Six New Constituents from the Fruit of Hypericum patulum and Their Anti-Inflammatory Activity.
Four new xanthone glucosides, 3-hydroxy-2-methoxyxanthone-4-O-β-D-glucopyranoside (1), 4,8-dihydroxy-2-methoxyxanthone-3-O-β-D-glucopyranoside (2), 2-methoxyxanthone-5-O-β-D-glucopyranoside (3), 4-hydroxy-2-methoxyxanthone-3-O-β-D-glucopyranoside (4), a new phenolic acid, 4,4'-dihydroxy-3,3'-imino-di-benzoic acid monomethyl ester (5), and a new isoquinoline, methyl 6-hydroxy-1-oxo-1,2,3,4-tetrahydroisoquinoline-4-carboxylate (6) were isolated from the fruit of Hypericum patulum. The structural elucidation of the isolated compounds was primarily based on HR-ESI-MS, UV, IR, 1D and 2D NMR. All compounds were evaluated for their inhibitory effect against LPS-induced NO production in RAW 264.7 cells. Compound 2, 3 exhibited moderate inhibitory activity against NO production.


Phoretic mite communities associated with bark beetles in the maritime and stone pine forests of Setúbal, Portugal.
The phoretic mite communities of prominent bark beetle pests associated with pine stands of southern Portugal were sampled to determine whether they vary across bark beetle species and stand type. Bark beetles were sampled for mites from two primary (aggressive) bark beetle species (Ips sexdentatus and Orthotomicus erosus) and the most common secondary species (Hylurgus ligniperda) in maritime pine (Pinus pinaster) and stone pine (Pinus pinea) in the Setúbal province of Portugal. Twelve mite species, spanning diverse ecological roles, are found associated with these bark beetle systems. The relative abundances of the 12 species that make up the phoretic mite communities of maritime and stone pine varied significantly between host beetle species as well as between stand type, indicating that the phoretic host and dominant tree type are important drivers of mite community composition. The functional roles of these mites are outlined and their ecological significance in pine forest ecosystems is discussed.


Distant supervision for relation extraction with hierarchical selective attention.
Distant supervised relation extraction is an important task in the field of natural language processing. There are two main shortcomings for most state-of-the-art methods. One is that they take all sentences of an entity pair as input, which would result in a large computational cost. But in fact, few of most relevant sentences are enough to recognize the relation of an entity pair. To tackle these problems, we propose a novel hierarchical selective attention network for relation extraction under distant supervision. Our model first selects most relevant sentences by taking coarse sentence-level attention on all sentences of an entity pair and then employs word-level attention to construct sentence representations and fine sentence-level attention to aggregate these sentence representations. Experimental results on a widely used dataset demonstrate that our method performs significantly better than most of existing methods.


The marginal cost of satellite versus in-center hemodialysis.
Despite increasing numbers of patients receiving hemodialysis in satellite units (SHD), the economic aspects have not been widely explored. A cost analysis of SHD and in-center hemodialysis (ICHD) from a societal perspective was performed to establish the efficiencies associated with shifting resources and patients from ICHD to SHD. Costs were classified as fixed or variable and placed into categories. The resources for operating a SHD unit are the sum of two components: total fixed costs (TFC) and average variable cost (AVC) times SHD patient volume (Q). Using the TFC of a specific-sized SHD unit and the difference in AVC between ICHD and SHD the number of patients needed (Q) in the SHD unit for financial viability was determined. The formula TFC = (AVC(ICHD) - AVC(SHD)) X Q was used to determine the number of patients (Q) needed in a specific-sized SHD unit such that the yearly cost of SHD treatment would be the same as ICHD treatment. Our results show that SHD fixed costs can be fully offset if the volume of SHD patients is seven per year in a six-station unit. SHD costs were lower for nursing and physician fees. Therefore, ICHD care variable costs were $11,374 more per patient year. SHD patients would also have lower travel costs, a mean cost saving of $12,364 per year. SHD can result in significant savings both to the health-care system and to patients. Using the cost categories and formula presented, the number of patients needed in a specific-sized satellite unit to realize cost savings was determined for our program. We found that these savings can offset the fixed investment needed to operate a SHD unit at modest patient volumes.


Prototype ventilator and alarm algorithm for the NASA space station.
An alarm algorithm was developed to monitor the ventilator on the National Aeronautics and Space Administration space station. The algorithm automatically identifies and interprets critical events so that an untrained user can manage the mechanical ventilation of a critically injured crew member. The algorithm was tested in two healthy volunteers by simulating 260 critical events in each volunteer while the volunteer breathed via the ventilator. Thirteen critical events were induced eight times in random order, for the five different modes of ventilation. These events included various ventilator tubing disconnects, leaks, and occlusions, as well as power and gas supply failures. The algorithm identified the critical events and generated alarms in response to 99.2% (516 of 520, total) of the events. The alarm textual messages were correct 98% (505 of 516 messages) of the time. The alarm algorithm is an improvement over current alarms found on most ventilators because its alarm messages specifically identify failures in the patient breathing circuit or ventilator. The system may improve patient care by helping critical care personnel respond more rapidly and correctly to critical events.


A comparison of supracostal and infracostal access approaches in treating renal and upper ureteral stones using MPCNL with the aid of a patented system.
There are still disagreements on which is a better approach to choose to establish percutaneous tract for percutaneous nephrolitotomy (PCNL), between supracostal and infracostal approaches. The aim of this study is to investigate the safety, efficacy and practicability of minimally invasive PCNL (MPCNL) with the aid of a patented system either through supracostal or through infracostal access. A retrospective study was carried out for 83 patients with renal or upper ureteral stones. Under the guidance of B ultrasound or C-arm, these patients were treated by MPCNL through either 12th rib infracostal (Group 1, 43 cases) or supracostal (Group 2, 40 cases) access approach. These 2 groups were compared for total number of percutaneous tracts, average time in establishing a given percutaneous tract, the number of percutaneous tract used for each case, the average stone clearance time, the clearance rate of all stones by one surgery, and the amount of bleeding using a single percutaneous tract. There was a significantly smaller total number of percutaneous tracts needed, a smaller number of cases that needed two percutaneous tracts to clear stones completely, a shorter average time in establishing a percutaneous tract, and a smaller average amount of bleeding in infracostal access group. At the same time, there were a significantly larger number of cases in which stones were cleared completely using a single percutaneous tract and a higher renal stone clearance rate by one surgery. There were several advantages of infracostal access. These included accuracy in establishing a percutaneous tract, safety, quickness, convenience and flexibility in moving the patented sheath, and higher renal and upper ureteral stone clearance rate by one surgery.


Studies on the functional significance of a C-terminal S-shaped motif in human arginase type I: essentiality for cooperative effects.
The functional significance of a C-terminal S-shaped motif (residues 304-322) in human arginase I was explored by examining the kinetic properties of the R308A mutant and truncated species terminating in either Arg-308 or Ala-308. Replacement of Arg-308 with alanine, with or without truncation, yielded monomeric species. All mutants were kinetically indistinguishable from the wild-type enzyme at the optimum pH of 9.5. At the more physiological, pH 7.5, hyperbolic kinetics was observed for all the mutants, in contrast with the cooperative behavior exhibited by the wild-type species. In the presence of 2mM guanidinium chloride (Gdn(+)), the single mutant R308A changed to a trimeric and kinetically cooperative form, whereas the other enzyme variants were not altered. The S-shaped motif is suggested as essential for the cooperative response of the enzyme to l-arginine at pH 7.5. Gdn(+) is suggested to mimic the guanidine group of Arg-308 at the monomer-monomer interface.


A girl with bilateral ovarian tumours: Frasier syndrome.
Frasier syndrome (FS) is characterised by male pseudohermaphroditism, slowly progressing nephropathy and frequent development of gonadoblastoma. The Wilms' tumour suppressor gene (WT1 gene) plays an important role in the development of the urogenital system and the gonads. A splice mutation in intron 9 of the WT1 gene was recently described in patients with FS. We analysed the WT1 gene of a Japanese patient with male pseudohermaphroditism, steroid resistant-nephr-opathy and gonadoblastoma by the polymerase chain reaction and direct sequencing and detected a heterozygous point mutation in intron 9. analysis of the Wilms' tumour suppressor gene in a patient with Frasier syndrome by the polymerase chain reaction and direct sequencing detected a + 5G -->A transition at a position of the second alternative splice region of exon 9, important for predicting the risk of the occurrence of Wilms' tumour.


Clinical signs of acute ocular inflammatory response to endotoxin are not altered by increasing antioxidant potency of intraocular fluids.
Plasma antioxidant activity is due in large part to the ferroxidase activity of the copper (Cu) transport protein, ceruloplasmin. Implantation of osmotic pumps containing copper into rabbits resulted in a doubling of Cu concentration, ferroxidase activity, and antioxidant activity in plasma. Blood-ocular barriers essentially prevent the entry of large molecules such as proteins from plasma into the intraocular fluid compartments. However, during ocular inflammation, when these barriers are disrupted, plasma proteins can enter. Twenty-four hours after the induction of ocular inflammation by intraocular injection of endotoxin, the Cu concentration and antioxidant activity of intraocular fluids from the Cu pump animals was twice that of control groups, reflecting the differences in plasma levels. This is the first direct demonstration that alterations in plasma levels of ceruloplasmin can influence the antioxidant potency of the extracellular fluids. Increased intraocular fluid antioxidant activity did not affect the acute anterior segment response to endotoxin. However, it is possible that the time course and resolution of the response is altered by changes in extracellular fluid antioxidant activity. This possibility is currently under investigation.


Genomic and epigenetic alterations deregulate microRNA expression in human epithelial ovarian cancer.
MicroRNAs (miRNAs) are an abundant class of small noncoding RNAs that function as negative gene regulators. miRNA deregulation is involved in the initiation and progression of human cancer; however, the underlying mechanism and its contributions to genome-wide transcriptional changes in cancer are still largely unknown. We studied miRNA deregulation in human epithelial ovarian cancer by integrative genomic approach, including miRNA microarray (n = 106), array-based comparative genomic hybridization (n = 109), cDNA microarray (n = 76), and tissue array (n = 504). miRNA expression is markedly down-regulated in malignant transformation and tumor progression. Genomic copy number loss and epigenetic silencing, respectively, may account for the down-regulation of approximately 15% and at least approximately 36% of miRNAs in advanced ovarian tumors and miRNA down-regulation contributes to a genome-wide transcriptional deregulation. Last, eight miRNAs located in the chromosome 14 miRNA cluster (Dlk1-Gtl2 domain) were identified as potential tumor suppressor genes. Therefore, our results suggest that miRNAs may offer new biomarkers and therapeutic targets in epithelial ovarian cancer.


Sexual and physical abuse and identity of the perpetrator: Associations with binge eating and overeating in Project EAT 2018.
Sexual and physical abuse are risk factors for binge eating and overeating, but few studies have examined association of the identity of the perpetrator with survivors' risk of binge eating and overeating. To examine the risk of binge eating and overeating by (1) type of abuse and identity of the perpetrators and (2) cumulative abuse experiences/perpetrators. Data came from Eating and Activity over Time (N = 1407; ages 18-30 during 2017-2018). Sexual abuse perpetrators included family members, non-family members, and intimate partners. Physical abuse perpetrators included family members and intimate partners. Cumulative abuse experiences were defined as the number of types of abuse experienced. Modified Poisson regressions were used to examine the risk of binge eating (overeating with loss of control) and overeating (without loss of control), by (1) abuse type and perpetrator and (2) cumulative abuse experiences. Abuse was more strongly associated with binge eating than overeating. For binge eating, risk factors included familial and intimate partner sexual abuse (RR = 1.48 [95% CI = 1.01-2.17] and 2.41, [95% CI = 1.70-3.41], respectively) and physical abuse (RR = 1.84, [95% CI = 1.33-2.53] and 1.95, [95% CI = 1.35-2.81], respectively), after adjustment for sociodemographic variables. For overeating, associations with physical abuse were close to the null, and those with sexual abuse were modest, with wide CIs that overlapped the null. Abuse experiences were cumulatively associated with binge eating, but not overeating. Assessment of identity of the perpetrator, and cumulative abuse experiences/perpetrators may assist in identifying people at the greatest risk of binge eating.


Stability of sterigmatocystin during the bread making process and its occurrence in bread from the Latvian market.
Sterigmatocystin (STC) is a carcinogenic and mutagenic mycotoxin produced by fungi of many Aspergillus species. The aim of this research was to test the stability of STC during the bread making process and to check bread samples from the Latvian market for STC contamination, using a previously developed electrospray positive ionisation (ESI(+)) liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Wheat grain naturally contaminated with STC was used for bread baking. STC was found to be stable during the bread-making process. In the food survey 17% of the analysed breads were positive for STC, with concentration levels of 2-7 μg kg(-1). One out of six rye bread samples, one out of nine rye-wheat bread samples and three out of 14 wheat bread samples were contaminated with STC. Four out of five contaminated samples contained whole grains as the main ingredient. We conclude that whole grain bread may be a possible source of STC, although even STC-positive bread samples identified in this study contained quite low toxin levels.


[Influence of low-intensity laser radiation on the formation of liquid crystalline structures in a solution of glycoproteins].
Liquid-crystalline structure formation in glycoprotein solutions irradiated by helium-neon laser in the presence of hydrogen peroxide was observed by both polarizing microscopy and spectrophotometry. High molecular weight (2.10(6) Da) and heavily glycosylated (about 80%) glycoprotein was isolated from the mucus layer of pig small intestine. Remarkable changes of both optic parameters of the solutions and the morphology of liquid-crystalline structures were detected in irradiated samples compared to the non-irradiated ones.


Metabolite secretions of Lactobacillus plantarum YYC-3 may inhibit colon cancer cell metastasis by suppressing the VEGF-MMP2/9 signaling pathway.
Colorectal cancer (CRC) is a major clinical challenge, and the gut microbiome plays important roles in the occurrence and metastasis of CRC. Lactobacillus and their metabolites are thought to be able to suppress the growth of CRC cells. However, the antimetastatic mechanism of Lactobacillus or their metabolites toward CRC cells is not clear. Therefore, the aim of this study was to assess the inhibitory mechanism of cell-free supernatants (CFSs) of L. rhamnosus GG, L. casei M3, and L. plantarum YYC-3 on metastasis of CRC cells. YYC-3 CFS showed the highest inhibitory effect on CRC cell growth, invasion and migration, and inhibited MMP2, MMP9, and VEGFA gene and protein expression, and protein secretion. Furthermore, it suppressed the activities of MMPs by gelatin zymography. Moreover, the effective compounds in these CFSs were analyzed by Q Exactive Focus liquid chromatography-mass spectrometry. Our results showed that metabolite secretions of YYC-3 may inhibited cell metastasis by downregulating the VEGF/MMPs signaling pathway. These data suggest that treatment of CRC cells with metabolites from L. plantarum YYC-3 may reduce colon cancer metastasis.


Clinical Efficacy, Safety, and Feasibility of Using Video Glasses during Interventional Radiologic Procedures: A Randomized Trial.
To evaluate the clinical efficacy, safety, and feasibility of implementing video glasses in a variety of interventional radiologic (IR) procedures. Between August 2012 and August 2013, 83 patients undergoing outpatient IR procedures were randomized to a control group (n = 44) or an experimental group outfitted with video glasses (n = 39). State-Trait Anxiety Inventory (STAI) scores, sedation and analgesia doses, mean arterial pressure (MAP), heart rate (HR), respiratory rate (RR), pain scores, and procedure times were obtained. Complications and adverse events related to the use of video glasses were recorded. Postprocedural staff surveys and patient satisfaction surveys were completed. Women had greater preprocedural anxiety than men (P = .0056), and patients undergoing vascular interventions had greater preprocedural anxiety than those undergoing nonvascular interventions (P = .0396). When assessed after the procedure, patients who wore video glasses had significantly reduced levels of anxiety (-7.7 vs -4.4, respectively; P = .0335) and average MAP (-6.3 vs 2.1, respectively; P = .0486) compared with control patients. There was no significant difference in amount of sedation and analgesia, HR, RR, pain score, or procedure time between groups. No significant adverse events related to the use of video glasses were observed. Postprocedural surveys showed that video glasses were not distracting and did not interfere or pose a safety issue during procedures. Patients enjoyed using the video glasses and would use them again for a future procedure. Video glasses can be safely implemented during IR procedures to reduce anxiety and improve a patient's overall experience.


A different TIPE of immune homeostasis.
Proteins with death effector domains (DED) are key signal transducers involved in cell death and inflammation. In this issue of Cell, Sun et al. (2008) describe TIPE2, a DED protein that negatively regulates both T cell receptor and Toll-like receptor signaling. These findings reveal a new element critical to the maintenance of homeostasis in both the adaptive and innate immune systems.


Increased subclinical atherosclerosis in young adults with metabolic syndrome: the Bogalusa Heart Study.
The purpose of this study was to investigate the association of metabolic syndrome (MetS) with subclinical atherosclerosis, determined by ultrasound carotid intima-media thickness (CIMT) measurements, in young adults. Metabolic syndrome is associated with subclinical atherosclerosis and increased cardiovascular risk in older and middle-aged adults; however, these associations have not been studied among young adults. Non-diabetic subjects from Bogalusa Heart Study, a longitudinal study of atherosclerosis in young adults, underwent B-mode ultrasonography of the carotid arteries. Metabolic syndrome was defined with the National Cholesterol Education Program Adult Treatment Panel III (MetSNCEP) and World Health Organization (MetSWHO) definitions. CIMT and MetS associations were evaluated with multivariable regression and area under receiver-operator characteristic curve (AUC) analyses. Of 507 subjects (29% black, 39% male, mean [SD] age 32 [3] years), 67 (13%) had MetSNCEP and 65 (13%) had MetSWHO. Common (mean = 0.70 [0.11] mm vs. 0.66 [0.08] mm, p = 0.002) and internal CIMT (0.72 [0.21] mm vs. 0.68 [0.12] mm, p = 0.020) were higher among those with MetS(NCEP) than those without MetS(NCEP). Common (0.69 [0.11] mm vs. 0.66 [0.08] mm, p = 0.020) and internal CIMT (0.73 [0.23] mm vs. 0.68 [0.12] mm, p = 0.012) also were higher among those with MetSWHO than those without MetSWHO. Composite CIMT increased with the number of MetS components present (MetSNCEP r = 0.997, p < 0.001; MetSWHO r = 0.946, p = 0.053). Metabolic syndromeNCEP (AUC = 0.557, 95% confidence interval [CI] 0.513 to 0.601) and MetSWHO (AUC = 0.539, 95% CI 0.495 to 0.584) both predicted composite CIMT > or =75th percentile. In young adults, MetS is associated with increased atherosclerotic burden, and therefore, increased cardiovascular risk. These results support the importance of screening and early intervention in this population.


Interactions between DNA polymerase and aminofluorene adducts that affect the recognition and possibly the mutagenicity of the lesions.
Adducts of 2-aminofluorene on the C-8 position of guanine block DNA synthesis and lead to mutation. N-acetylated adducts adopt the syn conformation such that in DNA the guanine is displaced from the helix by the fluorene ring while unacetylated adducts prefer the anti conformation allowing normal base pairing of the guanine with cytosine. In vitro synthesis by both E. coli DNA polymerase I and T4 DNA polymerase terminates predominantly one nucleotide before acetylated adducts but has an increased tendency to terminate opposite the unacetylated adducts apparently reflecting the preferred conformations of the two species of reacted nucleoside. In complete contrast AMV reverse transcriptase correctly inserts cytosine opposite the acetylated adducts but prefers to terminate one nucleotide before the unacetylated adducts. We interpret these results as indicating that the specific properties of a replicating polymerase can influence the conformation of a reacted nucleoside thus altering its recognition and possibly its mutagenic activity.


Psychological Dimensions That Explain Integrity in Older Adults: Examination of a Structural Model.
The aim of this research was to confirm whether internal coping strengths, depressive symptomatology and gratitude disposition are significant predictors of integrity in older adults. Participated 394 Ecuadorian older adults with ages between 60 and 91 years old. Self-report were applied to assess the different variables under study. Specifically, integrity, coping, resilience, self-efficacy, mood and gratitude were assessed. A confirmatory model was estimated to predict ego-integrity. A personal adjustment factor composed of problem-focused coping strategies, resilience and self-efficacy, and gratitude showed positive and significant relationships with ego-integrity, whereas negative mood negatively predicted integrity. Integrity is a key variable to strengthen a coherent view of one's life history and highly relevant during ageing. Having personal strengths and dispositions that facilitate adaptation to ageing and maintaining a positive mood predicts the attainment of integrity. Integrity is an adjustment factor that facilitates adaptation to the stressors of ageing and to major life changes and loss of control indifferent areas of life.


Peripheral Primitive Neuroectodermal Tumor (pPNET) of the Parotid: Report of a Rare Case.
Primitive neuroectodermal tumors (PNETs) are a family of highly malignant soft tissue neoplasms mostly occurring in children and young adults. PNETs usually develop in the thoracopulmonary region, abdomen, pelvis, and rarely in the head and neck region. Here, a case of PNET located in the parotid gland is reported.


Circadian regulation of c-MYC in mice.
The circadian clock is a global regulatory mechanism that controls the expression of 50 to 80% of transcripts in mammals. Some of the genes controlled by the circadian clock are oncogenes or tumor suppressors. Among these Myc has been the focus of several studies which have investigated the effect of clock genes and proteins on Myc transcription and MYC protein stability. Other studies have focused on effects of Myc mutation or overproduction on the circadian clock in comparison to their effects on cell cycle progression and tumorigenesis. Here we have used mice with mutations in the essential clock genes Bmal1, Cry1, and Cry2 to gain further insight into the effect of the circadian clock on this important oncogene/oncoprotein and tumorigenesis. We find that mutation of both Cry1 and Cry2, which abolishes the negative arm of the clock transcription-translation feedback loop (TTFL), causes down-regulation of c-MYC, and mutation of Bmal1, which abolishes the positive arm of TTFL, causes up-regulation of the c-MYC protein level in mouse spleen. These findings must be taken into account in models of the clock disruption-cancer connection.


Neospora 2001.
Infection with the protozoan parasite Neospora caninum is emerging as a major cause of reproductive loss in cattle worldwide. Recent advances in research with the ultimate goal of devising effective and sustainable control measures were discussed at Neospora 2001. Neospora 2001 was held, 13-15 September 2001, at the Moredun Research Institute, Edinburgh, UK. The international meeting covered many aspects of bovine neosporosis including diagnosis, epidemiology and immunology.


Conservation and Role of Electrostatics in Thymidylate Synthase.
Conservation of function across families of orthologous enzymes is generally accompanied by conservation of their active site electrostatic potentials. To study the electrostatic conservation in the highly conserved essential enzyme, thymidylate synthase (TS), we conducted a systematic species-based comparison of the electrostatic potential in the vicinity of its active site. Whereas the electrostatics of the active site of TS are generally well conserved, the TSs from minimal organisms do not conform to the overall trend. Since the genomes of minimal organisms have a high thymidine content compared to other organisms, the observation of non-conserved electrostatics was surprising. Analysis of the symbiotic relationship between minimal organisms and their hosts, and the genetic completeness of the thymidine synthesis pathway suggested that TS from the minimal organism Wigglesworthia glossinidia (W.g.b.) must be active. Four residues in the vicinity of the active site of Escherichia coli TS were mutated individually and simultaneously to mimic the electrostatics of W.g.b TS. The measured activities of the E. coli TS mutants imply that conservation of electrostatics in the region of the active site is important for the activity of TS, and suggest that the W.g.b. TS has the minimal activity necessary to support replication of its reduced genome.


Pyrosequencing analysis of the beta-tubulin gene in Spanish Teladorsagia circumcincta field isolates.
Benzimidazole (BZ) resistance in gastrointestinal nematodes has been associated with single nucleotide polymorphisms (SNPs) at codons 200, 167 and 198 in the beta-tubulin isotype 1 gene and, recently, these SNPs have also been found in macrocyclic lactone (ML) resistant strains of Haemonchus contortus. On this basis, we have studied the same putative SNPs in Spanish Teladorsagia circumcincta field isolates by pyrosequencing. Single L3 (infective 3rd stage larvae) from five sheep flocks were tested after confirming their BZ susceptibility and degree of ivermectin (IVM) resistance. According to the Faecal Egg Count Reduction Test (FECRT) one flock was classified as IVM susceptible, another one was resistant, and the rest had a suspicion of resistance to IVM. DNA extraction was carried out on 598 single L3 and 56% of these were identified as T. circumcincta after the amplification of a species-specific ITS2 fragment. The number of L3 analyzed for the SNPs 198/200 was 255 and for the SNP 167 was 187. Results clearly indicate no resistance-associated SNPs were present at any codon, before or after treatment. Therefore, all T. cicumcincta L3 were designated as susceptible homozygous genotypes for all SNPs. The absence of the mutations in these populations would argue against resistance haplotypes being present in the parasite population prior to drug treatment, at least in Spanish T. circumcincta.


Automated video analysis of age-related motor deficits in monkeys using EthoVision.
Previous studies comparing age-related changes in locomotor function in nonhuman primates have generally relied on subjective human observations or rudimentary infrared motion sensors. Here, we used the automated video-tracking system EthoVision to objectively quantify locomotor activity in 6 young, 6 middle-aged and 12 aged female rhesus monkeys. The video records were analyzed for distance traveled, movement speed and vertical activity. Our results showed that the young monkeys (4.9 +/- 0.1 years old) traveled twice the distance and moved 48% faster than the middle-aged monkeys (15.7 +/- 0.5 years old), and traveled thrice the distance and moved 67% faster than the aged monkeys (26.3 +/- 0.9 years old). In addition, young monkeys were vertically more active (20/60 min) than both the middle-aged (7/60 min) and the aged (1/60 min) monkeys. Furthermore, the locomotor performance of the individual animals significantly correlated with increasing age for all three measures. We conclude that EthoVision is a reliable and objective tracking method for detecting age-related differences in locomotor movements in rhesus macaques, and possibly in humans.


Cryo-EM structures of prefusion SIV envelope trimer.
Simian immunodeficiency viruses (SIVs) are lentiviruses that naturally infect non-human primates of African origin and seeded cross-species transmissions of HIV-1 and HIV-2. Here we report prefusion stabilization and cryo-EM structures of soluble envelope (Env) trimers from rhesus macaque SIV (SIVmac) in complex with neutralizing antibodies. These structures provide residue-level definition for SIV-specific disulfide-bonded variable loops (V1 and V2), which we used to delineate variable-loop coverage of the Env trimer. The defined variable loops enabled us to investigate assembled Env-glycan shields throughout SIV, which we found to comprise both N- and O-linked glycans, the latter emanating from V1 inserts, which bound the O-link-specific lectin jacalin. We also investigated in situ SIVmac-Env trimers on virions, determining cryo-electron tomography structures at subnanometer resolutions for an antibody-bound complex and a ligand-free state. Collectively, these structures define the prefusion-closed structure of the SIV-Env trimer and delineate variable-loop and glycan-shielding mechanisms of immune evasion conserved throughout SIV evolution.


The mycotoxin research in foodstuffs in the Czech Republic in the 90th Years.
Mycotoxins are naturally occurring secondary metabolites produced by several toxigenic microscopic fungi on a variety of crops, especially cereal grains and further foodstuffs. Series of experimental research projects on the determination of mycotoxins (aflatoxins, cyclopiazonic acid, ochratoxin A, patulin, deoxynivalenol, fumonisin B1, T-2 toxin, zearalenone, sterigmatocystin, alternaria toxins) in several foods were realized in the National Reference Centre for Microscopic Fungi and Mycotoxins in the 90(th) years. The aim of our work was an estimation of dietary exposure to mycotoxins and risk assessment. The method of a solid phase extraction (SPE), liquid - liquid extraction and immunoaffinity chromatography (f. e. R-Biopharm, VICAM) were used to elaborate for sample analyses of mycotoxins in our projects. The mycotoxins were detected most frequently by chromatographic methods (HPTLC, HPLC, GC) and immunochemical methods (ELISA). Average dietary exposure has been calculated by multiplying of concentration data for specific foods with their consumption rates per 1 kg of b. w. per day. The estimation of the dietary exposure dose of mycotoxins for the Czech population is presented.


Submandibular abscess caused by Salmonella.
Salmonella species represent uncommon causative agents in focal infections of the head and neck. This article presents a rare case of Salmonella submandibular abscess in a 59-year-old patient with diabetes mellitus, chronic hepatitis and liver cirrhosis. The patient was successfully treated with drainage of the abscess and oral administration of quinolone. An immunocompromised condition can be a critical predisposing factor for Salmonella submandibular abscess.


Animal- and herd-level risk factors for leptospiral seropositivity among sows in the Mekong delta, Vietnam.
In 1998, a total of 424 sows had sera collected in the Mekong delta in Vietnam. Of these, 283 sows were from 151 small-scale family farms in 19 villages, and 141 from seven large-scale state farms. The sera were subjected to the microscopic agglutination test (MAT) for antibodies to 13 Leptospira serovars. The overall leptospiral seroprevalence for titres > or =1:100 and > or =1:400, was 73 and 29%, respectively, and was higher (P=0.001) at small- than at large-scale farms. The highest seroprevalence was recorded for Leptospira interrogans serovar (sv) bratislava (52%). At small-scale farms, higher prevalences were found to serovars L. interrogans sv icterohaemorrhagiae (P=0.04) and L. interrogans sv pomona (P=0.02). Epidemiological information (at the individual-animal and herd-levels) was collected with a questionnaire. The data were analysed using logistic multiple regression. At the animal-level, sows seropositive for L. interrogans sv australis and sv autumnalis had less direct contact with sows in neighbouring pens (odds ratio (OR)=0.3 and 0.4, respectively) and sows seronegative for L. interrogans sv bratislava were of lower age (OR=0.1 for seropositivity). Also, sows seropositive for L. interrogans sv icterohaemorrhagiae had higher odds (OR=5.8) if they had not been born on the farm (had been introduced to it as gilts). Herds seropositive for sv javanica showed association with farms not taking measures to control the local rodent population (OR=7.8). Serovar pomona was also linked to the use of artificial insemination (AI), as opposed to natural-breeding services (OR=11.2). These results indicate that housing and management could affect the seroprevalence of Leptospira infection in pigs.


Involvement of the Receptor for Advanced Glycation End Products (RAGE) in high fat-high sugar diet-induced anhedonia in rats.
Clinical and basic science investigation indicates a link between insulin resistance and anhedonia. Previous results of this laboratory point to impaired nucleus accumbens (NAc) insulin signaling as an underpinning of diet-induced anhedonia, based on use of a glucose lick microstructure assay. The present study evaluated whether advanced glycation end products (AGEs) and their receptor (RAGE), known to mediate obesogenic diet-induced inflammation and pathological metabolic conditions, are involved in this behavioral change. Six weeks maintenance of male and female rats on a high fat-high sugar liquid diet (chocolate Ensure) increased body weight gain, and markedly increased circulating insulin and leptin, but induced anhedonia (decreased first minute lick rate and lick burst size) in males only. In these subjects, anhedonia correlated with plasma concentrations of insulin. Although the diet did not alter plasma or NAc AGEs, or the expression of RAGE in the NAc, marginally significant correlations were seen between anhedonia and plasma content of several AGEs and NAc RAGE. Importantly, a small molecule RAGE antagonist, RAGE229, administered twice daily by oral gavage, prevented diet-induced anhedonia. This beneficial effect was associated with improved adipose function, reflected in the adiponectin/leptin ratio, and increased pCREB/total CREB in the NAc, and a shift in the pCREB correlation with pThr34-DARPP-32 from near-zero to strongly positive, such that both phospho-proteins correlated with the rescued hedonic response. This set of findings suggests that the receptor/signaling pathway and cell type underlying the RAGE229-mediated increase in pCREB may mediate anhedonia and its prevention. The possible role of adipose tissue as a locus of diet-induced RAGE signaling, and source of circulating factors that target NAc to modify hedonic reactivity are discussed.


Detection of human herpesvirus 6 DNA and chromosomal integration after allogeneic hematopoietic stem cell transplantation: A retrospective single center analysis.
Human herpesvirus 6 (HHV-6) can reactivate after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and may be associated with significant morbidity and mortality. The epidemiology of HHV-6 infections and their impact on outcome after allo-HSCT were retrospectively analyzed in 689 adult allo-HSCT recipients (January 2015-December 2018). Chromosomal integration of HHV-6 (ciHHV-6) in the donor was retrospectively investigated to critically evaluate antiviral treatment strategies. HHV-6 DNA in any specimen was found in 89 patients. HHV-6 infections (encephalitis (one), gastroenteritis (44), dermatitis (two), hepatitis (one), or pneumonitis (five)) were diagnosed in 53/689 patients (7.7%). Elevated levels of HHV-6 DNA were found in 38 patients (5.5%). ciHHV-6, analyzed in patients with HHV-6 viral loads ≥104 copies/ml, was identified in four patients (10/38 patients; 10.5%). Two of those displayed copy numbers of HHV-6 ranging from ≥2 × 105 to 2.5 × 106 copies/ml (HHV-6A). Here, ciHHV-6 was integrated into donor and not into the patients' cells. In this series of allo-HSCT recipients, 10.5% of patients with blood viral loads of HHV-6 showed ciHHV-6. Screening of the donor for ciHHV-6 before initiation of antiviral therapy is recommended.


Oncologic Outcomes Following Surgical Management of Clinical Stage II Sex Cord Stromal Tumors.
To investigate the clinical history of patients with clinical stage II sex cord stromal tumors who underwent retroperitoneal lymph node dissection (RPLND) at our institution. Our prospectively maintained testicular cancer database was queried to identify patients who presented with or developed clinical stage II sex cord stromal tumors and underwent RPLND at our institution between 1980 and 2018. Demographic, clinical, and pathologic characteristics were reviewed. Kaplan-Meier curves were graphed to assess recurrence-free and overall survival. Fourteen patients were included in the study with a median age of 44.2years. Four patients presented with clinical stage II disease and 10 patients developed metastatic disease during follow-up of initial clinical stage I disease with a median time to metastasis of 2.7years (range: 0.4-19.5 years). Of the 10 patients with orchiectomy pathology data available, all patients had at least 1 risk factor on testis pathology (mean: 2.9 risk factors). Nine patients received treatment prior to referral to our institution. All patients recurred post-RPLND at Indiana University. Median recurrence-free survival was 9.8 months. Twelve patients died of disease with a median overall survival of 14.4 months. Metastatic sex cord stromal tumors are rare and are more resistant to standard treatment modalities than metastatic germ cell tumors. Patients presenting with sex cord stromal tumors should consider prophylactic primary RPLND in the setting of 1 or more pathologic predictor of malignancy.


Preparation of Polyurethane Monolithic Resins and Modification with a Condensed Tannin-Yielding Self-Healing Property.
Resins of polyurethane were prepared from vegetable oils (crambe and castor) and modified by adding green corrosion inhibitor (condensed tannin). The oils were characterized by gas chromatography with flame-ionization detection (GC-FID), attenuated total reflectance Fourier transform infrared spectroscopy (FTIR-ATR) and thermogravimetric analysis (TGA). The reaction was monitored by characterizing the intermediate products (polyester and prepolymer). The polyester was characterized by solubility in methanol, acidity index, hydroxyl groups and FTIR-ATR, and the prepolymer was characterized by solid content, solvent content, isocyanate (NCO) groups and FTIR-ATR. The formation of PU resins was confirmed by FTIR-ATR and TGA, and the presence of tannin particles incorporated in the coating can be observed by optical microscopy (OM). The absence of the band attributed to NCO in FTIR-ATR spectra of the resins confirmed the complete reaction between polyester and prepolymer. The OM observation and a video demonstrate that Polyurethane (PU)-modified with condensed tannin resin presents self-healing effect, probably through the formation of new hydrogen bonds when in contact with deionized water. Therefore, these results open possibilities for new synthetic routes aiming at improving the very important self-healing property for protecting metals and their alloys against corrosion, extending significantly the metallic materials lifetime as previously demonstrated by our group.


Laparoscopy with augmented reality adaptations.
One of the most promising applications of Optical See-Through Augmented Reality is minimally laparoscopic surgery, which currently suffers from problems such as surgeon discomfort and fatigue caused by looking at a display positioned outside the surgeon's visual field, made worse by the length of the procedure. This fatigue is especially felt on the surgeon's neck, as it is strained from adopting unnatural postures in order to visualise the laparoscopic video feed. Throughout this paper, we will present work in Augmented Reality, as well as developments in surgery and Augmented Reality applied to both surgery in general and laparoscopy in particular to address these issues. We applied user and task analysis methods to learn about practices performed in the operating room by observing surgeons in their working environment in order to understand, in detail, how they performed their tasks and achieved their intended goals. Drawing on observations and analysis of video recordings of laparoscopic surgeries, we identified relevant constraints and design requirements. Besides proposals to approach the ergonomic issues, we present a design and implementation of a multimodal interface to enhance the laparoscopic procedure. Our method makes it more comfortable for surgeons by allowing them to keep the laparoscopic video in their viewing area regardless of neck posture. Also, our interface makes it possible to access patient imaging data without interrupting the operation. It also makes it possible to communicate with team members through a pointing reticle. We evaluated how surgeons perceived the implemented prototype, in terms of usefulness and usability, via a think-aloud protocol to conduct qualitative evaluation sessions which we describe in detail in this paper. In addition to checking the advantages of the prototype as compared to traditional laparoscopic settings, we also conducted a System Usability Scale questionnaire for measuring its usability, and a NASA Task Load Index questionnaire to rate perceived workload and to assess the prototype effectiveness. Our results show that surgeons consider that our prototype can improve surgeon-to-surgeon communication using head pose as a means of pointing. Also, surgeons believe that our approach can afford a more comfortable posture throughout the surgery and enhance hand-eye coordination, as physicians no longer need to twist their necks to look at screens placed outside the field of operation.


Aging of the erythrocyte. VII. On the possible causes of inactivation of red cell enzymes.
The sequence of inactivation of five bovine erythrocyte enzymes (glucose-6-phosphate dehydrogenase, pyruvate kinase, lactate dehydrogenase, aspartate aminotransferase and aldolase) was compared during in vivo aging of the red blood cell and during treatment of the cells with heat, gamma-radiation and an enzymatic source of the superoxide radical anion. Amounts of oxygen free radicals comparable to those formed physiologically in the cells were able to induce considerable inactivation of some enzymes; in no case was the physiological sequence of inactivation reproduced. These results seem to argue for a multifactorial inactivation of red cell enzymes during intravascular aging of the erythrocyte.


Chemometrics and in-line near infrared spectroscopic monitoring of a biopharmaceutical Chinese hamster ovary cell culture: prediction of multiple cultivation variables.
In the present study near infrared (NIR) spectroscopy was used to monitor the cultivation of mammalian Chinese hamster ovary (CHO) cells producing a monoclonal antibody in a fed-batch cell culture process. A temperature shift was applied during the cultivation. The cells were incubated at 37 °C and 33 °C. The Fourier transform near infrared (FT-NIR) multiplex process analyzer spectroscopy was investigated to monitor cultivation variables of the CHO cell culture from 10 independent batches using two channels of the FT-NIR. The measurements were performed on production scale bioreactors of 12,500 L. The cell cultures were analyzed with the spectrometer coupled to a transflection sterilizable fiber optic probe inserted into the bioreactors. Multivariate data analysis (MVDA) employing unsupervised principal component analysis (PCA) and partial least squares regression methods (PLS) were applied. PCA demonstrated that 96% of the observed variability was explained by the process trajectory and the inter-batch variability. PCA was found to be a significant tool in identifying batch homogeneity between lots and in detecting abnormal fermentation runs. Seven different cell culture parameters such as osmolality, glucose concentration, product titer, packed cell volume (PCV), integrated viable packed cell volume (ivPCV), viable cell density (VCD), and integrated viable cell count (iVCC) were monitored inline and predicted by NIR. NIR spectra and reference analytics data were computed using control charts to evaluate the monitoring abilities. Control charts of each media component were under control by NIR spectroscopy. The PLS calibration plots offered accurate predictive capabilities for each media. This paper underlines the capability for inline prediction of multiple cultivation variables during bioprocess monitoring.


Disruption of anterior insula modulation of large-scale brain networks in schizophrenia.
Systems level modeling of functional magnetic resonance imaging data has demonstrated dysfunction of several large-scale brain networks in schizophrenia. Anomalies across multiple functional networks associated with schizophrenia could be due to diffuse pathology across multiple networks or, alternatively, dysfunction at converging control(s) common to these networks. The right anterior insula has been shown to modulate activity in the central executive and default mode networks in healthy individuals. We tested the hypothesis that right anterior insula modulation of central executive and default mode networks is disrupted in schizophrenia and associated with cognitive deficits. In 44 patients with schizophrenia and 44 healthy control subjects, we used seed-based resting state functional connectivity functional magnetic resonance imaging analysis to examine connectivity between right insular subregions and central executive/default mode network regions. We also performed two directed connectivity analyses of resting state data: Granger analysis and confirmatory structural equation modeling. Between-group differences in path coefficients were used to evaluate anterior insula modulation of central executive and default mode networks. Cognitive performance was assessed with the rapid visual information processing task, a test of sustained attention. With multiple connectivity techniques, we found compelling, corroborative evidence of disruption of right anterior insula modulation of central executive and default mode networks in patients with schizophrenia. The strength of right anterior insula modulation of these networks predicted cognitive performance. Individuals with schizophrenia have impaired right anterior insula modulation of large-scale brain networks. The right anterior insula might be an emergent pathophysiological gateway in schizophrenia.


Effects of aerobic exercise on depression-like behavior and TLR4/NLRP3 pathway in hippocampus CA1 region of CUMS-depressed mice.
The paper observes regulation of the expression levels of NLRP3 and TLR4 in hippocampal CA1 neurons in CUMS mice by aerobic exercise with constructing CUMS depression mouse model, in order to explore the neuroprotective mechanism of aerobic exercise on the hippocampus of depressed mice. 24 healthy male 8-week-old C57BL/6 mice were randomly divided into CG, MG and ME. Thirteen stress-stimulating factors were randomly formulated into a CUMS stress-stimulating program. The mice were underwent 28 days of CUMS depression model, referenced clinical means for experimental research. The study was approved by the Ethics Committee of Yichun University (YCUEC IRB number LSK NO.2022.18). After model preparation, ME mice were subjected to moderate-intensity treadmill exercise training for 8 weeks. TST, FST and SPT were used to detect the depression-like behaviors of the mice in each group. Nissl staining was used to compare the cell morphology in the CA1 region of the mouse hippocampus. Immunohistochemical staining and western blot were used to detect the changes in the expression levels of NLRP3, TLR4 and other proteins in the CA1 region of the mouse hippocampus. The results of neurobehavioral assessment showed that, the immobility time of TST and FST were significantly increased, and SPT index was significantly decreased of MG mice. Compared with MG, ME mice significantly improved depression-like behaviors such as TST, FST and SPT index. Nissl staining showed that the morphology of neurons in CA1 region of hippocampus of MG mice were mostly vacuolar-like, with severe nuclear pyknosis. Abnormal morphological changes such as vacuolar-like and pyknotic pyknosis of neurons in the hippocampal CA1 region of ME mice were significantly reduced. Protein expression test showed that the number of NLRP3, TLR4, IL-1β and IL-10 positive neurons in hippocampal CA1 region of MG mice increased significantly compared with CG, and the proportion of positive cells increased significantly, while NLRP3 and TLR4 positive neurons in the hippocampal CA1 region of ME mice were significantly reduced, the proportion of TLR4 positive cells was significantly reduced. Systematic moderate-intensity exercise can effectively improve the depression-like behavior of CUMS depressed mice through the expression of TLR4/NLRP3 inflammatory signaling pathway, and provide an effective experimental basis for the clinical rehabilitation treatment of depression.


Health Outcomes of HIV-Infected People with Mental Illness.
Improving outcomes for people with HIV and mental illness will be critical to meeting the goals of the US National HIV/AIDS Strategy. In a retrospective analysis of the 2008-2010 cycles of the locally representative Philadelphia Medical Monitoring Project, we compared the proportions of HIV-infected adults with and without mental illness: (1) retained in care (≥2 primary HIV visits separated by ≥90 days in a 12-month period); (2) prescribed antiretroviral therapy (ART) at any point in a 12-month period; and (3) virally suppressed (HIV-1 RNA ≤200 copies/mL at the last measure in the 12-month period). Multivariable regression assessed associations between mental illness and the outcomes, adjusting for age, gender, race/ethnicity, insurance, alcohol abuse, injection drug use, CD4 count, and calendar year. Of 730 HIV-infected persons, representative of 9409 persons in care for HIV in Philadelphia, 49.0 % had mental illness. In adjusted analyses, there were no significant differences in retention (91.3 vs. 90.3 %; AOR 1.30, 95 % CI 0.63-2.56) and prescription of ART (83.2 vs. 88.7 %; AOR 0.79, 95 % CI 0.49-1.25) between those with and without mental illness. However, mentally ill patients were less likely to achieve viral suppression than those without mental illness (65.9 vs. 74.4 %; AOR 0.64, 95 % CI 0.46-0.90). These findings argue for the need to optimize ART adherence in this population.


[Effect of various methods of administration of live poliovirus vaccine on the immunoglobulin levels in the blood serum and mouth mucosa secretion in children].
Oral vaccination with live trivalent poliomyelitis in the liquid form in drops exerts a considerable influence on the IgA content in oral cavity secretes. After immunization with same vaccine in capsules no significant changes in IgA levels could be observed in oral cavity secretes for a 18-week observation period. A considerable increase in IgA level in oral secretes in children vaccinated with vaccine in capsules was observed after a 4th administration of the vaccine given orally in drops. Inrrespective of the method of vaccine administrations in the blood serum there was a considerable increase in the levels of immunoglobulins of all classes in the course of vaccination.


Histopathological comparison of pathergy positive and negative areas of newly diagnosed Behçet patients.
Behcet's disease is an inflammatory vasculitis, and pathergy reaction is a compulsory element of the diagnostic criteria. In the literature, there is no study comparing the histopathology of negative pathergy tests and positive pathergy tests in newly diagnosed Behcet's patients. The primary goal was to investigate the frequency of vasculitis, which is an important finding of Behcet's disease, in pathergy histopathology. The histopathology of patients with positive and negative pathergy tests revealed vasculitis or vasculopathy. In histopathology, neutrophils, lymphocytes, eosinophils, and mast cells in the perivascular area were counted and recorded as a percentage. Of the 88 Behcet's patients who participated in the study, 45 were female and 43 were male. The rate of vasculitis in the histopathology of those who were positive for the pathergy test was not statistically different than those of those with negative pathergy tests (P = .14). In our study, although the relationship between the male gender and uveitis was significant at the border, and the relationship between pathergy positivity and uveitis was not found to be significant, a significant relationship was found between vasculitis in histopathology and uveitis (P = .023). Based on the results of our study, we think that a pathergy test of histopathology will contribute to the clinical diagnostic evaluation in cases with suspicious clinical pathergy tests and especially in cases with negative pathergy tests.


Medication-related emergency department visits and hospital admissions in pediatric patients: a qualitative systematic review.
To review and describe the current literature pertaining to the incidence, classification, severity, preventability, and impact of medication-related emergency department (ED) and hospital admissions in pediatric patients. A systematic search of PubMED, Embase, and Web of Science was performed using the following terms: drug toxicity, adverse drug event, medication error, emergency department, ambulatory care, and outpatient clinic. Additional articles were identified by a manual search of cited references. English language, full-reports of pediatric (≤18 years) patients that required an ED visit or hospital admission secondary to an adverse drug event (ADE) were included. We included 11 studies that reported medication-related ED visit or hospital admission in pediatric patients. Incidence of medication-related ED visits and hospital admissions ranged from 0.5%-3.3% and 0.16%-4.3%, respectively, of which 20.3%-66.7% were deemed preventable. Among ED visits, 5.1%-22.1% of patients were admitted to hospital, with a length of stay of 24-72 hours. The majority of ADEs were deemed moderate in severity. Types of ADEs included adverse drug reactions, allergic reactions, overdose, medication use with no indication, wrong drug prescribed, and patient not receiving a drug for an indication. Common causative agents included respiratory drugs, antimicrobials, central nervous system drugs, analgesics, hormones, cardiovascular drugs, and vaccines. Medication-related ED visits and hospital admissions are common in pediatric patients, many of which are preventable. These ADEs result in significant healthcare utilization.


Retrospective cross-sectional review of survival rates in critically ill children admitted to a combined paediatric/neonatal intensive care unit in Johannesburg, South Africa, 2013-2015.
Report on survival to discharge of children in a combined paediatric/neonatal intensive care unit (PNICU). Retrospective cross-sectional record review. All children (medical and surgical patients) admitted to PNICU between 1 January 2013 and 30 June 2015. Primary outcome-survival to discharge. Secondary outcomes-disease profiles and predictors of mortality in different age categories. There were 1454 admissions, 182 missing records, leaving 1272 admissions for review. Overall mortality rate was 25.7% (327/1272). Mortality rate was 41.4% (121/292) (95% CI 35.8% to 47.1%) for very low birthweight (VLBW) babies, 26.6% (120/451) (95% CI 22.5% to 30.5%) for bigger babies and 16.2% (86/529) (95% CI 13.1% to 19.3%) for paediatric patients. Risk factors for a reduced chance of survival to discharge in paediatric patients included postcardiac arrest (OR 0.21, 95% CI 0.09 to 0.49), inotropic support (OR 0.085, 95% CI 0.04 to 0.17), hypernatraemia (OR 0.16, 95% CI 0.04 to 0.6), bacterial sepsis (OR 0.32, 95% CI 0.16 to 0.65) and lower respiratory tract infection (OR 0.54, 95% CI 0.30 to 0.97). Major birth defects (OR 0.44, 95% CI 0.26 to 0.74), persistent pulmonary hypertension of the new born (OR 0.44, 95% CI 0.21 to 0.91), metabolic acidosis (OR 0.23, 95% CI 0.12 to 0.74), inotropic support (OR 0.23, 95% CI 0.12 to 0.45) and congenital heart defects (OR 0.29, 95% CI 0.13 to 0.62) predicted decreased survival in bigger babies. Birth weight (OR 0.997, 95% CI 0.995 to 0.999), birth outside the hospital (OR 0.21, 95% CI 0.05 to 0.84), HIV exposure (OR 0.54, 95% CI 0.30 to 0.99), resuscitation at birth (OR 0.49, 95% CI 0.25 to 0.94), metabolic acidosis (OR 0.25, 95% CI 0.10 to 0.60) and necrotising enterocolitis (OR 0.23, 95% CI 0.12 to 0.46) predicted poor survival in VLBW babies. Ongoing mortality review is essential to improve provision of paediatric critical care.


Purification and characterisation of intracellular alpha-galactosidases from Acinetobacter sp.
Two alpha-galactosidases (Ag-I & Ag-II) were purified from Acinetobacter sp. Both the enzymes were monomeric with pH optima of 7.0 and molecular weight of 65 kDa for Ag-I and 37 kDa for Ag-II. The temperature optima for Ag-I was between 50 and 60 °C and that of Ag-II was 40 °C. Both the enzymes were strongly inhibited by metal ions Ag2+ and Hg+, pCMB and SDS (1 %). The enzymes were found to be active on both natural and synthetic substrates. Artificial substrate, pNPGal, has shown more affinity to enzyme than natural substrate raffinose. The half-life (t 1/2) of Ag-I varied from 1.85 h at 90 °C to 7.6 h at 70 °C.


Kinetics of immune responses elicited after three mRNA COVID-19 vaccine doses in predominantly antibody-deficient individuals.
Mass vaccination campaigns reduced COVID-19 incidence and severity. Here, we evaluated the immune responses developed in SARS-CoV-2-uninfected patients with predominantly antibody-deficiencies (PAD) after three mRNA-1273 vaccine doses. PAD patients were classified based on their immunodeficiency: unclassified primary antibody-deficiency (unPAD, n = 9), common variable immunodeficiency (CVID, n = 12), combined immunodeficiency (CID, n = 1), and thymoma with immunodeficiency (TID, n = 1). unPAD patients and healthy controls (HCs, n = 10) developed similar vaccine-induced humoral responses after two doses. However, CVID patients showed reduced binding and neutralizing titers compared to HCs. Of interest, these PAD groups showed lower levels of Spike-specific IFN-γ-producing cells. CVID individuals also presented diminished CD8+T cells. CID and TID patients developed cellular but not humoral responses. Although the third vaccine dose boosted humoral responses in most PAD patients, it had limited effect on expanding cellular immunity. Vaccine-induced immune responses in PAD individuals are heterogeneous, and should be immunomonitored to define a personalized therapeutic strategies.


Reproductive abnormalities in canine fucosidosis.
Male English springer spaniel dogs affected with fucosidosis, a lysosomal storage disorder, were found to be infertile while females with the disease reproduced successfully. Ejaculates of semen collected from affected dogs had reduced total sperm output and morphologically abnormal spermatozoa. A high proportion of ejaculated spermatozoa had midpiece droplets, bent tails and poor motility. Severely vacuolated epididymal epithelial cells were observed by light microscopy. Electron microscopic examination revealed membrane-bound vacuoles of variable size containing scanty amounts of granular to fibrillar material in epididymal epithelial cells, smooth muscle, myoid cells and Sertoli cells. Male infertility is believed to result from lysosomal storage of fucosyl-linked substrates in cells of the reproductive system. The extensive lesions in the epididymis may have interfered with maturation and transport of spermatozoa. Also, deficiency of alpha-L-fucosidase activity could have impaired the shedding of cytoplasmic droplets from spermatozoa and altered the surface glycoprotein composition of the sperm during epididymal transit.


Tanshinone IIA derivatives induced S-phase arrest through stabilizing c-myc G-quadruplex DNA to regulate ROS-mediated PI3K/Akt/mTOR pathway.
Herein, a derivate from tanshinone IIA, 1,6,6-trimethyl-11-phenyl-7,8,9,10-tetrahydro-6H-furo[2',3':1,2]phenanthro[3,4-d]imidazole (TA25), has been synthesized and investigated as potential inhibitor against the proliferation, migration and invasion of lung cancer cells. MTT assay and cell colony formation assay results showed that TA25 exhibits acceptable inhibitory effect against the proliferation of lung cancer A549 cells, and the value of IC50 was about 17.9 μM. This result was further confirmed by the inhibition of TA25 against the growth of xenograft lung cancer cells on zebrafish bearing tumor (A549 lung cancer cells). The results of wound-healing assay and FITC-gelatin invasion assay displayed that TA25 could inhibit the migration and invasion of lung cancer A549 cells. Moreover, the studies on the binding properties of TA25 interact with c-myc G-quadruplex DNA suggested that TA25 can bind in the G-quarter plane formed from G7, G11, G16 and G20 with c-myc G-quadruplex DNA through π-π stacking. Further study of the potential anti-cancer mechanism indicated that TA25 can induce S-phase arrest in lung cancer A549 cells, and this phenomenon resulted from the promotion of the production of reactive oxygen species and DNA damage in A549 cells under the action of TA25. Further research revealed that TA25 could inhibit the PI3K/Akt/mTOR signal pathway and increase the expression of p53 protein. Overall, TA25 can be developed into a promising inhibitor against the proliferation, migration and invasion of lung cancer cells and has potential clinical application in the near future.


Responsiveness of the first combination band of water to the state in organic and polymeric medium.
We studied the influence of intermolecular interactions on the first combination band between the OH-stretching and -deformation vibrations of the water that is dissolved in organic and polymeric materials. The water has two types. The first type occurs in the matrix in which water-interactive functional groups are densely distributed. The water is symmetrically hydrogen-bonded through each OH to two functional groups and shows one clear combination band, of which the frequency depends on the strength of the hydrogen bond. The second type occurs in the matrix in which the functional groups are rarely distributed. One OH of this water is hydrogen-bonded to the functional group but the other OH is free. The water shows two combination bands, of which one is sharp and strong at about 5300 cm-1 and the other is weaker and broader, the frequency changing in the 5200-5100 cm-1 range. Thus, the dissolved water sensitively responds to the nature of a water-interactive functional group and its distribution density in a matrix, and shows the first combination band that is characteristic of these factors.


Secular trends of hip fractures in France: impact of changing characteristics of the background population.
Hip fractures are a societal burden because of their high morbidity and mortality and the cost they generate. With the aging of the population, worries grow about an increase of the incidence and incidence rate of hip fracture in the future. Controversial data have been provided in relation to the referencepopulation used. The aim of this study was to assess the impact of the choice of the reference population in the incidence rate of hip fracture. Data were extracted from the French National Hospital Database related to the hospitalizations for hip fracture in France between 2002 and 2013 in patients over 59 years and were classified by gender and age (59-74, 75-84, over 84 years, over 59 years). The crude incidence rates of hip fracture were calculated by dividing the number of hospitalizations for hip fracture by the corresponding populations. To assess the impact of the choice of the reference population, we then calculated the adjusted incidence rates using direct standardization on age for the 2013 reference population. From 2002 to 2013, the incidence of hip fracture rose by 4.8% in women (from 49,287 to 51,661) and 21.8% in men (from 12,716 to 15,482) aged over 59 years. Meanwhile, French population over 59 years increased more with a rise of 21.3% in women and 28.7% in men, resulting in a decrease in the crude incidence rates of 13.6% in women and 5.4% in men. However, this decrease was larger after direct standardization on the 2013 population of reference as 25.6% in women and 19.2% in men as a result of a difference in age-structure of the population. The incidence of hip fractures continues to grow despite a reduced incidence rate throughout a 12-year-period.


On the relationship between reproduction, age and survival in two carbid beetles: : Calathus melanocephalus L. and Pterostichus coerulescens L. (Coleoptera, Carabidae).
In this study the inverse relationship between the amount of reproduction in female carabid beetles and their survival until the next breeding season, suggested by Murdoch (1966), was examined in Pterostichus coerulescens L. and Calathus melanocephalus L. This was tested with individual and differently reproducing females, kept at the same temperature, as well as with groups of females kept at different temperatures, i.e. with very different overall levels of reproduction.No relationship was found between the numbers of eggs laid by individual females and their survival until the next breeding season.The numbers of eggs laid by individual females varies greatly, even under constant and optimal conditions,. In contrast to this, an individual female continues to lay a characteristic number of eggs over several succeeding weeks. The amount of reproduction is highly correlated with temperature when food is optimal. In P. coerulescens there was a positive correlation between the amount of reproduction of the same individuals in two succeeding years, whereas in C. melanocephalus no relationship could be found.Both in the field and in our experiments individuals of P. coerulescens generally live for 3 to 4 years, longer than C. melanocephalus which usually only lives for 2 years. The 3-4 year-old individuals of P. coerulescens die during the breeding season or shortly afterwards, but also during the winter, whereas the 2-3 year-old individuals of C. melanocephalus die during the winter. Males and females show a similar mortality after breeding. The level of reproduction of a population depends on the proportion of old beetles, since especially the very old ones die before the end of the breeding season.We reject the hypothesis of Murdoch because of these data. However, in a different way from that of Murdoch, we also suggest that old beetles are of great importance for the survival of a population, namely that this form of heterogeneity of a population may enhance its stability under varying environmental conditions.


Two-tier approach combining molecular and culture-based techniques for optimized detection of vancomycin-resistant enterococci.
Given constantly high or even rising incidences of both colonization and infection with vancomycin-resistant enterococci (VRE), timely and accurate identification of carriers in high-risk patient populations is of evident clinical importance. In this study, a two-tier approach consisting of PCR-based screening and cultural confirmation of positive results is compared to the conventional approach solely based on culture on selective media. The 2-tier strategy was highly consistent with the conventional approach, and was found to possess high sensitivity and specificity (93.1% and 100%, respectively). The introduction of the PCR-based combined VRE screening approach significantly (P<0.0001) reduced median overall time to result by 44.3hours. The effect was found to be most pronounced in VRE negative samples. Positive vanA PCR was highly consistent with culture (PPV: 92.0%, 95% CI: 72.5-98.6%, NPV: 99.6%, 95% CI: 98.9-99.6%), thus allowing for preliminary reporting of VRE detection. In contrast, a vanB positive PCR does not allow for preliminary reporting (PPV: 58.5%, 95% CI: 44.2-71.6%, NPV: 99.8%, 95% CI: 99.2-100%). The introduction of a molecular assay for rapid detection of VRE from rectal swabs combined with cultural confirmation proved to be reliable and time saving, especially in a setting of low VRE prevalence and predominance of vanA positive strains.


[Effects of dopamine and orciprenalin on human metabolism (author's transl)].
10 healthy probands were given on several subsequent days by intravenous infusion lasting 20' only orciprenalin and dopamine in therapeutic doses of 0.1 mg/kg bodyweight and 0.5 mg, respectively. In all cases application of orciprenalin resulted in a significant rise of glucose, insuline, lactate and free fatty acids. By contrast, no such changes were observed after infusion of dopamine. This suggests that a rise of free fatty acids in the serum may be important for the occurrence of cardiac arrythmia.


Genetical and biological control of cotton ashy stem caused by Macrophomina phaseolina in outdoor pot experiment.
Two outdoor pot experiments were carried out to evaluate the reaction of 11 commercial Egyptian cotton cultivars Macrophomina phaseolina, the incitant of ashy stem in cotton and to evaluate the antagonistic ability of 27 isolates of Trichoderma sp. against pathogen cotton cultivars Giza 85, Giza 87, Giza 89 and Giza 90 were resistant to M. phaseolina because both survival and plant height of these cultivars was not affected when the soil was infested with the pathogen. None of the cultivars were found to be immune to highly pathogenic of M. phaseolina isolate. Of the 27 isolate's of Trichoderma that were evaluated, the best antagonistic performance was given by isolates nos. 2, 10, and 16 were promising for commercialization because they significantly increased survival and improved plant height and dry weight of the surviving cotton seedlings.


Glutamate receptors and persistent pain: targeting forebrain NR2B subunits.
Glutamate is the fast excitatory transmitter in mammalian brains. It binds to two major classes of glutamate receptors: ionotropic and metabotropic receptors. Ionotropic receptors contain three subtype receptors, including N-methyl-d-aspartate (NMDA) receptors. Activation of NMDA receptors is important for initiating long-lasting changes in synapses. In the forebrain structures that are known to contribute to the formation and storage of information, NMDA receptors have an important role in persistent inflammatory pain by reinforcing glutamate sensory transmission. Mice with enhanced forebrain NMDA receptor function demonstrate selective enhancement of persistent pain and allodynia. Drugs targeting NMDA NR2B subunits in the forebrain could serve as a new class of medicine for controlling persistent pain in humans.


Role of protein kinase Cdelta in paraquat-induced glial cell death.
Paraquat (1,1'-dimethyl-4,4'-bipyridinium) is structurally similar to the neurotoxin 1-methyl-4-phenyl-4-phenylpyridium ion (MPP+), the active metabolite of the parkinsonism-inducing agent 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), which can induce the parkinsonism property in rodents, nonhuman primates, and human. In contrast to the neurotoxic effects of paraquat, little is known about its effects on glial cells. Here, we examined the mechanisms of paraquat toxicity in glial cells in culture. Paraquat treatment also reduced the viability of C6 glial cells in primary astrocyte cultures, and cell death was mostly apoptotic in nature. PKCdelta played a central role in the paraquat-induced glial cell death: (1) the PKCdelta-specific inhibitor rottlerin blocked paraquat-induced glial cell death; (2) paraquat induced tyrosine and threonine phosphorylation of PKCdelta; and (3) transfection of the dominant-negative mutant of PKCdelta attenuated paraquat toxicity. PKCdelta was also involved in the generation of reactive oxygen species (ROS), which mediated the paraquat toxicity. The nicotinamide adenine dinucleotide phosphate (reduced form) oxidase (NADPH oxidase) inhibitor diphenyleneiodonium blocked the paraquat-induced ROS production and subsequent cell death, indicating the involvement of NADPH oxidase in the cytotoxic action of paraquat in glia. PKCdelta was also important in glial cell death induced by MPP+ but not in that induced by rotenone. Last, Rac1 appeared to antagonize paraquat toxicity in glia. These results indicate a gliotoxic effect of paraquat and an opposing role of PKCdelta and Rac1 in paraquat-induced glial cell death.


Is fetal gender associated with adverse perinatal outcome in intrauterine growth restriction (IUGR)?
The purpose of this study was to determine if there is a difference in perinatal outcome by gender among growth-restricted fetuses. This was a retrospective cohort study of intrauterine growth restriction (IUGR) singleton pregnancies over a 5-year period. Clinical outcomes compared by gender included preterm delivery, perinatal mortality (PNM), respiratory distress syndrome (RDS), grade 3 or 4 intraventricular hemorrhage (IVH), necrotizing enterocolitis (NEC), and periventricular leukomalacia (PVL). Statistical analysis included bivariate and multivariable techniques. Seven hundred and twenty-seven singleton pregnancies with IUGR were identified. Three hundred and forty-six (47.6%) were males. Birth weight was similar between the groups. After adjusting for maternal demographics, medical history, gestational age, mode of delivery, and antenatal corticosteroids, adverse perinatal outcomes were similar between the groups. Severity of outcomes was also similar between males and females (P = .66). Male fetuses with IUGR have similar outcomes when compared with female IUGR fetuses. Gender does not play a role in perinatal outcome in the setting of fetal growth restriction.


A new familial 'giant platelet syndrome' with structural, metabolic and functional abnormalities of platelets due to a primary megakaryocyte defect.
We have investigated platelet morphology, function and biochemistry in a family affected by dominantly inherited congenital thrombocytopenia with giant platelets. A defect of aggregation was defined with evidence of disturbed fatty acid distribution in platelet membrane phospholipids and impaired arachidonic acid mobilization. Release of 5-hydroxytryptamine was normal. Platelet ultrastructure was grossly abnormal and volume analysis revealed small as well as giant forms. Bone marrow megakaryocytes were morphologically abnormal and the distribution of mean nuclear ploidy atypical, suggesting the production of abnormal platelets due to a primary megakaryocyte disorder. The features of this condition are distinct from those previously described in familial thrombocytopenia and constitute a new "giant platelet syndrome". The qualitative and quantitative platelet defects may be secondary to a disturbance of megakaryocyte cytoplasmic fragmentation.


Renal function at hospital admission and mortality due to acute kidney injury after myocardial infarction.
The role of an impaired estimated glomerular filtration rate (eGFR) at hospital admission in the outcome of acute kidney injury (AKI) after acute myocardial infarction (AMI) has been underreported. The aim of this study was to assess the influence of an admission eGFR<60 mL/min/1.73 m(2) on the incidence and early and late mortality of AMI-associated AKI. A prospective study of 828 AMI patients was performed. AKI was defined as a serum creatinine increase of ≥ 50% from the time of admission (RIFLE criteria) in the first 7 days of hospitalization. Patients were divided into subgroups according to their eGFR upon hospital admission (MDRD formula, mL/min/1.73 m(2)) and the development of AKI: eGFR ≥ 60 without AKI, eGFR<60 without AKI, eGFR ≥ 60 with AKI and eGFR<60 with AKI. Overall, 14.6% of the patients in this study developed AKI. The admission eGFR had no impact on the incidence of AKI. However, the admission eGFR was associated with the outcome of AMI-associated AKI. The adjusted hazard ratios (AHR, Cox multivariate analysis) for 30-day mortality were 2.00 (95% CI 1.11-3.61) for eGFR<60 without AKI, 4.76 (95% CI 2.45-9.26) for eGFR ≥ 60 with AKI and 6.27 (95% CI 3.20-12.29) for eGFR<60 with AKI. Only an admission eGFR of <60 with AKI was significantly associated with a 30-day to 1-year mortality hazard (AHR 3.05, 95% CI 1.50-6.19). AKI development was associated with an increased early mortality hazard in AMI patients with either preserved or impaired admission eGFR. Only the association of impaired admission eGFR and AKI was associated with an increased hazard for late mortality among these patients.


Age-related changes to triceps surae muscle-subtendon interaction dynamics during walking.
Push-off intensity is largely governed by the forces generated by the triceps surae (TS) muscles (gastrocnemius-GAS, soleus-SOL). During walking, the TS muscles undergo different fascicle kinematics and contribute differently to biomechanical subtasks. These differences may be facilitated by the Achilles tendon (AT), which is comprised of subtendons that originate from the TS muscles. We and others have revealed non-uniform displacement patterns within the AT-evidence for sliding between subtendons that may facilitate independent muscle actuation. However, in older adults, we have observed more uniform AT tissue displacements that correlate with reduced push-off intensity. Here, we employed dual-probe ultrasound imaging to investigate TS muscle length change heterogeneity (GAS-SOL) as a determinant of reduced push-off intensity in older adults. Compared to young, older adults walked with more uniform AT tissue displacements and reduced TS muscle length change heterogeneity. These muscle-level differences appeared to negatively impact push-off intensity-evidenced by between-group differences in the extent to which TS muscle length change heterogeneity correlates with mechanical output across walking tasks. Our findings suggest that the capacity for sliding between subtendons may facilitate independent TS muscle actuation in young adults but may restrict that actuation in older adults, likely contributing to reduced push-off intensity.


Postprandial Hemodynamic Changes in Hypertensives Treated with Calcium Antagonists or ACE Inhibitors.
Our aim was to investigate possible abnormalities in postprandial hemodynamic changes in hypertensives treated with different vasodilating drugs, calcium antagonists, or angiotensin-converting enzyme inhibitors. Eleven healthy subjects and 22 hypertensive patients effectively treated with an angio tensin-converting enzyme inhibitor (ACEI) (n = 9) or calcium antagonists (n = 13) were studied. Cardiac output and blood pressure were monitored every 20 min from 2 h before lunch to 3 h after using a computer-assisted impedance cardiograph coupled with an automatic blood pressure monitor. After meals, a significant decrease in mean arterial pressure (minus sign7.9% plus minus 2.1) was observed in ACEI-treated hypertensives when compared with the minor changes observed in calcium-antagonist-treated hypertensives (minus sign3.7% plus minus 1.5) and in normotensives (minus sign2.7% plus minus 1.5). When compared with normotensives, the patients treated with ACEI showed a larger postprandial fall in total peripheral resistance index (minus sign20.8% plus minus 3.4 versus minus sign15.3% plus minus 4.1) with a larger increase in heart rate (11.3% plus minus 2.3 versus 8.1% plus minus 1.3). In hypertensives treated with calcium antagonists, the postprandial hemodynamic changes appeared blunted and not significant. Different antihypertensive drugs appear to have different effects on the postprandial hemodynamic changes.


A diphenyl ether derived bidentate secondary phosphine oxide as a preligand for nickel-catalyzed C-S cross-coupling reactions.
A new bidentate secondary phosphine oxide (SPO) was synthesized from diphenyl ether via ortho-lithiation, phosphorylation with PhP(Cl)NEt2, and hydrolysis in an acidic medium. Nickel(0) species ligated with this new SPO was established as a more effective catalyst than Ni(0)-Ph2P(O)H for the cross-coupling of aryl iodides with aryl thiols.


Prognosis of diabetics with diabetes onset before the age of thirty-one. II. Factors influencing the prognosis.
In 307 patients with diabetes mellitus, developed prior to 1933 and before age 31 it was demonstrated that: (1) frequent contact with a specialized diabetes clinic from an early stage of the disease; (2) a good quality of "metabolic control"; (3) a low insulin dose; (4) a body weight of 10% less than ideal; and (5) a mean blood pressure below 100 mm Hg, all had significantly beneficial effects upon the survival. It was also found that patients domiciled in Copenhagen had a significantly better prognosis than patients domiciled outside Copenhagen. Frequent contact with a diabetes centre was accompanied by an appreciable decrease in disabling late diabetic complications.


Rectus abdominus tear in a collegiate cheerleader.
Acute muscle injuries can occur at all skill levels across multiple different sports. Imaging is often used for diagnosis and is necessary for recognizing the extent of injury, complications, and overall prognosis. Ultrasound can be used in diagnosing muscle tears. However, there are less than a dozen documented cases that use ultrasound in the evaluation and diagnosis of a rectus abdominus tear, though the actual number of cases are most likely underreported in the literature. This case discusses an 18-year-old college athlete that suffered a distal rectus abdominus tear after performing a stunt as a cheerleader and her subsequent treatment.


A Cross-Cultural Adaptation and Content Validity of COMFORTneo Scale into Brazilian Portuguese.
The instrument used to assess neonatal pain must be adequate regarding the type of pain, population, country, and language to provide the best evidence-based clinical strategies; however, few neonatal pain instruments have been translated and validated for the Brazilian population. The aim was to perform a cross-cultural adaptation of the COMFORTneo scale into Brazilian Portuguese and to evaluate the content validity of the adapted scale. The cross-cultural adaptation process followed six main steps: translation, synthesis of the translations, back-translation, submission to the expert committee, final version pretest, being that 65 individuals participated in this stage, including both healthcare professionals and students, and submission to the committee for process appraisal. Additionally, an equivalence form composed of a four-point Likert scale was sent to each committee participant to calculate the content validity index (CVI). The CVI was obtained as the sum of the items ranked as three or four by the experts divided by the total number of experts. No difficulties were reported in the production of translated versions. The CVI for the final version of the translated instrument was 0.99. The final version was reviewed to correct any possible grammatical errors. The layout was modified as necessary, and instructions on scale scoring were added to facilitate the application, resulting in the COMFORTneo Brazil scale. The COMFORTneo scale was properly and cross-culturally adapted into Brazilian Portuguese, reaching semantic, idiomatic, experimental, and conceptual equivalence with the original instrument, and a good CVI.


Transaldolase deficiency in a two-year-old boy with cirrhosis.
Transaldolase (TALDO) deficiency is a rare inborn error of the pentose phosphate pathway. We report the clinical presentation and laboratory findings of a new patient with TALDO deficiency. The two-year-old Arabic boy presented with neonatal onset of anemia and thrombocytopenia, tubulopathy, and rickets and was subsequently found to have cirrhosis and deafness. A comparison with other TALDO deficient patients is given.


Dopamine receptor blockade increases glandular kallikrein-like activity in the neurointermediate lobe of the rat pituitary.
The effect of dopamine receptor blockers on glandular kallikrein-like activity in the neurointermediate lobe of the rat pituitary was examined. Male rats were given daily injections of haloperidol (2.5 mg/kg), perphenazine (5 mg/kg) or sulpiride (60 mg/kg) for 7 days. Homogenates of the neurointermediate lobe were prepared. Latent proteases were activated with trypsin and proteolytic activity was measured at 37 degrees C, pH 8.0 using chromogenic peptide substrates. All three dopamine receptor blockers produced about a 100% increase in glandular kallikrein-like activity. The results suggest that glandular kallikrein-like activity in the neurointermediate lobe is under inhibitory control by dopaminergic mechanisms.


Establishment of growth factor-dependent MOPC 104E cell line in vitro.
The MOPC 104E cell line has been adapted to grow in vitro using a combination of feeder layer and growth factor(s). The growth of this myeloma cell line is dependent on the presence of growth factor(s). Growth-promoting activity generated from T-cell-mitogen-stimulated, Corynebacterium parvum-stimulated spleen cell culture supernatant, and peritoneal adherent cell culture supernatants gives dose-dependent proliferation. Generation of growth factors in the serum-free bovine serum albumin-substituted media and a rapid assay system based on [3H]thymidine uptake for the quantitation of growth promoting activity are described.


Aqueous penetration and biological activity of moxifloxacin 0.5% ophthalmic solution and gatifloxacin 0.3% solution in cataract surgery patients.
To measure the achievable perioperative aqueous concentration of the commercially available topically administered fourth generation fluoroquinolones, moxifloxacin 0.5% ophthalmic solution, and gatifloxacin 0.3% ophthalmic solution, and to correlate this concentration with the agents' biological efficacy in the aqueous humor of patients undergoing routine cataract surgery. Prospective, randomized, parallel, double-masked, clinical trial. Fifty patients undergoing cataract extraction. Patients (n = 25) were given perioperative topical moxifloxacin 0.5% or topical gatifloxacin 0.3% (n = 25). One drop of antibiotic was administered every 10 minutes for 4 doses beginning 1 hour prior to surgery. Aqueous humor was sampled via paracentesis and antibiotic concentrations were determined using validated high performance liquid chromatography (HPLC) procedures. Dilution analyses were performed to determine the biological efficacy of the agents in the aqueous against Staphylococcus epidermidis, the most common cause of postcataract endophthalmitis. Aqueous humor antibiotic concentrations were measured using HPLC and microdilution bioassay techniques. Biological activity was measured as minimal inhibitory dilution and minimal bactericidal dilution. Aqueous humor concentrations for moxifloxacin via HPLC analysis were 1.80 (+/-1.21) microg/ml, whereas those for gatifloxacin were 0.48 (+/-0.34) microg/ml. This 3.8-fold difference in aqueous humor antibiotic concentrations was statistically significant (P = 0.00003). Similarly, the biological dilution analysis of the aqueous humor samples showed that moxifloxacin attained an estimated activity of 2.1 microg/ml, whereas the gatifloxacin activity was approximately 0.4 mug/ml, which represented a 4.9-fold difference. This study demonstrated that after topically administered perioperative antibiotics with cataract surgery, moxifloxacin 0.5% ophthalmic solution achieved a statistically significantly higher concentration in aqueous humor compared with gatifloxacin (P = 0.00003). Results from the broth dilution analysis showed that moxifloxacin 0.5% was biologically more active against S. epidermidis than gatifloxacin 0.3% in aqueous humor after topical application. There were no adverse events reported, and incision wounds healed quickly and as expected.


Characterization of bifunctional sphingolipid Δ4-desaturases/C4-hydroxylases of trypanosomatids by liquid chromatography-electrospray tandem mass spectrometry.
Six genes encoding putative sphingolipid desaturases have been identified in trypanosomatid genomes: one in Trypanosoma brucei (TbSLdes protein), one in Trypanosoma cruzi (TcSLdes) and four in Leishmania major (LmSLdes1-4), tandemly arrayed on chromosome 26. The six amino acid sequences showed the three characteristic histidine boxes, with a long spacer between the first and second box, as in fungal desaturases and bifunctional desaturases/hydroxylases, to which they are phylogenetically related. We functionally characterized the trypanosomatid enzymes by their expression in Saccharomyces cerevisiae sur2Δ mutant, which lacks C4-hydroxylase activity. The sphingoid base profile (dinitrophenyl derivatives) of each yeast mutant transformed with each one of the different parasite genes was analyzed by HPLC, using a sur2Δ mutant expressing the Schyzosaccharomyces pombe sphingolipid desaturase (SpSLdes) as positive control. TbSLdes was capable of desaturating endogenous sphingolipids at levels comparable to those found in SpSLdes. By contrast, L. major and T. cruzi enzymes showed either no or negligible activities. Using the HPLC system coupled to electrospray tandem quadrupole/time of flight mass spectrometry we were able to detect significant levels of desaturated and hydroxylated sphingoid bases in extracts of all transformed yeast mutants, except for those transformed with the empty vector. These results indicate that S. pombe, T. brucei, T. cruzi and L. major enzymes are all bifunctional. Using the same methodology, desaturated and hydroxylated sphingoid bases were detected in T. cruzi epimastigotes and L. major promastigote cells, as described previously, and in T. brucei procyclic and bloodstream forms for the first time.


Habenula as a Possible Target for Treatment-Resistant Depression Phenotype in Wistar Kyoto Rats.
The mechanisms of treatment-resistant depression (TRD) are not clear and are difficult to study. An animal model resembling human TRD is the Wistar Kyoto rat strain. In the present study, we focused on selecting miRNAs that differentiate rats of the WKY strain from Wistar Han (WIS) rats in two divisions of the habenula, the lateral and medial (LHb and MHb, respectively). Based on our preliminary study and literature survey, we identified 32 miRNAs that could be potentially regulated in the habenula. Six miRNAs significantly differentiated WKY rats from WIS rats within the MHb, and three significantly differentiated WKY from WIS rats within the LHb. Then, we selected relevant transcripts regulated by those miRNAs, and their expression in the habenular nuclei was investigated. For mRNAs that differentiated WKY rats from WIS rats in the MHb (Cdkn1c, Htr7, Kcnj9, and Slc12a5), their lower expression correlated with a higher level of relevant miRNAs. In the LHb, eight mRNAs significantly differentiated WKY from WIS rats (upregulated Htr4, Drd2, Kcnj5, and Sstr4 and downregulated Htr2a, Htr7, Elk4, and Slc12a5). These data indicate that several important miRNAs are expressed in the habenula, which differentiates WKY rats from WIS rats and in turn correlates with alterations in the expression of target transcripts. Of particular note are two genes whose expression is altered in WKY rats in both LHb and MHb: Slc12a5 and Htr7. Regulation of KCC2 via the 5-HT7 receptor may be a potential target for the treatment of TRD.


Anti-inflammatory (M2) macrophage media reduce transmission of oligomeric amyloid beta in differentiated SH-SY5Y cells.
Neuroinflammation plays an influential role in Alzheimer's disease (AD), although the mechanisms underlying this phenomenon remain largely unknown. Microglia are thought to be responsible for the majority of these effects and can be characterized into resting (M0), proinflammatory (M1), or anti-inflammatory (M2) functional phenotypes. We investigated the effects of conditioned macrophage media, as an analogue to microglia, on the transfer of oligomeric amyloid beta (oAβ) between differentiated SH-SY5Y cells. We also investigated how the different inflammatory environments related to intercellular and intracellular changes. We demonstrate that M2 products decrease interneuronal transfer of oAβ, while recombinant interleukin (IL)-4, IL-10, and IL-13 increase transfer. There were no alterations to the mRNA of a number of AD-related genes in response to the combination of oAβ and M0, M1, or M2, but several intracellular proteins, some relating to protein trafficking and the endosomal/lysosomal system, were altered. Stimulating microglia to an M2 phenotype may thus slow down the progression of AD and could be a target for future therapies.


Structure and function of the two-component flavin-dependent methanesulfinate monooxygenase within bacterial sulfur assimilation.
Methyl sulfur compounds are a rich source of environmental sulfur for microorganisms, but their use requires redox systems. The bacterial sfn and msu operons contain two-component flavin-dependent monooxygenases for dimethylsulfone (DMSO2) assimilation: SfnG converts DMSO2 to methanesulfinate (MSI-), and MsuD converts methanesulfonate (MS-) to sulfite. However, the enzymatic oxidation of MSI- to MS- has not been demonstrated, and the function of the last enzyme of the msu operon (MsuC) is unresolved. We employed crystallographic and biochemical studies to identify the function of MsuC from Pseudomonas fluorescens. The crystal structure of MsuC adopts the acyl-CoA dehydrogenase fold with putative binding sites for flavin and MSI-, and functional assays of MsuC in the presence of its oxidoreductase MsuE, FMN, and NADH confirm the enzymatic generation of MS-. These studies reveal that MsuC converts MSI- to MS- in sulfite biosynthesis from DMSO2.


The amino acid sequence of fragment A, an enzymically active fragment of diphtheria toxin. II. The cyanogen bromide peptides.
Cyanogen bromide cleavage of Fragment A from diphtheria toxin at the four methionines present in each molecule resulted in five major peptides which were isolated and studied by sequence methods. These five peptides of 4, 11, 14, 63, and 101 residues account for all 193 residues in Fragment A and provide overlaps for the tryptic peptides from the maleylated protein. Two additional peptides were isolated and shown to be shorter forms (8 and 10 residues) of the COOH-terminal cyanogen bromide peptide (11 residues).