Biomedical subjects
A White
Publications and source records attributed to A White.
Central actions of CRF on thermogenesis are mediated by pro-opiomelanocortin products.
Corticotrophin-releasing factor (CRF) causes central activation of thermogenesis. The aim of this study was to investigate whether this action is mediated by ACTH or other peptides derived from the ACTH precursor pro-opiomelanocortin (POMC) within the CNS. Central (intracerebroventricular) injection of rat CRF caused dose-dependent increases in resting oxygen consumption (VO2) in conscious rats (maximal 26 +/- 5% at 2 nmol CRF). These responses were significantly attenuated by pretreatment (i.c.v.) with either a monoclonal antibody raised to gamma 1MSH or with naloxone which antagonises beta-endorphin (beta-EP) actions. The increases were not affected by pretreatment with monoclonal antibodies to ACTH or the N-terminal of POMC. Central injections of gamma 1-melanocyte-stimulating hormone (MSH) or beta-EP caused dose-dependent increases in VO2 (maximal at 0.5-1.5 pmol) and these were markedly inhibited by pretreatment with the anti-gamma 1-MSH antibody or naloxone respectively. Injection of ACTH or alpha MSH did not significantly affect VO2 at doses up to 2 nmol. These data indicate that the central actions of CRF on thermogenesis may be mediated, at least in part, by release of gamma MSH and/or beta-EP.
The specificity properties that distinguish members of a set of homologous anti-digoxin antibodies are controlled by H chain mutations.
Five murine A/J strain anti-digoxin mAb (35-20, 40-40, 40-120, 40-140, and 40-160) have highly homologous H and L chain V regions, only differing by somatic mutation, yet differ in affinity and specificity. The availability of the VH and VL genomic clones from one hybridoma, 40-140, has now allowed studies involving in vitro mutagenesis and chain recombination among these five hybridomas. To determine the relative contributions of the mutations found in either VH or VL to the overall binding properties of these antibodies, we recombined the 40-140VH with the VL of each hybridoma. The 40-140VH gene was transfected into hybridoma variants that produce only VL. The recombinant antibodies show that the mutations present in VH, rather than in VL, affect the fine specificity properties of these antibodies, whereas, the mutations among both VH and VL chains are important in determining antigen affinity. From mutations present in VH that affect fine specificity properties, the comparison of the antibody sequences, and from the previously measured binding properties, we predicted and tested selected VH mutations for their ability to alter specificity or affinity by doing site-directed in vitro mutagenesis. The results for the somatic mutations found in this group of antibodies show: 1) VH mutations control the fine specificity properties that distinguish different members of this group; 2) in particular, VH residues 54 and 55 in CDR2 control the distinguishing characteristics of specificities between these antibodies; and 3) by mutagenesis, we had the unusual result of being able to alter Ag specificity without affecting affinity. A computer model of the 40-140 antibody binding site was generated which indicates that VH residues 54 and 55 are highly accessible.
CDC7 protein kinase activity is required for mitosis and meiosis in Saccharomyces cerevisiae.
The product of the CDC7 gene of Saccharomyces cerevisiae has multiple cellular functions, being needed for the initiation of DNA synthesis during mitosis as well as for synaptonemal complex formation and commitment to recombination during meiosis. The CDC7 protein has protein kinase activity and contains the conserved residues characteristic of the protein kinase catalytic domain. To determine which of the cellular functions of CDC7 require this protein kinase activity, we have mutated some of the conserved residues within the CDC7 catalytic domain and have examined the ability of the mutant proteins to support mitosis and meiosis. The results indicate that the protein kinase activity of the CDC7 gene product is essential for its function in both mitosis and meiosis and that this activity is potentially regulated by phosphorylation of the CDC7 protein.
The molecular mechanism for the assembly and secretion of ApoB-100-containing lipoproteins.
We have reviewed the literature on the intracellular transport of ApoB-100 and the assembly of the ApoB-100-containing lipoproteins. ApoB-100 is a large molecule (4536 aa) that requires some 15 min to be completed. During the synthesis, the protein could take one of two pathways: a degradational pathway and a pathway that leads to secretion of the protein on mature lipoproteins. The degradational pathway starts with a cotranslational incorporation of ApoB-100 into the membrane of the endoplasmic reticulum in such a way that a relatively large portion of the sequence is exposed on the cytoplasmic surface of this membrane. The membrane bound ApoB-100 is retained in the ER and will eventually undergo intracellular degradation. To enter the pathway that leads to lipoprotein formation, ApoB-100 has to be cotranslationally translocated to the lumen of the ER. ApoB-100 will interact with the lipids during this translation-translocation process and the mature lipoprotein is released into the lumen of the secretory pathway when ApoB-100 is completed and leaves the ribosome. In addition to the mature lipoproteins, the secretory pathway contains an ApoB-100-containing lipoprotein with the density of a HDL particle. This particle is not secreted from the cells but is retained and eventually degraded. Of importance for the retention are sequences present in the C-terminal half of the protein. The mature lipoproteins rapidly leave the ER lumen and are transported to the Golgi apparatus, through which transfer takes considerably longer. The assembly process is a potential site for the regulation of the secretion of the ApoB-100-containing lipoproteins. This process is dependent on active synthesis of phosphatidylcholine and it is also highly dependent on the rate of triacylglycerol synthesis. On the other hand, ApoB-100 appears to be constitutively expressed. An increase in the rate of lipoprotein assembly induced by an increased triacylglycerol synthesis gives rise to an increased recruitment of ApoB-100 nascent polypeptides to interact cotranslationally with lipids. ApoB-100 that is not used for lipoprotein assembly is cotranslationally bound to the ER membrane and sorted to degradation.
Effects of transport and racing on ionic changes in thoroughbred race horses.
1. Packed cell volume (PCV), blood glucose, total plasma proteins (TPP) and plasma electrolytes, osmolality, cortisol and aldosterone alterations produced by transport and racing, were investigated in race horses. 2. Plasma cortisol, sodium and blood glucose, found after transport, were higher, while aldosterone was lower than control levels. 3. After racing, PCV, blood glucose, TPP and plasma cortisol, sodium and osmolality were higher than control, while chloride diminished and aldosterone returned to control values. 4. These results demonstrate that transport and racing are different kinds of stressors, suggesting that the sympathetic system and hypophysis-suprarenal cortex axis have a dissimilar contribution to the physiological response.
Reinke's oedema and thyroid function.
Reinke's oedema (RO) of the vocal folds is a condition of unknown aetiology. Cigarette smoking and vocal abuse may contribute to its development. Hypothyroidism has been described as an aetiological factor but, to date, no controlled study has been published confirming this association. This prospective, controlled study compared thyroid function in 61 consecutive RO patients with an age and sex matched control group (n = 65) without laryngeal disease. Thyroid function was assessed by measuring serum thyroid stimulating hormone (TSH), free thyroxine (T4) and tri-iodothyronine (T3). Hypothyroidism was diagnosed when TSH was above normal with a T3 and T4 below the normal range. Four RO patients were hypothyroid at the time of diagnosis, compared to five of the control group. Six RO patients had past or present hypothyroidism compared to seven of the controls. One RO patient and two controls were euthyroid with marginally elevated TSH levels. Although the incidence of hypothyroidism in this group of Reinke's oedema patients is higher than expected in a normal population, it is similar to that in an age and sex matched control group, reflecting the prevalence of hypothyroidism in middle aged women. This study suggests that hypothyroidism is not an aetiological factor in the development of Reinke's oedema.
Diptheroids and associated infections as a cause of failed instrument stabilization procedures in the lumbar spine.
Between February 1985 and October 1987, the authors identified seven patients with occult lumbar spine infections associated with the presence of spinal fixation hardware. Six of these infections were with organisms of low virulence; four of the seven patients had polymicrobial infections. All of the polymicrobial infections contained a Diptheroid as one of the isolates. Two of the seven patients studied had normal sedimentation rates. All had white blood cell counts less than 12,000 cells. Imaging studies were not helpful with the exception of one case with a positive gallium scan. The diagnoses were supported by clinical presentation, pathologic tissue changes, positive cultures, and response to therapy. Successful therapy was obtained by removal of hardware and treatment with antibiotics.
Laryngeal mask airway for coronary artery bypass grafting.
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Laryngeal mask airway for coronary artery bypass grafting.
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Episodic ACTH and cortisol secretion in normal children.
The aim of this study was to determine the normal relationship between ACTH and cortisol secretion in children. Fourteen children (nine male, five female; median age 11.3 years) were hospitalized and blood samples were taken every 20 min for 24 h. A circadian rhythm was observed with median 0900 h and midnight ACTH values of 1.80 and less than 0.97 pmol/l, and for cortisol 296 and 62 nmol/l respectively. The median (range) areas under the curve for ACTH and cortisol were 29.7 (9.0-53.8) pmol/l/h and 5114 (3562-8630) nmol/l/h respectively. There were no significant differences detected for ACTH and cortisol secretion between males and females, or between prepubertal (n = 9) and pubertal subjects (n = 5). Using a novel form of time series analysis we have shown that both ACTH and cortisol are secreted with a dominant periodicity of 0.7-1.0 h, representing 24-34 secretory episodes of ACTH and cortisol in 24 h. For cortisol, but not ACTH, there is a significant secondary periodicity of 2-3.2 h. To look for shared periodicities we have used the technique of coherency. This reveals that for six of the children ACTH and cortisol are secreted with a significant shared periodicity of 0.8-1.0 h, and for a further five children a similar secondary shared periodicity is present. Therefore in normal children ACTH and cortisol secretion are interdependent and episodic but are not influenced by either pubertal status or gender.
Corticotrophin releasing hormone (CRH1-41) stimulates the secretion of adrenocorticotrophin, vasopressin and oxytocin but not adrenocorticotrophin precursors: evidence from petrosal sinus sampling in man.
To examine the relationship between corticotrophin releasing hormone (CRH), arginine vasopressin (AVP) and oxytocin (OXT) we have studied the responses of adenohypophyseal and neurohypophyseal hormones to CRH in eight patients (age 26-64 years, six female) with suspected pituitary-dependent Cushing's syndrome during bilateral, simultaneous inferior petrosal sinus catheterization. Blood samples were taken from both petrosal sinuses and a peripheral vein before, and at 5-min intervals for 15 min after, an intravenous injection of 100 micrograms human CRH1-41. CRH increased sinus AVP concentrations in all eight patients and OXT concentrations in four of five patients studied. Although AVP concentrations often increased in both sinuses, the side of maximal AVP rise was termed side(max-AVP). CRH did not affect peripheral or petrosal sinus mean concentrations of LH, FSH, GH or TSH. While there was no change in mean peripheral concentrations of AVP, OXT, ACTH, ACTH precursors or prolactin after CRH, sinus concentrations of OXT, ACTH and prolactin on side(max-AVP) were markedly elevated over contralateral values. CRH did not increase mean sinus concentrations of ACTH precursors. In seven patients with either no radiological abnormality or the pituitary fossa or a small adenoma the mean ACTH precursor/ACTH ratio in blood sampled from all sites was 2.1 +/- 0.16 (mean +/- SEM, n = 50). In a patient with a large, locally invasive tumour the mean ACTH precursor/ACTH molar ratio was 32.1 +/- 1.3 (n = 12; P less than 0.001), suggesting that alterations in this molar ratio may reflect the biological properties of the tumour. The source of CRH-stimulatable AVP and OXT remains uncertain.(ABSTRACT TRUNCATED AT 250 WORDS)
Neurophysiological effects of left vagal stimulation in man.
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Regulation of ACTH secretory pathways in cultured pituitary cells.
Although chloroquine, an agent that disrupts regulated protein secretion, has previously been shown to decrease the adrenocorticotropic hormone (ACTH) secretory response to adenosine 3',5'-cyclic monophosphate or corticotropin-releasing factor (CRF) in AtT-20 and rat anterior pituitary cells, respectively, it has no effect on the response to vasopressin. The present study extended experiments with chloroquine to cultured sheep anterior pituitary cells, which have a greater maximum response to vasopressin. Chloroquine (200 microM) had no effect on basal ACTH secretion or on stimulation by vasopressin. In contrast to the rat, the net response to CRF was tripled by chloroquine in ovine cells. The effect of chloroquine on the response to CRF was more effective by coexposure of cells to CRF and chloroquine than by pretreatment with chloroquine. Monensin or vinblastine did not increase the ACTH response to CRF. The results indicate ACTH release in response to vasopressin is chloroquine insensitive in this way, can be dissociated from the mechanism that responds to CRF, and would be consistent with the CRF response mechanism involving pathways that can alter the secretory pool of ACTH. When chloroquine acts to increase the response to CRF, it is likely not to act by stabilizing the CRF-receptor complex.
Penetration of administered IgG into the maxillary sinus and long-term clinical effects of intravenous immunoglobulin replacement therapy on sinusitis in primary hypogammaglobulinaemia.
The influence of long-term intravenous immunoglobulin (i.v. IgG) replacement therapy on the clinical course of chronic sinusitis in patients with primary hypogammaglobulinaemia has not previously been reported. We have analysed the efficacy of i.v. IgG therapy and the penetration of administered i.v. IgG into the maxillary sinus. Seventeen patients with primary hypogammaglobulinaemia received i.v. IgG replacement therapy to maintain pre-infusion serum IgG concentrations above 4 g/l for periods of 12 to 58 (mean 36.7) months. Cases with established chronic sinusitis prior to therapy did not have symptomatic or radiological improvement at this dose, although no sinusitis developed de-novo in the 3 previously unaffected patients. The administered IgG penetrated into maxillary sinus antral lavage fluid in 3 patients from whom secretions were obtained at antroscopy. This indicates that poor clinical responses are not due to lack of penetration of antibodies to the required sites of action. Larger doses of i.v. IgG may be more effective in this situation, but the addition of antibiotics at high dosage may be a more economical therapeutic alternative. These findings highlight the importance of diagnosis of hypogammaglobulinaemia by measurement of serum IgG concentrations in patients who suffer from recurrent sinusitis, as the early institution of i.v. IgG therapy may prevent the development of sinusitis refractory to i.v. IgG therapy.
Recent advances in the diagnosis and treatment of infectious diseases.
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A sensitive radioimmunoassay using a monoclonal antibody that is equipotent for ercalcitriol and calcitriol (1,25-dihydroxy vitamin D2 and D3).
A monoclonal antibody has been used in a sensitive radioimmunoassay that measures 1,25-dihydroxyvitamin D2 (ercalcitriol) and 1,25-dihydroxyvitamin D3 (calcitriol) with equal potency. This important characteristic has not been reported for any other radioimmunoassay for 1,25-dihydroxyvitamin D. The two forms can be assayed in human serum together or individually after HPLC separation. Sample preparation entails acetonitrile extraction followed by C18-Sep-pak chromatography and HPLC. The assay measures 98% of added analyte, and achieves inter- and intra-assay coefficients of variation of 10.7% at 34 pg/ml and 7.8% at 81 pg/ml respectively. The limit of detection is 1.25 pg/tube and 50% displacement of bound ligand is achieved at 14 pg/tube. The reference interval is 20-50 pg/ml, mean 35. The correlation between results from the monoclonal radioimmunoassay and an established polyclonal antibody method was r = 0.98, slope 0.99. The assay has particular application in patients treated with vitamin D2 since 1,25-dihydroxyvitamin D2 can now be measured accurately in the presence of 1,25-dihydroxyvitamin D3.
Antibodies to neuropeptides: biological effects and mechanisms of action.
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