Effects of adjuvant on neuropeptide-like immunoreactivity in experimentally induced temporomandibular arthritis in rats.

Substance P (SP)-, neurokinin A (NKA)-, calcitonin gene-related peptide (CGRP)- and neuropeptide Y (NPY)-like immunoreactivities (-LI) were examined in cerebrospinal fluid (CSF), plasma and temporomandibular joint (TMJ) perfusates in rats 1 and 12 h after inoculation at the base of the tail (0.05 ml) or injection into the right TMJ (0.01 ml) of heat-killed Mycobacterium butyricum in paraffin oil. In the rats inoculated at the base of the tail (polyarthritic rats), there was a significant increase of CGRP-LI and NKA-LI. The changes in neuropeptide-LI were not as marked in the CSF of rats injected with adjuvant in one TMJ (monoarthritic rats) as in the polyarthritic group. Instead, the most significant changes in the monoarthritic rats were seen in the perfusates of both TMJs. The increases in SP-, NKA-, CGRP- and NPY-LI were significant for both TMJs and more pronounced than in the polyarthritic rats. The results show that inoculation of adjuvant at the base of the tail induces significant changes of neuropeptide-LI predominantly in CSF, whilst an intra-articular injection induces bilateral changes in neuropeptide-LI in joint perfusate. Therefore, two different neural mechanisms may be involved early in adjuvant-induced poly- and monoarthritis.

Capsaicin effects on substance P and CGRP in rat adjuvant arthritis.

The effects of capsaicin on the sensory neuropeptides substance P and calcitonin gene-related peptide were analyzed in the ankle joints and dorsal root ganglia (L2-L6) of adult female Lewis rats. The study included 23 normal rats and 23 arthritic rats, all injected subcutaneously with capsaicin (total dose 200 mg/kg bw). Another two groups of animals from a previous study, i.e., 23 normal rats and 23 arthritic rats not given capsaicin served as controls. Adjuvant arthritis was induced by inoculation with heat-killed mycobacteria. The morphological distribution of sensory neuropeptides was assessed by immunohistochemistry and the tissue concentrations were determined by radioimmunoassay. In normal rats, capsaicin significantly reduced the concentrations of substance P and calcitonin gene-related peptide in ankle joints (54 and 36%, respectively) as well as dorsal root ganglia (40 and 54%, respectively). In arthritic rats those pretreated with capsaicin had significantly lower concentrations of substance P and calcitonin gene-related peptide in dorsal root ganglia (19 and 42%, respectively) compared to the arthritic controls. In the ankle joints, however, only the SP concentration was reduced (42%). Notably, this was accompanied by a 40% reduction in inflammatory response as assessed by comparing the ankle joint weights of the experimental groups. In general, there was a good correlation between the neuropeptide concentrations in ipsilateral ankle joints and the corresponding dorsal root ganglia as assessed in individual rats. The present study of adjuvant induced arthritis shows that capsaicin administration reduces the otherwise up-regulated levels of sensory neuropeptides in dorsal root ganglia and ankle joints. However, capsaicin at the dose given can only mitigate, not completely prevent the development of joint inflammation. Nonetheless, the findings suggest that antineuronal therapy targeted against specific neurotransmitters may prove useful in inflammatory joint disease.

Effects of surgical denervation on substance P and calcitonin gene-related peptide in adjuvant arthritis.

The occurrence of substance P (SP) and calcitonin gene-related peptide (CGRP) was assessed in the ankle joints and dorsal root ganglia (L2-L6) by immunohistochemistry and radioimmunoassay (RIA) after unilateral sciatic nerve section in adjuvant arthritis. Nerve section in arthritic rats had no clear-cut effect on warmth, redness, and swelling of ankle joints, but significantly affected the occurrence of SP and CGRP. Immunohistochemical analysis showed an almost complete disappearance of SP-positive fibers in the ipsilateral ankle joint in normal rats after axotomy, whereas in arthritic rats occasional SP-positive fibers remained. In dorsal root ganglia, only occasional SP-positive cells could be detected in normal and arthritic rats after axotomy. A similar but somewhat less pronounced effect of axotomy was noted for neuronal CGRP-LI. RIA showed a decrease in SP in ankle joints by 45% in normal rats and 58% in arthritic rats; the decrease in CGRP was 41% and 47%, respectively. In dorsal root ganglia, the decrease in SP after surgical denervation was 25% in normal rats and 54% in arthritic rats; the decrease in CGRP was 18% and 27%, respectively. The tissue concentrations of SP and CGRP in ipsilateral ankle joints and dorsal root ganglia were consistently correlated in normal as well as arthritic rats. The present study shows that an interruption of the nerve supply to joints cannot fully prevent the development of arthritis, although it significantly reduces the occurrence of sensory neuropeptides.

Extraction and quantitation of neuropeptides in bone by radioimmunoassay.

The feasibility of extracting and quantifying neuropeptides in bone by radioimmunoassay was investigated in a study including 60 diaphyseal rat femora. Substance P, calcitonin gene-related peptide, neuropeptide Y and vasoactive intestinal polypeptide, previously identified in bone by immunohistochemistry, were extracted from separate homogenates of bone, periosteum and bone marrow in a solution of 4% EDTA and 2 M acetic acid. Measurable amounts of all four neuropeptides in bone, periosteum and bone marrow were obtained by radioimmunoassay in a reproducible manner. The neuropeptide immunoreactivities were characterized by reverse-phase high performance liquid chromatography. Among the four neuropeptides analyzed, neuropeptide Y consistently exhibited the highest concentrations in the different tissues. Overall, cortical bone showed the lowest neuropeptide concentrations. The concentration of vasoactive intestinal polypeptide was higher in periosteum than in bone marrow, whereas that of calcitonin gene-related peptide was uniform in these tissues. The distributional differences observed in bone tissue may be explained by a variety of physiological roles attributed to neuropeptides such as regulation of nociception, vasoactivity, immune function and local bone metabolism. The described methodology offers a new means of investigating a neuropeptidergic involvement in various disorders of the skeleton.

Extraction of neuropeptides from joint tissue for quantitation by radioimmunoassay. A study in the rat.

The feasibility of extracting neuropeptides from rat knee joints for quantitation by radioimmunoassay was tested. The investigation, based on 25 adult Lewis rats, focused on substance P, calcitonin gene-related peptide, neuropeptide Y, and vasoactive intestinal polypeptide. The relative recovery of the peptides in different extraction media was assessed Both knee joints including the articulating epiphysis were dissected and cut into small pieces. The series was divided into five subgroups, 10 joints in each, for extraction in five different media: 1) 1 M acetic acid in 4% EDTA, 2) 2 M acetic acid in 4% EDTA, 3) neutral water in 4% EDTA, 4) 2 M acetic acid in 4% EDTA and 95% alcohol, and 5) 2 M acetic acid without EDTA. Measureable concentrations of the four neuropeptides were reproducibly assessed by RIA. Although all extraction media provided measurable concentrations, 2 M acetic acid in 4% EDTA was found to give the highest overall yield of the four neuropeptides analyzed. Reverse-phase HPLC confirmed that the immunoreactivities assessed by RIA corresponded to the four neuropeptides of interest. Experimental and clinical evidence suggest a neurogenic involvement in the pathophysiology of inflammatory joint disease, e.g., rheumatoid arthritis. The extraction procedure described offers a means of determining neuropeptide concentrations in joint tissue under normal and pathologic conditions by RIA.

Endothelin concentrations in respiration-related structures of the medulla during the perinatal period of the rat.

Endothelin-like immunoreactivity (ET-LI) was quantified in the developing (foetal and postnatal) rat brain stem and cerebellum using radioimmunoassay. The brain stem structures chosen for this study were (a) dorsal medullary region (DMR) including the region of nucleus tractus solitarius where the peripheral chemoreceptor afferents are known to terminate, (b) ventral medullary region (VMR) where the central chemoreceptors are thought to be located and (c) cerebellum (CER), as a control area. Compared to the prenatal period, significantly elevated concentrations of ET-LI were detected in the early postnatal period and thereafter the concentrations decreased: DMR and VMR: in comparison to the prenatal concentrations, a two-fold increase was found on the day of birth which further increased significantly (P < 0.001) on postnatal day 1 only in the region of DMR; CER: low concentrations of ET-LI were found in the early postnatal period which were not significantly different from the prenatal values. No ET-LI could be detected in any of the three regions in the adult rats. The results are discussed in view of the hypothesis that (1) endothelin appears to play an important role in the perinatal period and (2) it is involved in the chemoreceptor pathway.

Electroconvulsive stimuli and brain peptides: effect of modification of seizure duration on neuropeptide Y, neurokinin A, substance P and neurotensin.

We studied the effects of modification of duration of seizures induced by electroconvulsive stimuli (ECS) on the changes in concentration of neuropeptide Y (NPY), neurokinin A (NKA), substance P (SP) and neurotensin (NT)-like immunoreactivity (-LI) in specific rat brain regions. Rats were divided into groups pretreated with saline, indomethacin, flurbiprofen or diazepam prior to either six sham ECSs or six ECSs. After sacrifice by focused microwave irradiation, brains were dissected into frontal cortex, occipital cortex, striatum, hippocampus, pituitary and hypothalamic sections. Peptides were extracted and measured in extract aliquots by specific radioimmunoassays. Repeated ECS increased NPY-LI and NKA-LI in the hippocampus and the occipital cortex. No effect on SP-LI or NT-LI was found. Indomethacin and flurbiprofen had no effect on the tonic seizure time following ECS, and they did not affect the ECS-induced alterations of the brain peptides. Diazepam pretreatment decreased the tonic seizure time following ECS in a dose-dependent manner. However, diazepam did not modify the ECS-induced increase in NPY-LI and NKA-LI concentrations. The results firmly establish that ECS leads to specific peptide increases in discrete rat brain regions and raise the possibility that such changes may not entirely be a consequence of seizures per se.

Community-based rehabilitation: an evaluation study.

Leprosy gives rise to two types of stigmatization, one from the disease and its neuropathetic manifestations, with their resultant disability and handicaps, and the other due to social ostracism. The process of rehabilitation should begin from the moment the disease is diagnosed, and the earlier its detection the better the prognosis for patients. The family unit to which the patient belongs plays a vital role in his social life, ensuring and enhancing his self-respect and dignity in society, and this fact must be recognized when evolving a strategy for rehabilitation. In no circumstances should a patient be removed from his natural home environment. It is important that the community is made leprosy conscious and gets more involved in hastening the social assimilation of patients. Communication plays an important role throughout the rehabilitation process. One of the major functions is the removal of the social stigma in the family and in the community and this involves communication skills to ensure interaction between the staff and patients' families and the education of the community. A highlight of community-based rehabilitation is the excellent rate of repayment of loans by the patients to whom they were made. Also of note is the extent to which former defaulters make repayments due to the continuous rapport and good interpersonal relationship between the staff and patients. Most of the subjects of this study were drawn from the lower economic strata of society and for them the most essential consideration is to make a living, however meagre.(ABSTRACT TRUNCATED AT 250 WORDS)