Risk factors for nosocomial pneumonia in critically ill trauma patients.
To determine risk factors for nosocomial pneumonia in critically ill trauma patients. Prospective cohort study. The trauma intensive care unit (ICU) of a 1500-bed tertiary-care hospital. All critically ill trauma patients (n = 103) admitted consecutively between November 1995 and October 1996. A comparison of data recorded at the time of ICU admission and during the clinical evolution in patients with (n = 23) and without (n = 80) nosocomial pneumonia was made. Data referred mainly to possible risk factors were recorded; they also included factors related to pneumonia etiology and evolutive factors. Predictors of nosocomial pneumonia were assessed by logistic regression analysis. The presence of significant growth on quantitative cultures of the protected specimen brush (> or = 103 colony forming units/mL) was required to accept pneumonia as microbiologically proven, as well as the concurrence of a cohort of clinical and radiologic signs. Twenty-three (22.3%) patients developed nosocomial pneumonia. The mean age of these patients was 41.7 yrs; 18 of them (78.3%) were men. The microorganisms isolated in significant concentrations were Acinetobacter baumanii (ten cases), Staphylococcus aureus (11 cases), Pseudomonas aeruginosa (five cases), Haemophilus influenzae (two cases), and Klebsiella pneumoniae, Citrobacter freundii, Serratia marcescens, Enterococcus spp., Enterobacter spp., coagulase-negative Staphylococcus, and Streptococcus intermedius (one case each one). Risk factors for pneumonia by univariate analysis included nasogastric tube; continuous enteral feeding; prolonged mechanical ventilation (>1 day); use of H2-receptor antagonist, sucralfate, muscle relaxants, corticosteroids, barbiturates, and inotropic agents; positive end-expiratory pressure; intense sedation; re-intubation; tracheotomy; urgent brain computed tomography (CT) scan; craniotomy; iatrogenic event; and hyperventilation. The mortality rate was 43.5% (10 of 23) in the nosocomial pneumonia group and 18.8% in patients without nosocomial pneumonia (p =.02). Also, the mean stay in the ICU, the therapeutic charge (measured with total and mean punctuation of the Therapeutic Intervention Scoring System) and the complications, infectious and noninfectious, of the clinical evolution were significantly more frequent in patients with nosocomial pneumonia than in those without pneumonia (p <.05). In the multivariate analysis, continuous enteral feeding, craniotomy, prolonged mechanical ventilation (>24 hrs), use of positive end-expiratory pressure, and corticotherapy were independent predictors of nosocomial pneumonia. It seems that factors related to the patient's clinical course, rather than variables registered on the first days of ICU admission, are those that would exert an influence on the development of nosocomial pneumonia in critically ill trauma patients. In this way, from our point of view, in our study the main risk factors are the use of prolonged mechanical ventilation (>4 hrs) and positive end-expiratory pressure. At the same time, we can conclude that the reduction of this infection incidence could decrease the mean stay in the ICU, the therapeutic charge, and the prognosis in terms of mortality and morbidity.


CD45 is essential for Fc epsilon RI signaling by ZAP70, but not Syk, in Syk-negative mast cells.
The ZAP70/Syk family of protein tyrosine kinases plays an important role in Ag receptor signaling. Structural similarity of Syk and ZAP70 suggests their functional overlap. Previously, it was observed that expression of either ZAP70 or Syk reconstitutes Ag receptor signaling in Syk-negative B cells. However, in CD45-deficient T cells, Syk, but not ZAP70, restores T cell receptor-signaling pathway. To study the function of Syk, ZAP70, and CD45 in mast cells, a Syk/CD45 double-deficient variant of RBL-2H3 cells was characterized. After transfection, stable cell lines were isolated that expressed ZAP70, Syk, CD45, ZAP70 plus CD45, and Syk plus CD45. IgE stimulation did not induce degranulation in parental double-deficient cells, nor in the cells expressing only CD45. ZAP70 expression did not restore Fc epsilon RI signaling unless CD45 was coexpressed in the cells. However, Syk alone restored the IgE signal transduction pathway. The coexpression of CD45 with Syk had no significant effects on the responses to FcepsilonRI-aggregation. There was much better binding of Syk than ZAP70 to the phosphorylated Fc epsilon RI gamma-ITAM. Furthermore, unlike Syk, ZAP70 required CD45 to display receptor-induced increase in kinase activity. Therefore, in mast cells, ZAP70, but not Syk, requires CD45 for Ag receptor-induced signaling.


Desirability-based multiobjective optimization for global QSAR studies: application to the design of novel NSAIDs with improved analgesic, antiinflammatory, and ulcerogenic profiles.
Up to now, very few reports have been published concerning the application of multiobjective optimization (MOOP) techniques to quantitative structure-activity relationship (QSAR) studies. However, none reports the optimization of objectives related directly to the desired pharmaceutical profile of the drug. In this work, for the first time, it is proposed a MOOP method based on Derringer's desirability function that allows conducting global QSAR studies considering simultaneously the pharmacological, pharmacokinetic and toxicological profile of a set of molecule candidates. The usefulness of the method is demonstrated by applying it to the simultaneous optimization of the analgesic, antiinflammatory, and ulcerogenic properties of a library of fifteen 3-(3-methylphenyl)-2-substituted amino-3H-quinazolin-4-one compounds. The levels of the predictor variables producing concurrently the best possible compromise between these properties is found and used to design a set of new optimized drug candidates. Our results also suggest the relevant role of the bulkiness of alkyl substituents on the C-2 position of the quinazoline ring over the ulcerogenic properties for this family of compounds. Finally, and most importantly, the desirability-based MOOP method proposed is a valuable tool and shall aid in the future rational design of novel successful drugs.


Whole genome sequencing for mutation discovery in a single case of lysosomal storage disease (MPS type 1) in the dog.
Mucopolysaccharidosis (MPS) is a metabolic storage disorder caused by the deficiency of any lysosomal enzyme required for the breakdown of glycosaminoglycans. A 15-month-old Boston Terrier presented with clinical signs consistent with lysosomal storage disease including corneal opacities, multifocal central nervous system disease and progressively worsening clinical course. Diagnosis was confirmed at necropsy based on histopathologic evaluation of multiple organs demonstrating accumulation of mucopolysaccharides. Whole genome sequencing was used to uncover a frame-shift insertion affecting the alpha-L-iduronidase (IDUA) gene (c.19_20insCGGCCCCC), a mutation confirmed in another Boston Terrier presented 2 years later with a similar clinical picture. Both dogs were homozygous for the IDUA mutation and shared coat colors not recognized as normal for the breed by the American Kennel Club. In contrast, the mutation was not detected in 120 unrelated Boston Terriers as well as 202 dogs from other breeds. Recent inbreeding to select for recessive and unusual coat colors may have concentrated this relatively rare allele in the breed. The identification of the variant enables ante-mortem diagnosis of similar cases and selective breeding to avoid the spread of this disease in the breed. Boston Terriers carrying this variant represent a promising model for MPS I with neurological abnormalities in humans.


Taurocholic acid metabolism by gut microbes and colon cancer.
Colorectal cancer (CRC) is one of the most frequent causes of cancer death worldwide and is associated with adoption of a diet high in animal protein and saturated fat. Saturated fat induces increased bile secretion into the intestine. Increased bile secretion selects for populations of gut microbes capable of altering the bile acid pool, generating tumor-promoting secondary bile acids such as deoxycholic acid and lithocholic acid. Epidemiological evidence suggests CRC is associated with increased levels of DCA in serum, bile, and stool. Mechanisms by which secondary bile acids promote CRC are explored. Furthermore, in humans bile acid conjugation can vary by diet. Vegetarian diets favor glycine conjugation while diets high in animal protein favor taurine conjugation. Metabolism of taurine conjugated bile acids by gut microbes generates hydrogen sulfide, a genotoxic compound. Thus, taurocholic acid has the potential to stimulate intestinal bacteria capable of converting taurine and cholic acid to hydrogen sulfide and deoxycholic acid, a genotoxin and tumor-promoter, respectively.


Rejecting a general check-up: case report.
65-old gentleman rejected the general check-up recommendation, because he felt healthy. As his chosen family doctor, I also perceive him as the healthy patient, not having any risks. In spite of my personal concerns, it was obvious that I had to respect his choice, because of Act on Patients Rights and the Physicians Ethical Code. But, at that moment I was not certain about the evidence on the effectiveness of general chech-ups and some professional dilemmas remain. Therefore, we did a limited literature search. A Cochrane systematic analysis as well as other systematic reviews has shown that there is no evidence on the effectiveness of general check-ups. In the professional recommendations general check-ups are replaced by preventive measures with proven effectiveness, such as those recommended by the Canadian Task Force on Preventive Health Care and the U.S. Preventive Task Force. It helps me to solve my professional dilemmas around this patient.


Outcome of caval division techniques for partial anomalous pulmonary venous connections to the superior vena cava.
Correction of a high partial anomalous pulmonary venous (PV) connection to the superior vena cava (SVC) may be complicated by sinus node dysfunction, or SVC or PV obstruction. We have used the caval division technique when the anomalous PV insert high above the cavoatrial junction and report our long-term outcome. Between 1990 and 2009, 178 consecutive patients underwent operations for a partial anomalous PV connection. Caval division was used in 41 patients (23%). Median age was 34 years (range, 1.3 to 76 years). Thirty patients (73%) had an atrial septal defect. Eighteen patients (44%) underwent Warden procedure in which the SVC was divided, the upper end was connected to the right atrial appendage, and the remaining SVC served as a conduit for PV drainage to the left atrium through the atrial septal defect. A modified Warden was performed in 17 patients (41%) using a short ringed interposition graft between the SVC and the right atrial appendage. Six patients (15%) with persistent left SVC underwent right SVC division without reimplantation. No early deaths occurred. One Warden group patient (5%) required early reoperation for SVC obstruction. One patient (2%) with atrial fibrillation underwent a concomitant Maze procedure and permanent pacemaker implantation for tachybrady syndrome. All other patients remained in sinus rhythm at follow-up. Follow-up was complete in all patients (mean, 2±2.6 years; maximum, 14 years). One noncardiac-related death occurred at 14 years. SVC obstruction developed in 3 Warden group patients (17%) and was managed with angioplasty and stenting. No patient had pulmonary vein stenosis on follow-up. Caval division techniques appear to be effective, with low morbidity and mortality. The modified Warden procedure may decrease the risk of SVC obstruction. Late arrhythmias or PV obstruction are uncommon.


Hydrogel-Based Bioinks for Cell Electrowriting of Well-Organized Living Structures with Micrometer-Scale Resolution.
Bioprinting has become an important tool for fabricating regenerative implants and in vitro cell culture platforms. However, until today, extrusion-based bioprinting processes are limited to resolutions of hundreds of micrometers, which hamper the reproduction of intrinsic functions and morphologies of living tissues. This study describes novel hydrogel-based bioinks for cell electrowriting (CEW) of well-organized cell-laden fiber structures with diameters ranging from 5 to 40 μm. Two novel photoresponsive hydrogel bioinks, that is, based on gelatin and silk fibroin, which display distinctly different gelation chemistries, are introduced. The rapid photomediated cross-linking mechanisms, electrical conductivity, and viscosity of these two engineered bioinks allow the fabrication of 3D ordered fiber constructs with small pores (down to 100 μm) with different geometries (e.g., squares, hexagons, and curved patterns) of relevant thicknesses (up to 200 μm). Importantly, the biocompatibility of the gelatin- and silk fibroin-based bioinks enables the fabrication of cell-laden constructs, while maintaining high cell viability post printing. Taken together, CEW and the two hydrogel bioinks open up fascinating opportunities to manufacture microstructured constructs for applications in regenerative medicine and in vitro models that can better resemble cellular microenvironments.


Stable or changing well-being? Daily hassles and life satisfaction of Czech adolescents over the last three decades.
While the assumption that the sociopolitical and economic situation affects adolescents' well-being, encompassing life satisfaction and a positive sense of self, is plausible, few studies have confirmed such macrosocial influences. The case of the Czech Republic offers an example of a society transitioning from totalitarian government (from 1989) to western democracy. Our study provides statistical description of Czech adolescents' well-being over the past 30 years in association with the subjective perception of everyday problems. These daily hassles represent experiences and conditions of daily living that have been appraised as salient and harmful or threatening to adolescents' well-being. We analyzed four samples of adolescents aged 14-17 years surveyed at four time points over the last three decades-1992, 2001, 2011, and 2019, total N = 4,005 (1992: 255, 2001: 306, 2011: 363, 2019: 3081; 54.6% females). The results show that life satisfaction, self-esteem, and self-reported daily hassles changed only marginally from 1992 to 2019 with small differences related to the post-revolution 1992 cohort. Adolescents reported increasing problems in school, relationships with parents, sports, and leisure time over the study period. A model linking daily hassles and self-esteem to life satisfaction across four cohorts showed that daily hassles strongly predicted life satisfaction except in the post-revolution cohort of 1992 when life satisfaction was also the lowest. The effect was slightly higher in females. Across the cohorts, gender differences in life satisfaction changed from males being more satisfied in 1992 to females being more satisfaction in 2019. Limitations stemming from sampling differences across cohorts are discussed.


A reverse transcriptase-PCR assay for detecting filarial infective larvae in mosquitoes.
Existing molecular assays for filarial parasite DNA in mosquitoes cannot distinguish between infected mosquitoes that contain any stage of the parasite and infective mosquitoes that harbor third stage larvae (L3) capable of establishing new infections in humans. We now report development of a molecular L3-detection assay for Brugia malayi in vectors based on RT-PCR detection of an L3-activated gene transcript. Candidate genes identified by bioinformatics analysis of EST datasets across the B. malayi life cycle were initially screened by PCR using cDNA libraries as templates. Stage-specificity was confirmed using RNA isolated from infected mosquitoes. Mosquitoes were collected daily for 14 days after feeding on microfilaremic cat blood. RT-PCR was performed with primer sets that were specific for individual candidate genes. Many promising candidates with strong expression in the L3 stage were excluded because of low-level transcription in less mature larvae. One transcript (TC8100, which encodes a particular form of collagen) was only detected in mosquitoes that contained L3 larvae. This assay detects a single L3 in a pool of 25 mosquitoes. This L3-activated gene transcript, combined with a control transcript (tph-1, accession # U80971) that is constitutively expressed by all vector-stage filarial larvae, can be used to detect filarial infectivity in pools of mosquito vectors. This general approach (detection of stage-specific gene transcripts from eukaryotic pathogens) may also be useful for detecting infective stages of other vector-borne parasites.


The Fibromyalgia Impact Questionnaire: a useful tool in evaluating patients with post-Lyme disease syndrome.
To determine the reliability and validity of a modified version of the Fibromyalgia Impact Questionnaire (FIQ) in evaluating patients with post-Lyme disease syndrome (PLDS). In this cross-sectional analysis 13 PLDS, 18 fibromyalgia (FM), and 16 healthy controls (n = 47) completed a modified FIQ containing items to evaluate physical impairment, symptom severity, and global well-being. Comparisons between groups were done using analysis of variance with a significance level set at 0.05. PLDS patients demonstrated statistically significantly greater levels of impairment than controls in physical functioning, FIQ total score, global well-being, joint pain, fatigue, depression, ability to perform activities of daily living, and memory/concentration. FM patients demonstrated a statistically significantly greater level of impairment than the control group in all categories, and the scores were significantly higher than the PLDS group in the measurement of physical impairment, FIQ total score, muscle pain, and joint pain. Overall, the instrument possesses good reliability and validity, although adequacy of this instrument to measure impairment in the male PLDS population needs further elucidation. The results of this study suggest that the modified FIQ may be a useful tool in evaluating PLDS patients. The findings suggest that there may be some differences in the etiopathology of the symptoms experienced by PLDS and FM patients.


Trp(250) -hK2 is defective in intracellular trafficking and activates the unfolded protein response.
hK2, a member of the kallikrein protease family encoded by KLK2, is expressed exclusively in prostate and is a putative adjunct tumor marker for prostate cancer screening. The T allele of rs198977, a single nucleotide polymorphism in exon 5 of KLK2, codes for W-hK2 and is associated with lower serum hK2 levels and higher risk of prostate cancer than the C allele encoding R-hK2. To elucidate the mechanism that underlies this SNP's function, we transfected plasmids expressing R-hK2 or W-hK2 into PC3, HeLa and HEK293A cells and measured the hK2 level in cell lysates and conditioned media. The level of W-hK2 was lower than R-hK2 in conditioned media but was not different from R-hK2 in cell lysates. W-hK2 was hardly colocalized with Golgi-targeted fluorescent protein whereas R-hK2 colocalized. Reporter assays related to the unfolded protein response (UPR) and phospho-eIF2α immunoblot showed that W-hK2 increased UPR activity more than R-hK2. These results indicated that W-hK2 had a defect in cellular trafficking from the ER to the Golgi complex due to its misfolding and that it activated the UPR, suggesting a mechanism to explain the association of the T allele with higher prostate cancer risk.


Assessment of gene flow across a hybrid zone in red-tailed chipmunks (Tamias ruficaudus).
The role of hybridization in animal speciation is controversial and recent research has challenged the long-standing criterion of complete reproductive isolation to attain species status. The speciation-with-gene-flow model posits that the genome is semi-permeable and hybridization may be a phase in the process of divergence. Here, we apply these concepts to a previously identified zone of mtDNA introgression between the two strongly morphologically differentiated subspecies of red-tailed chipmunk (Tamias ruficaudus) in the US Inland Northwest. Using multilocus genotype data from the southern, older contact zone, we demonstrate that neutral gene flow is unusually low between the subspecies across the Lochsa River. This is geographically congruent with the discontinuity in bacular morphology, indicating that the cline of mitochondrial DNA (mtDNA) haplotypes is displaced. Furthermore, we elucidate the evolutionary forces responsible by testing hypotheses of lineage sorting and hybridization. We determined that introgressive hybridization is the cause of mtDNA/morphology incongruence because there are non-zero levels of migration and gene flow. Although our estimate of the age of the hybrid zone has wide credibility intervals, the hybridization events occurred in the Late Pleistocene and the divergence occurred in the Middle Pleistocene. Finally, we assessed substructure within and adjacent to the hybrid zone and found that the hybrid zone constitutes a set of populations that are genetically differentiated from parental sets of populations; therefore, hybridization in this system is not likely an evolutionary sink, but has generated novel combinations of genotypes.


Molecular structures and metabolic characteristics of protein in brown and yellow flaxseed with altered nutrient traits.
The objectives of this study were to investigate the chemical profiles; crude protein (CP) subfractions; ruminal CP degradation characteristics and intestinal digestibility of rumen undegraded protein (RUP); and protein molecular structures using molecular spectroscopy of newly developed yellow-seeded flax (Linum usitatissimum L.). Seeds from two yellow flaxseed breeding lines and two brown flaxseed varieties were evaluated. The yellow-seeded lines had higher (P < 0.001) contents of oil (44.54 vs 41.42% dry matter (DM)) and CP (24.94 vs 20.91% DM) compared to those of the brown-seeded varieties. The CP in yellow seeds contained lower (P < 0.01) contents of true protein subfraction (81.31 vs 92.71% CP) and more (P < 0.001) extensively degraded (70.8 vs 64.9% CP) in rumen resulting in lower (P < 0.001) content of RUP (29.2 vs 35.1% CP) than that in the brown-seeded varieties. However, the total supply of digestible RUP was not significantly different between the two seed types. Regression equations based on protein molecular structural features gave relatively good estimation for the contents of CP (R(2) = 0.87), soluble CP (R(2) = 0.92), RUP (R(2) = 0.97), and intestinal digestibility of RUP (R(2) = 0.71). In conclusion, molecular spectroscopy can be used to rapidly characterize feed protein molecular structures and predict their nutritive value.


Rhizoma Dioscoreae extract protects against alveolar bone loss in ovariectomized rats via microRNAs regulation.
The aim of this study was to evaluate the osteoprotective effect of aqueous Rhizoma Dioscoreae extract (RDE) on the alveolar bone of rats with ovariectomy-induced bone loss. Female Wistar rats underwent either ovariectomy or sham operation (SHAM). The ovariectomized (OVX) rats were treated with vehicle (OVX), estradiol valerate (EV), or RDE. After treatments, the bone mineral density (BMD) and the three-dimensional microarchitecture of the alveolar bone were analyzed to assess bone mass. Microarrays were used to evaluate microRNA expression profiles in alveolar bone from RDE-treated and OVX rats. The differential expression of microRNAs was validated using real-time quantitative RT-PCR (qRT-PCR), and the target genes of validated microRNAs were predicted and further analyzed using Ingenuity Pathway Analysis (IPA). The key findings were verified using qRT-PCR. Our results show that RDE inhibits alveolar bone loss in OVX rats. Compared to the OVX rats, the RDE-treated rats showed upregulated expression levels of 8 microRNAs and downregulated expression levels of 8 microRNAs in the alveolar bone in the microarray analysis. qRT-PCR helped validate 13 of 16 differentially expressed microRNAs, and 114 putative target genes of the validated microRNAs were retrieved. The IPA showed that these putative target genes had the potential to code for proteins that were involved in the transforming growth factor (TGF)-β/bone morphogenetic proteins (BMPs)/Smad signaling pathway (Tgfbr2/Bmpr2, Smad3/4/5, and Bcl-2) and interleukin (IL)-6/oncostatin M (OSM)/Jak1/STAT3 signaling pathway (Jak1, STAT3, and Il6r). These experiments revealed that RDE could inhibit ovariectomy-induced alveolar bone loss in rats. The mechanism of this anti-osteopenic effect in alveolar bone may involve the simultaneous inhibition of bone formation and bone resorption, which is associated with modulation of the TGF-β/BMPs/Smad and the IL-6/OSM/Jak1/STAT3 signaling pathways via microRNA regulation.


Epidemiology of avian diseases.
A large number of diseases occur in domestic, farm-raised poultry. Only two of the many different diseases are notifiable and subject to governmental control: highly pathogenic avian influenza and Newcastle disease. Diagnosis and treatment or prevention of all other conditions are left to the skills of farmers and their veterinarians. Poultry production is aimed at providing more and tastier food for the ever growing human community. Infectious diseases and technical errors during production and processing need to be minimised. The concept of hazard analysis critical control point (HACCP) has already been introduced into food processing and quality assessment. The regulations laid down in ISO 9000 will soon become a powerful and practical tool for monitoring and improving the productivity of live poultry. Approved epidemiological concepts and tools will enable the poultry industry to achieve constant and safe production. Certification on the basis of ISO 9000 of all areas of poultry production is a new approach for maintaining the health of poultry, for tracing and subsequently eliminating breaks in productivity, and securing production without health hazards for the consumer.


[Update on cerebral monitoring and protective methods].
Clamping ischaemia is responsible for the 20-30% of the neurological complications during carotid surgery. It is impossible at present to determine preoperatively for certainty the patients who are at risk and the ones who are not at risk for clamping ischemia. Intraoperative monitoring is mandatory in all the cases to point out cerebral ischemia and to prevent neurological deficit with an adequate brain protection. Methods used to predict brain ischaemia include local anesthesia, EEG, SEP, stump pressure measurement, transcranial doppler, regional cerebral blood flow measurement and evaluation of venous blood gases from the internal jugular vein. Local anesthesia is a safe and simple method of assessing cerebral ischemia during carotid clamping but has some limits particularly in case of brain ischaemia for the technical difficulties to install an indwelling shunt in an awake and often troubled patient. Moreover the only possibility of cerebral protection under local anesthesia is an increase in systemic blood pressure with a resulting growth in cardiac morbidity and mortality. For these reasons operation should be performed under general anesthesia to assure a better farmacogical brain protection when preoperative evaluation demonstrates a reduction of cerebral vasoreactivity, with a consequent high risk of clamping ischemia. At present EEG is one of the most used technique of cerebral monitoring under general anesthesia with a sensibility and specificity in the diagnosis of brain ischaemia that is about 90%. The best cerebral protection is obtained with a selective shunting. Burst suppression, with high dose thiopentone, has recently demonstrated its reliability alone or in association with an indwelling shunt in patients at high risk for clamping ischaemia. Of course a systematic cerebral protection together with an accurate control of the blood pressure is necessary in all the patients. This behaviour allowed us to obtain good results in the last 500 patients with a morbidity and mortality respectively of 1% and 1.6% with no significant differences between patients who tolerated carotid clamping and those with clamping ischaemia.


Differential induction of gelatinase B (MMP-9) and gelatinase A (MMP-2) in T lymphocytes upon alpha(4)beta(1)-mediated adhesion to VCAM-1 and the CS-1 peptide of fibronectin.
Integrin alpha(4)beta(1) on the surface of T lymphocytes interacts with vascular cell adhesion molecule-1 (VCAM-1) and fibronectin during migration of lymphocytes from the blood to sites of inflammation. Migrating lymphocytes actively modify their environment through a number of mechanisms including proteolysis of the extracellular matrix by matrix metalloproteinases (MMP) synthesized by the cells. In this study, expression of MMP upon alpha(4)beta(1)-mediated adhesion of leukocytes to two major ligands, the IIICS-1 domain of fibronectin and VCAM-1, has been examined. Adhesion of T lymphoblastoid Jurkat cells to the CS-1 peptide induced expression of mRNA for two MMPs, gelatinase A (MMP-2) and gelatinase B (MMP-9). As evaluated by relative RT-PCR and Northern blot analyses, the level of mRNA was upregulated about 4- to 5-fold for both MMPs compared to control cells maintained in suspension. With time, both enzymes were detected in conditioned media and inside the cells, and their identities were verified by Western blotting and gelatin zymography. Adhesion of Jurkat cells to the second major alpha(4)beta(1) ligand, VCAM-1, upregulated mRNA for MMP-2 (3.5-fold) and failed to induce expression of mRNA for MMP-9. Accordingly, only MMP-2 protein was detected in conditioned media of cells adherent to VCAM-1. Occupancy of alpha(4)beta(1) on the surface of suspended cells with soluble CS-1 peptide or VCAM-1 did not upregulate synthesis and release of MMPs. A similar pattern of induction of MMPs after adhesion to CS-1 and VCAM-1 was observed in T lymphocytes isolated from human blood. These results demonstrate that adhesion of T lymphocytes through alpha(4)beta(1) to different ligands, which bind to similar or overlapping sites in the integrin, induces intracellular events leading to distinct patterns of MMPs biosynthesis.


Comparison of analytic techniques: chemical, isotopic, and microprobe analyses.
The development of new techniques and modifications to existing methods are providing important information about cellular water and electrolytes in relation to transepithelial transport. Information obtained for one epithelium, the toad urinary bladder, from chemical analyses, isotopic studies, and electron microprobe analysis, is reviewed. Chemical and isotopic analyses allow estimation of average cellular water and ion contents and determination of the origin of ions from the media bathing the mucosal and basolateral cellular membranes. The electron microprobe provides additional information about the distributions of ions between the cell types that constitute the epithelial layer and also allows some measurement of ion distributions within individual cells. In toad urinary bladder epithelial cells there is broad agreement between the results obtained with these different techniques and a summary is provided of possible relationships between transepithelial sodium transport and cellular composition.


The beneficial acclimation hypothesis versus acclimation of specific traits: physiological change in water-stressed Manduca sexta caterpillars.
Do organisms make beneficial physiological adjustments in response to environmental change? We examined this question by measuring the effects of short-term (12-36 h) and long-term (larval lifetime) hydric stress on the tobacco hornworm, Manduca sexta. Larvae were reared from the first instar on low-water (69%) or high-water (80%) artificial diets and then transferred early in the fifth instar to the same or opposite diet (2x2 design). Within the subsequent 36 h, we measured 24-h growth rates and three primary determinants of the water budget: water gain via consumption and water loss via evaporation and defecation. Larvae preexposed to low-water diet grew less rapidly on low-water diet than those switched acutely to low-water diet from high-water diet, showing that larvae preexposed to a particular environment do not necessarily acclimate beneficially to that environment. Our data on water fluxes to and from larvae, however, strongly suggest that water-stressed larvae did make beneficial physiological adjustments. Larvae responded to short-term hydric stress by minimizing rates of water excretion, primarily by increasing rates of rectal water absorption. Larvae responded to chronic water stress by significantly reducing rates of evaporative water loss; they also showed additional reductions in fecal water excretion, but these decreases were due to lowered consumption and not to further increases in rate of rectal water absorption. This mismatch between maladaptive acclimation of organismal performance and beneficial adjustment of suborganismal traits can be reconciled by recognizing that organismal physiology is hierarchical: fitness-related performance traits represent the aggregate outcome of numerous, more mechanistic physiological traits. Although chronic exposure to an environment may depress the aggregate effect of these mechanistic traits on performance, organisms are not precluded from making beneficial adjustments to individual traits contributing to performance.


Reinforcing saccadic amplitude variability.
Saccadic endpoint variability is often viewed as the outcome of neural noise occurring during sensorimotor processing. However, part of this variability might result from operant learning. We tested this hypothesis by reinforcing dispersions of saccadic amplitude distributions, while maintaining constant their medians. In a first experiment we reinforced the least frequent saccadic amplitudes to increase variability, and then reinforced the central part of the amplitude distributions to reduce variability. The target was placed at a constant distance from the fovea after the saccade to maintain the postsaccadic visual signal constant and an auditory reinforcement was delivered depending on saccadic amplitude. The second experiment tested the effects of the contingency. We reinforced high levels of variability in 4 participants, whereas 4 other participants were assigned to a yoked control group. On average, saccadic amplitude standard deviations were doubled while the medians remained mostly unchanged in the experimental participants in both experiments, and variability returned to baseline level when low variability was reinforced. In the control group no consistent changes in amplitude distributions were observed. These results, showing that variability can be reinforced, challenge the idea of a stochastic neural noise. We instead propose that selection processes constrain saccadic amplitude distributions.


A Six-SNP haplotype of ADAM33 is associated with asthma in a population of Cartagena, Colombia.
A disintegrin and metalloprotein-33 (ADAM33) participates in the bronchial remodeling process in asthma, and genetic analyses pointed it out as a candidate gene in asthma. To analyze the association between ADAM33 and asthma and total and mite-specific IgE levels in a population of the Caribbean Coast of Colombia, we genotyped 6 single-nucleotide polymorphisms of ADAM33 in 429 asthmatics, 401 controls and 116 family trios using fluorogenic probes. Total and specific IgE against Blomia tropicalis and Dermatophagoides pteronyssinus were determined by ELISA. Case-control and family-based analyses were performed. Case-control association analyses were corrected by population stratification using a set of 52 ancestry-informative markers. Eight common haplotypes were identified; among them, H4 (GCAGGG) was associated with asthma in the family group (Z score: -2.049, p = 0.04). We also found an association between the TT genotype of ST+7 and asthma in the case-control study (p = 0.05) that disappeared after correcting for multiple testing. In the family-based analysis, this genotype was a risk factor for asthma (p = 0.01), high total IgE (Z score: 2.546, p = 0.01) and high specific IgE against B. tropicalis (p = 0.02) and D. pteronyssinus (Z score: 2.414, p = 0.01). V4 was associated with specific IgE against B. tropicalis (p = 0.03); T2 with asthma (p = 0.03), high total IgE (p = 0.02) and IgE against D. pteronyssinus (p = 0.03) and T1 with high total IgE (p = 0.04). None of these associations was maintained after correction for multiple testing. Our findings suggest a relevant role of ADAM33 in thepathogenesis of asthma in this population.


Diagnostic accuracy of transbronchial lung cryobiopsy for interstitial lung disease diagnosis (COLDICE): a prospective, comparative study.
Transbronchial lung cryobiopsy (TBLC) is a novel technique for sampling lung tissue for interstitial lung disease diagnosis. The aim of this study was to establish the diagnostic accuracy of TBLC compared with surgical lung biopsy (SLB), in the context of increasing use of TBLC in clinical practice as a less invasive biopsy technique. COLDICE was a prospective, multicentre, diagnostic accuracy study investigating diagnostic agreement between TBLC and SLB, across nine Australian tertiary hospitals. Patients with interstitial lung disease aged between 18 and 80 years were eligible for inclusion if they required histopathological evaluation to aid diagnosis, after detailed baseline evaluation. After screening at a centralised multidisciplinary discussion (MDD), patients with interstitial lung disease referred for lung biopsy underwent sequential TBLC and SLB under one anaesthetic. Each tissue sample was assigned a number between 1 and 130, allocated in a computer-generated random sequence. Encoded biopsy samples were then analysed by masked pathologists. At subsequent MDD, de-identified cases were discussed twice with either TBLC or SLB along with clinical and radiological data, in random non-consecutive order. Co-primary endpoints were agreement of histopathological features in TBLC and SLB for patterns of definite or probable usual interstitial pneumonia, indeterminate for usual interstitial pneumonia, and alternative diagnosis; and for agreement of consensus clinical diagnosis using TBLC and SLB at MDD. Concordance and κ values were calculated for each primary endpoint. This study is registered with the Australian New Zealand Clinical Trials Registry, ACTRN12615000718549. Between March 15, 2016, and April 15, 2019, we enrolled 65 patients (31 [48%] men, 34 [52%] women; mean age 66·1 years [SD 9·3]; forced vital capacity 83·7% [SD 14·2]; diffusing capacity for carbon monoxide 63·4% [SD 12·8]). TBLC (7·1 mm, SD 1·9) and SLB (46·5 mm, 14·9) samples were each taken from two separate ipsilateral lobes. Histopathological agreement between TBLC and SLB was 70·8% (weighted κ 0·70, 95% CI 0·55-0·86); diagnostic agreement at MDD was 76·9% (κ 0·62, 0·47-0·78). For TBLC with high or definite diagnostic confidence at MDD (39 [60%] of 65 cases), 37 (95%) were concordant with SLB diagnoses. In the 26 (40%) of 65 cases with low-confidence or unclassifiable TBLC diagnoses, SLB reclassified six (23%) to alternative high-confidence or definite MDD diagnoses. Mild-moderate airway bleeding occurred in 14 (22%) patients due to TBLC. The 90-day mortality was 2% (one of 65 patients), following acute exacerbation of idiopathic pulmonary fibrosis. High levels of agreement between TBLC and SLB for both histopathological interpretation and MDD diagnoses were shown. The TBLC MDD diagnoses made with high confidence were particularly reliable, showing excellent concordance with SLB MDD diagnoses. These data support the clinical utility of TBLC in interstitial lung disease diagnostic algorithms. Further studies investigating the safety profile of TBLC are needed. University of Sydney, Hunter Medical Research Institute, Erbe Elektromedizin, Medtronic, Cook Medical, Rymed, Karl-Storz, Zeiss, and Olympus.


The Illinois HIV transmission statute: unconstitutionally vague or politically vogue?
This article does not challenge the prudence of enacting statutes criminalizing the transmission of the HIV virus. Instead, the author asks whether a particular statute, one already enacted in Illinois, is unconstitutionally vague because it does not give adequate warning to individuals that certain behavior is proscribed or because it is overbroad in that it prohibits constitutionally protected conduct. The author also offers a redrafted Illinois statute that she believes will pass constitutional muster.


Switching versus augmentation: a prospective, naturalistic comparison in depressed, treatment-resistant patients.
(1) To directly compare the effectiveness of switching antidepressants with augmenting them in depressed patients who do not respond to an initial adequate trial and (2) to determine whether there is a decreased likelihood of response to a second switch or augmentation trial in those patients who did not respond to the first intervention for treatment-resistant depression. In a naturalistic, open-label design, all depressed outpatients (DSM-IV criteria) who were treatment resistant were prospectively assessed. Short- and long-term outcomes of switching versus augmentation were compared using the Clinical Global Impressions scale. In the acute phase, 37 (50.0%) of 74 subjects responded to 1 of the 2 interventions for treatment-resistant depression. Forty-five percent (N = 17) and 56% (N = 20) of the patients who had their antidepressant switched or augmented, respectively, responded to that intervention. Nearly three fourths (71.4%) of the acute responders maintained their response through 6 months of follow-up. In 18 patients who did not respond to the first switch or augmentation, 9 (50.0%) responded to a second trial. Switching antidepressants was somewhat less effective than augmentation, although this difference was not statistically significant. For patients who do not respond to an augmentation or switch, our results suggest that a second trial for treatment-resistant depression may be as effective as the first.


[Rivastigmine as adjunctive therapy in the therapeutic dilemma for the treatment of hallucinations due to Parkinson disease].
We report three cases of patients with Parkinson's disease without dementia, admitted to our hospital because of hallucinations. The anti-Parkinson medication was adapted and the patients started with rivastigmine. As a result, hallucinations no longer occurred. A 79 years old man also required short-term quetiapine because of agitation and anti-Parkinson doses were without side effects, as a result of which mobility improved. An 84 years old woman reported mild side effects of rivastigmine, without consequences, whereas her mobility appeared to be good. A 72 years old woman reported mild memory problems upon admission, which improved during admission, as did her mobility after increasing the anti-Parkinson medication doses. Treatment of rivastigmine can be useful in the therapeutic dilemma in the treatment of hallucinations in patients with Parkinson's disease (start anti-psychotic or reduce anti-Parkinson medication). In addition to adapting anti-Parkinson doses and sometimes short-term treating with an anti-psychotic, treatment with rivastigmine appears to be a quick improvement, without serious side effects. Also, mobility can improve, due to the possibility of increasing the anti-Parkinson doses, if necessary. Because of the many remaining questions, prospective randomised trials are needed.


Comparative in-vitro and in-vivo activity of AM-1155 against anaerobic bacteria.
The in-vitro activity of AM-1155, a 6-fluoro-8-methoxy quinolone, was compared with those of temafloxacin, sparfloxacin, tosufloxacin, ciprofloxacin, ofloxacin and cefmetazole, a cephamycin, against a variety of anaerobic bacteria. Although AM-1155 demonstrated only modest activity against the Bacteroides fragilis group and Prevotella bivia (MIC90s > or =3.13 mg/mL), 76% of the B. fragilis strains tested were inhibited at AM-1155 concentrations of 0.78 mg/L. AM-1155 was highly active against Prevotella intermedia, Porphyromonas gingivalis, Fusobacterium spp., Clostridium perfringens and Mobiluncus spp. (MIC90s < or =0.39 mg/L). An in-vivo study using a mixed infection with AM-1155- and tosufloxacin-susceptible B. fragilis and Escherichia coli strains in rat granuloma pouch was performed. AM-1155 was effective against both organisms whereas tosufloxacin was effective only against E. coli. These results correlated well to the higher pouch levels of AM-1155 than those of tosufloxacin. Clostridium difficile overgrowth was found in the caecum of mice treated with ampicillin both 1 and 7 days after 5 days dosing, but not in AM-1155-treated mice. These results suggest that the clinical efficacy of AM-1155 against infections involving most anaerobic bacteria except for the B. fragilis group and P. bivia should be evaluated further.


Bioluminescence resonance energy transfer (BRET) imaging in plant seedlings and mammalian cells.
Bioluminescence resonance energy transfer (BRET) has become a widely used technique to monitor protein-protein interactions. It involves resonance energy transfer between a bioluminescent donor and a fluorescent acceptor. Because the donor emits photons intrinsically, fluorescence excitation is unnecessary. Therefore, BRET avoids some of the problems inherent in fluorescence resonance energy transfer (FRET) approaches, such as photobleaching, autofluorescence, and undesirable stimulation of photobiological processes. In the past, BRET signals have generally been too dim to be effectively imaged. Newly available cameras that are more sensitive coupled to image splitter now enable BRET imaging in plant and mammalian cells and tissues. In addition, new substrates and enhanced luciferases enable brighter signals that allow even subcellular BRET imaging. Here, we report methods for BRET imaging of (1) localization of COP1 dimerization in plant cells and tissues and (2) subcellular distributions of interactions of the CCAAT/Enhancer Binding Protein α (C/EBPα) in single mammalian cells. We also discuss methods for the correction of BRET images for tissues that absorb light of different spectra. This progress should catalyze further applications of BRET for imaging and high-throughput assays.


Outbreak of a Brucella canis infection in a beagle colony in West Germany.
Brucella infections in dogs were caused by the more frequent strains of brucellae i.e. Br. abortus, Br. suis, and Br. melitensis. Since 1966 it is known that in dog colonies, mostly beagle, in the United States, a new brucella species caused infections. This species was called Br. canis. The infection showed similar effects with dogs as the other brucellae with other domestic animals. With the dogs abortions occurred on the 40th to 50th day of gestation and the infection often resulted in sterility. In 1973 the author detected Br. canis for the first time in Europe; shortly afterwards the same bacteria was isolated in Japan. It is known how the disease spread to Japan, but how it came to Europe is still a mystery. Here it was first detected in a beagle colony with approximately 200 animals used for breeding. Within a few months the disease spread in the colony and finally the birthrate of inseminated bitches was only 29%; 50% had abortions and the rest had not conceived. In this communication characteristics of Br. canis are discussed.


Subcellular Localization of Rice Acyl-CoA-Binding Proteins ACBP4 and ACBP5 Supports Their Non-redundant Roles in Lipid Metabolism.
Acyl-CoA-binding proteins (ACBPs), conserved at the acyl-CoA-binding domain, can bind acyl-CoA esters as well as transport them intracellularly. Six ACBPs co-exist in each model plant, dicot Arabidopsis thaliana (thale cress) and monocot Oryza sativa (rice). Although Arabidopsis ACBPs have been studied extensively, less is known about the rice ACBPs. OsACBP4 is highly induced by salt treatment, but down-regulated following pathogen infection, while OsACBP5 is up-regulated by both wounding and pathogen treatment. Their differential expression patterns under various stress treatments suggest that they may possess non-redundant functions. When expressed from the CaMV35S promoter, OsACBP4 and OsACBP5 were subcellularly localized to different endoplasmic reticulum (ER) domains in transgenic Arabidopsis. As these plants were not stress-treated, it remains to be determined if OsACBP subcellular localization would change following treatment. Given that the subcellular localization of proteins may not be reliable if not expressed in the native plant, this study addresses OsACBP4:GFP and OsACBP5:DsRED expression from their native promoters to verify their subcellular localization in transgenic rice. The results indicated that OsACBP4:GFP was targeted to the plasma membrane besides the ER, while OsACBP5:DsRED was localized at the apoplast, in contrast to their only localization at the ER in transgenic Arabidopsis. Differences in tagged-protein localization in transgenic Arabidopsis and rice imply that protein subcellular localization studies are best investigated in the native plant. Likely, initial targeting to the ER in a non-native plant could not be followed up properly to the final destination(s) unless it occurred in the native plant. Also, monocot (rice) protein targeting may not be optimally processed in a transgenic dicot (Arabidopsis), perhaps arising from the different processing systems for routing between them. Furthermore, changes in the subcellular localization of OsACBP4:GFP and OsACBP5:DsRED were not detectable following salt and pathogen treatment, respectively. These results suggest that OsACBP4 is likely involved in the intracellular shuttling of acyl-CoA esters and/or other lipids between the plasma membrane and the ER, while OsACBP5 appears to participate in the extracellular transport of acyl-CoA esters and/or other lipids, suggesting that they are non-redundant proteins in lipid trafficking.


Monte Carlo dose enhancement studies in microbeam radiation therapy.
A radical radiation therapy treatment for gliomas requires extremely high absorbed doses resulting in subsequent deleterious side effects in healthy tissue. Microbeam radiation therapy (MRT) is an innovative technique based on the fact that normal tissue can withstand high radiation doses in small volumes without any significant damage. The synchrotron-generated x-ray beam is collimated and delivered to an array of narrow micrometer-sized planar rectangular fields. Several preclinical experiments performed at the Brookhaven National Laboratory (BNL) and at the European Synchrotron Radiation Facility (ESRF) confirmed that MRT yields a higher therapeutic index than nonsegmented beams of the same characteristics. This index can be greatly improved by loading the tumor with high atomic number (Z) contrast agents. The aim of this work is to find the high-Z element that provides optimum dose enhancement. Monte Carlo simulations (PENELOPE/penEasy) were performed to assess the peak and valley doses as well as their ratio (PVDR) in healthy tissue and in the tumor, loaded with different contrast agents. The optimization criteria used were maximization of the ratio between the PVDR values in healthy tissue respect to the PVDR in the tumor and minimization of bone and brain valley doses. Dose enhancement factors, PVDR, and valley doses were calculated for different high-Z elements. A significant decrease of PVDR values in the tumor, accompanied by a gain in the valley doses, was found in the presence of high-Z elements. This enables the deposited dose in the healthy tissue to be reduced. The optimum high-Z element depends on the irradiation configuration. As a general trend, the best outcome is provided by the highest Z contrast agents considered, i.e., gold and thallium. However, lanthanides (especially Lu) and hafnium also offer a satisfactory performance. The remarkable therapeutic index in microbeam radiation therapy can be further improved by loading the tumor with a high-Z element. This study reports quantitative data on several dosimetric magnitudes in order to find the optimum contrast agent. Although the final choice of the element will also depend on possible cytotoxicity, three elements were found to be worthy of mention: gold, thallium, and lutetium.


Clinical characteristics in early Parkinson's disease in a central California population-based study.
There is considerable variation in the phenotypic appearance of individuals with idiopathic Parkinson's disease (PD), which may translate into differences in disease progression in addition to underlying disease etiology. In this publication, we report on the demographic and clinical characteristics of 162 individuals diagnosed with clinically probable PD from January 1998 to June 2003 who resided in predominantly rural communities in central California. The majority of the subjects were Caucasian, male, and between 60 and 79 years of age. The akinetic-rigid and tremor-dominant subtypes were more common than the mixed subtype. The majority of subjects displayed motor signs of rigidity (92.0%), bradykinesia (95.7%), and gait problems (87.0%), whereas less than half (43.3%) of the subjects displayed a tremor. Three fourths of patients received a Hoehn and Yahr Scale score of Stage 2 or higher. One third of the patients were treated with levodopa, and patients under 60 years of age were more likely to be treated with dopamine agonists. Within 3 years after first diagnosis, 13% of subjects showed some signs of depression and 17% of subjects met criteria for mild dementia. Among our subjects, 17.3% reported a family history of PD in first- or second-degree relatives,15.4% a family history of essential tremor, and 14.2% of Alzheimer's disease. This study represents the most extensive phenotypic description of rural U.S. residents in the initial stages of PD who were recruited in a population-based manner; future follow-up may provide valuable information regarding the prognostic indication of these symptoms/signs and improve our understanding of the underlying etiology of PD.


Blue-phase liquid crystal cored optical fiber array with photonic bandgaps and nonlinear transmission properties.
Blue-phase liquid crystal (BPLC) is introduced into the pores of capillary arrays to fabricate fiber arrays. Owing to the photonic-crystals like properties of BPLC, these fiber arrays exhibit temperature dependent photonic bandgaps in the visible spectrum. With the cores maintained in isotropic as well as the Blue phases, the fiber arrays allow high quality image transmission when inserted in the focal plane of a 1x telescope. Nonlinear transmission and optical limiting action on a cw white-light continuum laser is also observed and is attributed to laser induced self-defocusing and propagation modes changing effects caused by some finite absorption of the broadband laser at the short wavelength regime. These nonlinear and other known electro-optical properties of BPLC, in conjunction with their fabrication ease make these fiber arrays highly promising for imaging, electro-optical or all-optical modulation, switching and passive optical limiting applications.


Desomorphine Screening Using Commercial Enzyme-Linked Immunosorbent Assays.
Desomorphine ("Krokodil") is a semi-synthetic opioid that has drawn attention as a recreational drug, particularly in Russia, neighboring former Soviet Republics, Eastern and Central Europe. It has no accepted medicinal uses and is currently a schedule I drug in the United States. In clandestine environments, desomorphine is synthesized from codeine using red phosphorous, hydroiodic acid and gasoline. Residual starting materials in illicit preparations have been associated with severe dermatological effects and extensive tissue necrosis. Desomorphine is not well studied, and there are limited reports concerning its pharmacology or detection in biological matrices. Immunoassays are widely relied upon for both antemortem and postmortem toxicology screening. Although desomorphine is an opioid of the phenanthrene-type, its ability to bind to conventional opioid antibodies has not been described. In this report we describe the cross-reactivity of desomorphine using six commercially available enzyme-linked immunosorbent assays (Immunalysis Opiates Direct ELISA, Immunalysis Oxycodone/Oxymorphone Direct ELISA, Randox Opiate ELISA, OraSure Technologies OTI Opiate Micro-plate EIA, Neogen Opiate Group ELISA and Neogen Oxycodone/Oxymorphone ELISA). Cross-reactivites were highly variable between assays, ranging from 77 to <2.5%. In general, assays directed towards morphine produced greater cross-reactivity with desomorphine than those directed towards oxycodone. The Immunalysis Opiates Direct ELISA produced the greatest cross-reactivity, although several of the assays evaluated produced cross-reactivity of a sufficient magnitude to be effective for desomorphine screening.


Inducible GBP5 Mediates the Antiviral Response via Interferon-Related Pathways during Influenza A Virus Infection.
Guanylate binding protein (GBP) 5 belongs to the GBP family, which is involved in important cellular processes, including signal transduction, translation, vesicle trafficking, and exocytosis. Structurally, GBPs display a high degree of homology and share highly conserved GTP-binding or hydrolysis domains. GBP5 was reported to be a critical cellular factor in inflammasome assembly. However, little is known about its role in the host antiviral innate immune response. In this study, we found that GBP5 expression was significantly elevated in influenza patients and influenza A virus-infected A549 human lung epithelial cells. The overexpression of GBP5 inhibited virus replication by enhancing the expression of virus-induced interferon (IFN) and IFN-related effectors. Knockdown of GBP5 had the opposite effect. Moreover, GBP5 enhanced endogenous IFN expression by interacting with the NF-κB-essential modulator complex and stimulating NF-κB signaling. Additionally, the expression of proinflammatory factors, such as IL-6, IL-8, tumor necrosis factor-α, cyclooxygenase-2, and inducible nitric oxide synthase, was also activated by GBP5. Taken together, our results reveal that GBP5 inhibited virus replication through the activation of IFN signaling and proinflammatory factors.


Outcomes of cardiac rehabilitation participants and nonparticipants in a rural area.
Nationally, only 11%-20% of cardiac patients, on average, enroll in cardiac rehabilitation programs after their cardiac events. The purpose of this study was to examine: (a) differences in functional health outcomes, clinical risk factor outcomes, and lifestyle behaviors between patients who participated in cardiac rehabilitation (CR) and those who did not during the first year after their cardiac event; and (b) to examine predictors of and reasons for CR participation and non-participation in a Midwestern, rural clinical population. Green's health education framework guided the study. A cross-sectional, comparative design was used to mail surveys to 538 cardiac patients who were hospitalized over a 1-year period at a regional medical center; 255 surveys were returned, and the final sample numbered 222. Of these, 154 (69%) attended CR. Compared to nonparticipants (n = 68), participants reported significantly higher levels of functioning on 7 of the 8 subscales of the Medical Outcome Study Short Form-36 (SF-36). In relation to clinical risk factor outcomes, participants had a significantly lower body mass index than nonparticipants. Patients who attended CR reported that they had switched to low-fat foods, started an exercise program, lost weight, lowered stress, lowered blood pressure, and reduced blood cholesterol at significantly higher rates than non-participants. Patients were more likely to participate in CR if their physician explained its benefits, if they were told about it during their hospitalization, and if they lived close to a CR program. Patients with coronary heart disease need to be referred to CR for improved physical and psychosocial functioning and successful risk factor modification.


Spatially-selective amplified spontaneous emission source derived from an ultrahigh gain solid-state amplifier.
An investigation is made into the performance of a high power solid-state amplified spontaneous emission (ASE) source with near-diffraction-limited beam quality. The radiation from this ASE source has high spatial quality and power, but unlike a laser it has a high misalignment tolerance and does not require a precisely aligned cavity. The source is based on a diode-pumped Nd:YVO4 laser crystal in a bounce amplifier geometry with a uniquely ultra-high gain (~104-105). Double-pass ASE radiation with high power levels (>6W) is achieved in a near-diffraction-limited spatial quality. We further demonstrate that the double-pass ASE source also displays high spatial selectivity and capability to compensate for a phase diffuser, inserted in the double-pass arm, with only a small degradation in beam quality and power.


Zika Virus Infection, Reproductive Organ Targeting, and Semen Transmission in the Male Olive Baboon.
Zika virus (ZIKV) infection in pregnant women is a serious threat to the development and viability of the fetus. The primary mode of ZIKV transmission to humans is through mosquito bites, but sexual transmission has also been well documented in humans. However, little is known of the short- and long-term effects of ZIKV infection on the human male reproductive system. This study examines the effects of ZIKV infection on the male reproductive organs and semen and the immune response of the olive baboon (Papio anubis). Nine mature male baboons were infected with ZIKV (French Polynesian strain) subcutaneously. Six animals were euthanized at 41 days, while three animals were euthanized at 10 or 11 days postinfection (dpi). Viremia and clinical evidence of infection were present in all nine baboons. ZIKV RNA was present in the semen of five of nine baboons. ZIKV was present in the testes of two of three males euthanized at 10 or 11 dpi, but in none of six males at 41 dpi. Immunofluorescence of testes suggested the presence of ZIKV in sperm progenitor cells, macrophage penetration of seminiferous tubules, and increased tumor necrosis factor alpha (TNF-α), particularly in vascular walls. These data demonstrate that male olive baboons approximate the male human ZIKV response, including viremia, the adaptive immune response, and persistent ZIKV in semen. Although gross testicular pathology was not seen, the demonstrated breach of the testes-blood barrier and targeting of spermatogenic precursors suggest possible long-term implications in ZIKV-infected primates.IMPORTANCE Zika virus (ZIKV) is an emerging flavivirus spread through mosquitoes and sexual contact. ZIKV infection during pregnancy can lead to severe fetal outcomes, including miscarriage, fetal death, preterm birth, intrauterine growth restriction, and fetal microcephaly, collectively known as congenital Zika syndrome. Therefore, it is important to understand how this virus spreads, as well as the resulting pathogenesis in translational animal models that faithfully mimic ZIKV infection in humans. Such models will contribute to the future development of efficient therapeutics and prevention mechanisms. Through our previous work in olive baboons, we developed a nonhuman primate model that is permissive to ZIKV infection and transfers the virus vertically from mother to fetus, modeling human observations. The present study contributes to understanding of ZIKV infection in male baboon reproductive tissues and begins to elucidate how this may affect fertility, reproductive capacity, and sexual transmission of the virus.


Transcription from the thyroid hormone-dependent promoter of the Xenopus laevis thyroid hormone receptor betaA gene requires a novel upstream element and the initiator, but not a TATA Box.
The thyroid hormone receptor (TR) beta genes in Xenopus laevis are regulated by thyroid hormone in all organs of an animal during metamorphosis. This autoregulation appears to be critical for systematic transformations of different organs as a tadpole is transformed into a frog. To understand this autoregulation, we have previously identified a thyroid hormone response element in the hormone-dependent promoter of the X. laevis TRbetaA gene. We report here the detailed characterization of the promoter. We have now mapped the transcription start site and demonstrated the existence of an initiator element at the start site critical for promoter function. More important, our deletion and mutational experiments revealed a novel upstream DNA element that is located 125 base pairs upstream of the start site and that is essential for active transcription from the promoter. Promoter reconstitution experiments showed that this novel element does not function as an enhancer, but acts as a core promoter element, which, together with the initiator, directs accurate transcription from the promoter. Finally, we provide evidence for the existence of a protein(s) that specifically recognizes this element. Our studies thus demonstrate that the TRbetaA promoter has a unique organization consisting of an initiator and a novel upstream promoter element. Such an organization may be important for the ubiquitous but tissue-dependent temporal regulation of the gene by thyroid hormone during amphibian metamorphosis.


Willingness to receive a COVID-19 booster vaccine and its associated factors among adults with chronic disease: A cross-sectional study in Putrajaya, Malaysia.
Booster vaccination has been shown to reduce transmission and serious infection with COVID-19. This study examined the willingness to receive a COVID-19 booster vaccine and its associated factors among high-risk patients at Klinik Kesihatan Putrajaya Presint 9. A cross-sectional study was conducted among patients aged >18 years attending Klinik Kesihatan Putrajaya Presint 9 with a high risk of contracting COVID-19 recruited via systematic random sampling. Data were collected using a self-administered questionnaire. A multiple logistic regression analysis was performed to identify the associated factors. The response rate for this study was 97.4% (N=489). The median patient age was 55 years. Approximately 51.7% were men, and 90.4% were Malays. Approximately 81.2% were willing to receive a COVID-19 booster vaccine. The patients who perceived COVID-19 as a serious illness (Adjusted Odd Ratio, AOR=2.414), those who perceived COVID-19 booster vaccines as beneficial (AOR=7.796), those who disagreed that COVID-19 booster vaccines have many side effects (AOR=3.266), those who had no doubt about the content of COVID-19 vaccines (AOR=2.649) and those who were employed (AOR=2.559) and retired (AOR=2.937) were more likely to be willing to receive a booster vaccine than those who were unemployed and those who did not have close friends or family members who contracted severe COVID-19 (AOR=2.006). The majority of the participants were willing to receive a COVID-19 booster vaccine. Healthcare authorities should take initiatives to design targeted public intervention programmes to increase the willingness for COVID-19 booster vaccination.


Dissecting the inflammatory response in polymyalgia rheumatica: the relative role of IL-6 and its inhibition.
The efficacy of tocilizumab (TCZ), a monoclonal antibody to the interleukin (IL)-6 receptor, in suppressing disease activity in glucocorticoid-naïve patients with new-onset polymyalgia rheumatica (PMR) was studied. Its effect on a panel of cytokines and growth factors was evaluated. Three patients, fulfilling the PMR ACR/EULAR criteria, received TCZ at the dosage of 8 mg/kg every 4 weeks for three times followed by prednisone 0.2 mg/kg in case of inefficacy. Concentrations of IL-10, IL-6, tumour necrosis factor (TNF)-α, IL-1β, IL-10, IL-17, interferon (IFN)-γ, vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and leukaemia inhibitory factor (LIF) were measured at baseline, after 72 h of the first TCZ infusion and then at weeks 1, 4, 5, 8, 9, 12, 13, 14, 16, and 22. A slight clinical improvement was seen only after the first TCZ infusion, but was largely inferior to that of conventional doses of GC administered subsequently. An ischaemic visual accident suggestive of GCA occurred in one patient during TCZ treatment. IL-6 was increased at baseline compared to controls, further increased after the first TCZ infusion, and was suppressed by GC. IL-17 production decreased during TCZ treatment and reverted to pre-treatment levels after GC. VEGF e PDGF showed a less constant pattern, but an increase of VEGF concentration antedated visual symptoms. The other cytokines were not detectable in patients and controls. In our small sample, TCZ was not able to suppress inflammation at the same degree as GC. As a result, monotherapy with TCZ in PMR cannot be recommended, although its efficacy as adjunctive treatment in GC-resistant patients should be further evaluated.


Is demand-side financing equity enhancing? Lessons from a maternal health voucher scheme in Bangladesh.
Demand-side financing (DSF) is used in the less-developed countries of the world to improve access to healthcare and to encourage market supply. Under DSF, households receive vouchers that can be used to pay for healthcare services. This study evaluated the effects of a universal DSF on maternal healthcare service utilization in Bangladesh. A household survey was conducted in and around the voucher scheme area one year after the initiation of the project. Women who gave birth within a year prior to the survey were interviewed. The utilization rates of maternal health services were found to be higher for all socioeconomic groups in the project area than in the comparison areas. Voucher recipients in the project area were 3.6 times more likely to be assisted by skilled health personnel during delivery, 2.5 times more likely to deliver the baby in a health facility, 2.8 times more likely to receive postnatal care (PNC), 2.0 times more likely to get antenatal care (ANC) services and 1.5 times more likely to seek treatment for obstetric complications than pregnant women not in the program. The degree of socioeconomic inequality in maternal health service utilization was also lower in the project area than in the comparison area. The use of vouchers evidenced much stronger demand-increasing effects on the poor. Poor voucher recipients were 4.3 times more likely to deliver in a health facility and two times more likely to use skilled health personnel at delivery than the non-poor recipients. Contrary to the inverse equity hypothesis, the voucher scheme reduced inequality even in the short run. Despite these improvements, socioeconomic disparity in the use of maternal health services has remained pro-rich, implying that demand-side financing alone will be insufficient to achieve the Millennium Development Goal for maternal health. A comprehensive system-wide approach, including supply-side strengthening, will be needed to adequately address maternal health concerns in poor developing countries.


Sensory temporal thresholds and interhemispheric transfer times in multiple sclerosis: a preliminary study of a new outcome measure.
This preliminary study investigated the potential of a cerebral lateralization technique that measures both, temporal judgements to sensory stimuli and interhemispheric transfer time (IHTT), as an outcome measure in multiple sclerosis (MS). Tactile stimulation was delivered to one or both hands by mechanical tactile stimulators. Pairs of light emitting diodes were presented to hemifields for visual stimulation. Response consisted of a binary forced-choice (YES/NO) judgement as to the simultaneity of the onset of pairs of stimuli. Both tactile and visual temporal thresholds were significantly higher in MS patients than controls in every presentation condition. IHTT estimates (threshold differences between unilateral and bilateral presentations) for the tactile and visual tasks were also significantly longer in patients than controls. Age correlated with sensory temporal thresholds for the control group only. These findings suggest that this technique may be a useful outcome measure in MS. We hypothesize that myelin injury slows central conduction therefore impairing the ability to judge the onset of sensory stimuli and increasing IHTTs.


Trace metal concentrations and water solubility in size-fractionated atmospheric particles and influence of road traffic.
The abundance and the behavior of metals (Al, Ti, Mn, Fe, Co, Ni, Cu, Zn, Se, Ag, Cd, Sn, Ba, Pt, Hg, and Pb) and ions (Na+ K+ Mg2+ Ca2+, NH4+, Cl-, NO3-, SO4(2-), PO4(3-), and oxalate) in size-fractionated atmospheric particulate matter (PM) were studied in the U. K. and Ireland at four observation sites simulating extreme degrees of vehicular-traffic influence in the environment. Trace metals in urban PM showed distinct types of size-fractionated behavior depending on the particle sources from which they originate. In coarse PM (1.5 < Dp < 3.0 microm) the concentrations of copper, barium, and iron correlated closely across over 2 orders of magnitude in urban air, which is seen as evidence that major portions of transition metals (Cu, Ba, Fe, and Mn) are released through abrasive vehicular emissions, particularly the wear of brake linings. Further results are strongly indicative of a decoupling of coarse iron and calcium, the former arising predominantly from vehicles, the latter from soil resuspension. In fine PM (Dp < 0.5 microm), several combustion and secondary sources of particulates were identified, but these were much less unique in terms of elemental fingerprints. An analysis of the water solubility of trace metals yielded that solubility varies considerably with element and, to a lesser extent, with particle size. Notable differences were found to the elemental water solubilities determined in previous work, partially explained by differences in extraction procedures.


Blueberry as a Potential Radiosensitizer for Treating Cervical Cancer.
Cervical cancer (CC) is a leading cause of death in women worldwide. Radiation therapy (RT) for CC is an effective alternative, but its toxicity remains challenging. Blueberry is amongst the most commonly consumed berries in the United States. We previously showed that resveratrol, a compound in red grapes, can be used as a radiosensitizer for prostate cancer. In this study, we found that the percentage of colonies, PCNA expression level and the OD value of cells from the CC cell line SiHa were all decreased in RT/Blueberry Extract (BE) group when compared to those in the RT alone group. Furthermore, TUNEL+ cells and the relative caspase-3 activity in the CC cells were increased in the RT/BE group compared to those in the RT alone group. The anti-proliferative effect of RT/BE on cancer cells correlated with downregulation of pro-proliferative molecules cyclin D and cyclin E. The pro-apoptotic effect of RT/BE correlated with upregulation of the pro-apoptotic molecule TRAIL. Thus, BE sensitizes SiHa cells to RT by inhibition of proliferation and promotion of apoptosis, suggesting that blueberry might be used as a potential radiosensitizer to treat CC.


Drug-induced retroperitoneal fibrosis: short aetiopathogenetic note, from the past times of ergot-derivatives large use to currently applied bio-pharmacology.
Among the secondary forms of retroperitoneal fibrosis (RPF), that drug-induced shows very intriguing aspects given both the broad range of involved pharmaceuticals and the considerable interest arisen from the related pathogenetic mechanisms. The particular incidence, in the last four decades past century, of the RPF due to long-term use of ergot alkaloid derivatives (ergotamine, methysergide, pergolide, bromocriptine, cabergoline) and specific L-dopa derived agents, such as methyldopa, as well as to different analgesics, came progressively down given that their long-term use for either the prevention of migraine attacks or the therapy of chronic pathologies (Parkinson's disease, prolactinoma, pain management, etc) has been, year after year, supplanted or even made unavailable in many countries. More recently, instead, the occurrence of the RPF has been sometimes identified with the use of antitumoral chemotherapeutics, such as carboplatin and methotrexate, and, just lately, as an unusual side-effect of certain biological agents, about which it is timely to go into specific pathogenetic problems in more depth.


Female and male sexual responses in female cats with ventromedial hypothalamic lesions.
The effects of lesions in the ventromedial hypothalamus (VMH) on male and female sexual behavior in female cats were examined. Following preoperative testing for receptivity, proceptivity, and male mating behavior, 27 female cats received either lesions in the anterior or posterior portion of the VMH or sham lesions. Neither of the VMH lesion placements reduced proceptivity and receptivity scores. However, the female mating pattern was significantly altered in that although females in both lesion groups initially allowed a stud male to mount, they usually did not allow the male to intromit. As in rodents, the VMH in the cat appears to be an important neural area for the display of normal female sexual behavior. Low levels of male sexual behavior were seen in the three groups preoperatively, and no changes were observed in the levels of male behavior following lesion placement. Thus, because lesions that disrupted female behavior did not affect male sexual behavior, it appears that the neural areas controlling homotypical and heterotypical sexual behaviors are not necessarily neurally linked.


Massive perivillous fibrin deposition: Diagnosis, obstetrical features, and treatment.
MPVFD (Massive perivillous fibrin deposition) is placental lesion characterized by extensive massive deposits of fibrin in the intervillous space, extending over at least 25 % of the placental volume. Currently, this pathology can only be detected through histopathological examination of the placenta after a pregnancy has ended. The underlying mechanisms are poorly studied, there is no biomarker available for the diagnosis of MPVFD and treatment protocols are experimental and still lacking. The objective of this study is to systematically review the literature on the associated clinicopathologic features, treatment, and prognosis of MPVFD. We ended up with 17 studies, of these 12 studies were considered relevant for this article and included in the final analysis. All studies reporting MPVFD are retrospective. MPVFD is associated with recurrent miscarriage, intra uterine fetal death (IUFD), intra uterine growth restriction (IUGR) and preterm delivery. The prevalence in pregnancies with a delivery after 22 weeks of gestation was at 1.1 % and even higher to 2.7 % in recurrent early miscarriages. The reported risk of fetal death in MPVFD ranges mainly from 15 to 80 %. Preterm delivery is spontaneous in 50 to 70 % of cases and induced by of a severe intrauterine growth restriction (IUGR) in 30 to 50 % of cases depending on the study. Its causes and treatment are still poorly understood, although several avenues have been explored. This review summarizes current understanding of the prevalence, diagnostic features, clinical consequences, immune pathology, and potential prophylaxis against recurrence in this chronic inflammatory placental syndrome.


Diacerein inhibits the synthesis of resorptive enzymes and reduces osteoclastic differentiation/survival in osteoarthritic subchondral bone: a possible mechanism for a protective effect against subchondral bone remodelling.
Subchondral bone alterations represent an essential component of osteoarthritis (OA). Modifying the abnormal subchondral bone metabolism may be indicated to treat OA. We investigated the effect of diacerein and rhein on the changes occurring in subchondral bone during OA. To this end, we determined the drugs' effects on metalloprotease-13 (MMP-13) synthesis on subchondral bone and on the osteoblast signalling pathways. In osteoclasts, we studied MMP-13 and cathepsin K production as well as cell differentiation, proliferation, and survival. The effect of diacerein/rhein on the production of subchondral bone MMP-13 was determined by enzyme-linked immunosorbent assay. Signalling pathways were evaluated on osteoblasts by Western blot. Osteoclast experiments were performed using cells from the pre-osteoclastic murine cell line Raw 264.7. Osteoclast MMP-13 and cathepsin K activities were determined by specific bioassays and differentiation of these cells quantified by tartrate-resistant acid phosphatase staining. Diacerein and rhein reduced, in a dose-dependent manner, the interleukin-1-beta (IL-1beta)-induced MMP-13 production in OA subchondral bone. This effect occurred through the inhibition of ERK1/2 (extracellular signal-regulated kinase-1/2) and p38. In osteoclasts, they significantly reduced the activity of MMP-13 and cathepsin K. Moreover, these drugs effectively blocked the IL-1beta effect on the osteoclast differentiation process and the survival of mature osteoclasts. Altogether, these data suggest that diacerein/rhein could impact the abnormal subchondral bone metabolism in OA by reducing the synthesis of resorptive factors and osteoclast formation.


Limited diversity of Anopheles darlingi in the Peruvian Amazon region of Iquitos.
Anopheles darlingi is the most important malaria vector in the Amazon basin of South America, and is capable of transmitting both Plasmodium falciparum and P. vivax. To understand the genetic structure of this vector in the Amazonian region of Peru, a simple polymerase chain reaction (PCR)-based test to identify this species of mosquito was used. A random amplified polymorphic DNA-PCR was used to study genetic variation at the micro-geographic level in nine geographically separate populations of An. darlingi collected in areas with different degrees of deforestation surrounding the city of Iquitos. Within-population genetic diversity in nine populations, as quantified by the expected heterozygosity (H(E)), ranged from 0.27 to 0.32. Average genetic distance (F(ST)) among these populations was 0.017. These results show that the nine studied populations are highly homogeneous, suggesting that strategies can be developed to combat this malaria vector as a single epidemiologic unit.


The Risk Factors of Exacerbation in Interstitial Pneumonia With Autoimmune Features: A Single-Center Observational Cohort Study.
To investigate the long-term outcomes, including risk factors, for exacerbation between monotherapy and combination therapy in patients with interstitial pneumonia with autoimmune features (IPAF). We assessed 672 patients between April 2009 and March 2019 who were evaluated using high-resolution computed tomography (HRCT) of the chest. We applied the IPAF criteria. Fifty-two patients who visited our department for at least 6 months were diagnosed with IPAF. Clinical, laboratory, and imaging data were collected from medical records and statistically analyzed. Among the 52 cases of IPAF, we compared the characteristics at diagnosis between treated (n = 28) and untreated patients (n = 24). The exacerbation rates were 42.9% (n = 12) and 8.3% (n = 2) (P = 0.0051), respectively. Among the treated patients, smoking history, high titer of KL-6, and the duration from diagnosis to the start of treatment were significant risk factors for exacerbation (P = 0.0062, 0.011, and 0.019, respectively). The number of risk factors was significantly and positively associated with exacerbation rate (P = 0.0053). Among the treated patients, glucocorticoid (GC) monotherapy was used in 13 cases, and GC and immunosuppressant (IS) combination therapy was used in 14 patients. There was no significant difference in the treatment methods between patients with and without risk factors (P = 0.47). When comparing the long-term outcomes between the monotherapy and combination therapy groups, the 3-year non-exacerbation rates were 72.9 and 45.9% (P = 0.020), respectively. IPAF patients with risk factors had a high exacerbation rate, regardless of the type of treatment. New interventions aimed at preventing exacerbations in these patients are required.


Running Mechanics and Variability with Aging.
As the elderly population in the United States continues to grow, issues related to maintenance of health become increasingly important. Physical activity has positive benefits for healthy aging. Running, a popular form of exercise, is associated with the risk of developing injury, especially in older runners. Initial differences between older and younger runners have been observed, but these were observed without consideration of other differences between groups, such as running mileage. This study aims to compare running mechanics and lower-extremity coordination variability in matched groups of healthy younger and healthy older runners. Three-dimensional kinetics and kinematics were collected while 14 older adults (45-65 yr) and younger adults (18-35 yr) ran overground at 3.5 m·s. Knee, ankle, and hip joint angles and moments were determined. Discrete measures at foot strike (maximum and minimum) were determined and compared between groups. Segment angles during stance were utilized to calculate segment coordination variability between pelvis and thigh, thigh and shank, and shank and foot, using a modified vector coding technique. Knee and ankle joint angles were similar between groups (P > 0.05). Older runners had greater hip range of motion (P = 0.01) and peak hip flexion (P = 0.001) at a more extended hip position than younger runners. Older runners had smaller ankle plantarflexion moment (P = 0.04) and hip rotational moment (P = 0.005) than younger runners. There were no between-group differences in any of the variability measures (P > 0.05). Runners appear to maintain movement patterns and variability during running with increasing age, indicating that running itself may be contributing to maintenance of health among older runners in the current study.


The effects of HIV and systolic blood pressure on mortality risk in rural South Africa, 2010-2019: a data note.
South Africa is experiencing both HIV and hypertension epidemics. Data were compiled for a study to identify effects of HIV and high systolic blood pressure on mortality risk among people aged 40-plus in a rural South African area experiencing high prevalence of both conditions. We aim to release the replication data set for this study. The research data comes from the 2010-11 Ha Nakekela (We Care) population-based survey nested in the Agincourt Health and socio-Demographic Surveillance System (AHDSS) located in the northeast region of South Africa. An age-sex-stratified probability sample was drawn from the AHDSS. The public data set includes information on individual socioeconomic characteristics and measures of HIV status and blood pressure for participants aged 40-plus by 2019. The AHDSS, through its annual surveillance, provided mortality data for nine years subsequent to the survey. These data were converted to person-year observations and linked to the individual-level survey data using participants' AHDSS census identifier. The data can be used to replicate Houle et al. (2022) - which used discrete-time event history models stratified by sex to assess differential mortality risks according to Ha Nakekela measures of HIV-infection, HIV-1 RNA viral load, and systolic blood pressure.


Methylphenidate, dextroamphetamine, and levamfetamine. Effects on schizophrenic symptoms.
Methylphenidate hydrochloride dextroamphetamine sulfate, and levamfetamine succinate have potential as pharmacologic tools for the indirect evaluation of the role of neurotransmitters in schizophrenia. In actively ill schizophrenic patients, methylphenidate administered intravenously causes a brief but clear intensification of preexisting psychotic symptoms, such as hallucinations and delusions. In our study, methylphenidate, dextroamphetamine, and levamfetamine were administered in equimolar doses to schizophrenic patients. Methylphenidate was a more effective activator of symptoms than dextroamphetamine, which in turn was more effective than levamfetamine. Levodopa (L-dopa) given orally also reportedly produces a temporary worsening of schizophrenic symptoms. While these findings augment a body of information suggesting that dopamine and norepinephrine may play a role in the activation of schizophrenic symptoms, our findings with methylphenidate (reportedly weak in eliciting stereotyped behaviour in rat) and our review of the literature indicate complexities that remain to be resolved. There is some utility of the procedure for differential diagnosis and selective therapy, but this is still of occasional and limited potential.


True hemangiopericytoma of the nasal cavity.
Two cases of nasal tumors with pericytic myoid differentiation are reported. The tumors occurred in a 77-year-old woman and a 60-year-old man as polypoid lesions covered by normal mucosa. Histologically, the tumors were composed of uniform short spindle or stellate cells with indistinct cell borders arranged in narrow and short fascicles. Numerous blood vessels of various sizes were common in both cases. The tumor cells of both cases stained intensely with anti-vimentin and anti-actin antibodies, but not with anti-desmin, CD34, or anti-high-molecular-weight caldesmon antibodies. Ultrastructural examination revealed well-developed actin thin filaments with dense bodies, subplasmalemmal plaques, intercellular junctions, and irregular discontinuous basement membranes. These histopathologic features suggest true pericytic differentiation of the tumors (true hemangiopericytoma), unlike soft tissue-type hemangiopericytoma. Generally, sinonasal hemangiopericytomas are subdivided into soft tissue-type hemangiopericytomas and true hemangiopericytomas identical to the cases presented here. Soft tissue-type hemangiopericytomas are frequently highly aggressive, whereas true hemangiopericytomas show localized benign behavior. Sinonasal true hemangiopericytomas should be strictly differentiated from soft tissue-type hemangiopericytomas.


Preclinical development of human granulocyte-macrophage colony-stimulating factor-transfected melanoma cell vaccine using established canine cell lines and normal dogs.
Tumor vaccines and gene therapy have received significant attention as means of increasing cellular and humoral immune responses to cancer. We conducted a pilot study of seven research dogs to determine whether intradermal injection of canine tumor cells transfected via the Accell particle-mediated gene transfer device with the cDNA for human granulocyte-macrophage colony-stimulating factor (hGM-CSF) would generate biologically relevant levels of protein and result in demonstrable histological changes at sites of vaccination. Tumor cell vaccines of 10(7) irradiated canine melanoma cells were nontoxic, safe, and well tolerated. No significant alterations in blood chemistry values or hematological profiles were detected. A histological review of control vaccine sites revealed inflammatory responses predominated by eosinophils, whereas vaccine sites with hGM-CSF-transfected tumor cells had an influx of neutrophils and macrophages. Enzyme-linked immunosorbent assays of skin biopsies from vaccine sites had local hGM-CSF production (8.68-16.82 ng/site of injection) at 24 hours after injection and detectable levels (0.014-0.081 ng/site) for < or =2 weeks following vaccination. Flow cytometric analysis of hGM-CSF-transfected cells demonstrated < or =25% transfection efficiency, and hGM-CSF levels obtained during time-course assays demonstrated biologically relevant levels for both irradiated and nonirradiated samples. These data demonstrate the in vivo biological activity of irradiated hGM-CSF-transfected canine tumor cells and help provide evidence for a valid translational research model of spontaneous tumors.


Quantitative analysis of proton-linked transport system. beta-Galactoside exit in Escherichia coli.
The exit of lactose and thiomethyl-beta-D-galactoside from Escherichia coli ML308-225 has been studied to determine the role of carrier-dependent (zero-trans efflux) and carrier-independent (leak) processes. On the basis of its sensitivity to p-chloromercuribenzene sulphonate the exit of lactose was found to be almost wholly mediated by the carrier. Consistent with this conclusion was the finding that the rate of exit of this sugar was dependent on the external pH, being considerably slower at acid pH. On the other hand exit of thiomethyl-beta-D-galactoside was found to be composed of both carrier-dependent and carrier-independent processes. Both processes exhibited first-order kinetics with the rate constants for zero-trans efflux and leak being 0.137 min-1 and 0.079 min-1, respectively. The relevance of these findings for out earlier proposal for the methods of attenuation of solute accumulation is discussed [Booth, Mitchell & Hamilton (1979) Biochem. J. 182, 687--696].


Microsporidial infections in humans: current practice and developments in laboratory diagnosis.
Microsporidia are small, intracellular parasites that infect a wide range of hosts, including vertebrates, invertebrates and fish. They were discovered more than a century ago. The first well documented human case, however, was not reported until 100 years later. Since the first case of intestinal microsporidiosis was reported in 1985, numerous cases of microsporidiosis have been reported in immunocompromised patients, especially those in the later stages of human immunodeficiency virus (HIV) infection. Microsporidia also have been described in various other clinical conditions, including keratoconjunctivitis, sinusitis, peritonitis and myositis. The numbers of cases reported have risen dramatically since 1985, which can be explained partly by the acquired immune deficiency syndrome (AIDS) pandemic and partly by increased laboratory awareness. Some studies have shown that up to 50% of selected AIDS patients are infected with microsporidia. Diagnosis depended initially on the use of invasive techniques, namely histological examination of biopsy material. Since then, however, there have been important advances in the detection of microsporidial spores in clinical samples. Recent developments in the diagnosis of microsporidiosis are described, including light microscopy staining methods, fluorescent staining, electron microscopy and molecular techniques.


Developing a simulation-based training curriculum in transesophageal ultrasound with the use of the endobronchial ultrasound-endoscope.
There is an increasing need to focus on how best to train respiratory physicians to perform EUS with bronchoscope-guided fine-needle aspiration biopsy (EUS-B-FNA). At current, training is mostly performed in the clinical environment under expert supervision; however, the advent of simulation-based education now provides a low-risk setting for novice trainees to learn and practice basic endosonography skills from identifying and understanding normal anatomy as well as pathology, maneuvering of endoscope, interpretation of images, and mastering of sampling techniques. In this descriptive educational paper, we used a six-step approach as a framework to describe the development of a structured training program combining EUS-B-FNA with the already well-established certification training program in endobronchial ultrasound transbronchial needle aspiration. This comprehensive training curriculum includes a theoretical course to achieve foundational knowledge, followed by simulation-based training until mastery standards are met, and supervised clinical apprenticeship. All steps should end with an objective assessment to achieve certification. This systematic development will hopefully encourage endosonography leaders and educators to collaborate and implement an evidence-based comprehensive endosonography curriculum that aims to provide the trainee with the essential EUS-B competencies to ensure that lung cancer patients are diagnosed and staged correctly.


Bronchoscopic intrabullous autologous blood instillation: a novel approach for the treatment of giant bullae.
The current standard therapy for patients with giant bullae is surgical bullectomy; however, high operative risk and comorbidities preclude surgical procedures in many patients. Autologous blood instilled directly into bullae can induce an inflammatory reaction, leading to scarring, fibrosis, and ultimately volume loss. We have treated 5 patients with this minimally invasive approach as day-case procedures using moderate sedation. Three of the 5 patients had shrinkage of the bullae, leading to large and clinically meaningful improvements in lung function, exercise capacity, and quality of life 3 months after treatment.


Cytochrome P450-catalyzed dealkylation of atrazine by Rhodococcus sp. strain NI86/21 involves hydrogen atom transfer rather than single electron transfer.
Cytochrome P450 enzymes are responsible for a multitude of natural transformation reactions. For oxidative N-dealkylation, single electron (SET) and hydrogen atom abstraction (HAT) have been debated as underlying mechanisms. Combined evidence from (i) product distribution and (ii) isotope effects indicate that HAT, rather than SET, initiates N-dealkylation of atrazine to desethyl- and desisopropylatrazine by the microorganism Rhodococcus sp. strain NI86/21. (i) Product analysis revealed a non-selective oxidation at both the αC and βC-atom of the alkyl chain, which is expected for a radical reaction, but not SET. (ii) Normal (13)C and (15)N as well as pronounced (2)H isotope effects (εcarbon: -4.0‰ ± 0.2‰; εnitrogen: -1.4‰ ± 0.3‰, KIEH: 3.6 ± 0.8) agree qualitatively with calculated values for HAT, whereas inverse (13)C and (15)N isotope effects are predicted for SET. Analogous results are observed with the Fe(iv)[double bond, length as m-dash]O model system [5,10,15,20-tetrakis(pentafluorophenyl)porphyrin-iron(iii)-chloride + NaIO4], but not with permanganate. These results emphasize the relevance of the HAT mechanism for N-dealkylation by P450.