Two mechanisms were considered as reasonable possibilities, a Michael addition mechanism and an enamine mechanism. On the basis of a variety of chemical studies, including tedious radiolabeling experiments, it was concluded that inactivation by 7 proceeds by a Michael addition mechanism. Here, a crystal structure of 7 bound to pig liver GABA-AT is reported, which clearly demonstrates that the adduct formed is derived from an enamine mechanism. This represents another example of how crystallography is an important tool for elucidation of inactivation mechanisms.


Fiber Composition of the Grasscutter (Thryonomys swinderianus, Temminck 1827) Thigh Muscle: An Enzyme-histochemical Study.
The aim of this study was to describe de fiber composition in the thigh muscles of grass cutter (Thryonomys swinderianus, Temminck 1827). Ten 4 to 6-month-old (3 to 4 kg) male grasscutter were used in this study. Eleven skeletal muscles of the thigh [M. biceps femoris (BF), M. rectus femoris (RF), M. vastus lateralis (VL), M. vastus medialis (VM), M. tensor fasciae latae (TFL), M. semitendinosus (ST), M. semimembranosus (SM), M. semimembranosus accessorius (SMA), M. Sartorius (SRT), M. pectineus (PCT), M. adductor magnus (AM)] were collected after animals euthanasia and examined by light microscopy. Three muscle fiber types (I, IIB and IIA) were found in these muscles using enzyme histochemical techniques [myosine adenosine triphosphatase (ATPase) and nicotinamide adenine dinucleotide tetrazolium reductase (NADH-TR)]. Ten of these eleven muscles are composed by 89% to 100% of fast contracting fibers (types IIA and IIB), while the SMA was almost exclusively formed by slow contracting fibers.


Cost-effectiveness of cholesterol lowering. Results from the Scandinavian Simvastatin Survival Study (4S)
An analysis of the cost-effectiveness of simvastatin was conducted, based on the Scandinavian Simvastatin Survival Study (4S). The total cost of hospitalization in the placebo group was 52.8 million Swedish kronor (SEK) (5.15 million pounds), compared with SEK 36.0 million (3.51 million pounds) in the simvastatin group. This amounts to a 32% reduction, or a saving of SEK 16.8 million (1.6 million pounds) or SEK 7560 (738 pounds) per patient. The net cost per patient for the duration of the study (5.4 years) was SEK 13,540 (1324 pounds). Simvastatin treatment saved an estimated 0.377 undiscounted life years (0.240 life years discounted at 5% per annum). The cost of simvastatin therapy per discounted life-year saved was therefore SEK 56,400 (5502 pounds). Sensitivity analysis, examining the effect of different life expectancies, costs of initiation and monitoring of simvastatin therapy, and discount rates, showed the results to be stable. Conclusion. The cost per life-year saved of simvastatin in the treatment of post-myocardial infarction and angina patients, as determined from 4S data, is well within the range normally considered cost-effective.


Pyrroloquinoline quinone prevents knee osteoarthritis by inhibiting oxidative stress and chondrocyte senescence.
Accumulating evidence suggests that oxidative stress plays an important role in the progression of osteoarthritis (OA), and pyrroloquinoline quinone (PQQ) is considered a strong antioxidant. However, it is unclear whether PQQ can prevent the progression of OA by inhibiting oxidative stress. In this study, anterior cruciate ligament transection (ACLT)-induced OA mice received a diet supplemented with/without PQQ, and were compared with each other and with sham-operated mice. Our results showed that in PQQ-untreated OA mice, articular surfaces collapsed, while the thickness of articular cartilage and the abundance of cartilage matrix protein decreased significantly, whereas PQQ supplementation largely prevented these alterations. We also found that oxidative stress, DNA damage, cellular senescence and the secretion of senescence-associated inflammatory cytokines were increased in PQQ-untreated OA mice compared with sham-operated mice. However, these parameters were obviously rescued in PQQ-treated OA mice. This study demonstrated that PQQ supplementation can prevent ACLT-induced OA by inhibiting oxidative stress, DNA damage, cell senescence and the development of the senescence-associated secretory phenotype.


Primary tonsillar tuberculosis: a case report.
A 67-year-old man with an enlarged left tonsil underwent a tonsillectomy for the suspicion of malignancy. He had had a dental extraction six months earlier. Clinical and histopathological investigations established the diagnosis of primary tonsillar tuberculosis.


Process Fingerprint in Micro-EDM Drilling.
The micro electrical discharge machining (micro-EDM) process is extensively used in aerospace, automotive, and biomedical industries for drilling small holes in difficult-to-machine materials. However, due to the complexity of the electrical discharge phenomena, optimization of the processing parameters and quality control are time-consuming operations. In order to shorten these operations, this study investigates the applicability of a process fingerprint approach in micro-EDM drilling. This approach is based on the monitoring of a few selected physical quantities, which can be controlled in-line to maximize the drilling speed and meet the manufacturing tolerance. A Design of Experiments (DoE) is used to investigate the sensitivity of four selected physical quantities to variations in the processing parameters. Pearson's correlation is used to evaluate the correlation of these quantities to some main performance and hole quality characteristics. Based on the experimental results, the potential of the process fingerprint approach in micro-EDM drilling is discussed. The results of this research provide a foundation for future in-line process optimization and quality control techniques based on machine learning.


Effects of birth order and maternal age on infant and child mortality in rural Nepal.
"The present study examines the effects of woman's age at childbirth (maternal age) and birth order on infant and child mortality in rural Nepal." Data are from the Nepal Fertility Survey of 1976, carried out as part of the World Fertility Survey. The results show that the risk of infant death is higher for extreme birth orders. "The higher risk of infant death to first order births is mainly due to the fact that such births occur predominantly to young women. The importance of the synergistic effect of maternal age and birth order is also demonstrated: for example, fifth order births to women aged 20-24 and third order births to women under 20 have a risk of about one infant death in every four live births."


Notch signaling specifies prosensory domains via lateral induction in the developing mammalian inner ear.
During inner ear morphogenesis, the process of prosensory specification defines the specific regions of the otic epithelium that will give rise to the six separate inner ear organs essential for hearing and balance. The mechanism of prosensory specification is not fully understood, but there is evidence that the Notch intercellular signaling pathway plays a critical role. The Notch ligand Jagged1 (Jag1) is expressed in the prosensory domains, and mutation of Jag1 impairs sensory formation. Furthermore, pharmacological inhibition of Notch in vitro during prosensory specification disrupts the prosensory process. Additionally, activation of Notch by cDNA electroporation in chick otocysts results in formation of ectopic sensory patches. Here we test whether Notch activity is sufficient for prosensory specification in the mouse, using a Cre-/loxP approach to conditionally activate the Notch pathway in nonsensory regions of the inner ear epithelia during different stages of otic vesicle morphogenesis. We find that broad ectopic activation of Notch at very early developmental stages causes induction of prosensory markers throughout the entire otic epithelium. At later stages of development, activation of Notch in nonsensory regions leads to induction of sensory patches that later differentiate to form complete ectopic sensory structures. Activation of Notch in isolated nonsensory cells results in lateral induction of Jag1 expression in neighboring cells and spreading of prosensory specification to the adjacent cells through an intercellular mechanism. These results support a model where activation of Notch and propagation through lateral induction promote prosensory character in specific regions of the developing otocyst.


Prediction of Rhizoma Drynariae Targets in the Treatment of Osteoarthritis Based on Network Pharmacology and Experimental Verification.
Rhizoma Drynariae has been widely used for the treatment of osteoarthritis (OA), but its potential targets and molecular mechanisms remain to be further explored. Targets of Rhizoma Drynariae and OA were predicted by relevant databases, and a protein-protein interaction (PPI) network was constructed to identify key targets. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed to obtain related pathways and then select significant pathways associated with OA. The OA chondrocyte model was established by inflammatory factor-induced SW1353 chondrocytes, and molecular docking was conducted to verify the above theoretical prediction. The results showed that a total of 86 Rhizoma Drynariae-OA interaction targets were identified, among which IL-6 and AKT1 were the key targets in the PPI network. Luteolin was the most critical component of Rhizoma Drynariae. KEGG results indicated that the effects of Rhizoma Drynariae on OA are associated with the PI3K/AKT, TNF, IL-17, apoptosis, and HIF-1 signaling pathway. The PI3K/AKT pathway can activate the downstream NF-κB pathway and further regulate the transcription and expression of downstream IL-6, IL-17, HIF-1α, Bax, and TNF, suggesting that the PI3K/AKT/NF-κB pathway is the critical pathway in the treatment of OA with Rhizoma Drynariae. Active components of Rhizoma Drynariae and key proteins of the PI3K/AKT/NF-κB signaling pathway were subjected to molecular docking, whose results showed that luteolin and IKK-α played a critical role. In vitro experiments indicated that both aqueous extracts of Rhizoma Drynariae (AERD) and luteolin inhibited the expression of IL-6 and HIF-1α and suppressed the activation of PI3K/AKT/NF-κB, IL-17, and TNF pathways. The measurement of mitochondrial membrane potential (Δψm) indicated that AERD and luteolin can decrease the LPS-induced early apoptotic cells. Luteolin had a more prominent inhibitory effect than AERD in the abovementioned in vitro experiments. In conclusion, the therapeutic mechanism of Rhizoma Drynariae against OA may be closely related to the inhibition of the PI3K/AKT/NF-κB pathway and downstream pathways, and luteolin plays a vital role in the treatment.


Visible light-based stereolithography bioprinting of cell-adhesive gelatin hydrogels.
Stereolithography-based bioprinting offers advantages in resolution and rapid printing time, and thus has received major attention in recent years. However, traditional stereolithography-based bioprinting utilizes an ultraviolet light which may cause mutagenesis and carcinogenesis of cells. In this paper, we present a new visible light crosslinkable bioink that is based on cell-adhesive gelatin. The bioink consists of Eosin Y (EY) based photoinitiator and gelatin methacrylate (GelMA) pre-polymer solution. We examined the feasibility of using visible light from a commercial beam projector to pattern the EY-GelMA bioink. We measured the absorbance of bioink to characterize its sensitivity to visible light and performed bioprinting to test its ability to promote cell adhesion. It is found that the EY-GelMA bioink has an absorption peak at roughly 522 nm, and that it can be successfully crosslinked by visible light from the commercial projector. We performed the bioprinting experiments and visualized the cell morphology using nuclei/F-actin staining. Experimental results show that most of the cells attached to the EY-GelMA bioink after five days' culturing. Ultimately, the EY-GelMA bioink can support both visible light crosslinking and cell adhesion, offering great potential in bioprinting and tissue engineering.


Neuroprotection in glaucoma.
Neuroprotective therapies in glaucoma may play a role in preventing ischemia and oxidative damage that results in apoptosis of retinal ganglion cells and optic nerve damage. Although intraocular pressure (IOP) is the only known modifiable risk factor for glaucoma, disease progression commonly occurs despite IOP control, suggesting that factors other than IOP play a role in its pathogenesis and can potentially act as targets for neuroprotection. Factors including mediators of apoptosis, ischemic changes, poor ocular blood flow and neurotoxins have been hypothesized to play a role in glaucoma progression. Neuroprotective targets include glutamate-induced neurotoxicity, nitric oxidase synthetase, neurotropins, calcium channel receptors, free radicals, vascular insufficiency, the rho-kinase pathway, and more. Drugs related to these factors are being evaluated for their role in neuroprotection, although this area of investigation faces several challenges including limited evidence for these agents' efficacy in clinical studies. Additionally, while IOP-lowering therapies are considered neuroprotective as they generally slow the progress of glaucoma progression, they are limited by the extent of their effect beyond IOP control. The aim of this article is to review the current treatment options available for neuroprotection and to explore the drugs in the pipeline.


Chronic binge alcohol administration impairs glucose-insulin dynamics and decreases adiponectin in asymptomatic simian immunodeficiency virus-infected macaques.
Alcohol use disorders (AUDs) frequently exist among persons living with HIV/AIDS. Chronic alcohol consumption, HIV infection, and antiretroviral therapy (ART) are independently associated with impairments in glucose-insulin dynamics. Previous studies from our laboratory have shown that chronic binge alcohol (CBA) administration decreases body mass index, attenuates weight gain, and accentuates skeletal muscle wasting at end-stage disease in non-ART-treated simian immunodeficiency virus (SIV)-infected male rhesus macaques. The aim of this study was to investigate whether CBA and ART alone or in combination alter body composition or glucose-insulin dynamics in SIV-infected male rhesus macaques during the asymptomatic phase of SIV infection. Daily CBA or sucrose (SUC) administration was initiated 3 mo before intrarectal SIV inoculation and continued until the study end point at 11 mo post-SIV infection. ART or placebo was initiated 2.5 mo after SIV infection and continued until study end point. Four treatment groups (SUC/SIV ± ART and CBA/SIV ± ART) were studied. CBA/SIV macaques had significantly decreased circulating adiponectin and resistin levels relative to SUC/SIV macaques and reduced disposition index and acute insulin response to glucose, insulin, and C-peptide release during frequently sampled intravenous glucose tolerance test, irrespective of ART status. No statistically significant differences were observed in homeostatic model assessment-insulin resistance values, body weight, total body fat, abdominal fat, or total lean mass or bone health among the four groups. These findings demonstrate CBA-mediated impairments in glucose-insulin dynamics and adipokine profile in asymptomatic SIV-infected macaques, irrespective of ART.


Brain death and organ donation.
Brain death is the irreversible cessation of all brain functions. Brainstem death is the 'physiological core' of brain death. The Indian Parliament has given legal recognition to brain death though it applies only in the context of performance of organ transplantation. Brain death is diagnosed if there is irreversible loss of consciousness, absence of brainstem reflexes and apnoea. Care and diligence in the application of the criteria for brain death provide important safeguards for Individual patients and the community in general. These criteria also allow death to be diagnosed with certainty prior to the occurrence of circulatory arrest. Solid organ transplantation has become possible through the diagnosis of brain death but is not the primary consideration; the management of a potential organ donor, who is brain dead, is also vital. If optimal preservation of organs for transplantation is to be achieved the clinician needs to understand the pathophysiology and consequences of changes occurring in various organs after brain death and active management is required to reverse or control these changes. Discussions about organ donation with relatives of brain deed patients are never easy. These should always be frank and sympathetic. It has been suggested that those whose interests lie in transplantation must bear the responsibility of educating the general public. This will help intensivists who expose themselves knowingly to the unpleasant aspects of organ donation.


Chiral Lagrangian from Duality and Monopole Operators in Compactified QCD.
We show that there exists a special compactification of QCD on R^{3}×S^{1} in which the theory has a domain where continuous chiral symmetry breaking is analytically calculable. We give a microscopic derivation of the chiral Lagrangian, the chiral condensate, and the Gell-Mann-Oakes-Renner relation m_{π}^{2}f_{π}^{2}=-m_{q}⟨q[over ¯]q⟩. Abelian duality, monopole operators, and flavor-twisted boundary conditions play the main roles. The flavor twisting leads to the new effect of fractional jumping of fermion zero modes among monopole instantons. Chiral symmetry breaking is induced by monopole-instanton operators, and the Nambu-Goldstone pions arise by color-flavor transmutation from gapless "dual photons." We also give a microscopic picture of the "constituent quark" masses. Our results are consistent with expectations from chiral perturbation theory at large S^{1}, and yield strong support for adiabatic continuity between the small-S^{1} and large-S^{1} regimes. We also find concrete microscopic connections between N=1 and N=2 supersymmetric gauge theory dynamics and nonsupersymmetric QCD dynamics.


Current issues in the physician-patient relationship.
This review is one of a series of articles reflecting the sections of the Code of Ethics of the American Society for Surgery of the Hand. The physician-patient relationship has been subjected to numerous stressors over the past several decades. This report reviews historical elements of the relationship and discusses current issues related to it: asymmetry, challenges to the physician, and interpersonal, systemic, and societal challenges.


A methodology for meeting context-specific information needs related to nursing orders.
Nurse information needs at the point of care exist; however, once provided with access to resources, nurses seek evidence-based answers. The success of answering an information need through an automated Infobutton relies on the program's ability to anticipate a clinician's question. This study's six process methodology aims to meet context-specific information needs related to clinical information system (CIS) nursing orders. Context-specific questions were derived from nurse generic questions generated from nurse focus group data, observational data, and an established taxonomy. The context-specific questions, were filtered by nursing order semantic type within a logical clinical context cluster (LCCC), which is a clinical association identified from the nurse perspective. Nursing orders of the same semantic type are associated with similar types of nurse information needs and are used to assess Internet-based resources for the capability to answer those information needs. LCCCs help identify resources and CIS areas ripe for an Infobutton.


Identification and Quantitative Analysis of Indole-3-Acetyl-l-Aspartate from Seeds of Glycine max L.
Indole-3-acetyl-l-aspartate (IAAsp) was isolated from seeds of Glycine max L. cv. Hark and its identity established by its chromatographic performance and its mass spectral fragmentation. Following acid hydrolysis, the aspartate moiety was shown to be the l-enantiomer by reverse phase high performance liquid chromatographic retention time of the bisethyl ester derivatized with 2,3,4,6-tetra-O-acetyl-beta-d-glycopyranosyl isothiocyanate. Isotope dilution analysis using [(14)C]IAAsp as internal standard showed that soybean seed contained 10 mumol/kg IAAsp and this accounted for one-half of the total indoleacetic acid of the seed.


The Safety of Anti-SARS-CoV-2 Vaccines: Vigilance Is Still Required.
The opinion I put forward in this paper is that attention must continue to be paid to clinical observations compatible with a detrimental effect of anti-SARS-CoV-2 in certain diseases of immunological nature. Using the example of the atypical thrombocytopenic thromboses caused by adenoviral-vector-based vaccines, I argue that usual post-marketing pharmacovigilance programs may fail in identifying very rare vaccine-related disorders. Since the robust protective immunity induced by mRNA vaccines is related to their distinct capacity to induce strong stimulation of T follicular helper cells, I suggest that the safety of mRNA vaccines should be further assessed by appropriately designed epidemiological and mechanistic studies focusing on lymphoproliferative and autoimmune diseases in which T follicular helper cells were found to play a key role.


Dynactin is necessary for synapse stabilization.
We present evidence that synapse retraction occurs during normal synaptic growth at the Drosophila neuromuscular junction (NMJ). An RNAi-based screen to identify the molecular mechanisms that regulate synapse retraction identified Arp-1/centractin, a subunit of the dynactin complex. Arp-1 dsRNA enhances synapse retraction, and this effect is phenocopied by a mutation in P150/Glued, also a dynactin component. The Glued protein is enriched within the presynaptic nerve terminal, and presynaptic expression of a dominant-negative Glued transgene enhances retraction. Retraction is associated with a local disruption of the synaptic microtubule cytoskeleton. Electrophysiological, ultrastructural, and immunohistochemical data support a model in which presynaptic retraction precedes disassembly of the postsynaptic apparatus. Our data suggests that dynactin functions locally within the presynaptic arbor to promote synapse stability.


[The state of diopter and visual acuity in central serous chorioretinopathy].
To investigate the state of diopter and best corrected visual acuity in active central serous chorioretinopathy (CSC). Twenty-three unilateral active CSC patients (23 eyes) were observed with Integrated Optometry Apparatus. The period of follow-up was from 3 - 12 months (mean, 7.2 months). Of the 23 patients, 15 eyes (65.2%) of active CSC and 5 fellow eyes (21.7%) showed hyperopic state. There were significant differences in hyperopic state between the active eyes and fellow eyes (P < 0.01). After follow-up, there were only 5 eyes of CSC (21.7%) showed slight hyperopia. There were significant differences in hyperopic state between before and after follow-up of the eyes of CSC (P < 0.01) Among the eyes of the best corrected visual acuities were increased 3 row or above, 90.9% eyes of the best corrected visual acuities were resumed to 1.0. Most of the active CSC demonstrated temporary hyperopic state due to the neuroretinal detachment of macula. The best corrected visual acuities with the Integrated Optometry Apparatus in active CSC are useful in evaluating the final visual acuities of the eyes.


Potential Epigenetic-Based Therapeutic Targets for Glioma.
Glioma is characterized by a high recurrence rate, short survival times, high rates of mortality and treatment difficulties. Surgery, chemotherapy and radiation (RT) are the standard treatments, but outcomes rarely improve even after treatment. With the advancement of molecular pathology, recent studies have found that the development of glioma is closely related to various epigenetic phenomena, including DNA methylation, abnormal microRNA (miRNA), chromatin remodeling and histone modifications. Owing to the reversibility of epigenetic modifications, the proteins and genes that regulate these changes have become new targets in the treatment of glioma. In this review, we present a summary of the potential therapeutic targets of glioma and related effective treating drugs from the four aspects mentioned above. We further illustrate how epigenetic mechanisms dynamically regulate the pathogenesis and discuss the challenges of glioma treatment. Currently, among the epigenetic treatments, DNA methyltransferase (DNMT) inhibitors and histone deacetylase inhibitors (HDACIs) can be used for the treatment of tumors, either individually or in combination. In the treatment of glioma, only HDACIs remain a good option and they provide new directions for the treatment. Due to the complicated pathogenesis of glioma, epigenetic applications to glioma clinical treatment are still limited.


The role of melatonin in post-partum psychosis and depression associated with bipolar disorder.
Recent data has highlighted the association of a bipolar disorder (BD) with an increased risk of post-partum psychosis and depression. It is suggested that genetic- and environmental-induced decrease in the levels of melatonin in BD contributes to post-partum disorders. Melatonin may also have some efficacy in the treatment of BD, especially in decreasing the side-effects associated with lithium and the neuroleptics. It is proposed that the optimization of melatonin levels, perhaps in conjunction with optimized vitamin D3 level, would decrease post-partum psychosis and depression associated with BD.


Sleep laboratory evaluation of the effects and efficacy of trazodone in depressed insomniac patients.
Trazodone (150 mg to 400 mg) was administered to six depressed patients with significant sleep disturbances in an 8-week single-blind study design. Patients were evaluated psychologically by means of the Hamilton Rating Scales for Anxiety and Depression. Polysomnographic monitoring in the sleep laboratory was conducted at each of the time points corresponding to the psychiatric evaluations. Five of the six subjects completed treatment. Patients showed a significant improvement in symptoms of depression and in their polysomnographic-determined sleep architecture. There was a 44% improvement in persistent sleep latency, decreasing from a mean +/- SD of 51.0 +/- 59.3 minutes at baseline to 28.5 +/- 24.2 minutes after 5 weeks of active treatment. Total sleep time improved 14% from 387.1 +/- 59.2 minutes at baseline to 441.3 +/- 23.7 minutes after 5 weeks. Stage IV sleep more than doubled with an increase of 153% from 1.9 +/- 3.0% at baseline to a more normal 4.8 +/- 5.5%. There was no change in percentage of rapid eye movement (REM); however, REM latency increased 28% from a mean of 74.6 +/- 35.9 minutes at baseline to a mean of 95.6 +/- 28.8 minutes. Sleep efficiency improved from 80.6 +/- 12.3%, considered clinically significant insomnia, to 91.9 +/- 4.9%, which is well within normal sleep patterns.


Novel ATPase of SNF2-like protein family interacts with androgen receptor and modulates androgen-dependent transcription.
Nuclear receptors, including the androgen receptor (AR), regulate target cell transcription through interaction with auxiliary proteins to modify chromatin structure. We describe herein a novel AR-interacting protein, termed ARIP4, that has structural features typical of the SNF2-like protein family. With regard to the Snf2 domain, the closest homolog of ARIP4 is the ATRX protein. ARIP4 is a nuclear protein and comprises 1466 amino acids. It interacts with AR in vitro and in cultured yeast and mammalian cells. ARIP4 can be labeled with 8-azido-[gamma-32P]ATP and exhibits DNA-dependent ATPase activity. Like several ATP-dependent chromatin remodeling proteins, ARIP4 generates superhelical torsion within linear DNA fragments in an ATP-dependent manner. With a stably integrated target promoter, ARIP4 elicits a modest enhancement of AR-dependent transactivation. In transient cotransfection assays, ARIP4 modulates AR function in a promoter-dependent manner; it enhances receptor activity on minimal promoters, but does not activate more complex promoters. ARIP4 mutants devoid of ATPase activity fail to alter DNA topology and behave as trans-dominant negative regulators of AR function in transient assays.


Discovering the Role of FZD4 Gene in Human Cutaneous Squamous Cell Carcinoma.
Frizzled 4 (FZD4) is an important receptor for Wnt proteins that stimulate several downstream signaling pathways. It has been known that the FZD4-Wnt interaction is involved in many types of cancers. However, the role of FZD4 in cutaneous squamous cell carcinoma (CSCC) has not been well studied. We sought to investigate the association between FZD4 expression level and tumor cell proliferation and apoptosis rates in CSCC. Expression of FZD4 at mRNA level in CSCC tissues and controls was measured. Colo16 cell proliferation and viability were measured by CCK-8 assay and flow cytometry respectively after siRNA and plasmid transfection. We discovered a significant downregulation of FZD4 expression in CSCC tissues and cell lines compared to controls. Furthermore, our data suggested that over expression of FZD4 inhibited proliferation and promoted apoptosis of Colo16 cells. The results indicated that FZD4 may play as a tumor suppressor gene in the pathogenesis of CSCC.


Substitutions in the Glycogenin-1 Gene Are Associated with the Evolution of Endothermy in Sharks and Tunas.
Despite 400-450 million years of independent evolution, a strong phenotypic convergence has occurred between two groups of fish: tunas and lamnid sharks. This convergence is characterized by centralization of red muscle, a distinctive swimming style (stiffened body powered through tail movements) and elevated body temperature (endothermy). Furthermore, both groups demonstrate elevated white muscle metabolic capacities. All these traits are unusual in fish and more likely evolved to support their fast-swimming, pelagic, predatory behavior. Here, we tested the hypothesis that their convergent evolution was driven by selection on a set of metabolic genes. We sequenced white muscle transcriptomes of six tuna, one mackerel, and three shark species, and supplemented this data set with previously published RNA-seq data. Using 26 species in total (including 7,032 tuna genes plus 1,719 shark genes), we constructed phylogenetic trees and carried out maximum-likelihood analyses of gene selection. We inferred several genes relating to metabolism to be under selection. We also found that the same one gene, glycogenin-1, evolved under positive selection independently in tunas and lamnid sharks, providing evidence of convergent selective pressures at gene level possibly underlying shared physiology.


A technique to remove epulis: case reports.
The purpose of this study was to describe an alternative surgical approach to removing epulis while preserving and improving the mucogingival complex. After scaling and root planing, six patients underwent surgical treatment to remove the growths. A subepithelial connective tissue graft was interposed between the bone and epulis using a pedicle flap. The lesion was always preserved. The epulis disappeared after several months, with minor variations in healing time among patients. At 60-month follow-up visits, all patients exhibited good gingival health, with no recurrence.


Metal Artifact Reduction in Dental Computed Tomography Images Based on Sinogram Segmentation Using Curvelet Transform Followed by Hough Transform.
In X-ray computed tomography (CT), the presence of metal objects in a patient's body produces streak artifacts in the reconstructed images. During the past decades, many different methods were proposed for the reduction or elimination of the streaking artifacts. When scanning a patient, the projection data affected by metal objects (missing projections) appear as regions with high intensities in the sinogram. In spiral fan beam CT, these regions are sinusoid-like curves on sinogram. During the first time, if the metal curves are detected carefully, then, they can be replaced by corresponding unaffected projections using other slices or opposite views; therefore, the CT slices regenerated by the modified sonogram will be imaged with high quality. In this paper, a new method of the segmentation of metal traces in spiral fan-beam CT sinogram is proposed. This method is based on a sinogram curve detection using a curvelet transform followed by 2D Hough transform. The initial enhancement of the sinogram using modified curvelet transform coefficients is performed by suppressing all the coefficients of one band and applying 2D Hough transform to detect more precisely metal curves. To evaluate the performance of the proposed method for the detection of metal curves in a sinogram, precision and recall metrics are calculated. Compared with other methods, the results show that the proposed method is capable of detecting metal curves, with better precision and good recovery.


The Stanford Hodgkin's disease (HD) studies--an update.
Successive, prospective clinical trials for adults with HD have been conducted from 1962-1991. With approximately 75% of patients in continuous remission as a result of current therapy, attention may be focused on reducing treatment complications for the majority of patients and improving efficacy in selected, high risk populations. This overview will describe recently completed and ongoing clinical trials at Stanford University which address these therapeutic objectives.


Complete and Incomplete Resection for Progressive Glioblastoma Prolongs Post-Progression Survival.
The role of resection in progressive glioblastoma (GBM) to prolong survival is still controversial. The aim of this study was to determine 1) the predictors of post-progression survival (PPS) in progressive GBM and 2) which subgroups of patients would benefit from recurrent resection. We have conducted a retrospective bicentric cohort study on isocitrate dehydrogenase (IDH) wild-type GBM treated in our hospitals between 2006 and 2015. Kaplan-Maier analyses and univariable and multivariable Cox regressions were performed to identify predictors and their influence on PPS. Of 589 patients with progressive IDH wild-type GBM, 355 patients were included in analyses. Median PPS of all patients was 9 months (95% CI 8.0-10.0), with complete resection 12 months (95% CI 9.7-14.3, n=81), incomplete resection 11 months (95% CI 8.9-13.1, n=70) and without resection 7 months (95% CI 06-08, n=204). Multivariable Cox regression demonstrated a benefit for PPS with complete (HR 0.67, CI 0.49-0.90) and incomplete resection (HR 0.73, 95% CI 0.51-1.04) and confirmed methylation of the O6-methylguanine-DNA-methyltransferase (MGMT) gene promoter, lower age at diagnosis, absence of deep brain and multilocular localization, higher Karnofsky Performance Status (KPS) and recurrent therapies to be associated with longer PPS. In contrast, traditional eloquence and duration of progression-free survival had no effect on PPS. Subgroup analyses showed that all subgroups of confirmed predictors benefited from resection, except for patients in poor condition with a KPS <70. Out data suggest a role for complete and incomplete recurrent resection in progressive GBM patients regardless of methylation of MGMT, age, or adjuvant therapy but not in patients with a poor clinical condition with a KPS <70.


Accurate Follicle Enumeration in Adult Mouse Ovaries.
Sexually reproducing female mammals are born with their entire lifetime supply of oocytes. Immature, quiescent oocytes are found within primordial follicles, the storage unit of the female germline. They are non-renewable, thus their number at birth and subsequent rate of loss largely dictates the female fertile lifespan. Accurate quantification of primordial follicle numbers in women and animals is essential for determining the impact of medicines and toxicants on the ovarian reserve. It is also necessary for evaluating the need for, and success of, existing and emerging fertility preservation techniques. Currently, no methods exist to accurately measure the number of primordial follicles comprising the ovarian reserve in women. Furthermore, obtaining ovarian tissue from large animals or endangered species for experimentation is often not feasible. Thus, mice have become an essential model for such studies, and the ability to evaluate primordial follicle numbers in whole mouse ovaries is a critical tool. However, reports of absolute follicle numbers in mouse ovaries in the literature are highly variable, making it difficult to compare and/or replicate data. This is due to a number of factors including strain, age, treatment groups, as well as technical differences in the methods of counting employed. In this article, we provide a step-by-step instructional guide for preparing histological sections and counting primordial follicles in mouse ovaries using two different methods: [1] stereology, which relies on the fractionator/optical dissector technique; and [2] the direct count technique. Some of the key advantages and drawbacks of each method will be highlighted so that reproducibility can be improved in the field and to enable researchers to select the most appropriate method for their studies.


Peak blood alcohol prediction: an empirical test of two computer models.
Two computer programs (Computerized Blood Alcohol Calculator and Blood Alcohol Content Estimator) were compared for reliability in predicting peak blood alcohol concentration (BAC). Subjects were middle-aged volunteers (N = 40; 20 men, 20 women) who each consumed a constant amount of alcohol (40 ml of 95% alcohol or 30 g ethanol) prior to undergoing breath testing to determine peak BAC. The observed BACs were compared with the predicted BACs obtained with each of the computer programs. The Computerized Blood Alcohol Calculator provided marginally better BAC predictions than did the Blood Alcohol Content Estimator. However, both computer models seriously underestimated the peak BACs for this group of subjects. Results are discussed with particular reference to the need for additional studies of age, gender, and body composition in predicting peak BACs for heterogeneous subject groups.


Expression of Fas ligand in newly diagnosed childhood acute lymphoblastic leukemia.
Immunocytochemistry was used to analyze the expression of the Fas ligand (FasL) in sixty-four children with newly diagnosed acute lymphoblastic leukemia (ALL). FasL expression was detected in 55 of 64 children (86%). Children with newly diagnosed ALL and FasL expression exhibited longer relapse-free and overall survival times under chemotherapy than did patients who did not express the Fas ligand. These data suggest that Fas/FasL mediated signaling may play a general role in the cytotoxicity of anticancer drugs.


Nicotinamide phosphoribosyltransferase inhibitors, design, preparation, and structure-activity relationship.
Existing pharmacological inhibitors for nicotinamide phosphoribosyltransferase (NAMPT) are promising therapeutics for treating cancer. By using medicinal and computational chemistry methods, the structure-activity relationship for novel classes of NAMPT inhibitors is described, and the compounds are optimized. Compounds are designed inspired by the NAMPT inhibitor APO866 and cyanoguanidine inhibitor scaffolds. In comparison with recently published derivatives, the new analogues exhibit an equally potent antiproliferative activity in vitro and comparable activity in vivo. The best performing compounds from these series showed subnanomolar antiproliferative activity toward a series of cancer cell lines (compound 15: IC50 0.025 and 0.33 nM, in A2780 (ovarian carcinoma) and MCF-7 (breast), respectively) and potent antitumor in vivo activity in well-tolerated doses in a xenograft model. In an A2780 xenograft mouse model with large tumors (500 mm(3)), compound 15 reduced the tumor volume to one-fifth of the starting volume at a dose of 3 mg/kg administered ip, bid, days 1-9. Thus, compounds found in this study compared favorably with compounds already in the clinic and warrant further investigation as promising lead molecules for the inhibition of NAMPT.


Porous polymer monolith assisted electrospray.
Coupling low-flow analytical separation instrumentation such as capillary electrophoresis, capillary electrochromatography, nano-HPLC, and microfluidic-based devices with electrospray ionization mass spectrometry has yielded powerful analytical tools. However, conventional coupling methodologies such as nanospray suffer from limitations including poor conductive coating robustness, constant clogging, complicated fabrication processes, and incompatibility with large flow rate regimes. This study demonstrates that robust nanospray emitters can be fabricated through the formation and utilization of a porous polymer monolith (PPM) at the end of a fused-silica capillary. Stable electrosprays can be produced from capillaries (75-100-microm i.d.) at a variety of flow rates (50-1000 nL/min) without the need to taper the capillaries by etching or pulling. The PPM is photopatterned to be present only near the capillary exit aperture using conditions that generate pore sizes similar to those seen with nanospray tips. The porous nature of the PPM aids in developing a stable electrospray generating a single clearly visible Taylor cone at relatively high flow rates while at low flow rates (<100 nL/min) a mist, presumably from multiple small Taylor cones, develops. The hydrophobic nature of the PPM should limit problems with band broadening associated with droplet spreading at the capillary exit, while the multiple flow paths inherent in the PPM minimize clogging problems associated with conventional nanospray emitters. Total ion current traces for a constant infusion of standard PPG and cytochrome c solutions are very stable with deviations ranging from only 3 to 8%. The PPM-assisted electrospray produces mass spectra with excellent signal-to-noise ratios from only a few femtomoles of material.


Chronic illness and compliance.
For a person with a chronic illness, life is altered in some way. Whether simple or complex, changes are nevertheless permanent. A patient with a chronic disease assesses recommended treatments on how well they can be integrated into his life. Evidence suggests that an individual's perception of his situation will determine whether or not he will comply with a medical regimen. Health professionals neglect the patient's point of view if they believe that patients regard the health professional as an absolute authority, thereby contributing to noncompliance. The contingency contract provides a model of patient-provider transaction, which requires input from both parties. The health professional can work with the person to make the medical regimen compatible with the individual's lifestyle.


Evidence for intracellular cleavage of plasminogen activator inhibitor type 2 (PAI-2) in normal epidermal keratinocytes.
Plasminogen activator inhibitor type 2 (PAI-2) is a serine proteinase inhibitor (serpin), present in high quantities in stratified squamous epithelia. Detergent extracts of human epidermis or cultured keratinocytes contain primarily active, nonglycosylated PAI-2. In keratinocytes, the vast majority of PAI-2 is retained within the cell, supporting the hypothesis that PAI-2 may serve specific intracellular function(s) through interaction with an unknown cytoplasmic proteinase. During interaction with the target proteinase, cleavage of PAI-2 within its reactive site loop leads to the formation of a more stable, "relaxed" conformation (PAI-2r). Using a monoclonal antibody specific for PAI-2r, we demonstrate here that PAI-2r is present in keratinocytes of the granular and basal layers of normal human epidermis. In addition, PAI-2r is detectable in cultured human epidermal keratinocytes, where it is concentrated in a detergent-insoluble fraction within differentiating cells. These data provide evidence for the presence of an endogenous, keratinocyte-derived proteinase that constitutively cleaves intracellular PAI-2 in normal human epidermal keratinocytes. Cleavage of PAI-2 by this proteinase may reflect specific intracellular action of PAI-2 in normal cells. Finally, we demonstrate that a commercially available anti-PAI-2 monoclonal antibody (#3750, American Diagnostica, Greenwich, CT), under native experimental conditions, preferentially recognizes the uncleaved, active form of PAI-2 and does not efficiently detect PAI-2r.


[A clinico-anatomical correlation study of logorrhoea].
The degree of verbal production necessary to be considered logorrhoea has still not been defined, and no clear correlation has been established between the topography of the dysfunction and this symptom. To provide quantitative data about normal verbal production and to identify the location within the brain of the alterations observed in neuroimage of patients with logorrhoea. The oral verbal production of 60 control subjects between 20 and 80 years of age was quantified by analysing five speeches. Ten patients who exceeded the 75th percentile in at least two of the five speeches underwent structural and functional neuroimaging tests. RESULTS. The data on verbal production of normal subjects are reported. Age, sex and habits (smoking, coffee, alcoholic drinks) did not exert an influence, but the degree of schooling was seen to have an effect. All the patients were diagnosed with frontotemporal degeneration, although in one case there were also coexisting vascular risk factors and subcortical vascular lesions, which reduce the degree of certainty of the diagnosis. Cortical atrophy is located in the right anterior temporal lobes (100% anteromedial, 100% anteroinferior, 70% anterolateral), left anterior temporal (90% anteromedial, 90% anteroinferior, 60% anterolateral), right prefrontal (30% basal, 50% dorsolateral, 20% medial) and left prefrontal (20% basal, 30% dorsolateral, 20% medial). Oral verbal production is influenced by level of education and, in a sample of patients with probable frontotemporal degeneration and logorrhoea, all the patients showed alterations in the anteroinferior and anteromedial regions of the right temporal lobe.


Fetal programming of cardiovascular function through exposure to maternal undernutrition.
A substantial and robust body of epidemiological evidence indicates that prenatal dietary experience may be a factor determining cardiovascular disease risk. Retrospective cohort studies indicate that low birth weight and disproportion at birth are powerful predictors of later disease risk. This prenatal influence on non-communicable disease in later life has been termed programming. Maternal nutritional status has been proposed to be the major programming influence on the developing fetus. The evidence from epidemiological studies of nutrition, fetal development and birth outcome is, however, often weak and inconclusive. The validity of the nutritional programming concept is highly dependent on experimental studies in animals. The feeding of low-protein diets in rat pregnancy results in perturbations in fetal growth and dimensions at birth. The offspring of rats fed low-protein diets exhibit a number of metabolic and physiological disturbances, and are consistently found to have high blood pressure from early postnatal life. This experimental model has been used to explore potential mechanisms of programming through which maternal diet may programme the cardiovascular function of the fetus. Indications from this work are that fetal exposure to maternally-derived glucocorticoids plays a key role in the programming mechanism. Secondary to this activity, the fetal hypothalamic-pituitary-adrenal axis may stimulate renin-angiotensin system activity, resulting in increased vascular resistance and hypertension.


Characterization of a nuclear protein conferring brefeldin A resistance in Schizosaccharomyces pombe.
The fungal metabolite brefeldin A disrupts protein secretion and causes the redistribution of the Golgi complex to the endoplasmic reticulum. Previously we isolated six genes that, when present in multiple copies, confer brefeldin A resistance to wild type Schizosaccharomyces pombe. Here we describe the characterization of one of these genes, hba1. This gene encodes an essential protein that shares homology with the mammalian protein RanBP1 and the protein encoded by the Saccharomyces cerevisiae gene YRB1 and contains a peptide motif present in several proteins found within the nuclear pore complex. The protein encoded by hba1 is localized to the nucleus, and it was determined that this protein is phosphorylated in vivo. The characterization of hba1 thus demonstrates a novel mechanism of drug resistance in S. pombe.


Glycolysis-associated enzymes existing in the follicular lumen of the thyroid may interfere with energy metabolism.
Synthesis and storage of the thyroid hormone precursor, thyroglobulin (TG), occurs within the follicular lumen of the thyroid and the TG is then absorbed into cells for further processing before release into the blood. However, the mechanism of energy metabolism in the follicular lumen of the thyroid remains unknown. In the present study, the three dimensional structure of thyroid follicles was constructed using a primary culture of swine cells and the follicular protein was identified via matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Three glycolysis-associated enzymes, enolase, pyruvate kinase and phosphoglyceraldehyde dehydrogenase were identified in addition to TG. These results support the hypothesis that anaerobic glycolysis of glucose exists in the follicle and supports energy consumption for hormone synthesis.


Glycosylated Artificial Virus-Like Hybrid Vectors for Advanced Gene Delivery.
The major obstacle facing efficient gene therapy is the development of reliable delivery vehicles, which are both nontoxic and biocompatible and possess efficient cell-specific gene delivery. Previously, hybrid delivery vehicles comprising anionic liposomes and cationic polymers have been used successfully for gene therapy. In this study, hybrid vectors based on glycosylated artificial viral envelopes (including two novel compositions mimicking HIV and HSV envelopes) and polyethylenimine were morphologically and physiologically characterised. Transfection studies showed that the hybrid vectors based on the control liposomes, and their glycosylated modifications, had significantly higher transfection rates compared to the polyplexes. Improvement in the transfection efficiency was observed with the glycosylated HIV- and HSV-mimicking hybrid vectors, which also showed a safe biocompatibility profile based on the cytotoxicity and haemocompatibility assays. These glycosylated artificial viral envelope-based hybrid vectors could be used as safe gene delivery systems with potential to become new compositions for efficient nonviral gene therapy.


The dual role of wild phages for horizontal gene transfer among Salmonella strains.
Salmonella bacteriophages seem to mediate horizontal transfer of virulence functions among Salmonella strains in two different ways: by general transduction and also by lysogenic conversion. The majority of wild phages isolated from Salmonella strains belong to the P22 like phages and were able to transduce. Our data show that the lysogenic conversion is generally accompanied by changes in the susceptibility to the typing phages used for epidemiological purposes. Similar phage type conversions to S. Typhimurium DT104 could be detected upon lysogenization with two other S. Typhimurium strains. For some S. Typhimurium strains the typical phage pattern is actually associated with alterations of virulence characteristics. For example, all tested wild type isolates of phage types DT49 and DT204 were found to be SopE phi-lysogens. The Anderson typing phages interfere with the prophages and/or cryptic phages and so the complex genetic short-term evolution can be demonstrated in the lab. This is one reason for the successful application of phage typing in Salmonella epidemiology since the 50s.


Ca2+ binding to bovine lactoferrin enhances protein stability and influences the release of bacterial lipopolysaccharide.
Bovine lactoferrin (bLf) is known to damage the outer membrane of Gram-negative bacteria by binding to bacterial lipopolysaccharide (LPS). We report that LPS is released from bacterial outer membranes also when apo- or metal-saturated Lf is separated from bacterial cells by a dialysis membrane. This process occurs in phosphate-buffered saline with no added Ca2+ and Mg2+ and is hindered by addition of these cations. The effect of bLf is similar to that induced by EDTA and has been ascribed to chelation of Ca2+. In fact, it may be envisaged that Ca2+-binding sites on LPS have different affinities and that bLf can remove those ions that are more weakly bound. Ca2+ binding does not alter Lf iron-binding properties significantly or its UV and CD spectral features but brings about changes in the FT-IR bands due to carboxylate residues. Ca2+ binding is characterized by an apparent dissociation constant of 6 microM and a stoichiometry of 1.55 Ca2+ per Lf molecule; it enhances bLf stability towards chemical and thermal denaturation. The increase in stability takes place in both the apo- and iron-saturated forms but not in the desialilated protein, indicating that the carboxylate groups of the sialic acid residues present on two of the glycan chains are involved in Ca2+ binding.