Compartmental fat distribution in the abdomen of dogs relative to overall body fat composition.

BACKGROUND: Adipose tissue may have different metabolic and endocrine functions depending on the region of the body in which it is located. While visceral or intra-abdominal fat has been found to contribute to leptin concentrations, insulin resistance and obesity-related diseases, there are only a few imaging studies documenting the preferential distribution of body fat to either the intra-abdominal or subcutaneous compartments in dogs. This study aimed to determine if CT-measured abdominal fat distributed preferentially to the visceral space (V) relative to the subcutaneous space (SQ), with increasing DXA-determined total body fat percentage; and if ultrasound measurements of the ventral midline subcutaneous (SAT) and visceral adipose thickness (VAT) can be used to estimate the distribution of fat to the subcutaneous and visceral abdominal spaces, in a sample of 22 dogs with variable body condition. RESULTS: Multivariate analysis showed no statistically significant correlation between visceral to subcutaneous fat ratio (V/SQ) and increasing total body fat percentage (ß = - 0.07, p = 0.733), but strong correlation with age (ß = 0.71 p = 0.002). A substantial amount of variation for the ultrasound visceral adipose thickness to subcutaneous fat thickness (VAT/SAT) could be explained by both CT V/SQ and sex (R2Adjusted = 0.477, p = 0.001), with female dogs having significant lower VAT/SAT ratios compared to the male dogs (p = 0.047). The ultrasound fat measurements appeared moderately reliable, but a larger sample number is required to confirm this. CONCLUSIONS: The findings suggest that dogs with a relatively healthy to slightly overweight body condition score, distribute fat relatively similarly between their peritoneal (visceral) and subcutaneous abdominal compartments with increasing total body fat percentage. However, there was increased fat distribution to the peritoneal space relative to the subcutaneous space with increasing age. Further, abdominal ultrasound may be useful in estimating the ratio of fat distribution to both the abdominal visceral and subcutaneous spaces.

Abdominal volume computed tomography assessment of body composition in dogs.

BACKGROUND: Computed tomography (CT) has been used to estimate body composition and determine tissue distribution in dogs, despite limited validation. This may introduce error into estimates of body composition studies and its effect on health in dogs. Further, the modality has not been validated against dual-energy X-ray absorptiometry (DXA) or over a wide range of dog breeds, ages and sexes. The objective of this study was to validate the use of semi-automated, abdominal volume CT for estimating total body composition of dogs relative to DXA. Twenty-two staff-owned dogs (weighing between 5.1-60 kg) were sedated and underwent full body DXA scan and abdominal CT. Abdominal tissue composition was estimated by CT using semi-automated volume segmentation, over predetermined tissue Hounsfield threshold values. Abdominal tissue composition determined by the various CT threshold ranges was compared to total body composition determined by DXA. RESULTS: Abdominal tissue composition estimated by CT strongly correlated with the estimates derived from DXA with a small Bland-Altman mean percentage differences in values: total body mass (- 250/2000HU: r2 = 0.985; - 1.10%); total fat mass (- 250/-25HU: r2 = 0.981; - 1.90%); total lean tissue mass (- 25/150HU: r2 = 0.972; 3.47%); and total bone mineral content (150/2000HU: r2 = 0.900; - 0.87%). Although averaged CT values compared well to DXA analysis, there was moderate variation in the individual predicted values. There was near perfect inter- and intra-observer agreement in segmentation volumes for abdominal fat. CONCLUSIONS: Abdominal volume computed tomography (CT) accurately and reliably estimates total body composition in dogs, but greater variations may be observed in dogs weighing less than 10 kg.

HE4 suppresses the expression of osteopontin in mononuclear cells and compromises their cytotoxicity against ovarian cancer cells.

Ovarian cancers are known to evade immunosurveillance and to orchestrate a suppressive immune microenvironment. Here we examine the role of human epididymis protein 4 (HE4), an ovarian cancer biomarker, in immune evasion. Through modified subtractive hybridization analyses we have characterized the gene targets of HE4 in human peripheral blood mononuclear cells (PBMCs), and established a preliminary mechanism for HE4-mediated immune failure in ovarian tumours. Upon exposure of purified PMBCs to HE4, osteopontin (OPN) and dual-specificity phosphatase 6 (DUSP6) emerged as the most suppressed and up-regulated genes, respectively. SKOV3 and OVCAR8, human ovarian carcinoma cell lines, exhibited enhanced proliferation in conditioned media from HE4-exposed PBMCs, an effect that was attenuated by the addition of recombinant OPN or OPN-inducible cytokines [interleukin (IL)-12 and interferon (IFN)-Ɣ]. Additionally, upon co-culture with PBMCs, HE4-silenced SKOV3 cells were found to be more susceptible to cytotoxic cell death. The relationship between HE4 and OPN was reinforced further through the analysis of serous ovarian cancer patient samples. In these biopsy specimens, the number of OPN+ T cells correlated positively with progression free survival (PFS) and inversely with serum HE4 level. Taken together, these findings show that HE4 enhances ovarian cancer tumorigenesis by compromising OPN-mediated T cell activation.

Assessment of the clinical usefulness of ultrasound-guided cytological specimens obtained from gastrointestinal lesions in dogs and cats.

OBJECTIVES: Cytological biopsies are an integral additional test to an abdominal ultrasound when a lesion is identified, but there is little published on factors that that may impact achieving a clinically useful sample of gastrointestinal lesions obtained by ultrasound-guided fine-needle cytologic biopsy. This retrospective, descriptive study aimed to assess factors that may influence the clinical usefulness of submitted cytological samples collected from gastrointestinal lesions by ultrasound-guided percutaneous fine-needle cytologic biopsy. MATERIAL AND METHODS: Gastrointestinal cytological samples obtained from 25 dogs and 19 cats over 2.5 years were reviewed and determined as clinically useful or clinically useless as per the cytology report. Variables dependent on the ultrasound exam that were used in the analysis included lesion location, lesion thickness, loss of gastrointestinal layering, and the number of slides submitted. RESULTS: Thirty (30/44) of the submitted cytological samples were considered clinically useful. Factors associated with achieving a clinically useful sample in univariable models included the number of slides submitted and the thickness of the lesion. However, these two variables appear inter-related, as a weak correlation existed between them. Where histologic biopsies were obtained, a clinically useful sample had a partial or complete agreement with histology in three of 12 and eight of 12 cases, respectively. CLINICAL SIGNIFICANCE: Ultrasound-guided fine-needle cytological biopsies of gastrointestinal masses provided a clinically useful sample in two-third of the cases, especially if more slides were provided to the cytologist and thicker lesions were sampled.

L-Lactic acid: a mosquito attractant isolated from humans.

L-Lactic acid was the major component in material isolated from humans that was active as an attractant for female yellow fever mosquitoes, Aedes aegypti (L.). The L(+)-isomer was several times as attractive as the D-isomer. Good correlation was found between the attractiveness of an individual to mosquitoes and the quantity of lactic acid present in an acetone washing of his hand.

Effect of probiotic on innate inflammatory response and viral shedding in experimental rhinovirus infection - a randomised controlled trial.

Ingestion of probiotics appears to have modest effects on the incidence of viral respiratory infection. The mechanism of these effects is not clear; however, there is evidence from animal models that the probiotic may have an effect on innate immune responses to pathogens. The purpose of this randomised, placebo-controlled study was to determine the effect of administration of Bifidobacterium animalis subspecies lactis Bl-04 on innate and adaptive host responses to experimental rhinovirus challenge. The effect on the response of chemokine (C-X-C motif) ligand 8 (CXCL8) to rhinovirus infection was defined as the primary endpoint for the study. 152 seronegative volunteers who had been supplemented for 28 days, 73 with probiotic and 79 with placebo, were challenged with RV-A39. Supplement or placebo administration was then continued for five days during collection of specimens for assessment of host response, infection, and symptoms. 58 probiotic and 57 placebo-supplemented volunteers met protocol-defined criteria for analysis. Probiotic resulted in higher nasal lavage CXCL8 on day 0 prior to virus challenge (90 vs 58 pg/ml, respectively, P=0.04, ANCOVA). The CXCL8 response to rhinovirus infection in nasal lavage was significantly reduced in the probiotic treated group (P=0.03, ANCOVA). Probiotic was also associated with a reduction in nasal lavage virus titre and the proportion of subjects shedding virus in nasal secretions (76% in the probiotic group, 91% in the placebo group, P=0.04, Fisher Exact test). The administration of probiotic did not influence lower respiratory inflammation (assessed by exhaled nitric oxide), subjective symptom scores, or infection rate. This study demonstrates that ingestion of Bl-04 may have an effect on the baseline state of innate immunity in the nose and on the subsequent response of the human host to rhinovirus infection. Clinicaltrials.gov registry number: NCT01669603.

NMR structure of stem-loop SL2 of the HIV-1 psi RNA packaging signal reveals a novel A-U-A base-triple platform.

The genome of the human immunodeficiency virus type-1 (HIV-1) contains a stretch of approximately 120 nucleotides known as the psi-site that is essential for RNA packaging during virus assembly. These nucleotides have been proposed to form four stem-loops (SL1-SL4) that have both independent and overlapping functions. Stem-loop SL2 is important for efficient recognition and packaging of the full-length, unspliced viral genome, and also contains the major splice-donor site (SD) for mRNA splicing. We have determined the structure of the 19-residue SL2 oligoribonucleotide by heteronuclear NMR methods. The structure is generally consistent with the most recent of two earlier secondary structure predictions, with residues G1-G2-C3-G4 and C6-U7 forming standard Watson Crick base-pairs with self-complementary residues C16-G17-C18-C19 and A12-G13, respectively. However, residue A15, which is located near the center of the stem, does not form a predicted bulge, and residues A5 and U14 do not form an expected Watson-Crick base-pair. Instead, these residues form a novel A5-U14-A15 base-triple that appears to be stabilized by hydrogen bonds from A15-H61 and -H62 to A5-N1 and U14-O2, respectively; from A5-H61 to U14-O2, and from C16-H42 to U14-O2'. A kink in the backbone allows the aromatic rings of the sequential U14-A15 residues to be approximately co-planar, adopting a stable "platform motif" that is structurally similar to the A-A (adenosine) platforms observed in the P4-P6 ribozyme domain of the Tetrahymena group I intron. Platform motifs generally function in RNA by mediating long-range interactions, and it is therefore possible that the A-U-A base-triple platform mediates long-range interactions that either stabilize the psi-RNA or facilitate splicing and/or packaging. Residue G8 of the G8-G9-U10-G11 tetraloop is stacked above the U7-A12 base-pair, and the remaining tetraloop residues are disordered and available for potential interactions with either other RNA or protein components.

NMR structure of the HIV-1 nucleocapsid protein bound to stem-loop SL2 of the psi-RNA packaging signal. Implications for genome recognition.

The RNA genome of the human immunodeficiency virus type-1 (HIV-1) contains a approximately 120 nucleotide Psi-packaging signal that is recognized by the nucleocapsid (NC) domain of the Gag polyprotein during virus assembly. The Psi-site contains four stem-loops (SL1-SL4) that possess overlapping and possibly redundant functions. The present studies demonstrate that the 19 residue SL2 stem-loop binds NC with affinity (K(d)=110(+/-50) nM) similar to that observed for NC binding to SL3 (K(d)=170(+/-65) nM) and tighter than expected on the basis of earlier work, suggesting that NC-SL2 interactions probably play a direct role in the specific recognition and packaging of the full-length, unspliced genome. The structure of the NC-SL2 complex was determined by heteronuclear NMR methods using (15)N,(13)C-isotopically labeled NC protein and SL2 RNA. The N and C-terminal "zinc knuckles" (Cys-X(2)-Cys-X(4)-His-X(4)-Cys; X=variable amino acid) of HIV-1 NC bind to exposed guanosine bases G9 and G11, respectively, of the G8-G9-U10-G11 tetraloop, and residues Lys3-Lys11 of the N-terminal tail forms a 3(10) helix that packs against the proximal zinc knuckle and interacts with the RNA stem. These structural features are similar to those observed previously in the NMR structure of NC bound to SL3. Other features of the complex are substantially different. In particular, the N-terminal zinc knuckle interacts with an A-U-A base triple platform in the minor groove of the SL2 RNA stem, but binds to the major groove of SL3. In addition, the relative orientations of the N and C-terminal zinc knuckles differ in the NC-SL2 and NC-SL3 complexes, and the side-chain of Phe6 makes minor groove hydrophobic contacts with G11 in the NC-SL2 complex but does not interact with RNA in the NC-SL3 complex. Finally, the N-terminal helix of NC interacts with the phosphodiester backbone of the SL2 RNA stem mainly via electrostatic interactions, but does not bind in the major groove or make specific H-bonding contacts as observed in the NC-SL3 structure. These findings demonstrate that NC binds in an adaptive manner to SL2 and SL3 via different subsets of inter and intra-molecular interactions, and support a genome recognition/packaging mechanism that involves interactions of two or more NC domains of assembling HIV-1 Gag molecules with multiple Psi-site stem-loop packaging elements during the early stages of retrovirus assembly.

Ineffectiveness of intranasal zinc gluconate for prevention of experimental rhinovirus colds.

Zinc has generally been administered by the oral route in studies of prevention or treatment of the common cold. The purpose of these studies was to evaluate the effectiveness of intranasal zinc gluconate for prevention of experimental rhinovirus infection and illness. Ninety-one volunteers, 41 treated with active medication and 50 treated with placebo, received study medication for 3 days, were inoculated with rhinovirus, and then were treated with study medication for an additional 6 days. Rhinovirus infection was documented in 37 (74%) of the 50 placebo-treated volunteers and in 32 (78%) of the 41 volunteers treated with active medication. Zinc treatment had no effect on total symptom score, rhinorrhea, nasal obstruction, or the proportion of infected volunteers who developed clinical colds. These data do not support a role for intranasal zinc gluconate for prevention or treatment of the common cold.

The role of oxidative stress in rhinovirus induced elaboration of IL-8 by respiratory epithelial cells.

A direct correlation has been reported between the severity of symptoms associated with rhinovirus infection and the concentration of interleukin-8 in nasal secretions. The purpose of these studies was to examine the mechanism of rhinovirus-induced IL-8 elaboration. Rhinovirus infection induced oxidative stress in Beas-2b cells and the concentration of H2O2 in supernatant media from rhinovirus challenged cells was 12.5 +/- 6.1 microM 1 h after challenge compared to 0.7 +/- 0.3 microM in supernatant from control cells. N-acetyl cysteine inhibited RV-induced NF-kappaB activation and IL-8 elaboration. IL-8 concentrations were 36 +/- 2 pg/ml and 10 +/- 1 pg/ml 6 h after virus challenge in untreated and NAC-treated (30 mM NAC) cells, respectively. Despite the effects of NAC on IL-8 elaboration and NF-kappaB activation, RV stimulated increases in supernatant H2O2 were not altered by NAC. These data suggest that RV stimulation of IL-8 in respiratory epithelium is mediated through production of oxidative species and the subsequent activation of NF-kappaB.

Cytomat-R: a computer-controlled multiple laser source multiparameter flow cytophotometer system.

A multiple illumination wavelength multiparameter flow cytophotometer system, using laser sources and controlled by a small, general-purpose digital computer, has been produced for use in the development of new flow cytometric techniques. Three different laser wave-lengths can be used simultaneously to illuminate different regions of the flow chamber; as many as five measurements of light scattering at various angles, extinction, and fluorescence at one or more wavelengths can be made at each illuminated station. Cells in suspension may be examined at rates of 1000 cells/sec, with seven correlated optical measurements being recorded for each cell. A library of programs for data manipulation and statistical analysis make it possible to use the system to develop and implement cell characterization, counting and classification procedures for basic and clinical research applications.

Counterimmunoelectrophoresis of urine for diagnosis of bacterial pneumonia in pediatric outpatients.

Thirty-eight pediatric outpatients with pneumonia were studied by counterimmunoelectrophoresis for the presence of Haemophilus influenzae type b or pneumococcal antigenuria. Of the 38 patients eight (21%) had H influenzae type b antigenuria and two (5%) had pneumococcal antigenuria. H influenzae type b antigenuria was detected more frequently in patients less than 2 years of age than in older children. Urine counterimmunoelectrophoresis appears to be a useful tool for the etiologic diagnosis of bacterial pneumonia and should facilitate further studies of the epidemiology, pathogenesis, and clinical spectrum of this disease.

Effect of topical adrenergic decongestants on middle ear pressure in infants with common colds.

BACKGROUND: Otitis media is frequently a complication of the common cold. Obstruction of the eustachian tube ostia by nasopharyngeal edema has been suggested as a cause of this complication. OBJECTIVE: To determine the effect of a topical adrenergic decongestant on middle ear pressure in infants with common cold symptoms. METHODS: The study was conducted with a randomized, double blinded, placebo-controlled design. Middle ear pressure was determined in infants 6 to 18 months old who had common cold symptoms. Infants with abnormal middle ear pressure (< or = -100 mm H2O) in either ear were treated with intranasal phenylephrine drops or placebo. The effect of the treatment on middle ear pressure in ears with abnormal pressure at baseline was determined 1 h later. RESULTS: Twenty-three of 44 infants with abnormal middle ear pressures received intranasal phenylephrine drops and 21 received placebo. Middle ear pressure remained abnormal after treatment with phenylephrine in 29 of 33 (88%) ears and after treatment with placebo in 26 of 34 (76%). The mean change in middle ear pressure after treatment was +23 mm H2O in the active group and +40 mm H2O in the placebo group. CONCLUSIONS: Treatment of nasal obstruction with topical adrenergic decongestants does not improve abnormal middle ear pressures during the common cold.

Passive immunization for prevention of rotavirus illness in healthy infants.

Infant formula supplemented with bovine antibody (BCIg) to human rotavirus has been reported to prevent or modify rotavirus infection and illness. The purpose of this field trial was to determine the safety and feasibility of passive immunization with bovine antibody for the prevention of rotavirus infection and illness in healthy infants. Sixty-four infants, 31 of whom received BCIg and 33 placebo, participated in the study. Rotavirus infection was detected in 11 (35%) and 14 (42%) infants in the 2 groups, respectively. Symptomatic rotavirus infection was documented in 1 (3%) of the infants receiving BCIg and in 6 (18%) of the infants who received the placebo (P > 0.05). The number of days with diarrhea/1000 days of observation was significantly less in the BCIg group, 4.2, than in the placebo group, 16.8 (P < 0.01). Similarly the number of days with rotavirus-associated diarrhea/1000 days was less in the BCIg group, 1.0, than in the placebo group, 13 (P < 0.01). This study establishes the feasibility of providing passive immunity for the prevention of rotavirus illness in healthy infants.

Historical cohort evaluation of ribavirin efficacy in respiratory syncytial virus infection.

Evaluation of ribavirin therapy for respiratory syncytial virus infection of the lower respiratory tract is problematic because of multiple risk factors and variable severity of illness in respiratory syncytial virus-infected patients. To address these difficulties we used multivariate analysis and performed a historical concurrent cohort study in two children's hospitals one of which had used ribavirin since licensing and one that had not utilized ribavirin therapy. The medical records of 158 patients who satisfied the American Academy of Pediatrics inclusion criteria for receiving ribavirin were analyzed for three seasons (1988 to 1991). No significant benefit of ribavirin therapy could be appreciated for the whole group in length of stay (median, 5.0 vs. 6.5 days), days on oxygen therapy (median, 5 vs. 3), progression to ventilator status (3.8 vs. 3.9%) or mortality (1.9% vs. 1.9%) for ribavirin treatment vs. supportive care. Multivariate analysis failed to uncover a beneficial effect of ribavirin when all risk factors were considered. No significant differences were noted when ventilated and nonventilated patients were examined separately. Our data raise questions about the efficacy of ribavirin when used in common practice and suggest that further prospective study, with appropriate analysis, is needed to justify the continued widespread use of this drug.

Assessment of adherence with medications in human immunodeficiency virus-infected children.

The purpose of this study was to determine the reliability of screening for medication adherence in HIV-infected children. The results suggest that caregivers who are unable to describe the medication regimen or who are nonadherent with appointments are unlikely to adhere to the medication regimen. Adherence with at least 90% of medication doses was associated with a virologic response.

Prospective comparison of the immune response of infants to three Haemophilus influenzae type b vaccines.

The antibody response to three Haemophilus influenzae type b vaccines was examined in 134 infants 17 to 22 months of age. Sera were collected from each subject before and 1 month after vaccination with either purified H. influenzae type b capsular polysaccharide (polyribosyl-ribitol phosphate (PRP] or one of two protein-conjugated vaccines (PRP-D or HbOC). Comparison of the two conjugate vaccines revealed that HbOC produced an antibody response greater than or equal to 1.0 micrograms/ml in 96% compared with 72% who were vaccinated with PRP-D (P less than 0.05). The isotype distribution of the antibodies produced by the two vaccines was similar. While all of the vaccines resulted in higher concentrations of anticapsular IgG1 than IgG2, the IgG1:IgG2 ratio was significantly higher in subjects immunized with HbOC. The IgG1:IgG2 ratio was similar in subjects immunized with PRP or PRP-D. The clinical significance of these observations remains to be determined.

The treatment of rhinovirus infections: progress and potential.

The common cold is an important illness both as a result of the economic impact of this common disease and because of the morbidity associated with the complications of the illness. Recent attempts to develop antiviral treatments for the common cold represent a substantial advance over previous efforts. Formidable barriers remain to be overcome, however, before any of these new products will be proven to be clinically useful. Recent advances in our understanding of the pathogenesis of common cold symptoms have provided insights into potential new targets for the treatment of this illness.

The high rate of blood donor exposure for critically ill neonates.

The purpose of this study was to determine the number and volume of red blood cell (RBC) transfusions and the number of donors a newborn is exposed to during his or her newborn intensive care unit (NICU) stay. On one day at the Medical University of South Carolina (MUSC) and two days at the University of Virginia Hospital (UVH) all babies who had or were receiving RBCs comprised the study group. Patient records were reviewed at discharge. Fifty-two (70%) of the 75 NICU babies had or were receiving RBCs and were enrolled. The average number of RBC transfusions was nine (range 1 to 28, median 7) and the average transfusion volume was 16.5 ml (range 5 to 60) for a total volume of 148 ml transfused during a NICU stay. Each baby was exposed to an average of 6.9 donors (range 1 to 25, median 6.5). The practice of splitting RBC packs to share among different infants and of giving multiple small volume transfusions maximizes donor exposure and transfusion-related infectious risks in this population.

The outcome of candiduria in pediatric patients.

The presence of Candida in the urine of a seriously ill, pediatric patient presents a management problem because of a lack of information concerning the natural history of candiduria and its relationship to disseminated candidiasis. In this retrospective study, the outcome of candiduria was examined in a group of 54 pediatric patients to determine any predictors of disseminated candidiasis. Medical records were reviewed to identify urine collection methods, Candida colony counts, results of cultures from other body sites, antifungal therapy, and clinical course. Six (11%) of the 54 patients had evidence of systematic Candida infection. In only two of these patients was candiduria the first evidence of disseminated candidiasis. Invasive infection was associated with candiduria more frequently in neonates and patients with central venous catheters and/or immunosuppressive therapy. Urine colony counts were not helpful for assessing the risk of invasive disease. Candiduria appears to be of little consequence in patients who are generally healthy. However, candiduria in high-risk patients, even in the presence of perineal candidal infection or an indwelling urinary catheter, should prompt a careful evaluation for disseminated infection.