Additional reports of failure to respond to treatment after rabies exposure in Thailand.
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Biomedical subjects
Publications and source records attributed to T Hemachudha.
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Three cases of idiopathic hypertrophic cranial pachymeningitis are presented. The diagnosis was based on the CT scan or MRI findings (or both) of thickened enhancing dura. In all cases, meningeal biopsies were performed and microscopic findings were compatible with nonspecific inflammation. All cases presented with subacute and chronic localized headache. Two cases had associated chronic meningitis. One cases presented with a syndrome of multiple cranial nerve involvement (polyneuritis cranialis). Corticosteroids, in the form of prednisolone 60 mg/day, were effective in all cases. Two cases with less severe pachymeningitis received corticosteroids for 2 weeks, then were tapered off in 4 to 6 weeks. A case with extensive lesions needed a long-term low dosage of prednisolone, 5 to 10 mg/day for maintenance therapy. Idiopathic hypertrophic cranial pachymeningitis may be related to the Tolosa-Hunt syndrome, the syndrome of polyneuritis cranialis, and multifocal fibrosclerosis.
Rabies is a complex disease. We still do not understand the mechanisms of clinically diverse furious and dumb types and its fatal course. Moreover clinical symptomatology, once believed to be unique, may be variable, particularly in those patients who develop disease after exposure to virus of the insectivorous or frugivorous bat origin. This review summarizes classic and nonclassic clinical features associated with canine and bat rabies variants and also atypical presentations of rabies survivors. Difference in cellular tropism either at the inoculation site or in the central nervous system or differences in route of spread, or both, may account for these discrepancies. Furthermore, these may affect different sets of neurotransmitters that in turn modulate variable neurobehavioural patterns and neuroendocrine-immune cascades.
Functions of the muscarinic acetylcholine receptor (mAChR) were studied in rabid dog brains using [3H]quinuclidinyl benzilate (QNB) as a radioligand. Of various brain regions, hippocampus and brainstem were the areas mostly affected in terms of impaired specific binding to [3H]QNB, as compared to other regions, as well as to those of controls. Saturation studies of the hippocampus revealed significantly elevated dissociation equilibrium constant (K(d)) values in both furious (n = 5) (9.80 + or - 2.77 nM) and dumb (n = 6) (6.01 + or - 1.08 nM) types of rabies as compared to 11 controls (2.15 + or - 0.31 nM), whereas the maximum number of receptor sites (B (max)) values were comparable among all subgroups of normal (1.38 + or - 0.10 pmol/mg protein), dumb (1.43 + or - 0.17 pmol/mg protein) and furious (1.28 + or - 0.12 pmol/mg protein) rabies types. Hippocampal K(d) values were comparable between high (fluorescent antibody test-FAT and polymerase chain reaction-PCR positive; n = 4) (7.47 + or - 3.27 nM), and low (FAT-negative and PCR-positive; n = 4) virus amount (8.34 + or - 3.93 nM) but these were significantly higher than controls (n = 4) (1.58 + or - 0.17 nM). Our data suggest a functional derangement of mAChR at specific sites of hippocampus and brainstem which is not dependent on the amount of virus.
The increase in world-wide travel means that physicians everywhere require an understanding of rabies and its prevention, to advise intending travellers, or to follow-up on treatment begun overseas. In this article, we discuss measures to prevent rabies.
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A simple, sensitive, and specific polymerase chain reaction (PCR) protocol for detection of rabies virus is described. The process consists of sample preparation, reverse transcription, two-step DNA amplification, and detection of the amplified product. RNA was extracted from animal and human brain by phenol-chloroform using guanidinium thiocyanate. Viral RNA was then amplified in a two-step PCR that used two sets of nested primers designed to amplify rabies nucleocapsid (N) sequence. Rabies nucleocapsid sequence was amplified from all brain samples from 95 dogs and 3 humans with rabies confirmed by fluorescent antibody (FAT) and mouse inoculation tests (MIT). Rabies-negative brain samples (110 dogs, 2 humans) were PCR-negative. The process requires < 24 h. Detection of viral RNA was still possible in brain material that was left at room temperature for 72 h. As little as 8 pg of rabies virus RNA could be detected. This technique could have practical applications as a confirmatory test to FAT at busy rabies diagnostic centers.
Cytotoxic lymphocyte function in 13 patients with rabies was studied by counting the number of CD56 cells and assessing natural killer (NK) cell activity. There was no significant difference in the number of killer cells between rabies patients and 31 normal controls (P greater than 0.05). Two of six non-fatal encephalitic patients due to causes other than rabies had reduced CD56 numbers. Base-line NK cell responses versus K562 cell targets did not differ among the normal control and rabies groups (P greater than 0.05). Study of the non-rabies encephalitis group showed heterogeneous results with wide variation. Significant enhancement of NK activity was seen in four rabies patients and in 10 normal control subjects tested after interferon-alpha (IFN-alpha) and IL-2. None of the four patients with encephalitis due to causes other than rabies showed such enhancement. Our results suggest that NK cells of rabies patients are not fully stimulated and that this might contribute to the virulence of rabies. The cause of this phenomenon remains unknown.
Serum neutralizing antibody to rabies virus was determined in previously unvaccinated Thai pet dogs after receiving one subcutaneous dose of inactivated tissue culture rabies vaccine (Rabdomun, Coopers Animal Health Company, Germany, 4.55 IU ml-1 potency). Geometric mean titres on days 14, 30, 60, 180 and 360 were 2.14, 2.30, 0.45, 0.14 and 0.05 IU ml-1, respectively, by the rapid immunofluorescent focus inhibition test. Titres of neutralizing antibody to rabies virus did not correlate with the age of the dog at the time of vaccination or with the presence or absence of anaemia or blood parasites. Six out of 50 (12%), 11 out of 43 (25.6%) and 13 out of 31 (42%) dogs had no detectable rabies antibody in serum 60, 180 and 360 days, respectively, after vaccination. Three of these antibody-negative dogs were given another dose of vaccine. Antibody reappeared on day 14 but rapidly declined within 60 days. These data suggest that one dose of tissue culture vaccine in dogs by the subcutaneous route of injection is not adequate to maintain rabies neutralizing antibody in serum for 1 year.
The 2-1-1 rabies postexposure treatment schedule is an abbreviated regimen in which a tissue culture rabies vaccine is administered intramuscularly at two sites on day 0, and at one site on days 7 and 21. Compared to the standard five-dose intramuscular regimen, the 2-1-1 schedule reduces the number of clinic visits from five to three and the amount of vaccine used by 20%. One hundred Thai patients, who were severely exposed to rabies, were treated with rabies immune globulin and the 2-1-1 regimen using purified Vero cell rabies vaccine. They were followed for 1 year. Rabies antibody titres were measured in 10% of this group. All patients survived and adverse reactions were mild. A satisfactory antibody response (a titre greater than 0.5 IU ml-1) occurred in all ten patients studied at day 14, but persisted for 90 days in 80% and for 360 days in only 50%. The authors therefore do not recommend use of the 2-1-1 schedule in severely exposed patients who also need to receive rabies immune globulin.
Antibodies to two components of rabies virus, nucleocapsid (N) and glycoprotein (G), were compared in 11 rabies patients with those in nine recipients of Vero cell rabies vaccine. All rabies vaccinees had antibodies to N and G components by day 10 after the first vaccine injection. A similar but not identical response was observed in three out of 11 rabies patients. Serum antibodies appeared in rabies patients as early as 3 days after onset of the first symptoms of the disease. In these antibody-positive rabies patients, levels of both antibodies, but particularly of anti-N antibody, were lower than in the vaccinated group. Our results suggest that the process of immune recognition and of antibody development in human rabies is more likely to occur early in the pre-clinical phase, and that reactivity to N protein may be crucial for elicitation of neutralizing antibody.
Plasma C3c levels were examined in 56 patients with immune (27) and non-immune (29) mediated neurological diseases by crossed immunoelectrophoresis. Plasma samples were collected during the active phase of illness in both groups, usually within 7 days of admission. 11 patients (4 Guillain-Barré Syndrome-GBS, 3 chronic inflammatory demyelinating polyneuropathy-CIDP, 4 myasthenia gravis-MG) had their plasma saved sequentially during the active and the recovery phase. Plasma C3c levels were elevated in the group with immune mediated diseases when compared with those of non-immune mediated diseases. The sensitivity and specificity of C3c as a diagnostic test for immune mediated neurological diseases were 61.4 and 100% respectively with a positive and negative predictive value of 100 and 41%. the C3c levels in plasma correlated well with disease severity in MG and GBS patients. Such a correlation was also evident in all CIDP patients except one that had persistent elevation in the presence of clinical improvement. Results suggest that the plasma C3c level may be useful for differentiating immune from non-immune mediated neurological diseases. Plasma C3c may also be used for monitoring disease severity, particularly in myasthenia gravis.
We reported the occurrence of anticardiolipin antibodies (ACA) by using an enzyme-linked immunosorbent assay (ELISA) in sera of patients with neurologic complications from Semple rabies vaccination. There was a correlation between the presence of ACA and the disease severity. Sixteen of 25 patients (64%) with major neurological complications, 2 of 21 patients (10%) with minor complications, and non of the normal vaccinees had an ACA response. In comparison to this, ACA was found in 10/43 (24%) of patients with post-infectious encephalitis (PIE), Guillain-Barré syndrome (GBS) and multiple sclerosis (MS), 5/22 (23%) and 4/31 (13%) of patients with degenerative neurological diseases and central nervous system (CNS) infections, respectively. There was no specific restriction to any particular isotype. Frequency difference of ACA responses was unremarkable in systemic lupus erythematosus (SLE) patients with (3/9) and without (3/10) CNS involvement. It is not conclusive about the pathogenetic role of ACA. This remains to be determined.
Development of neurologic complications after Semple rabies vaccine is closely linked to development of antibody to myelin basic protein (MBP). The portions of MBP against which the antibodies are directed were analyzed by enzyme immunoassay in sera and cerebrospinal fluid from 27 patients with vaccine complications. Most of the antibody was directed to regions of MBP peptides 45-89 and 90-170. There was no apparent correlation between antibody specificity for MBP peptides 1-44, 45-89 and 90-170 and the type of post-vaccinal neurologic complication. We conclude that the immunoglobulin repertoire in human B lymphocytes for responding to human MBP favors the portion of the MBP molecule containing residues 45-170.
We studied the distribution of rabies viral antigen in the brain and spinal cord of 7 patients with rabies by immunohistochemical techniques. Four patients presented with encephalitis, the remaining 3 had paralysis. Neither the rabies viral antigen distribution nor inflammation paralleled clinical presentations. Patients who had survival times of 7 days or less (4/7) had a greater amount of antigen-positive neurons in brainstem and spinal cord regardless of the clinical type. Neuroglial cells were also found to contain rabies antigen. Our findings suggest that virus localization may not account for the difference in clinical manifestations.
Three failures of postexposure rabies treatment using imported purified duck embryo cell and Vero cell rabies vaccines are reported from Thailand. Reference is made to eight additional previously reported Thai patients, six of whom had received human diploid cell vaccine. An analysis of these cases reveals that there were serious flaws in management in all of these patients. It is stressed that 45% of human rabies deaths in Thailand occur within 20 days of being bitten and 71% are dead within 28 days. This short incubation period does not allow much time to start immunotherapy. Of Bangkok dogs found to have rabies at autopsy, approximately 8% have a rabies immunization history. Once a dog has bitten a patient immunotherapy should not be delayed in countries with a high incidence of dog rabies. Patients with chronic disease, alcoholics and drug addicts may have an impaired immune response to postexposure rabies vaccines.
Lymphocyte subsets of 7 patients with encephalitic and paralytic rabies were determined by immunocytochemical techniques using mouse monoclonal antibodies. Almost all patients had diminished mononuclear cells of Leu 7 phenotype (natural killer cells). Cells of Leu 12 marker (B cells) were decreased in 3 paralytic rabies patients compared with those of 4 patients in the encephalitic group.