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Biomedical subjects

B L Smith

Publications and source records attributed to B L Smith.

At least 163 records · Page 9Linked to original sources

Concurrent expression of erythroid and renal aquaporin CHIP and appearance of water channel activity in perinatal rats.

Major phenotypic changes occur in red cell membranes during the perinatal period, but the underlying molecular explanations remain poorly defined. Aquaporin CHIP, the major erythroid and renal water channel, was studied in perinatal rats using affinity-purified anti-CHIP IgG for immunoblotting, flow cytometry, and immunofluorescence microscopy. CHIP was not detected in prenatal red cells but was first identified in circulating red cells on the third postnatal day. Most circulating red cells were positive for CHIP by the seventh postnatal day, and this proportion rose to nearly 100% by the 14th day. The ontogeny of red cell CHIP correlated directly with acquisition of osmotic water permeability and inversely with Arrhenius activation energy. Only minor alterations in the composition of red cell membrane lipids occurred at this time. Immunohistochemical analysis of perinatal kidneys demonstrated a major induction of CHIP in renal proximal tubules and descending thin limbs at birth, coincident with the development of renal concentration mechanisms. Therefore, water channels are unnecessary for oxygen delivery or survival in the prenatal circulation, however CHIP may confer red cells with the ability to rehydrate rapidly after traversing the renal medulla, which becomes hypertonic after birth.

Aging↗

Sequential changes in the development of the pancreatic lesion of zinc toxicosis in sheep.

Forty-two 10-month-old castrated male sheep were dosed with zinc oxide to study the pathogenesis of the pancreatic lesion. For 4 weeks, the sheep were dosed three times per week with 240 mg Zn (as ZnO)/kg body weight/dose, and seven groups of six sheep each were necropsied at 4, 7, 14, 21, 28, 56, and 112 days after the start of dosing. Plasma zinc concentrations rose rapidly to 2.0-2.5 micrograms Zn/ml over the dosing period and fell rapidly to less than 1 microgram Zn/ml within 2 weeks after dosing ceased. Organ zinc levels in liver, kidney, and pancreas fell from concentrations above 800 micrograms Zn/g (dry matter basis) to less than 200 micrograms/g within 4 weeks after dosing ceased. Although no animals showed any clinical signs of zinc toxicity, many sheep dosed with zinc oxide developed pancreatic lesions. Pancreatic lesions took up to 4 weeks to develop fully. The early pancreatic lesions involved necrosis of the pancreatic duct epithelium, periductular inflammation, and interlobular fat necrosis, all evident at 7 days but not at 4 days. These early lesions were followed by edema, lobular cystic change, atrophy, fibrosis, and a ductular hyperplasia. The initial lesion of the pancreas was ductular injury, and the subsequent pancreatic lesions may have developed in those lobules whose excretory ducts were obstructed by inflammatory debris.

Animals↗

Psychological tests don't think: an appreciation of Schafer's Psychoanalytic Interpretation in Rorschach Testing.

Roy Schafer 's Psychoanalytic Interpretation in Rorschach Testing has, for nearly 40 years, represented a hallmark of psychodynamically informed psychological assessment. In this article, Schafer's contribution is reviewed in light of recent contributions to the Rorschach literature. Following a summary of the main sections of the work, Schafer's approach to the Rorschach is critically re-examined from the perspective of modem assessment practice. Differences between the epistemological bases of psychoanalytic and empirical methods of Rorschach interpretation are highlighted, and the prospects for an integration of these two approaches are considered. It is concluded that Schafer's contribution remains as fresh today as when written 40 years ago.

Journal Article↗

A simple gag for the intragastric dosing of guinea pigs (Cavia porcellus).

A simple gag has been designed and tested which enables the intragastric intubation of the guinea pig by one person without the use of sedation or light anaesthesia. The system described was used successfully for the daily dosing of materials to 7 guinea-pigs for 14 days without discomfort, aspiration or any injury to the upper gastro-intestinal tract.

Animals↗

Inhibition of protein kinase C function by injection of intracellular receptors for the enzyme.

We tested the hypothesis that the translocation and function of protein kinase C (PKC) requires the binding of PKC to its intracellular receptors (RACKs), using insulin-induced maturation of Xenopus oocytes. We show that after exposure of oocytes to insulin, PKC translocated from the cytosol to the particulate fraction. PKC is also required for insulin-induced oocyte maturation: microinjection of a PKC inhibitory peptide delayed maturation. To determine whether translocation of PKC was a result of the binding of PKC to the RACKs in the particulate fraction, we microinjected purified rat brain RACKs into oocytes before insulin exposure. Microinjection of RACKs, but not inactive phosphorylated RACKS, inhibited PKC translocation and delayed oocyte maturation. These results suggest an in vivo role for RACKs in a function mediated by PKC.

Animals↗

p65 fragments, homologous to the C2 region of protein kinase C, bind to the intracellular receptors for protein kinase C.

Receptors for activated protein kinase C (RACKs) have been isolated from the particulate cell fraction of heart and brain. We previously demonstrated that binding of protein kinase C (PKC) to RACKs requires PKC activators and is via a site on PKC that is distinct from the substrate binding site. Here, we examine the possibility that the C2 region in the regulatory domain of PKC is involved in binding of PKC to RACKs. The synaptic vesicle-specific p65 protein contains two regions homologous to the C2 region of PKC. We found that three p65 fragments, containing either one or two of these PKC C2 homologous regions, bound to highly purified RACKs. Binding of the p65 fragments and PKC to RACKs was mutually exclusive; preincubation of RACKs with the p65 fragments inhibited PKC binding, and preincubation of RACKs with PKC inhibited binding of the p65 fragments. Preincubation of the p65 fragments with a peptide resembling the PKC binding site on RACKs also inhibited p65 binding to RACKs, suggesting that PKC and p65 bind to the same or nearby regions on RACKs. Since the only homologous region between PKC and the p65 fragments is the C2 region, these results suggest that the C2 region on PKC contains at least part of the RACK binding site.

Amino Acid Sequence↗

Reconstitution of functional water channels in liposomes containing purified red cell CHIP28 protein.

Water rapidly crosses the plasma membranes of red blood cells (RBCs) and renal tubules through highly specialized channels. CHIP28 is an abundant integral membrane protein in RBCs and renal tubules, and Xenopus laevis oocytes injected with CHIP28 RNA exhibit high osmotic water permeability, Pf [Preston et al. (1992) Science 256, 385-387]. Purified CHIP28 from human RBCs was reconstituted into proteoliposomes in order to establish if CHIP28 is itself the functional unit of water channels and to characterize its physiological behavior. CHIP28 proteoliposomes exhibit Pf which is up to 50-fold above that of control liposomes, but permeability to urea and protons is not increased. Like intact RBC, the Pf of CHIP28 proteoliposomes is reversibly inhibited by mercurial sulfhydryl reagents and exhibits a low Arrhenius activation energy. The magnitude of CHIP28-mediated water flux (11.7 x 10(-14) cm3/s per CHIP28) corresponds to the known Pf of intact RBCs. These results demonstrate that CHIP28 protein functions as a molecular water channel and also indicate that CHIP28 is responsible for most transmembrane water movement in RBCs.

Aquaporin 1↗

Staff perceptions of reinforcer responsiveness and aberrant behaviors in people with mental retardation.

Perceptions by staff of the classes of reinforcers and aberrant behaviors of a sample of 470 people with predominantly severe or profound mental retardation were explored. Principal components analysis of a 45-item survey suggested eight classes of reinforcers: consumable, verbal-speaker, visual-motor, social, physical-contact, passive-observer, play, and academic reinforcers. Stepwise multiple regression was used to predict five classes of maladaptive behaviors as measured by the Aberrant Behavior Checklist (irritability, lethargy, stereotypy, hyperactivity, and inappropriate speech) from the eight classes of reinforcers. Each class of psychopathology was related to a unique set of predictors. All classes of psychopathology could be predicted by staff perceptions of underresponsiveness to social reinforcers and overresponsiveness to consumable reinforcers. The findings of organized structures of reinforcers and their covariation with pathological behaviors have implications for research and intervention as well as theoretical value in defining aberrant behaviors in people with mental retardation.

Adolescent↗

Parenchymal injury and biliary obstruction in relation to photosensitization in sporidesmin-intoxicated lambs.

The hepatic changes were compared in lambs photosensitized or not photosensitized after exposure to sporidesmin. Injury to both the parenchyma and the biliary system was more severe in the photosensitized than in the non-photosensitized lambs. The activities of gamma-glutamyltransferase and glutamate dehydrogenase, and the total, conjugated and unconjugated bilirubin concentrations were significantly higher in sera from the photosensitized than from the non-photosensitized lambs. Hepatic glycogen levels were decreased in both the photosensitized and the non-photosensitized lambs, but were significantly lower in the former. Hence it is possible that lesions in hepatocytes contribute to retention of phylloerythrin and so to photosensitization.

Animals↗

Nucleotide sequences and biologic properties of toxic shock syndrome toxin 1 from ovine- and bovine-associated Staphylococcus aureus.

Toxic shock syndrome toxin (TSST) 1 was purified from ovine (TSST-ovine) and bovine (TSST-bovine) mastitis-associated Staphylococcus aureus. These toxins were previously reported to have molecular weights identical to that of human TSST-1. However, TSST-ovine was reported as having an isoelectric point (pI) of 8.5, whereas TSST-bovine has the same pI (7.2) as TSST-1. Nucleotide sequence analysis revealed that TSST-bovine was identical to TSST-1 and that TSST-ovine had 14 nucleotide differences that changed 9 amino acid residues. Only 1 nucleotide difference, at position 514, was predicted to cause an amino acid charge difference, as glutamic acid at position 132 of TSST-1 was changed to lysine in TSST-ovine. Like TSST-1, TSST-ovine was mitogenic, but unlike TSST-1, it was not pyrogenic, was unable to enhance endotoxic shock, and was unable to induce TSS in a rabbit model. Also, TSST-ovine was less reactive to certain monoclonal antibodies raised against TSST-1.

Amino Acid Sequence↗

Evaluation of the contralateral breast. The role of biopsy at the time of treatment of primary breast cancer.

Ninety-five women who underwent blind contralateral breast biopsy during surgical treatment of a known breast cancer primary were studied prospectively. All biopsies were performed between 1981 and 1989. Patients with palpable or mammographic abnormalities prompting the contralateral biopsy were excluded so that the study sample included only truly blind contralateral biopsies. Only two infiltrating carcinomas were found, resulting in a positive biopsy rate of 2.1% for invasive disease. Three additional biopsies showed only lobular carcinoma in situ, a finding that usually does not alter clinical management. One patient with a negative contralateral biopsy developed invasive carcinoma in that breast within 2 years of the biopsy. The authors were unable to identify any subgroup of patients at increased risk of a positive contralateral biopsy. These results suggest that blind biopsy of the contralateral breast performed at the time of the initial treatment of breast carcinoma is not an efficient method of cancer detection. Alternative management strategies are discussed.

Adult↗

Relationships between duration and temporal variability in children's speech.

Various studies of speech production development have observed that duration and temporal variability tend to decrease as children get older. Because both of these factors are commonly viewed as general indicators of neuromotor maturation of children's speech skills, it would seem reasonable to assume that they should be rather closely correlated with one another. However, an examination of a variety of temporal data from children ranging between approximately 2 1/2 and 9 1/2 years of age indicates that there is not necessarily as close a correspondence between these two variables as might be expected. It also appears that children's duration measures tend to be more adultlike than measures of their intrasubject variability.

Adult↗

Failure of predicted creatinine clearance equations in HIV-seropositive patients.

The Cockcroft and Gault (CG) [1976] method of predicted creatinine clearances (CCR) accurately predicts measured 24-hour CCR values in healthy volunteers. The present study compared the relationship between measured and predicted CCR through 5 methods: CG, J1 [Jelliffe 1971], J2 [Jelliffe 1973], M [Mawer et al. 1972], and H [Hull et al. 1981], in 42 HIV-seropositive patients: 21 ARC/21 AIDS, 35M/7F, 26 homosexual/16 intravenous drug users, age: 37 +/- 7 years, actual body weight: 74 +/- 14 kg, CD4: 0.286 +/- 0.185 x 10(9) cells per liter (mean +/- SD). Measured CCR values poorly correlated with serum creatinine levels (r = -0.35; p < 0.01). The average measured CCR was 106 +/- 29 ml/min compared with 94 +/- 21 (CG; r = 0.49), 78 +/- 13 (J1; r = 0.41), 77 +/- 14 (J2; r = 0.44), 97 +/- 21 (M; r = 0.51) and 95 +/- 17 ml/min (H; r = 0.32). Standardization to body surface area or lean body weight or stratification by patient factors (gender, disease stage, risk factors, drug treatment) did not improve correlations. However, patients with normal microalbumin excretion rates had more predictable CCR values compared with those who had excess excretion, suggesting the influence of HIV-associated nephropathy on CCR estimation. Since all predicted CCR equations consistently underestimated actual values, these equations should be used with caution in estimating measured CCR in HIV-seropositive patients.

Adolescent↗

Intracellular receptors for activated protein kinase C. Identification of a binding site for the enzyme.

Protein kinase C (PKC) isozymes comprise a family of cytosolic enzymes that translocate to different intracellular sites on activation. We have recently characterized at least two intracellular receptor proteins for PKC (termed RACKs for receptors for activated C-kinase) in the Triton-insoluble material of the particulate fraction from neonatal rat heart. Here, we identify a sequence that appears to resemble the PKC binding site on these RACKs. A peptide (peptide I) with the sequence KGDYEKILVALCGGN bound PKC, and binding was markedly increased in the presence of PKC activators. Furthermore, peptide I inhibited PKC binding to RACKs in a dose-dependent manner. These data suggest that these RACKs have a common PKC binding sequence. Since peptide I inhibited PKC binding to RACKs in vitro, it may be a useful tool to inhibit PKC translocation and subsequent function in vivo.

Amino Acid Sequence↗

Erythrocyte Mr 28,000 transmembrane protein exists as a multisubunit oligomer similar to channel proteins.

A novel Mr 28,000 erythrocyte transmembrane protein was recently purified and found to exist in two forms, "28kDa" and "gly28kDa," the latter containing N-linked carbohydrate (Denker, B. M., Smith, B. L., Kuhajda, F. P., and Agre, P. (1988) J. Biol. Chem. 263, 15634-15642). Although 28kDa protein resembles the Rh polypeptides biochemically, structural homologies were not identified by immunoblot or two-dimensional iodopeptide maps. The NH2-terminal amino acid sequence for the first 35 residues of purified 28kDa protein is 37% identical to the 26-kDa major intrinsic protein of lens (Gorin, M. B., Yancey, S. B., Cline, J., Revel, J.-P., and Horwitz, J. Cell 39, 49-59). Antisera to a synthetic peptide corresponding to the NH2-terminus of 28kDa protein gave a single reaction of molecular mass 28kDa on immunoblots of erythrocyte membranes. Selective digestions of intact erythrocytes and inside-out membrane vesicles with carboxypeptidase Y indicated the existence of a 5-kDa COOH-terminal cytoplasmic domain. Multiple studies indicated that 28kDa and gly28kDa proteins exist together as a multisubunit oligomer: 1) similar partial solubilizations in Triton X-100; 2) co-purification during ion exchange and lectin affinity chromatography; 3) cross-linking in low concentrations of glutaraldehyde; and 4) physical analyses of purified proteins and solubilized membranes in 1% (v/v) Triton X-100 showed 28kDa and gly28kDa proteins behave as a large single unit with Stokes radius of 61 A and sedimentation coefficient of 5.7 S. These studies indicate that the 28kDa and gly28kDa proteins are distinct from the Rh polypeptides and exist as a multisubunit oligomer. The 28kDa protein has NH2-terminal amino acid sequence homology and membrane organization similar to major intrinsic protein and other members of a newly recognized family of transmembrane channel proteins.

Amino Acid Sequence↗

Haemolytic activity and nephrotoxicity of 2-hydroxy-1,4-naphthoquinone in rats.

The short-term toxicity of 2-hydroxy-1,4-naphthoquinone (lawsone) and 2-methyl-1,4-naphthoquinone (menadione) has been compared in rats. 2-Methyl-1,4-naphthoquinone has been shown previously to cause haemolytic anaemia in animals, and this was confirmed in the present experiment. 2-Hydroxyl-1,4-naphthoquinone was found also to cause haemolysis, in a dose-dependent manner, as reflected by decreased blood packed cell volumes and haemoglobin levels and by histopathological changes in spleen, liver and kidney. With both naphthoquinones, the haemolysis was of the oxidative type, characterized by the presence of Heinz bodies within erythrocytes. Haemolysis was the only toxic change identified in rats dosed with 2-methyl-1,4-naphthoquinone. In contrast, 2-hydroxyl-1,4-naphthoquinone was not only a haemolytic agent but also a nephrotoxin, causing renal enlargement, elevated plasma levels of urea and creatinine and histologically-identified tubular necrosis, largely confined to the distal segment of the proximal convoluted tubules. The relationship between the in vivo toxic effects of these naphthoquinones and previously-reported data on their in vitro cytotoxic action is discussed.

Animals↗