Identification of Helicobacter pylori infection in symptomatic patients in Surabaya, Indonesia, using five diagnostic tests.

SUMMARY The prevalence of Helicobacter pylori infection in Indonesia is controversial. We examined the H. pylori infection rate in 78 patients in a hospital in Surabaya using five different tests, including culture, histology, immunohistochemistry, rapid urease test, and urine antibody test. Furthermore, we analysed virulence factors in H. pylori strains from Indonesia. The H. pylori infection rate was only 11.5% in all patients studied, and 2.3% of Javanese patients and 18.0% of Chinese patients were infected (P = 0.01). Although severe gastritis was not observed, activity and inflammation were significantly higher in patients positive for H. pylori than in patients negative for H. pylori. Among genotypes identified from five isolated strains, cagA was found in four; two were vacA s1m1. All cagA-positive strains were oipA 'on' and iceA1 positive. We confirmed both a low H. pylori infection rate and a low prevalence of precancerous lesions in dyspeptic patients in a Surabaya hospital, which may contribute to the low incidence of gastric cancer in Indonesia.

Double plating technique for fixing tibial plateau leveling osteotomy and modified cranial closing wedge ostectomy of the tibia in a dog with cranial cruciate ligament disease and excessive plateau angle: case report / [Técnica de placa dupla para fixação concomitante da osteotomia de nivelamento do platô tibial e ostectomia modificada em cunha de fechamento cranial da tíbia em um cão com doença do ligamento cruzado cranial e ângulo de platô tibial excessivo: relato de caso]

Tibial plateau leveling osteotomy (TPLO) associated to cranial wedge closing ostectomy (CCWO) has been one of the best options to manage cranial cruciate ligament (CCL) disease and excessive tibial plateau angle (TPA) in large dogs, however, the complication rate is potentially high. It is believed that a more robust fixation is necessary to stabilize them and decrease the risk of implant failure. A 6-year-old male American Pit Bull, weighing 36kg, with 90-day history of right hind limb lameness, was diagnosed with CCL disease. Due to the excessive tibial plateau angle (42°), TPLO was associated with a modified CCWO using a double plating technique. A final TPA of 12° was accomplished, and a restricted level of exercises and physiotherapy were recommended. The patient was followed monthly until the fifth month postoperatively, when radiographic bone consolidation and no lameness were observed. By the date of this submission, 3 years after the procedure, the owner has reported no complications. The double plating technique for fixing TPLO and modified CCWO proved to be effective for the treatment of CrCL deficiency in a large dog with an excessive TPA.(AU)

A osteotomia de nivelamento do platô tibial (TPLO) associada à ostectomia modificada em cunha de fechamento cranial da tíbia (CCWO) tem sido uma das melhores opções para tratamento de cães grandes com doença do ligamento cruzado cranial (DLCCr) e ângulo excessivo do platô tibial, mas o índice de complicações é alto. Acredita-se haver necessidade de fixação mais robusta para reduzir as chances de falha nos implantes. Um cão macho, seis anos, da raça American Pit Bull, 36kg, com histórico de claudicação em membro pélvico direito há 90 dias, foi diagnosticado com DLCCr. Devido ao ângulo excessivo do platô tibial (42°), a osteotomia de nivelamento do platô tibial foi associada à ostectomia modificada em cunha de fechamento cranial da tíbia (CCWO) por meio da técnica de placa dupla. No pós-operatório imediato, identificou-se TPA de 12°; exercícios controlados e fisioterapia foram recomendados e a evolução do quadro foi analisada mensalmente até o quinto mês pós-cirurgia. Com 150 dias de evolução, não houve alterações de locomoção e havia ocorrido completa consolidação radiográfica das osteotomias. Até o momento da submissão deste artigo, três anos pós-procedimento, o tutor relata ausência de complicações, via contato telefônico. Portanto, a técnica modificada mostrou-se eficaz no tratamento da DLCCr.(AU)

Comparison of radiographic and tomographic evaluations for measurement of the Canal Flare Index in dogs / [Comparação entre avaliação radiográfica e tomográfica para mensuração do Canal Flare Index em cães]

The outcome of total hip arthroplasty (THA) in dogs is directly related to surgical planning. Templating of radiographs prior to THA should help the surgeon anticipate prosthesis size and femoral shape allowing canal fill of the proximal metaphysis by the implant ensuring primary stable fixation. The canal flare index (CFI) obtained from radiograph has been used as a measure of risk of complications for the technique in human beings and dogs. However, standard radiographs only provide limited data for the selection of cementless prostheses and the assessment of their fit within the femoral canal, due to factors like radiographic magnification and femoral rotation. Therefore, three-dimensional evaluation based on computed tomography (CT) may be a better tool for CFI measurement. The aim of this study was to compare anatomical measurement with CFI values obtained from craniocaudal radiography and CT. Craniocaudal radiographs using a horizontal radiographic beam (CR), CT, and anatomical macroscopic measurements (A) were obtained from 45 femurs from 23 canine cadavers. The differences between the values of CFI obtained from radiograph (CFI-R), computed tomography on transverse (CFI- TT) and longitudinal axis (CFI-TL) compared to the CFI obtained from macroscopic measurements - gold standard - (CFI-A), and 95% limits of agreement (LOA) between the values, were evaluated by the Bland-Altman method. Dimensions obtained from CT techniques had a greatest mean difference from anatomical and CFI values were also different (P=0.032). Under the experimental conditions, the craniocaudal radiograph, provided the most accurate measurement of the CFI (mean difference: 0.087 ± 0.42).(AU)

O resultado da artroplastia total do quadril (ATQ) em cães está diretamente relacionado ao planejamento cirúrgico. O templating radiográfico pré-operatório da ATQ deve ajudar o cirurgião a prever o tamanho da prótese e o formato do fêmur, o que permitirá um preenchimento ideal da metáfise proximal pelo implante, garantindo, assim, fixação primária estável. O índice de alargamento do canal (Canal Flare Index - CFI) obtido em radiografias tem sido utilizado como fator de risco de complicações para a técnica em humanos e cães. No entanto, as radiografias podem fornecer apenas dados limitados para a seleção de próteses não cimentadas e a avaliação do seu encaixe no canal femoral, devido a fatores como ampliação radiográfica e rotação femoral. Portanto, a avaliação tridimensional baseada na tomografia computadorizada (TC) pode ser uma ferramenta vantajosa para a mensuração do CFI. O objetivo deste estudo foi comparar a medida anatômica com os valores de CFI obtidos na radiografia craniocaudal e na TC. Radiografias craniocaudais utilizando feixe radiográfico horizontal (CR), tomografia computadorizada e medidas macroscópicas anatômicas (A) foram obtidas de 45 fêmures de 23 cadáveres caninos. As diferenças entre os valores de CFI obtidos na radiografia (CFI-R), na tomografia computadorizada no eixo transversal (CFI-TT) e no eixo longitudinal (CFI-TL), em comparação com os valores de CFI obtidos nas medições macroscópicas - padrão-ouro - (CFI-A) e os limites de concordância de 95% (LOA) entre os valores, foram avaliadas pelo método de Bland-Altman. As dimensões obtidas pelas técnicas de TC apresentaram maior diferença média dos valores anatômicos, e as do CFI também foram diferentes (P=0,032). Nas condições experimentais, a radiografia craniocaudal forneceu a medida mais precisa do CFI (diferença média: 0,087 ± 0,42) para representar o padrão-ouro deste estudo.(AU)

Biocompatibilidade do gel de quitosana associado ao glicerol fosfato na reparação de defeitos ósseos induzidos experimentalmente no rádio de coelhos (Oryctolagus cuniculus) / [Biocompatibility of chitosan gel associated with glycerol phosphate for the healing of experimentally induced radial defects in rabbits (Oryctolagus cuniculus)]

A engenharia de tecidos caracteriza-se como ciência interdisciplinar, a qual vem desenvolvendo biomateriais para a regeneração do tecido ósseo no âmbito das medicinas humana e veterinária. O objetivo desta pesquisa foi avaliar a regeneração óssea obtida da aplicação do hidrogel de quitosana associado ao glicerol fosfato em falha óssea experimentalmente induzida no rádio de coelhos. Foram utilizados 15 coelhos adultos, distribuídos aleatoriamente em dois grupos, representados por cada um dos rádios de cada animal, sendo um grupo tratado com hidrogel de quitosana associado ao glicerol fosfato (grupo biomaterial - GB) e um grupo que não recebeu tratamento com o biomaterial (grupo controle - GC). Os animais foram avaliados radiograficamente, por densitometria óptica e análise histológica, nos períodos 30, 60 e 90 dias pós-operatórios. Houve superioridade estatística na média geral das avaliações radiográficas do GB (2,33±0,48) sobre o GC (1,77±0,06). As médias gerais de avaliação densitométrica do GB foram superiores às do GC, sendo 6,207±1,374 e 5,71±1,512, respectivamente. A avaliação histopatológica do GB foi superior à do GC nos períodos de 30, 60 e 90 dias. Assim, é possível afirmar que o hidrogel de quitosana constitui biomaterial de características desejáveis, promovendo consolidação óssea mais rápida e eficiente, sem causar reações adversas.(AU)

Tissue engineering is an interdisciplinary science that has been developing biomaterials for bone regeneration in medicine and veterinary medicine, following an imminent need. The aim of this study was to evaluate bone regeneration after use of chitosan hydrogel associated with glycerol phosphate in experimentally induced bone gap in the radius of rabbits. Fifteen adult rabbits were randomly distributed in two experimental groups, represented by each radius of every single animal. The animals in the Biomaterial Group (GB) were treated with a glycerol phosphate-associated chitosan hydrogel and in the Control Group (GC) they received no treatment with the biomaterial. The animals were evaluated clinically, radiographically, histologically and by optic densitometry at 30, 60 and 90 days postoperatively. There was statistical superiority in the general average of the radiographic estimates of GB (2.33 ± 0.48) over the CG (1.77 ± 0.06). The general averages of GB densitometric evaluation were higher than the CG, being 6.207 ± 1.374 and 5.71 ± 1.512, respectively. Histopathological evaluation of GB was superior to CG in periods of 30, 60 and 90 days. Chitosan hydrogel constitutes a biomaterial of desired characteristics, promoting faster and more efficient bone repair when compared to GC.(AU)

The roles of Ca2+/calmodulin- and cGMP-dependent pathways in gametogenesis of a rodent malaria parasite, Plasmodium berghei.

The induction mechanism of gamete formation (gametogenesis) in a rodent malaria parasite, Plasmodium berghei, was investigated using Ca2+ antagonists, protein kinase inhibitors and amiloride, an inhibitor of monovalent cation/H+ exchange. Treatment with 3,4,5-trimethoxybenzoic acid 8-(diethylamino)octyl ester (TMB-8, a Ca2+ release inhibitor) and W-7/W-66 (calmodulin inhibitors) blocked formation of male gametes by inhibiting DNA synthesis from 1.5C to 8C level. In contrast, inhibitors of cAMP/cGMP-dependent protein kinases such as H-8, H-87, H-89 and staurosporine also ceased the development of gametocytes, but DNA synthesis in male gametocytes occurred as in the controls. Electron microscopy revealed that male gametocytes treated with TMB-8 and W-7 failed to enlarge nuclei and to form axonemes in the cytoplasm. In female gametocytes, treatment with both Ca2+ antagonists resulted in a dramatic morphological change in the endoplasmic reticulum (ER), which is thought to be a Ca2+ store. The ER network condensed near nuclei and was laminated by the abnormal attachment of ribosomes between two ER membranes. On the other hand, male gametocytes treated with protein kinase inhibitors or amiloride had enlarged nuclei and axonemes, but failed to develop further. The ER network in female gametocytes treated with these inhibitors was similar to that in the controls.

Possible roles of Ca2+ and cGMP as mediators of the exflagellation of Plasmodium berghei and Plasmodium falciparum.

The roles of Ca2+ and cyclic nucleotides as secondary, intracellular messengers for exflagellation of Plasmodium berghei and Plasmodium falciparum were investigated. Treatment with Ca2+ antagonists such as TMB-8 (an inhibitor of intracellular Ca2+ release) or W-7 (a calmodulin inhibitor) strongly inhibited exflagellation induced by alkaline medium at pH 8.0 whereas EGTA (a Ca2+ chelator) or nicardipine and nifedipine (Ca2+ channel inhibitors) had no effect. These results may indicate that mobilization of parasites' internal resources of Ca2+ is a prerequisite for exflagellation. Agents which increase cAMP levels did not induce exflagellation at the non-permissive pH of 7.3, and had no significant inhibitory effect at the permissive pH of 8.0. IBMX (cAMP/cGMP-phosphodiesterase inhibitor), however, enhanced exflagellation at pH 7.3, indicating the possibility that cGMP, but not cAMP, may be involved in the induction of exflagellation. Furthermore, cGMP or agents which increase cGMP levels such as nitroprusside (a potent activator of guanylate cyclase), enhanced exflagellation at pH 7.3, whereas N-methyl-hydroxylamine (guanylate cyclase inhibitor) inhibited the exflagellation at pH 8.0. From these results, it may be concluded that the induction of exflagellation requires both Ca2+ mobilization and an increase in cGMP levels.

Allelic diversity at the merozoite surface protein-1 locus of Plasmodium falciparum in clinical isolates from the southwestern Brazilian Amazon.

Nucleotide sequences of each variable block in the Plasmodium falciparum merozoite surface protein-1 gene (PfMSP-1) may be grouped into one of two or three possible allelic types, named after the reference isolates MAD20, K1, and RO33. Allelic diversity at this locus basically results from different combinations of allelic types in variable blocks. We used a polymerase chain reaction (PCR)-based strategy to type the variable blocks 2, 4a, 4b, and 10 of the PfMSP-1 gene of P. falciparum isolates from 54 symptomatic malaria patients living in Rondonia, a hypoendemic area in the southwestern Brazilian Amazon. Ten different PfMSP-1 gene types, defined as unique combinations of allelic types in variable blocks, were identified among the 54 isolates. Twenty-one isolates (39%) harbored more than one gene type and two had at least three genetically distinct clones. Hybrid sequences, with a MAD20-type sequence in the 5' segment (4a) and a K1-type sequence in the 3' segment (4b), were quite common in block 4. Direct sequencing of block 4 PCR products revealed a new putative recombination site in four isolates. In contrast with previous studies, the observed distribution of gene types does not deviate significantly from that expected under the null hypothesis of random association between allelic types detected in each variable block. These contradictory data are discussed with reference to the immunoepidemiologic features prevailing in distinct malaria-endemic areas.

In vitro susceptibility of Plasmodium falciparum isolates from Myanmar to antimalarial drugs.

In vitro drug susceptibility profiles were assessed in 75 Plasmodium falciparum isolates from 4 sites in Myanmar. Except at Mawlamyine, the site closest to the Thai border, prevalence and degree of resistance to mefloquine were lower among the Myanmar isolates as compared with those from Thailand. Geometric mean concentration that inhibits 50% (IC50) and 90% (IC90) of Mawlamyine isolates were 51 nM (95% confidence interval [CI], 40-65) and 124 nM (95% CI, 104-149), respectively. At the nearest Thai site, Maesod, known for high-level multidrug resistance, the corresponding values for mefloquine IC50 and IC90 were 92 nM (95% CI, 71-121) and 172 nM (95% CI, 140-211). Mefloquine susceptibility of P. falciparum in Myanmar, except for Mawlamyine, was consistent with clinical-parasitological efficacy in semi-immune people. High sensitivity to artemisinin compounds was observed in this geographical region. The data suggest that highly mefloquine-resistant P. falciparum is concentrated in a part of the Thai-Myanmar border region.

In vitro susceptibility of Plasmodium falciparum isolates in Vietnam to artemisinin derivatives and other antimalarials.

We performed in vitro drug susceptibility assays of ten Plasmodium falciparum isolates collected from Vict Nam in 1995. All isolates were found to be highly sensitive to artesunate, dihydro-artemisinin and artemisinin. They were also sensitive to quinine. All of them were resistant to chloroquine and mefloquine in vitro. This study provides the baseline estimates of in vitro susceptibility levels of the Vietnamese isolates to artermisinin and their derivatives because the data were collected soon after these drugs were introduced countrywide. It also describes some quantitative profiles of the in vitro response of other standard antimalarial drugs in Viet Nam, which is presently limited.

A comparative study of the spicule venom of Euproctis caterpillars.

Caterpillar spicule venoms were extracted and studied for the following activities: arginine ester (BAEE) hydrolase, tyrosine ester (ATEE) hydrolase, protease (casein digestion) and phospholipase A (indirect hemolytic activity). Crude spicule venom of E. chrysorrhoea preferably hydrolyzed BAEE in contrast to E. subflava venom, which hydrolyzed ATEE in preference to BAEE. This difference was confirmed by Sephadex G-100 elution profiles. The esterase activity in E. chrysorrhoea venom was separated into two peaks with average mol. wts. of 96,000 and 44,000. The first peak demonstrated optimal BAEE hydrolysis at pH 8.6 and 37 degrees C, whereas the second peak optimally hydrolyzed both BAEE and ATEE at pH 8.45 and pH 8.6 at 45 degrees C respectively. The esterase activity in E. subflava venom was separated into two peaks with average mol. wts of 63,000 and 32,000 showing optimal hydrolysis of BAEE at pH 8.9 and 37 degrees C, and of ATEE at pH 7.75 and pH 8.5 at 40 degrees C. The column fractions showed comparable proteolytic activity, irrespective of differences between their esterase activities. The presence of phospholipase A (PLA) enzyme in crude spicule venom of both species was evident from their indirect hemolytic activities. The PLA activity eluted with the void volume and seems to be associated with some high molecular weight protein. Under the assay conditions used, E. subflava venom contained 50-100 times less PLA activity than E. chrysorrhoea venom.

Protease activities in the spicule venom of Euproctis caterpillars.

The spicule venoms of Euproctis chrysorrhoea and Euproctis subflava were investigated for their capacity to hydrolyze chromogenic tripeptide substrates with selective affinities for various serine proteases. Seven substrates were assayed with affinities for trypsin and thrombin, trypsin and urokinase, serine proteases, chymotrypsin, glandular kallikrein, plasma kallikrein and plasmin. Venom material has a broad spectrum of affinities for the substrates with relative high plasma kallikrein activities. In E. chrysorrhoea venom, trypsin-like activities predominated, whereas E. subflava venom hydrolyzed, in preference, substrates with an affinity for chymotrypsin. The venoms were fractionated on Sephadex G-100, leading to three fractions, all having serine protease activity. The ratios of substrate specificities were markedly different, indicating that in both caterpillar venom preparations at least two separate serine proteases are present. In addition, in human plasma, inhibitor activity could be detected to the kallikrein activity of E. chrysorrhoea, but not of E. subflava. The trypsin-like activity was not inhibited by human plasma. These and earlier studies warrant the assumption that serine proteases, particularly kallikrein, are major factors in the elicitation of clinical symptoms observed after contact with caterpillar spicules.

Cyclical transmission of Plasmodium berghei (Coccidiida: plasmodiidae) by Anopheles omorii (Diptera: Culicidae).

Anopheles omorii, a tree-hole breeding anopheline collected in Japan, transmitted a rodent malaria, Plasmodium berghei ANKA strain, in the laboratory. Mice with 0.2-15% parasitemia, 0.01-1.5% gametocytemia, and 0.001-0.5% exflagellations were used as infective hosts. Oocyst numbers ranged from 2 to 171 (mean 44) on the midgut 8-16 d after the blood meal. Several to hundreds of sporozoites were detected in the salivary glands 14 d after feeding. The mosquitoes were infective to mice from 13 to 40 d after feeding. An. omorii occurs naturally at temperatures less than 25 degrees C and therefore is a suitable laboratory vector for the cyclic transmission of P. berghei, a malaria parasite that completes sporogony at 18-21 degrees C.

Allelic diversity in the merozoite surface protein-1 and epidemiology of multiple-clone Plasmodium falciparum infections in northern Tanzania.

Allelic diversity in the merozoite surface protein-1 (MSP-1) of Plasmodium falciparum, a major malaria vaccine candidate, was examined in clinical isolates from holoendemic northern Tanzania. The variable blocks 2, 4a, 4b, 6, and 10 of the MSP-1 gene were typed by allelic type-specific polymerase chain reaction. Twenty-four possible MSP-1 gene types were defined as unique combinations of allelic types detected in each variable block. Thirteen gene types were identified, and 187 P. falciparum populations were fully typed among 79 isolates. In contrast with recent findings in Vietnam, we were unable to detect nonrandom associations between allelic types in the typed variable blocks. Most patients (60%) harbored more than 1 genetically distinct parasite population (average: 2.37 populations per isolate) and, in 1 patient, 6 different versions of this single-copy gene were found. Statistical analysis suggests that parasites carrying different MSP-1 gene types are not independently distributed in the host population. The epidemiological consequences of these findings are discussed.

Sequence diversity of serine repeat antigen gene exon II of Plasmodium falciparum in worldwide collected wild isolates.

Field isolates of Plasmodium falciparum collected from endemic areas of Southeast Asia, Solomon Islands, tropical African countries and Brazil were analyzed for the genetic diversity of the exon II of serine repeat antigen gene (SERA) by sequencing of genomic DNA. Of sixty-nine isolates, as compared to the reported FCR3, K1 and Honduras-1 types of exon II sequences, 5, 9 and 20 new allelic forms were found in 23 isolates of the FCR3 type, 36 of the K1 type and 10 of the Honduras-1 type. A group of novel non-synonymous substitutions, 4 new insertions and 3 new deletions of octamer units were found in the octamer repeat region (OR) of the exon II, and most of them clustered within a 40-residues domain. An octamer "SNPVSSEP" revealed in the OR was confirmed as a new repeat unit. Based on the sequences of the serine repeat region (SR) of the exon II, the allelic forms of the Honduras-1 type were conjectured to be the recombinant forms between the K1 type and FCR3 type. The allelic forms of K1 type with less or more repeat serine residues in the serine stretch of the SR than the reported 21 serine residues had most of the variations in the OR. Moreover, a biased geographical distribution of allelic forms was observed. Isolates from African and Southeast Asian countries accounted for most of the new allelic forms (29/33). All of the three types were detected in Southeast Asia but none of the FCR3 type in Africa. One of two groups of FCR3 new allelic forms was found solely in Brazil while another was mainly in Solomon Islands.

How prevalent is Plasmodium malariae in Rondônia, western Brazilian Amazon?

We have compared results of Plasmodium species identification obtained with conventional on-site microscopy of Giemsa-stained thick smears (GTS) and a semi-nested polymerase chain reaction (PCR) in 96 malaria patients from Rondônia, Western Brazilian Amazon. Mixed-species infections were detected by PCR in 30% patients, but no such case had been found on GTS. Moreover, P. malariae infections were detected in 9 of 96 patients (10%) by PCR, but were not identified by local microscopists. The potential impact of species misidentification on malaria treatment and control is discussed.

[A case of uveitis due to gnathostoma migration into the vitreous cavity].

We report a 26 year-old male patient who had floaters and hyperemia in his left eye following uveitis due to gnathostoma that had migrated into the vitreous cavity. Severe iridocyclitis and mild opacity of the vitreous body were observed, together with whitish-yellow subretinal tracks accompanied by dot and blot hemorrhages in the fundus. Slit lamp microscopic examination revealed a worm which writhed in the vitreous cavity. We performed vitrectomy to remove the worm from the anterior vitreous uneventfully, followed by prompt subsidence of the inflammatory signs. The worm was identified as a third instar larva of Gnathostoma doloresi. Eosinophilia and creeping eruption did not appear throughout the follow-up period. The patient was accustomed to eat live roaches and whitebait, as well as sliced raw beef liver.