Biomedical subjects
R E Davis
Publications and source records attributed to R E Davis.
Sensitive detection and identification of mycoplasma-like organisms in plants by polymerase chain reactions.
DNA amplification by polymerase chain reactions (PCR) was employed to detect host plant infection by several mycoplasmalike organisms (MLOs), including the aster yellows (AY), dwarf aster yellows (DAY), and periwinkle little leaf (0-1) MLOs. For PCR, two pairs of oligonucleotide primers, designated AY18pm and AY19pm, respectively, were synthesized on the basis of partial sequences of cloned AY MLO DNA fragments AY18 and AY19. Reaction mixtures containing primer pair AY18pm yielded a DNA product of 1.6Kbp, when template consisted of DNA extracted from AY MLO- or DAY MLO-infected Catharanthus roseus (periwinkle). A DNA product of 1.0Kbp was obtained with primer pair AY19pm, when template consisted of DNA extracted from C. roseus infected by AY MLO, DAY MLO, or periwinkle little leaf (strain O-1) MLO. MLO-specific bands were observed when reaction mixtures contained as little as 5 pg total nucleic acid from infected plants. No PCR product was observed when reaction mixtures contained only DNA from healthy plants or DNA from plants infected by western X MLO or by tomato big bud MLO. The findings indicated that the PCR system is useful for sensitive detection and differentiation of MLOs in infected hosts.
Type 1 diabetes and latent pernicious anaemia.
OBJECTIVE: To determine the prevalence of latent pernicious anaemia (PA) in Type 1 diabetics. DESIGN: Patients with Type 1 diabetes were screened at two yearly intervals on a continuing basis. SETTING: Diabetic Unit, Royal Perth Hospital, Western Australia. PATIENTS: Three hundred and seventy-one patients, 156 females and 215 males, attending a diabetic clinic. They were classified as having Type 1 diabetes on the basis of age of onset less than 40 years and requiring insulin from the start. MAIN OUTCOME MEASURES: Serum cobalamin levels were measured and studies of thyroid function and a blood count were done. Patients with a reduced serum level of cobalamin had tests for cobalamin absorption. RESULTS: Six patients had a low serum cobalamin level; five showed malabsorption of the vitamin which could be corrected by the addition of intrinsic factor. Four of these patients had no clinical signs of PA. The fifth was mildly anaemic (haemoglobin level 111 g/L) and had megaloblastic bone marrow. He was classified as having frank PA. The sixth patient was not available for further testing. These results give a prevalence of latent PA of 11 per 1000 in Type 1 diabetics, compared with 3.9 per 1000 in diabetics with clinically manifest disease and 1.27 per 1000 in the general population. All four patients with latent PA had hypothyroidism, based on low thyroxine levels, increased levels of thyroid stimulating hormone and the presence of thyroid antibodies. CONCLUSION: Latent PA is not uncommon in Type 1 diabetics. It has a long preclinical course and occurs in those patients with thyroid disease. The screening of Type 1 diabetics for latent PA has worthwhile benefits as a "preventive" health measure.
Glutamic acid-insensitive [3H]kainic acid binding in goldfish brain.
Kainic acid is supposed to be a specific agonist for a subclass of excitatory glutamate receptors in the vertebrate CNS. An investigation of (2 nM) [3H]kainic acid binding sites in goldfish brain, using quantitative autoradiography, has revealed evidence for two types of kainic acid receptors which differ in sensitivity to glutamic acid. L-Glutamic acid (0.1-1 mM) displaced over 95% of specific [3H]kainic acid binding elsewhere in the brain but only 10-50% in the cerebellum and cerebellar crest. These structures apparently contain [3H]kainic acid binding sites that are extremely insensitive to glutamic acid. The glutamic acid-insensitive [3H]kainic acid binding was not displaced by quisqualic acid, kynurenic acid, alpha-amino-3-hydroxy-5-methylisoxazolepropionic acid (AMPA), or N-methyl-D-aspartatic acid, but was completely displaced by the kainic acid analogue domoic acid. The data indicate that two types of high affinity binding sites for [3H]kainic acid exist in the goldfish brain: glutamic acid-sensitive and glutamic acid-insensitive. High affinity [3H]kainic acid binding may therefore not always represent binding to subsets of glutamic acid receptors.
Cholinergic agents: effect of methyl substitution in a series of arecoline derivatives on binding to muscarinic acetylcholine receptors.
Arecoline, arecaidine, and a series of derivatives, differing by the presence or absence of methyl groups at positions on the periphery of the molecule, were prepared, and their binding to muscarinic acetylcholine receptors was tested. On the basis of this study, muscarinic agonism for arecoline series is governed by strict structure-activity relationships, as previously observed for other agonist series. Only minor changes in nitrogen substitution were tolerated in the present series of arecoline derivatives.
Extracellular recordings from the motor nervous system of the nematode, Ascaris suum.
1. The close association of muscle and neurons in Ascaris suum makes it difficult to determine whether spikes recorded from nerve cords originate in muscle or neurons. We have developed criteria that distinguish muscle and neuronal activity. There are two categories of extracellular spikes. 2. The first category consists of spikes with a wide range of amplitudes, marked by large spikes. These spikes, which can be recorded over lateral muscle and over the dorsal and ventral nerve cords, are abolished when muscle is disrupted or removed, or when curare is applied. Large spikes are relatively infrequent, are correlated with intracellularly recorded muscle events, and respond to polarizations of motor neurons, implying that they originate in muscle. 3. The second spike category, small amplitude spikes, is exclusive to the ventral nerve cord, occurs more frequently than large spikes and displays patterned firing. Small spikes are not affected by muscle removal or by curare, and are correlated with motor neuronal post-synaptic potentials, but not with intracellularly recorded muscle events. We infer that they originate in neurons. 4. Low level activity recorded extracellularly over nerve cords may represent muscle activity due to tonic motor neuronal synaptic transmission. It responds to motor neuronal polarization and is suppressed by curare or muscle removal.
N-methyl-D-aspartate receptor antagonist MK-801 impairs learning but not memory fixation or expression of classical fear conditioning in goldfish (Carassius auratus).
The amnestic effects of the noncompetitive antagonist MK-801 on visually mediated, classic fear conditioning in goldfish (Carassius auratus) was examined in 5 experiments. MK-801 was administered 30 min before the training session on Day 1 to look for anterograde amnestic effects, immediately after training to look for retrograde amnestic effects, and before the training or test session, or both, to look for state-dependence effects. The results showed that MK-801 produced anterograde amnesia at doses that did not produce retrograde amnesia or state dependency and did not impair the expression of conditioned or unconditioned branchial suppression responses (BSRs) to the conditioned stimulus. The results indicate that MK-801 disrupts the mechanism of learning of the conditioned stimulus-unconditioned stimulus relation. Evidence is also presented that the learning processes that are disrupted by MK-801 occur during the initial stage of BSR conditioning.
Angiotensin II regulates the metabolism but not the secretion of vasoactive intestinal peptide in the rabbit.
1. We have previously demonstrated that the metabolism and secretion of vasoactive intestinal peptide are affected by both acute and chronic dietary sodium. Sodium concentrations in portal and systemic plasma were unaffected by differing levels of sodium intake or administration of an acute gastric sodium load. We sought, therefore, to determine whether other hormones involved in sodium homoeostasis (such as angiotensin II) might be involved in regulating the metabolism and secretion of vasoactive intestinal peptide. We determined the metabolic clearance rate and theoretical secretion rate of vasoactive intestinal peptide in rabbits on low sodium (high circulating angiotensin II) and high sodium (low circulating angiotensin II) diets with and without simultaneous angiotensin II infusion. 2. The metabolic clearance rate of vasoactive intestinal peptide was significantly higher in rabbits on the high sodium diet during both vehicle control (P < 0.01) and angiotensin II (P < 0.05) infusion. Angiotensin II infusion decreased the metabolism of vasoactive intestinal peptide in rabbits on both low (P < 0.01) and high (P < 0.01) sodium diets. 3. Although there was a significant difference in secretion rates between the two dietary groups (P < 0.025) under basal conditions, infusion of angiotensin II did not alter the secretion rate significantly in either group. 4. We conclude that angiotensin II regulates the metabolism of vasoactive intestinal peptide in the rabbit, but does not regulate its secretion.
The effects of a high sodium diet on the metabolism and secretion of vasoactive intestinal peptide in the rabbit.
1. In view of previous observations that the metabolism of vasoactive intestinal peptide (VIP) is significantly increased in sodium-depleted rabbits, we wished to determine whether a high sodium intake also leads to alterations in VIP metabolism. We performed metabolic clearance studies in rabbits maintained on a high sodium diet and normal control diets. These studies were performed both before and after the administration of 1.5 mmol kg-1 of sodium intravenously to observe the effects of an acute increase in body sodium. 2. The rabbits maintained on the high sodium diet had a significantly lower basal plasma VIP level (P less than 0.025), a lower metabolic clearance rate (MCR) of the peptide (P less than 0.025) and a lower secretion rate (P less than 0.005), compared with the normal control animals. These differences were maintained following the intravenous sodium infusion. 3. The administration of the intravenous sodium infusion resulted in a further decrease in MCR in the rabbits on the high sodium diet (P less than 0.05). 4. These results confirm that VIP metabolism is affected by high dietary intake of sodium, as well as a low sodium intake, adding further support to the hypothesis that VIP may be involved in sodium homeostasis.
Identification and analysis of a genomic strain cluster of mycoplasmalike organisms associated with Canadian peach (eastern) X disease, western X disease, and clover yellow edge.
Genetic interrelatedness among 13 strains of mycoplasmalike organisms (MLOs) from various sources was evaluated by dot hybridization and restriction fragment length polymorphism analyses using cloned DNA probes derived from Canadian peach X (CX) and western X (WX) MLOs. Dot hybridization analysis indicated that CX, WX, and clover yellow edge MLOs are closely related and form a distinct strain cluster that is only distantly related to the 10 other MLOs. Similarity coefficients derived from restriction fragment length polymorphism analysis revealed that CX, WX, and clover yellow edge MLOs represent three distinct genomic types.
Pharmacologic activity of CI-977, a selective kappa opioid agonist, in rhesus monkeys.
CI-977 is a selective, nonpeptide kappa opioid agonist. In rhesus monkeys, CI-977 is a potent antinociceptive agent against thermal stimuli after i.m. administration. Increasing the intensity of the nociceptive stimulus can reduce the analgesic activity of CI-977. Antinociceptive activity also was seen when PD 126212, containing CI-977 as the (-)-enantiomer, was administered sublingually. Naloxone antagonized the antinociceptive action of CI-977, demonstrating opiate receptor involvement in this activity. Monkeys treated with CI-977 also showed sedation at doses close to those required to produce antinociception. As with morphine, the sedative properties of CI-977 were associated with impaired cognitive performance. Aged monkeys appeared more sensitive than young monkeys to the performance-impairing effect of CI-977. Tolerance developed to the antinociceptive and response-suppressing effects. CI-977 was approximately 1000 times more potent than morphine as an analgesic when tested against a moderate (50 degrees C) thermal stimulus but less effective than morphine against a strong (55 degrees C) thermal stimulus.
T lymphocytes expressing HECA-452 epitope are present in cutaneous acute graft-versus-host disease and erythema multiforme, but not in acute graft-versus-host disease in gut organs.
Lymphocytes in formalin-fixed skin biopsies from patients with cutaneous acute graft-versus-host disease (aGVHD) were studied with HECA-452 (an antibody recognizing lymphocytes with skin-homing properties) and a panel of antibodies recognizing pan-B (L26 [CD20]), pan-T (L60 [CD43] and A6 [CD45RA]), and T-helper subset (OPD4) antigens in paraffin sections. Biopsies from patients with erythema multiforme (EM) were similarly studied for comparison. In both conditions, T lymphocytes stained by OPD4 were predominantly confined to the dermis, whereas those stained by HECA-452 were concentrated in the epidermis; however, there was considerable variation between cases, and overlap between findings in the dermis and epidermis. Lymphocytes similarly studied in paraffin sections of liver, salivary gland, and gut affected by aGVHD were essentially unreactive with HECA-452, although they were largely stained by pan-T markers and showed some comparable reactivity with OPD4. The findings suggest that aGVHD of the skin is mediated by a different set of lymphocytes than in gut organs, and may have a similar immunologic mechanism to EM.
Diagnosis and management of perilymph fistula: the University of North Carolina approach.
Perilymph fistula is a potentially disabling leakage of perilymphatic fluid from the otic capsule. The occurrence of perilymph fistula is now attributed to a wide variety of causes and the diagnosis is frustrated by a lack of characteristic signs or symptoms. A brief review of the literature and the University of North Carolina philosophy for the diagnosis and management of perilymph fistula is presented. Emphasis on timely evaluation and surgical technique is provided.
Joint task force "forward care" multicomponent health service support for an Army Reserve separate infantry brigade (mechanized)--Part III.
At the conclusion of 4 years' careful study of the health services support of a separate infantry brigade (mechanized) during the unit's annual training periods, the authors report on the effectiveness of a support team consisting of Army Reserve medical elements, an Active Army field unit, and a Public Health Service Clearing/Staging unit joining forces in a field environment to provide real world medical care to the same unit in a follow-on annual training period. The emphasis of the team created was on validating the forward care concept of field medical support. The result of this effort was "state of the art" medical service to the troops in the most forward areas, and a savings of 0.66 training days per soldier out of 10 days possible field training time. The cross-training of joint elements was enhanced by providing hands-on treatment of soldiers in a tactical environment, training that cannot be adequately replaced by simulated training.
Utility of molecular genetic analysis for the diagnosis of neoplasia in morphologically and immunophenotypically equivocal hematolymphoid lesions.
The authors reviewed retrospectively the results of molecular genetic analysis of 175 hematolymphoid lesions to assess the diagnostic utility of genotyping (analysis for immunoglobulin or beta-chain T-cell receptor gene rearrangements). All cases had been genotyped, and most were also immunophenotyped. Cases were assigned to control (90 cases) or problem groups (85 cases), depending on the absence or presence respectively of diagnostic uncertainty remaining after conventional (morphologic and immunophenotypic) analysis. All control cases had unequivocal morphologic features of non-Hodgkin's lymphoma (NHL) and had appropriate lineage-specific gene rearrangements, but immunostaining was almost as sensitive in demonstrating phenotypes that were diagnostically abnormal as well as lineage-specifying. However, genotyping was clearly diagnostically useful in the problem cases, which included a heterogeneous mixture of nodal and extranodal biopsy specimens of malignant (mostly NHL) and benign lesions. Genotyping demonstrated the appropriate absence or presence of gene rearrangements in 58 of 81 problem cases (72%) ultimately diagnosed as benign or malignant respectively, excluding four cases of Hodgkin's disease; in 25 cases (31%) this was judged to be essential to diagnosis. In the remaining problem cases genotyping did not contribute positively to diagnosis, but in no case was genotyping misleading when cautiously interpreted with primary reliance on conventional analysis and with knowledge of known causes of potentially misleading results, both positive and negative. It is concluded that although genotyping is highly sensitive and specific for hematolymphoid neoplasia, it provides little or no benefit in cases that are unequivocally malignant by conventional analysis, in cases of suspected Hodgkin's disease (HD), or for the sole purpose of lineage assignment. However, in cases whose diagnosis is uncertain after conventional analysis, the high yield of useful information from genotyping with careful interpretation warrants its application.
Neuropeptides in the nematode Ascaris suum.
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Inflammatory pseudotumor of lymph nodes. Additional observations and evidence for an inflammatory etiology.
Inflammatory pseudotumor of lymph nodes (IPT), a recently described benign cause of lymphadenopathy, was studied in 14 patients using paraffin-section immunohistochemistry. All biopsies showed a proliferation of spindle cells (small blood vessels, histiocytes, and "activated" fibroblasts resembling myofibroblasts) containing a mixture of inflammatory cells without atypia and involving the connective tissue framework (hilum and sinuses) of the lymph node. Ten patients had additional histologic features of IPT originally described (extranodal extension, obliterative vasculitis, and endothelial infiltration), and nine of these had associated fever of unknown origin, which in some was relieved by biopsy alone. Additional features observed focally in some cases but not previously described included lymph node parenchymal infarction, fibrinoid vascular necrosis, karyorrhexis, and involvement of only part of the lymph node. Immunostaining showed the lymphoid infiltrate to be predominantly of T-lineage (except for plasma cells), only a minority of which marked as T-helper cells. Numerous mononuclear cells resembling histiocytes were identified, some of which had a spindled shape but reacted with an antibody (KP1) of myelomonocytic specificity. Large fibroblastic cells expressed alpha-muscle actin but not desmin, similar to myofibroblasts in granulation tissue. The morphologic and immunohistologic features were similar to those in inflammatory pseudotumors of spleens and livers also studied, but the lack of simultaneous lymph node involvement argued against a common etiology. The findings suggest that the mass lesion of IPT is produced in response to localized lymph node inflammation or injury and further exclude hematolymphoid or mesenchymal neoplasia as a cause.
An NMDA intervention strategy in schizophrenia with "low-dose" milacemide.
Drugs (e.g., PCP) which interfere with glutamatergic transmission at the N-methyl D-aspartate (NMDA) subclass of glutamate receptors precipitate both positive and negative symptoms of psychosis in humans. Based on a proposed "glutamatergic deficiency" in schizophrenia, pharmacologic facilitation of NMDA-mediated neural transmission by direct stimulation of the strychine-insensitive glycine binding site was attempted with "low-dose" milacemide, an acylated "prodrug" of glycine that readily crosses the blood brain barrier and is converted into glycine in the brain. In a prior study, "high-dose" milacemide proved to have no therapeutic utility in schizophrenia. The failure was thought, possibly, to be related to higher doses of milacemide having antagonist actions at the NMDA receptor complex. In the current study, "low-dose" milacemide (400 mg/day), as the sole pharmacotherapeutic agent, was also without significant clinical benefit. Despite our negative findings for milacemide, other strategies for facilitating NMDA-mediated neural transmission in schizophrenia might be worth pursuing.