Potent gp120-like neurotoxic activity in the cerebrospinal fluid of HIV-infected individuals is blocked by peptide T.

The envelope protein of the human immunodeficiency virus (gp120) causes neuronal death in developing murine hippocampal cultures or rat retinal ganglion cells. In HIV-infected individuals, gp120 released from HIV-infected macrophages or other cells in the brain has been proposed as the etiology for the pathophysiology of AIDS central nervous system (CNS) disease by diffusing to act at a distance to cause damage and/or death to neighboring neurons. In this study, 28 cerebrospinal fluid (CSF) samples from HIV-infected individuals (79% were WR stage 1 and 2) and neurological disease controls were tested, blind to the investigator, for the presence of in vitro neuronal killing activity. Neurotoxic activity was detected with peak effects at a 1:10(5) dilution in CSF from 9/18 HIV-infected individuals and 1/10 neurological disease controls. Thus half of CSF from early stages of HIV disease are characterized by the presence of neurotoxic activity which is not present in control CSF (Fischers exact test, P < 0.05). The neuronal toxicity by patient CSF could be prevented by peptide T (1 nM). A monoclonal antibody to mouse CD4, RL.172, also attenuated or prevented CSF-induced neuronal killing in all four CSF samples tested. In addition, an antiserum to peptide T previously shown to bind gp120 and neutralize both infectively and direct gp120 neurotoxicity, neutralized the CSF factor. gp120, or a modified small fragment, is suggested to be the responsible toxic molecular entity. These results may be relevant to the pathophysiology of HIV-related CNS disease and the mechanism by which peptide T causes improvements.     

A VIP antagonist distinguishes VIP receptors on spinal cord cells and lymphocytes

Vasoactive intestinal peptide (VIP) is a neuropeptide which also interacts with cells of the immune system. The paucity of specific VIP receptor antagonists has hampered studies of possible receptor heterogeneity and of VIP function. To aid in achieving these goals, a new VIP antagonist, a hybrid between neurotensin and VIP, has been synthesized. This peptide interacted with VIP receptors on spinal cord cells with an affinity 10-fold greater than VIP itself. In contrast, 1000-fold higher concentrations of the antagonist were required to displace labeled VIP from its receptor on lymphoid cells as compared to VIP itself, suggesting VIP receptor heterogeneity between immune and spinal cord cells.     

Learning and sexual deficiencies in transgenic mice carrying a chimeric vasoactive intestinal peptide gene

The molecular mechanisms responsible for behavior are largely unknown. A state of the art model, paving the path from genes to behavior, is offered by transgenic animals. Candidate molecules are classic neuropeptides, such as vasoactive intestinal peptide (VIP). Transgenic mice harboring a chimeric VIP gene driven by the polyoma promoter were produced. Behavioral studies revealed learning impairment and prolonged retardation in memory acquisition in the genetically altered animals. Furthermore, reduced performance was observed when the male transgenic mice were tested for sexual activity in the presence of receptive females. Surprisingly, radioimmunoassays showed an approx 20% decrease in the VIP content of the transgenic mice brains. To directly assess genetically reduced VIP content as a cause for learning impairment, transgenic mice carrying diphtheria toxia-encoding sequences driven by the rat VIP promoter were created. These animals had reduced brain VIP and exhibited deficiencies in learning abilities, strongly supporting an important neurobiological function for VIP in vivo.     

Deletion of Brca2 exon 27 causes hypersensitivity to DNA crosslinks,chromosomal instability,and reduced life span in mice

The Brca2 tumor-suppressor gene contributes to genomic stability, at least in part by a role in homologous recombinational repair. BRCA2 protein is presumed to function in homologous recombination through interactions with RAD51. Both exons 11 and 27 of Brca2 code for domains that interact with RAD51; exon 11 encodes eight BRC motifs, whereas exon 27 encodes a single, distinct interaction domain. Deletion of all RAD51-interacting domains causes embryonic lethality in mice. A less severe phenotype is seen with BRAC2 truncations that preserve some, but not all, of the BRC motifs. These mice can survive beyond weaning, but are runted and infertile, and die very young from cancer. Cells from such mice show hypersensitivity to some genotoxic agents and chromosomal instability. Here, we have analyzed mice and cells with a deletion of only the RAD51-interacting region encoded by exon 27. Mice homozygous for this mutation (called brca2(lex1)) have a shorter life span than that of control littermates, possibly because of early onsets of cancer and sepsis. No other phenotype was observed in these animals; therefore, the brca2(lex1) mutation is less severe than truncations that delete some BRC motifs. However, at the cellular level, the brca2(lex1) mutation causes reduced viability, hypersensitivity to the DNA interstrand crosslinking agent mitomycin C, and gross chromosomal instability, much like more severe truncations. Thus, the extreme carboxy-terminal region encoded by exon 27 is important for BRCA2 function, probably because it is required for a fully functional interaction between BRCA2 and RAD51.     

Investigation of whether on-site general practice counsellors have an impact on psychotropic drug prescribing rates and costs.

BACKGROUND: Counselling services are now widespread within general practice. Although the cost-effectiveness of such services has yet to be fully investigated, benefits could include a reduction in prescribing of psychotropic drugs and of other drugs. AIM: A study set out to determine whether practices with counsellors differed from those without in terms of their prescribing rates and costs of psychotropic drugs. METHOD: Prescribing analyses and cost (PACT) level two data reports for the quarter to November 1991 ending January 1992, as appropriate, were sought from 354 practices with counsellors and a matched sample of 216 practices without counsellors which had participated in a previous national survey of counselling in general practice. The drug groups examined were: hypnotics and anxiolytics; antidepressants; analgesics; all central nervous system drugs; and all drugs apart from central nervous system drugs. For each group of drugs, the numbers of prescribed items, total prescribing costs, and costs per item were expressed as a proportion of the practice's number of prescribing units (that is, the age-adjusted number of registered patients) and as a percentage of the average for similar practices in its family health services authority. Practice characteristics were compared between practices with an on-site counsellor and those without. Practices with and without counsellors were compared with respect to their prescribing indicators. RESULTS: PACT reports were obtained from 214 practices (response rate 38%)--126 with counsellors and 88 without. Practices with counsellors and practices without counsellors were well matched in terms of location, list size, proportion of elderly patients, training and fundholding status, and number of health promotion clinics. No significant differences were found between practices with and without counsellors in the prescribing indicators for any group of psychotropic drugs examined or for central nervous system drugs as a whole. CONCLUSION: There were no appreciable differences found in this study between practices with and without counsellors in terms of psychotropic drug prescribing rates or costs. The reasons for this are unclear; more indepth studies of individual counselling services are required.     

Telephone versus postal surveys of general practitioners: methodological considerations.

BACKGROUND. High response rates to surveys help to maintain the representativeness of the sample. AIM. In the course of a wider investigation into counselling services within general practice it was decided to assess the feasibility of increasing the response rate by telephone follow up of non-respondents to a postal survey. METHOD. A postal survey was undertaken of a random sample of 1732 general practitioners followed by telephone administration of the questionnaire to non-respondents. The identical questionnaire was administered by telephone to a separate random sample of 206 general practitioners. RESULTS. Of 1732 general practitioners first approached by mail, 1683 were still in post of whom 881 (52%) completed the postal questionnaire and a further 494 (29%) the telephone interview. Of 206 general practitioners first contacted by telephone, 197 were still in post of whom 167 (85%) completed interviews. Compared with doctors first approached by mail, those first approached by telephone were significantly more likely to report having a partner with a special interest in psychiatry (P < 0.01); and a general practitioner, practice nurse or health visitor who worked as a counsellor (P < 0.01 in each case). A comparison of doctors first approached by telephone with those who completed telephone interviews after failing to respond to the postal questionnaire showed that postal non-respondents were significantly less likely to report having a general practitioner, practice nurse, health visitor or community psychiatric nurse who worked as a counsellor (P < 0.01 in each case). CONCLUSION. These findings suggest that non-response to the postal survey was associated with lack of activity in the study area. Telephone administration of questionnaires to postal non-respondents increased response rates to above 80% but, as telephone administration enhanced the reporting of counsellors, a social desirability bias may have been introduced.     

Inhibition of experimental autoimmune encephalomyelitis in Lewis rats by nasal administration of encephalitogenic MBP peptides: synergistic effects of MBP 68-86 and 87-99

Induction of mucosal tolerance by inhalation of soluble peptides with defined T cell epitopes is receiving much attention as a means of specifically down-regulating pathogenic T cell reactivities in autoimmune and allergic disorders. Experimental autoimmune encephalomyelitis (EAE) induced in the Lewis rat by immunization with myelin basic protein (MBP) and Freund's adjuvant (CFA) is mediated by CD4+ T cells specific for the MBP amino acid sequences 68-86 and 87-99. To further define the principles of nasal tolerance induction, we generated three different MBP peptides (MBP 68-86, 87-99 and the non- encephalitogenic peptide 110-128), and evaluated whether their nasal administration on day -11, -10, -9, -8 and -7 prior to immunization with guinea pig MBP (gp-MBP) + CFA confers protection to Lewis rat EAE. Protection was achieved with the encephalitogenic peptides MBP 68-86 and 87-99, MBP 68-86 being more potent, but not with MBP 110-128. Neither MBP 68-86 nor 87-99 at doses used conferred complete protection to gp-MBP-induced EAE. In contrast, nasal administration of a mixture of MBP 68-86 and 87-99 completely blocked gp-MBP-induced EAE even at lower dosage compared to that being used for individual peptides. Rats tolerized with MBP 68-86 + 87-99 nasally showed decreased T cell responses to MBP reflected by lymphocyte proliferation and IFN-gamma ELISPOT assays. Rats tolerized with MBP 68-86 + 87-99 also had abrogated MBP-reactive IFN-gamma and tumor necrosis factor-alpha mRNA expression in lymph node cells compared to rats receiving MBP 110-128 nasally, while similar low levels of MBP-reactive transforming growth factor-beta and IL-4 mRNA expressing cells were observed in the two groups. Nasal administration of MBP 68-86 + 87-99 only slightly inhibited guinea pig spinal cord homogenate-induced EAE, and passive transfer of spleen mononuclear cells from MBP 68-86 + 87-99-tolerized rats did not protect naive rats from EAE. Finally, we show that nasal administration of MBP 68-86 + 87-99 can reverse ongoing EAE induced with gp-MBP, although higher doses are required compared to the dosage needed for prevention. In conclusion, nasal administration of encephalitogenic MBP peptides can induce antigen-specific T cell tolerance and confer incomplete protection to gp-MBP-induced EAE, and MBP 68-86 and 87-99 have synergistic effects. Non-regulatory mechanisms are proposed to be responsible for tolerance development after nasal peptide administration.