Self-perceived upper extremity motor function predicts health-related quality of life in chronic stroke survivors.

PURPOSE: To examine whether the Upper Extremity Functional Index (UEFI) score independently contributes to the Stroke Impact Scale (SIS) score and quantified its relative contribution to SIS scores in chronic stroke survivors. MATERIALS AND METHODS: A cross-sectional study in a university-based rehabilitation centre with people with chronic stroke (N = 95) aged ≥ 50 years. The outcome measures included paretic hand grip strength, Fugl-Meyer Upper Extremity Assessment (FMA-UE), Wolf Motor Function Test (WMFT), UEFI, and SIS. RESULTS: Correlation analysis revealed that paretic hand grip strength, FMA-UE, UEFI, and WMFT scores exhibited a significant moderate positive correlation with SIS scores (r = 0.544-0.687, p < 0.001). The results of a regression model indicated that after adjustment for demographic factors and stroke-related impairments, the UEFI scores remained independently associated with SIS scores, accounting for 18.8% of the variance. The entire model explained 60.3% of the variance in SIS scores. CONCLUSIONS: Self-perceived UE motor function is a crucial component to be included in rehabilitation programmes aimed at enhancing quality of life and participation among chronic stroke survivors.

Observation-based outcome measures, e.g., Fugl­Meyer Assessment for Upper Extremity (FMA-UE), Wolf Motor Function Test (WMFT) could not predict the health-related quality of life (Stroke Impact scale (SIS)) in chronic stroke survivors in our study, which was contradictory with current studies.A self-perceived outcome measure to evaluate upper extremity function (Upper Extremity Functional Index (UEFI)) could independently predict the health-related quality of life (SIS), accounting for 18.8% of the variance.Our study demonstrated that self-perceived UE motor function would be an important component to optimize the rehabilitation programmes aimed at enhancing quality of life and social participation among chronic stroke survivors.

Effects of mirror therapy with electrical stimulation for upper limb recovery in people with stroke: a systematic review and meta-analysis.

PURPOSE: To provide updated evidence about the effects of MT with ES for recovering upper extremities motor function in people with stroke. METHODS: Systematic review and meta-analysis were completed. Methodological quality was assessed using the version 2 of the Cochrane risk-of-bias tool. The GRADE approach was employed to assess the certainty of evidence. RESULTS: A total of 16 trials with 773 participants were included in this review. The results demonstrated that MT with ES was more effective than sham (standardized mean difference [SMD], 1.89 [1.52-2.26]) and ES alone (SMD, 0.42 [0.11-0.73]) with low quality of evidence, or MT alone (SMD, 0.47[0.04-0.89]) with low quality of evidence for improving upper extremity motor control assessed using Fugl-Meyer Assessment. MT with ES had significant improvement of (MD, 6.47 [1.92-11.01]) the upper extremity gross gripping function assessed using the Action Research Arm Test compared with MT alone with low quality of evidence. MT combined with ES was more effective than sham group (SMD, 1.17 [0.42-1.93) for improving the ability to perform activities of daily living with low quality of evidence assessed using Motor Activity Log. CONCLUSION: MT with ES may be effective in improving upper limb motor recovery in people with stroke.

Combining Mirror Therapy (MT) and Electrical Stimulation (ES) modality could improve upper limb motor control, gross gripping function, and performance in ADLs based on ICF for people with stroke.Those individuals with subacute stroke are recommended as the optimal target group for the combined MT and ES.

Muscle-specific androgen receptor deletion shows limited actions in myoblasts but not in myofibers in different muscles in vivo.

The aim of this study was to investigate the direct muscle cell-mediated actions of androgens by comparing two different mouse lines. The cre-loxP system was used to delete the DNA-binding activity of the androgen receptor (AR) in mature myofibers (MCK mAR(ΔZF2)) in one model and the DNA-binding activity of the AR in both proliferating myoblasts and myofibers (α-actin mAR(ΔZF2)) in another model. We found that hind-limb muscle mass was normal in MCK mAR(ΔZF2) mice and that relative mass of only some hind-limb muscles was reduced in α-actin mAR(ΔZF2) mice. This suggests that myoblasts and myofibers are not the major cellular targets mediating the anabolic actions of androgens on male muscle during growth and development. Levator ani muscle mass was decreased in both mouse lines, demonstrating that there is a myofiber-specific effect in this unique androgen-dependent muscle. We found that the pattern of expression of genes including c-myc, Fzd4 and Igf2 is associated with androgen-dependent changes in muscle mass; therefore, these genes are likely to be mediators of anabolic actions of androgens. Further research is required to identify the major targets of androgen actions in muscle, which are likely to include indirect actions via other tissues.

Can short-term residential care for stroke rehabilitation help to reduce the institutionalization of stroke survivors?

BACKGROUND: Stroke survivors may not be receiving optimal rehabilitation as a result of a shortage of hospital resources, and many of them are institutionalized. A rehabilitation program provided in a short-term residential care setting may help to fill the service gap. OBJECTIVES: The primary objectives of this study were, first, to examine whether there were significant differences in terms of rehabilitation outcomes at 1 year after admission to the rehabilitation program (defined as baseline) between those using short-term residential care (intervention group) and those using usual geriatric day hospital care (control group), and, second, to investigate whether lower 1-year institutionalization rates were observed in the intervention group than in the control group. PARTICIPANTS: 155 stroke survivors who completed at least the first follow-up at 4 months after baseline. INTERVENTION: The intervention group was stroke survivors using self-financed short-term residential care for stroke rehabilitation. The control group was stroke survivors using the usual care at a public geriatric day hospital. MEASUREMENTS: Assessments were conducted by trained research assistants using structured questionnaires at baseline, 4 months, and 1 year after baseline. The primary outcome measures included Modified Barthel Index score, Mini-Mental Status Examination score, and the institutionalization rate. RESULTS: Cognitive status (as measured by Mini-Mental Status Examination score) of patients in both groups could be maintained from 4 months to 1 year, whereas functional status (as measured by Modified Barthel Index score) of the patients could be further improved after 4 months up to 1 year. Meanwhile, insignificant between-group difference in rehabilitation outcomes was observed. The intervention participants had a significantly lower 1-year institutionalization rate (15.8%) than the control group (25.8%). CONCLUSION: Short-term residential care for stroke rehabilitation promoted improvements in rehabilitation outcomes comparable with, if not better than, the usual care at geriatric day hospital. Furthermore, it had a significantly lower 1-year institutionalization rate. This type of service could be promoted to prevent institutionalization.

Antigenic, microbicidal and antiparasitic properties of an l-amino acid oxidase isolated from Bothrops jararaca snake venom.

Venoms from the bee Apis mellifera, the caterpillar Lonomia achelous, the spiders Lycosa sp. and Phoneutria nigriventer, the scorpions Tityus bahiensis and Tityus serrulatus, and the snakes Bothrops alternatus, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni, Bothrops neuwiedi, Crotalus durissus terrificus, and Lachesis muta were assayed (800mug/mL) for activity against Staphylococcus aureus. Venoms from B. jararaca and B. jararacussu showed the highest S. aureus growth inhibition and also against other Gram-positive and Gram-negative bacteria. To characterize the microbicidal component(s) produced by B. jararaca, venom was fractionated through gel exclusion chromatography. The high molecular weight, anti-S. aureus P1 fraction was further resolved by anion exchange chromatography through Mono Q columns using a 0-0.5M NaCl gradient. Bactericidal Mono Q fractions P5 and P6 showed significant LAAO activity using l-leucine as substrate. These fractions were pooled and subjected to Heparin affinity chromatography, which rendered a single LAAO activity peak. The anti-S. aureus activity was abolished by catalase, suggesting that the effect is dependent on H(2)O(2) production. SDS-PAGE of isolated LAAO indicated the presence of three isoforms since deglycosylation with a recombinant N-glycanase rendered a single 38.2 kDa component. B. jararaca LAAO specific activity was 142.7 U/mg, based on the oxidation of l-leucine. The correlation between in vivo neutralization of lethal toxicity (ED(50)) and levels of horse therapeutic antibodies anti-LAAO measured by ELISA was investigated to predict the potency of Brazilian antibothropic antivenoms. Six horses were hyperimmunized with Bothrops venoms (50% from B. jararaca and 12.5% each from B. alternatus, B. jararacussu, B. neuwiedii and B. moojeni). To set up an indirect ELISA, B. jararaca LAAO and crude venom were used as antigens. Correlation coefficients (r) between ED(50) and ELISA antibody titers against B. jararaca venom and LAAO were 0.846 (p<0.001) and 0.747 (p<0.001), respectively. The hemolytic and leishmanicidal (anti-Leishmania amazonensis) activity of LAAO was also determined.

Distribution and molecular characterization of tetracycline resistance in Laribacter hongkongensis.

OBJECTIVES: Laribacter hongkongensis is a newly discovered bacterium associated with gastroenteritis and found in freshwater fish. Although isolates resistant to tetracycline have been described, their resistance mechanisms have not been studied. PATIENTS AND METHODS: We describe the distribution and molecular characterization of tetracycline resistance in 48 L. hongkongensis isolates from humans and fish. RESULTS: Three human isolates and one fish isolate were resistant to tetracycline (MIC 128 mg/L) and doxycycline (MIC 8-16 mg/L) and had reduced susceptibility to minocycline (MIC 1-4 mg/L). A 3566 bp gene cluster, which contains tetR and tetA, was cloned from one of the tetracycline-resistant strains, HLHK5. While the flanking regions and 3' end of the tetA of HLHK5 were identical to the corresponding regions of a tetC island in Chlamydia suis, the tetA gene was almost identical to that of transposon Tn1721 and plasmids of gram-negative bacteria, suggesting that the tetA/tetR of HLHK5 may have arisen from illegitimate recombination. PCR and DNA sequencing showed the presence of tetA in the other three tetracycline-resistant L. hongkongensis strains. Sequencing and characterization of a 15,665 bp plasmid, pHLHK22, from strain HLHK22 revealed the presence of a similar tetA/tetR gene cluster. This novel plasmid also confers tetracycline resistance when transformed to Escherichia coli and other L. hongkongensis isolates. CONCLUSIONS: Horizontal transfer of genes, especially through Tn1721 and related plasmids, is likely an important mechanism for acquisition and dissemination of tetracycline resistance in L. hongkongensis. The present study is the first report on identification of tetA genes in bacteria of the Neisseriaceae family.

Plasmid profile and construction of a small shuttle vector in Laribacter hongkongensis.

Among 21 human strains of Laribacter hongkongensis, small plasmids were observed in four strains, and large ones in six strains. The smallest, 3264-bp plasmid, pHLHK19, has only one ORF that encodes a putative replication initiator protein and a predicted origin of replication (ori) with a DnaA box, three 18-bp direct repeats and five pairs of inverted repeats. An Escherichia coli-L. hongkongensis shuttle vector was constructed by ligating the HindIII-digested pHLHK19, containing the replication initiator protein and ori of pHLHK19, to HindIII-digested pBK-CMV. This shuttle vector can propagate in E. coli and L. hongkongensis with good transformation efficiencies.

Puberdade em touros Nelore criados em pasto no Brasil: características corporais, testiculares e seminais e de índice de capacidade andrológica por pontos / Puberty in Nellore bulls raised at pasture in Brazil: body, testicular and seminal characteristics and breeding soundness evaluation

Foram utilizados 23 touros da raça Nelore entre os 10 e 20 meses de idade, para caracterizar o desenvolvimento puberal. Os animais foram criados em condições extensivas com alimentação em pasto e suplementação mineral. A cada duas ou quatro semanas foram realizadas colheitas de dados que incluíram medidas de peso corporal, circunferência torácica, comprimento e largura testicular e circunferência escrotal. Foram, também, colhidos ejaculados pela eletroejaculação e avaliados os aspectos físicos e morfológicos. No momento da colheita de sêmen foram aferidas as porcentagens de desprendimento entre pênis e prepúcio e determinadas as idades médias ao desprendimento total (IDPPRE). Foram determinadas as idades médias ao aparecimento dos primeiros espermatozóides (ISEM1) e dos primeiros espermatozóides móveis (ISEM2) no ejaculado, e a puberdade seminal (IDPUB). Estabeleceram-se os índices de capacidade andrológica por pontos (ICAP). Os touros Nelore apresentaram idades médias ISEM1, ISEM2 e IDPUB de 13,1±2,2, 13,6±2,3 e 14,8±1,8 meses, respectivamente, e IDPPRE de 18,1±1,9. Aos 16 meses de idade, o grupo apresentou 91,4 por cento, 82,6 por cento e 73,9 por cento dos animais com ISEM1, ISEM2 e IDPUB manifestada, respectivamente. Observou-se correlação positiva entre idade dos touros e as características de crescimento corporal, testicular, aspectos físicos dos ejaculados e correlação negativa entre a idade dos touros e os defeitos espermáticos. Observou-se correlação negativa entre a idade dos touros e a pontuação para circunferência escrotal no ICAP à puberdade (-0,77; P<0,001). Touros da raça Nelore, criados a pastos, manifestaram a puberdade precocemente antes dos 15 meses de idade. Houve um período de 51 dias desde o aparecimento dos espermatozóides no ejaculado, até a puberdade seminal. A puberdade foi mais precoce nos touros jovens com maiores testículos.

Twenty three 10-months-old Nellore bulls, raised under pasture conditions in Brazil, were used to study puberty. Monthly measurement of scrotal circumference and testicular width and length were recorded. Semen was collected by eletroejaculation and evaluated according to physical and morphological aspects. Penile and prepuce detachment percentages were also evaluated. The ages of the presence of first spermatozoa in the ejaculate (ISEM1), first motile spermatozoa in the ejaculate (ISEM2), seminal puberty (IDPUB) and detachment between penile and prepuce (IDPPRE) were calculated. Scores of breeding soundness were measured. ISEM1, ISEM2, IDPUB and IDPPRE were 13.1±2.2, 13.6±2.3, 14.8±1.8 and 18.1±1.9 months, respectively. At 16 months of age, the animals presented 91.4 percent, 82.6 percent and 73.9 percent with ISEM1, ISEM2 and IDPUB reached, respectively. High positive correlation were observed between age and corporal and testicular growth or seminal physical aspects. Negative correlation was verified between age and spermatic defects. High negative correlation was observed between age and score of scrotal circumference at puberty (-0.7;7 P<0.001). Precocious puberty was observed before 15 months of age. From ISEM1 to IDPUB, 51 days were elapsed. Young bulls with larger testicles reached puberty earlier.

Antibodies against synthetic epitopes inhibit the enzymatic activity of mutalysin II, a metalloproteinase from bushmaster snake venom.

Mutalysin II (mut-II), a 22.5kDa zinc endopeptidase isolated from bushmaster (Lachesis muta muta) snake venom, is a direct acting fibrin(ogen)olytic proteinase. It induces monoclonal and polyclonal antibodies which efficiently neutralize the hemorrhagic effect of L. muta and several Bothrops whole venoms. To characterize epitopes of protective antibodies we have used the Spot method of multiple peptide synthesis to prepare 64 overlapping dodecapeptides frameshifted by three residues, covering the complete amino acid sequence of mut-II. The rabbit anti-mut-II antibodies binding pattern to peptides revealed several continuous antigenic regions: one in the N-terminal part, two in the central region and the other in the C-terminal of mut-II. By using homology modelling, a three-dimensional model of mut-II was built which showed that epitopes are surface exposed. Anti-peptide antibodies were raised against three peptides (one representative of each epitope region) covalently coupled as a mixture to keyhole limpet hemocyanin. Purified IgG from the resulting anti- peptide antibodies cross-reacted with mut-II and induced a dose-dependent inhibition of the mut-II catalyzed proteolysis of fibrinogen.

Intermittent versus indwelling urinary catheterization in older female patients.

OBJECTIVES: To compare the use of intermittent and indwelling catheterization in older female patients with urinary retention. METHODS: A randomized, 2-week prospective study in a geriatric rehabilitation ward. Female patients of age 65 years and older with post-voiding residual urine volume (PVRU) persistently > or = 300 ml were randomly assigned to one of the two groups: intermittent catheterization (IMC group, n=36) and indwelling catheterization (IDC group, n=45). The primary outcome was the proportion of subjects being catheter-free and had a PVRU < 150 ml on day 14. The secondary outcomes were the time to become catheter-free and the rate of bacteriuria on day 14. RESULTS: Sixteen out of 27 (59.3%) in the IMC group versus 27 out of 39 (69.2%) in the IDC group achieved the primary outcome on day 14 (P=.403) without significant difference in the PVRU. The IMC and IDC groups took a mean of 8.6+/-3.3 and 9.2+/-4.0 days to become catheter-free, respectively (P=.609). Fourteen out of 22 (63.6%) in the IMC group versus 21 out of 34 (61.8%) in the IDC group had bacteriuria on day 14 (P=.888). CONCLUSION: Given the similar success rate of regaining bladder voiding function, the similar rate of bacteriuria and considering that the IMC group only underwent a median of 3 times of intermittent catheterization, we believe that the approach of intermittent urinary catheterization when required would be justified in managing elderly female urinary retention in rehabilitation ward as the presence of indwelling catheters would hinder rehabilitation and adversely affect patient quality of life.

Construction of an inducible expression shuttle vector for Laribacter hongkongensis, a novel bacterium associated with gastroenteritis.

An Escherichia coli-Laribacter hongkongensis shuttle vector (pPW380) was constructed by ligating the 4701-bp EcoRI digested fragment of pHLHK8 to EcoRI digested pBK-CMV. An E. coli-L. hongkongensis inducible expression shuttle vector was further constructed by ligating a 2105-bp fragment that contains the tetracycline repressor and tetracycline-inducible promoter region of pALC2084 to the 8897-bp fragment of pPW380, deletion of the green fluorescent protein gene, and insertion of a multiple cloning site. This inducible expression system was able to express two commonly used reporter genes, the green fluorescent protein gene and the glutathione S-transferase gene, efficiently in E. coli and L. hongkongensis.

Localization of epitopes in the toxins of Tityus serrulatus scorpions and neutralizing potential of therapeutic antivenoms.

Overlapping pentadecapeptides covering the complete amino acid sequence of TsII, TsVII and TsIV toxins from the venom of scorpion Tityus serrulatus (Ts), were prepared by use of the Spot method of multiple peptide synthesis. Horse anti-Ts antisera for therapeutic use were tested for their binding to peptides. All nine antisera tested showed reactivity with several peptides from the three toxins. Three antigenic regions, one in the very N-terminal, the second in the central part and the other in the C-terminal part of the three toxins were frequently, but not constantly recognized, with an intensity that seemed to be related to the neutralizing potency of the tested antivenom. Thus the corresponding peptides (residues 1-15 and 48-62 of TsII; residues 1-15, 16-30 and 48-62 of TsIV and residues 1-15 and 47-61 of TsVII) were synthesized, coupled to KLH and used as antigens to coat the microtitration plates to determine any relationship between their ELISA reactivity with therapeutic horse antivenoms and the neutralizing potential of these antivenoms. The mixture of the N-terminal peptide of TsII, of the N-terminal TsVII peptide and of the C-terminal of TsIV was found to give a linear relationship with the neutralizing titer of horse serum of low neutralizing potency (< or =1 mg/ml). However, high neutralizing antivenoms did not show the expected response in peptide ELISA. This observation is discussed in the context of the occurrence of continuous and discontinuous epitopes on toxins.

Cloning and characterization of a chromosomal class C beta-lactamase and its regulatory gene in Laribacter hongkongensis.

Laribacter hongkongensis, a newly discovered bacterium recently shown to be associated with community-acquired gastroenteritis, is generally resistant to most beta-lactams except the carbapenems. We describe the cloning and characterization of a novel chromosomal class C beta-lactamase and its regulatory gene in L. hongkongensis. Two genes, ampC and ampR, were cloned by inserting restriction fragments of genomic DNA from L. hongkongensis strain HLHK5 into pBK-CMV to give the recombinant plasmid pBK-LHK-5. The ampR and ampC genes and their promoters were divergently oriented, with the ampR gene immediately upstream of the ampC gene and an intercistronic Lys-R motif, typical of inducible ampC-ampR regulatory systems. The deduced amino acid sequence of the cloned AmpC beta-lactamase (pI 8.1) contained consensus motifs characteristic of class C beta-lactamases but had identities no greater than 46% to known class C beta-lactamases. The kinetic properties of this AmpC were also compatible with those of a class C beta-lactamase. PCR of 20 clinical isolates of L. hongkongensis, including HLHK5, showed the presence of both ampC and ampR genes in all isolates. Southern hybridization suggested that the ampC gene of HLHK5 was chromosomally encoded. Subcloning experiments showed that the expression of the ampC gene of HLHK5 was regulated by its ampR gene, which acts as a repressor. The beta-lactamase characterized from strain HLHK5 was named LHK-5 (gene, bla(LHK-5)) and represents the first example of AmpC beta-lactamase in the beta subdivision of proteobacteria.

Epitope mapping of the antigenic protein TsNTxP from Tityus serrulatus scorpion venom using mouse, rabbit and sheep antibodies.

In the present investigation we used native and recombinant TsNTxP to elicit antibodies in three different animal models (mouse, rabbit and sheep). Differences among anti-TsNTxP antibodies were analyzed using sets of overlapping pentadecapeptides of the TsNTxP amino acid sequence and also modified peptides to reveal key residues in antibody-peptide binding. Despite the identification of similar peptides by the antibodies in the C and N-terminal, peculiarities of each system were observed including the level of reactivity and also the number and type of key residues in the continuous epitopes of TsNTxP. In addition, in vitro neutralization assays indicated that sheep are an alternative and efficient model for the production of anti-Tityus serrulatus venom.

Use of cefoperazone MacConkey agar for selective isolation of Laribacter hongkongensis.

A new selective medium, cefoperazone MacConkey agar (CMA), was developed for primary isolation of Laribacter hongkongensis from stool. Its performance in quantitative recovery and in a clinical evaluation of 4,741 human diarrheal stool specimens was superior to that of charcoal cefoperazone deoxycholate agar. In addition, with CMA, Arcobacter butzleri was unexpectedly isolated from the stools of six patients.

Determination of the neutralizing potency of horse antibothropic and anticrotalic antivenoms in blood samples collected on filter paper.

The correlation coefficients between in vivo neutralization of lethal toxicity (ED(50)) and levels of antibodies measured by enzyme-linked immunosorbent assay (ELISA) in blood samples collected on filter paper were investigated to test the potency of horse antibothropic and anticrotalic antivenoms. Sixteen horses were hyperimmunized with Bothrops venom (50% from B. jararaca and 12.5% each from B. alternatus, B. jararacussu, B. neuwiedii and B. moojeni) and 12 horses with Crotalus durissus terrificus venom. Crude venom of C. d. terrificus and the lethal fraction of B. jararaca venom were used as antigens to set up an indirect ELISA. The correlation coefficient between ED(50) and ELISA antibodies titers against C. d. terrificus and the lethal fraction of B. jararaca venom was r = 0.8 (P<0.001) and r = 0.78 (P<0.001), respectively.

Sandwich-ELISA detection of venom antigens in envenoming by Phoneutria nigriventer spider.

Enzyme linked immunosorbent assays (ELISA) were developed to detect antigens from Phoneutria nigriventer spider venom. Horse anti-P. nigriventer immunoglobulins were prepared by immunoaffinity chromatography and used to set up a sandwich-type ELISA. The specificity of the assay was demonstrated by its capacity to correctly discriminate between the circulating antigens in mice that were experimentally inoculated with P. nigriventer venom from those in mice inoculated with Lycosa sp. and Loxosceles intermedia spider venoms, Tityus serrulatus scorpion venom and Apis mellifera bee venom. Measurable absorbance signals were obtained with 0.8ng of venom per assay. The ELISA was used to follow the kinetic distribution of antigens in experimentally envenomed mice and to detect antigens in the sera of patients envenomed by P. nigriventer.

Neutralizing potency of horse antibothropic Brazilian antivenom against Bothrops snake venoms from the Amazonian rain forest.

Neutralization of lethal toxicity (50% effective dose; ED(50)), hemorrhagic (minimum hemorrhagic dose; MHD) and hemolytic activity (PLA(2)) and levels of antibodies, measured by enzyme-linked immunosorbent assay (ELISA), were investigated to test the potency of horse antibothropic serum (ABS) against Bothrops venoms from the Amazonian rain forest. ABS neutralized the lethal activity with a potency (mg of venom neutralized per 1 ml of antivenom) of 5.5, 3.7, 1.6, 1.3 and 6.5, respectively, for B. jararaca (reference venom for assessing the ABS potency in Brazil), B. atrox, B. brazili, B. bilineatus smaragdinus and B. taeniatus venoms. The volume of antivenom (microl) that neutralized one MHD of B. jararaca, B. atrox, B. brazili, B. bilineatus smaragdinus and B. taeniatus venoms was 5, 7.71, 7.76, 8.3 and 5, respectively. ABS neutralized the PLA(2) activity with a potency of 6.2, 3.2, 1.4, 2.6 and 5 respectively, for B. jararaca, B. atrox, B. brazili, B. bilineatus smaragdinus and B. taeniatus venoms. ELISA reactivity of ABS against the separate venoms was found to be quite variable. The reactivity against B. jararaca venom was higher than against other Bothrops venoms. In conclusion, the assays described here suggest that Brazilian Bothrops polyspecific antivenom is not very efficient in neutralizing the effects of venom from some Amazonian Bothrops species.

Neutralizing potency of horse antibothropic antivenom. Correlation between in vivo and in vitro methods.

The correlation coefficients between in vivo neutralization of lethal toxicity (ED50), neutralization of the hemolytic activity (PLA2) and levels of antibodies measured by ELISA, was investigated to test the potency of horse anti-bothropic antivenom. Twenty six horses were hyperimmunized with Bothrops venoms (B. alternatus, B. jararaca, B. jararacussu, B. neuwiedii and B. moojeni). To set up an indirect ELISA, for neutralization of PLA2 activity and for determination of ED50 in Swiss mice, the whole Bothrops jararaca venom (reference venom for assessing the bothropic antivenom potency in Brazil) was used. The toxic fraction (purified from B. jararaca venom by Sephadex G-100 chromatography) was also used as antigen for ELISA. All antivenoms analyzed effectively neutralized the lethal activity in the range of 1.6 to 9.6 mg/ml of antivenom. The correlation coefficient between ED50 and ELISA antibody titers against the crude venom and toxic fraction was r = 0.65 (P < 0.001) and r = 0.85 (P < 0.0001), respectively. Correlation between ED50 and neutralization of PLA2 activity was r = 0.52 (P < 0.01), and the correlation between ELISA antibody titers and neutralization of PLA2 activity was r = 0.58 (P < 0.002). Thus, the ELISA which measures only the antibody against the major toxic fraction of the B. jararaca venom should be most suitable for use as an in vitro assay of bothropic antivenom potency.