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Biomedical subjects

J R Day

Publications and source records attributed to J R Day.

At least 19 recordsLinked to original sources

The maximally attainable VO2 during exercise in humans: the peak vs. maximum issue.

The quantification of maximum oxygen uptake (V(O2 max)), a parameter characterizing the effective integration of the neural, cardiopulmonary, and metabolic systems, requires oxygen uptake (VO2) to attain a plateau. We were interested in whether a VO2 plateau was consistently manifest during maximal incremental ramp cycle ergometry and also in ascertaining the relationship between this peak VO2 (V(O2 peak)) and that determined from one, or several, maximal constant-load tests. Ventilatory and pulmonary gas-exchange variables were measured breath by breath with a turbine and mass spectrometer. On average, V(O2 peak) [3.51 +/- 0.8 (SD) l/min] for the ramp test did not differ from that extrapolated from the linear phase of the response in 71 subjects. In 12 of these subjects, the V(O2 peak) was less than the extrapolated value by 0.1-0.4 l/min (i.e., a "plateau"), and in 19 subjects, V(O2 peak) was higher by 0.05-0.4 l/min. In the remaining 40 subjects, we could not discriminate a difference. The V(O2 peak) from the incremental test also did not differ from that of a single maximum constant-load test in 38 subjects or from the V(O2 max) in 6 subjects who undertook a range of progressively greater discontinuous constant-load tests. A plateau in the actual VO2 response is therefore not an obligatory consequence of incremental exercise. Because the peak value attained was not different from the plateau in the plot of VO2 vs. work rate (for the constant-load tests), the V(O2 peak) attained on a maximum-effort incremental test is likely to be a valid index of V(O2 max), despite no evidence of a plateau in the data themselves. However, without additional tests, one cannot be certain.

Adult↗

The distribution of neuropeptide Y gene expression in the chicken brain.

Neuropeptide Y (NPY) is demonstrated to play an important role in central control of voluntary feed intake (FI) of a variety of species. The commercial broiler chicken has been intensively selected over generations for increased body weight, achieved largely through increased FI. This has resulted in a contemporary animal that does not regulate FI to maintain energy balance, and represents a model for hyperphagia and obesity if allowed unrestricted access to feed. In the present study, the distribution of NPY mRNA was mapped in the brain of juvenile, broiler-strain chicken, and results interpreted in the context of previous data for strains that do not exhibit hyperphagia. NPY mRNA was widely distributed in the broiler brain, and highly expressed in the hippocampus, nucleus commissurae pallii, infundibular hypothalamic nucleus, nucleus pretectalis pars ventralis and neurons around the nucleus rotundus. Moderately labeled neurons were found in the lateral septal organ, nucleus periventricularis hypothalamus and nucleus paraventricularis magnocellularis. The pallium exhibited only sparse labeling. Generally, the distribution of cell groups expressing NPY mRNA was consistent with those regions exhibiting NPY immunoreactivity, and also matches the distribution of receptor binding sites reported in the literature for the chicken brain. This suggests that NPY may be involved in functions controlled by these regions. The observation of NPY gene expression in brain regions involved in appetite regulation is consistent with the recognized importance of NPY in FI regulation in a variety of species, and with the chronic hyperphagia, characteristic of the broiler.

Animals↗

A spontaneous ductal aneurysm presenting with left recurrent laryngeal nerve palsy.

A 76-year-old man was found to have a left recurrent laryngeal nerve palsy, due to a spontaneous patent ductus arteriosus aneurysm. Ortner's syndrome (left recurrent laryngeal nerve palsy caused by cardiovascular pathology) secondary to patent ductus arteriosus aneurysm has not previously been described in the literature. We present a case, illustrated with surface rendering computer tomogram reconstructions.

Aged↗

Development of a dual monoclonal antibody immunoassay for total human kallikrein 2.

BACKGROUND: Human kallikrein 2 (hK2) shares 80% sequence identity with prostate-specific antigen (PSA). Because both hK2 and hK2-alpha(1)-antichymotrypsin (hK2-ACT) complexes have been identified in patient sera, we devised an immunoassay for total hK2 [(thK2); hK2 and hK2-ACT] and evaluated it in healthy subjects and patients with prostate disease. METHODS: We developed monoclonal antibodies (mAbs) with high specificity for hK2 and hK2-ACT and minimal cross-reactivity to PSA. Using these mAbs, a sandwich assay was developed and its specificity for forms of hK2 was assessed. Serum samples (n = 1035) from healthy volunteers, patients with increased PSA, and men who had undergone radical prostatectomy were assayed for thK2. We also measured thK2 in samples before and after storage under common laboratory conditions. RESULTS: The minimum detectable concentration in the thK2 assay was 0.008 microg/L, and PSA cross-reactivity was <0.001%. The assay detected prohK2 and three different hK2-serum protease complexes. The median serum concentration of thK2 in control samples (0.013 microg/L) was significantly lower than the median in samples from patients with increased PSA concentrations (0.085 microg/L). Immunoreactive hK2 changed little in samples stored for up to 1 month at -70 degrees C. CONCLUSIONS: The thK2 assay recognizes all forms of hK2 that have been found in bodily fluids to date.

Antibodies, Monoclonal↗

Expression of the lipocalin-type prostaglandin D synthase gene in the reproductive tracts of Holstein bulls.

The aim of this study was to localize expression of the prostaglandin D synthase gene in the reproductive tracts of Holstein bulls using northern blotting and in situ hybridization. For northern blotting, a digoxigenin-labelled prostaglandin D synthase cDNA probe was used to probe blots containing RNA isolated from the testes, epididymides, vas deferens, ampullae, seminal vesicles, prostate and bulbourethral glands of bulls. The digoxigenin-labelled cDNA for the bovine homologue of prostaglandin D synthase hybridized to a single band (approximately 0.9 kb) to RNA samples from the caput, corpus and cauda epididymides, as well as RNA samples from the vas deferens and the ampulla. The probe also detected a single band in testis samples, although the transcript size was slightly larger (approximately 1.0 kb) than the transcript found in the other tissues. The highest expression of prostaglandin D synthase was observed in the testes and caput epididymides. Prostaglandin D synthase transcripts were not found in the seminal vesicles or the prostate or bulbourethral glands using northern blotting. For in situ hybridization, antisense and sense riboprobes were synthesized and used to hybridize to cryosections obtained from the reproductive tissues of bulls. In situ hybridization of bull testes showed that prostaglandin D synthase transcripts were present within the germ cells in the adluminal compartment of the seminiferous tubules containing round and elongated spermatids, indicating that expression varied with stage of development of the seminiferous tubules. Prostaglandin D synthase expression was observed in the epithelial cells of the epididymides with greatest expression occurring in the caput epididymidis. Some expression was also observed in the epithelial cells of the vas deferens and a few cells of some lobules in the prostate and bulbourethral glands. Expression of the prostaglandin D synthase gene was not detected in ampullae or seminal vesicles by in situ hybridization.

Animals↗

Evidence of a role for neuropeptide Y and monoamines in mediating the appetite-suppressive effect of GH.

Among the many responses to GH administration is suppression of voluntary feed intake (FI) in some species, attributed to improvement in the efficiency of nutrient utilization and, therefore, reduced need for ingested substrates. Commercial broiler chickens have been genetically selected for generations for rapid growth, realized largely via the major correlated response of increased voluntary feed consumption. Neuropeptide Y (NPY) and monoamines play very important roles in the central regulation of feeding. Preliminary studies from our laboratory suggest that the appetite-suppressive effect of GH may be independent of its actions as a repartitioning agent, and may involve alterations in NPY expression at the pre-translational level. The purpose of this investigation was to explore the dose-response nature of the appetite-suppressive effect of GH in juvenile broilers, and the possible involvement of NPY and monoamines in this process. A GH dose-response study was conducted using 8-week-old female broilers infused i.v. with GH in a pulsatile pattern for 7 days at 0, 10, 50, 100 or 200 microgram/kg body weight per day. Hypothalamic NPY and epinephrine (EP) concentrations decreased in a dose-related manner with GH. At the highest dosage, voluntary FI decreased 19% (P<0.05) and hypothalamic NPY mRNA decreased approximately 50% in the infundibular nuclei and midline region (P<0.0001). In contrast, birds pairfed to the high-GH dosage group did not differ from controls, verifying that changes in NPY and monoamines were not secondary to reduced FI. We conclude that hypothalamic NPY and EP are likely candidates to explore further as mediators of the appetite-suppressive effect of GH.

Animals↗

Iron deficiency alters H- and L-ferritin expression in rat brain.

Ferritin (Ft) H and L subunits are independently regulated proteins with both transcriptional and translational regulation in response to cellular iron levels. While the heterogeneous distribution of ferritin and iron in the brain is now well established, the relative response of each subunit to iron deficiency and iron supplementation, is not well defined. Weanling male Sprague-Dawley rats (n=12 per group) were randomly assigned to an iron deficient (3.5 mg Fe/kg diet), control (35 mg Fe/kg diet) or supplemented (350 mg Fe/kg diet) diet for six weeks. The H-/L-ferritin subunit ratio and mRNA levels were determined. Overall, the protein ratio in control rats of H to L was approximately 45:1 compared to a ratio >60:1 in iron deficiency but the absolute amounts of each subunit varied greatly from one brain region to another. The ratio of H-:L-ferritin mRNA was 6:1 and was not affected by dietary iron deficiency in contrast to a potent effect on mRNA levels in liver. Severe iron deficiency reduced brain ferritin H protein levels significantly in all regions, whereas only ferritin L levels in striatum, substantia nigra and pons were affected by iron deficiency. Supplemental dietary iron increased both ferritin subunits, with the largest increase (50%) in the hippocampus. These data indicate that ferritin H and L subunits within the brain respond differently to iron status and suggest post transcription regulation as a key event.

Animals↗

Polyclonal and monoclonal antibodies to prostate-specific antigen can cross-react with human kallikrein 2 and human kallikrein 1.

OBJECTIVES: The human tissue kallikrein family contains three closely related proteases: human kallikrein 1 (hK1), human kallikrein 2 (hK2), and prostate-specific antigen (PSA). The structural homology between these three proteins suggests potential cross-reactivity interference when different immunologic techniques are used. This study evaluated PSA and hK2 monoclonal antibody (mAb) and polyclonal antibody (pAb) reactivities to hK1, hK2, and PSA. METHODS: mAbs and pAbs to hK2 and PSA were evaluated using Western blot analysis on hK1, hK2, PSA, and seminal plasma. RESULTS: pAbs to PSA and hK2 recognized all three human kallikreins, as well as fragments of hK2 and PSA. An mAb with minimal (less than 0.4%) cross-reactivity between PSA and hK2 and a cross-reactive mAb were found. mAbs specific to PSA or hK2 did not cross-react with the less homologous hK1 protein. A PSA mAb raised specifically to PSA fragments recognized both PSA and hK2 but did not cross-react with hK1. pAbs to hK1 cross-reacted slightly with PSA and not at all with hK2. CONCLUSIONS: Both pAbs and mAbs to hK2 and PSA may exhibit immunocross-reactivity. pAbs to PSA or hK2 react with all three human tissue kallikreins. The potential for cross-reactivity should be considered in any clinical or research procedures that use hK1, hK2, and PSA antibodies.

Antibodies, Monoclonal↗

New insights into the mechanism and actions of growth hormone (GH) in poultry.

Despite well documented anabolic effects of GH in mammals, a clear demonstration of such responses in domestic poultry is lacking. Recently, comprehensive dose-response studies of GH have been conducted in broilers during late post-hatch development (8 to 9 weeks of age). GH reduced feed intake (FI) and body weight gain in a dose-dependent manner, whereas birds pair-fed to the level of voluntary FI of GH-infused birds did not differ from controls. The reduction in voluntary FI may involve centrally mediated mechanisms, as hypothalamic neuropeptide Y protein and mRNA were reduced with GH, coincident with the maximal depression in FI. Growth of breast muscle was also reduced in a dose-dependent manner. Circulating IGF-I was not enhanced by GH, despite evidence that early events in the GH signaling pathway were intact. A GH dose-dependent increase in circulating 3,3',5-triiodothyronine(T3) paralleled decreases in hepatic 5D-III monodeiodinase activity, whereas 5'D-I activity was not altered. This confirms that a marked hyperthyroid response to GH occurs in late posthatch chickens, resulting from a decrease in the degradative pathway of T3 metabolism. This secondary hyperthyroidism would account for the decreased skeletal muscle mass (52) and lack of enhanced IGF-I (53) in GH-treated birds. Based upon these studies, it is now evident that GH does in fact have significant effects in poultry, but metabolic responses may confound the anabolic potential of the hormone.

Animals↗

Age-related expression of renal thrombospondin 1 mRNA in F344 rats: resemblance to diabetes-induced expression in obese Zucker rats.

Age-related increases occurred in renal thrombospondin 1 (TSP1) mRNA in F344 rats, resembling diabetes-induced TSP1 mRNA in the obese Zucker rat. TSP1 mRNA was 3.5-fold higher in 24-month-old than in 3-month-old F344 rats. TSP1 mRNA increased similarly in 5-month-old obese Zucker rats as compared with lean littermates and correlated positively with the extent of proteinuria (r = 0.71). In situ hybridization identified elevated TSP1 mRNA levels in epithelial cells of distended tubules as well as in interstitium near dilated tubules of both 24-month-old F344 rats and 5-month-old obese Zucker rats. Furthermore, thrombin increased TSP1 mRNA in mesangial and epithelial cells in culture, indicating that thrombin may contribute to elevated TSP1 expression in renal disease. Thrombin increased TSP1 mRNA within 30 min after treatment which required de novo synthesis of protein. The thrombin receptor tethered ligand peptide, SFLLRN, increased TSP1 mRNA, indicating that the thrombin-induced increase in TSP1 mRNA was due to direct thrombin receptor (PAR1) stimulation. These results show that increased TSP1 mRNA levels are a component of interstitial fibrosis seen in aged and diabetic kidneys and suggest that similar pathological changes occur in kidneys of aging and diabetic rats.

Aging↗

The effect of age and testosterone on the expression of glial fibrillary acidic protein in the rat cerebellum.

Testosterone reversed the age-related increase in glial fibrillary acidic protein (GFAP) in the male rat cerebellum, a brain region not generally associated with gonadal steroid hormone sensitivity. This supports the hypothesis that a decrease in circulating testosterone contributes to age-related increase in GFAP. These data also suggest that reductions in circulating gonadal steroids during aging could render the brain more susceptible to neurodegeneration and that hormone replacement therapy might have value in neurodegenerative disease intervention.

Aging↗

Development of monoclonal antibodies specific for human glandular kallikrein (hK2): development of a dual antibody immunoassay for hK2 with negligible prostate-specific antigen cross-reactivity.

OBJECTIVES: Human glandular kallikrein (hK2) is a protein that is 80% homologous to prostate-specific antigen (PSA), and, like PSA, is localized to the prostate. We developed a specific immunoassay for hK2 that can be used to evaluate its clinical diagnostic utility. METHODS: We developed monoclonal antibodies (mAbs) specific for hK2 by immunizing with hK2 and screening for clones reactive with hK2 and not PSA. Prototype sandwich assays using these mAbs were tested, and the optimum pair selected. Purified hK2 was used as standard and PSA cross-reactivity was assessed in the assay. Both hK2 and hK2-alpha1-antichymotrypsin (ACT) complexes have been identified in sera of patients with prostate cancer (PCa). Serum samples (n = 671) from healthy volunteers and patients with prostate disease were assayed for hK2 and PSA levels. RESULTS: The assay had a detection limit of less than 0.12 ng/mL and a less than 0.5% cross-reactivity with PSA. The assay preferentially detected free hK2 with a 3.5-fold higher molar response than with hK2-ACT. The mean serum concentration of hK2 in normal control samples was low (0.33 and 0.37 ng/mL for normal healthy men and women, respectively) but was elevated in patients with prostate disease (0.86 and 6.77 ng/mL for patients with benign prostatic hyperplasia and PCa, respectively). Negligible cross-reactivity to hK2 was measured by Tandem PSA assays (Hybritech). CONCLUSIONS: Significant concentrations of hK2, relative to PSA, were detected in human serum, especially in patients with prostate disease. Serum hK2 concentrations were not proportional to PSA concentration. Therefore, hK2 has the potential to be an independent and clinically useful marker for PCa.

Antibodies, Monoclonal↗

Effects of microgravity and bone morphogenetic protein II on GFAP in rat brain.

This study evaluated effects of bone morphogenetic protein II (BMP) on glial fibrillary acidic protein (GFAP) in the brain of female Fischer 344 rats during 14 days of spaceflight. GFAP mRNA decreased in vehicle-implanted rats flown on the space shuttle by 53 and 48% in the stratum moleculare and stratum lacunosum moleculare hippocampal subregions, respectively. GFAP mRNA was not significantly affected by BMP implantation during spaceflight. Rats returning from space exhibited a 56% increase in serum corticosterone. BMP treatment did not additively increase corticosterone elevations in microgravity but appeared to increase serum corticosterone and reduce GFAP mRNA in the stratum moleculare in control rats. These data suggest that exposure to microgravity reduces GFAP expression in hippocampal astrocytes.

Animals↗

The age-related increase in renal clusterin mRNA is accelerated in obese Zucker rats.

Clusterin is a multifunctional glycoprotein associated with development and tissue injury. Because renal function decreases with advancing age in the obese Zucker rat, clusterin mRNA expression was examined in the kidney of young adult Zucker rats and compared with age-related changes in renal clusterin mRNA expression in Fischer 344 (F344) rats. Renal clusterin mRNA levels in the obese Zucker rat were 2.5-fold higher by 3 mo of age and fourfold higher at 5 mo of age compared with the lean strain. In comparison, renal clusterin mRNA in 12-mo-old F344 rats was twofold higher than in 3-mo-old animals and was tenfold higher at 24 mo of age. Clusterin mRNA was positively correlated with urinary protein excretion and negatively correlated with creatinine clearance in Zucker rats. Clusterin was increased in select nephrons of the obese Zucker rat kidney and in 24-mo-old F344 rat kidney as assessed by in situ hybridization. Increased expression of clusterin mRNA occurred mostly in the tubular epithelium of dilated, convoluted proximal tubules. These data indicate that renal clusterin mRNA levels increase as a function of age and that age-related increases in renal clusterin and the associated tubular abnormalities are accelerated in obese Zucker rats.

Aging↗

Alterations in hippocampal expression of SNAP-25, GAP-43, stannin and glial fibrillary acidic protein following mechanical and trimethyltin-induced injury in the rat.

A set of well-defined antisera against neuronal and glial proteins were used to characterize patterns of protein expression in rat hippocampus following transection of the fimbira-fornix and perforant pathways or after administration of the selective neurotoxicant trimethyltin (8 mg/kg, i.p.). SNAP-25 (synaptosomal protein, mol. wt 25,000) is a neuron-specific, developmentally regulated presynaptic protein, stannin is a protein enriched in cells sensitive to trimethyltin, and GAP-43 (growth-associated protein, mol. wt 43,000) is associated with axonal growth and regeneration. Glial fibrillary acidic protein is an astrocyte-specific intermediate filament protein and a marker for reactive gliosis. SNAP-25 immunoreactivity was altered following both neurotoxicant and mechanical injury. Three days after fimbria-fornix/perforant path lesions, there was a loss of SNAP-25 immunoreactivity in hippocampal efferent pathways and in the lesioned entorhinal cortex. By day 12, there was evidence of reinnervation of hippocampal subfields by SNAP-25-immunopositive commissural afferent fibers. On day 3, immunoblots showed the appearance of SNAP-25a, a developmental isoform produced by alternative splicing of nine amino acids in exon 5, in lesioned tissues. This isoform declined by day 12 and was not found in contralateral control hippocampus or non-lesioned brain regions. Stannin immunoreactivity was unchanged, while GAP-43 was prominent on day 12 post-lesion. Glial fibrillary acidic protein immunoreactivity indicated gliosis near the site of pathway transection. In contrast, trimethyltin induced a marked loss of stannin immunoreactivity in hippocampal neurons seven days after injection. Trimethyltin increased glial fibrillary acidic protein staining in the hippocampus and other damaged regions. SNAP-25 immunoreactivity was markedly increased in mossy fibers and other hippocampal fields seven days following trimethyltin. Immunoblot analysis showed that only the adult SNAP-25b isoform was expressed after trimethyltin intoxication. These data suggest that SNAP-25 is a useful marker for presynaptic damage. Furthermore, reexpression of developmental isoforms of SNAP-25a may precede functional reinnervation when the postsynaptic target remains intact.

Animals↗

Using neutralizing routines to reduce problem behaviors.

Establishing operations can alter problem behaviors by changing the momentary value of reinforcers associated with those problem behaviors. If establishing operations (EOs) precede the presentation of discriminative stimuli (SDs) for problem behaviors, it may be possible to introduce neutralizing routines that both reduce the value of reinforcers associated with problem behaviors and decrease the occurrence of problem behaviors. The present study examined this logic with 3 adolescents with severe intellectual disabilities. Initial functional analyses indicated that problem behaviors were motivated by either escape or tangible items. Functional assessment interviews identified possible establishing operations that were associated with the occurrence of problem behavior and indicated that these establishing operations occurred over 1 hr before presentation of the SD for problem behaviors. We used an alternating treatments design to examine problem behaviors during instruction under four conditions: EO + SD, SD only, EO only, and neither SD nor EO. For all 3 participants, problem behaviors occurred almost exclusively during the EO + SD condition. A further analysis compared the EO + SD condition when neutralizing routines were embedded between the EO and the SD. Results from an ABAB reversal design supported the effectiveness of neutralizing routines to reduce these problem behaviors. Applied and theoretical implications are discussed.

Adolescent↗

Microwave superheating enhances immunocytochemistry in the freshly frozen rat brain.

The goal of this study was to evaluate the effects of microwave irradiation on immunocytochemical staining in freshly frozen rat brain tissue using an antibody previously shown to successfully stain only formalin-fixed materials. GFAP immunoreactivity was markedly increased in sections exposed to microwave heating when compared with control tissue. Uniform staining enhancement was mainly a result of increased numbers of immunoreactive soma and large astrocytic processes. The integrity of the tissue was not affected by this treatment. Other antibodies need to be tested with this method, but our results suggest that microwave irradiation is useful for rapid antigen retrieval in freshly frozen central nervous system tissue.

Animals↗

Functional expression of human and mouse plasma phospholipid transfer protein: effect of recombinant and plasma PLTP on HDL subspecies.

The molecular cloning of mouse plasma phospholipid transfer protein (PLTP) and the eukaryotic cell expression of complementary DNA for mouse and human PLTP are described. Mouse PLTP was found to share 83% amino acid sequence identity with human PLTP. PLTP was produced in baby hamster kidney cells. Conditioned medium from BHK cells expressing PLTP possessed both phospholipid transfer activity and high density lipoprotein (HDL) conversion activity. PLTP mRNA was detected in all 16 human tissues examined by Northern blot analysis with ovary, thymus, and placenta having the highest levels. PLTP mRNA was also examined in eight mouse tissues with the highest PLTP mRNA levels found in the lung, brain, and heart. The effect of purified human plasma-derived PLTP and human recombinant PLTP (rPLTP) on the two human plasma HDL subspecies Lp(A-I) and Lp(A-I/A-II) was evaluated. Plasma PLTP or rPLTP converted the two distinct size subspecies of Lp(A-I) into a larger species, an intermediate species, and a smaller species. Lp(A-I/A-II) particles containing multiple size subspecies were significantly altered by incubation with either plasma or rPLTP with the largest but less prominent subspecies becoming the predominant one, and the smallest subspecies increasing in concentration. Thus, PLTP promoted the conversion of both Lp(A-I) and Lp(A-I/A-II) to populations of larger and smaller particles. Also, both human PLTP and mouse rPLTP were able to convert human or mouse HDL into larger and smaller particles. These observations suggest that PLTP may play a key role in extracellular phospholipid transport and modulation of HDL particles.

Amino Acid Sequence↗