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Alkaline phosphatase (AP) is a membrane bound enzyme and when it is elevated in blood, it is mostly due to either hepatobiliary or bone diseases. We report isolated intestinal hyperphosphatasemia (IAP) in two sisters. Both sisters presented with identical trends of isolated AP elevation. Both underwent extensive workup for liver diseases including cholangiograms, and none was identified. Subsequent isoenzyme electrophoresis showed that 45%-56% of the elevated AP was due to IAP. This elevation of the intestinal AP is consistent with a rare hereditary biochemical abnormality, benign familial intestinal hyperphosphatemia. This condition should be considered in the differential diagnosis of otherwise isolated serum AP levels to avoid unnecessary investigations.

The isoenzyme creatine kinase muscle/brain (CK-MB) still plays an important role for the differential diagnosis of CK elevations and the clarification of their origin from heart or skeletal muscle. Therefore, it is necessary to know the diagnostic pitfalls in interpreting CK-MB results. We demonstrate a case of macro-CK type 2 in a 75-year-old patient with metastatic castration-resistant prostate cancer and its identification by isoenzyme electrophoresis, which can be typical for cancer diseases.

Species of the genus Meloidogyne are limiting factors in vegetable crop production. Studies have been conducted in Brazil on the occurrence of root-knot nematodes in growing vegetable crop areas without a reliable information on the etiology of root-knot nematode infestation, based on advanced identification procedures. Using modern techniques such as biochemical and molecular methods is possible to improve the accuracy of identifying species of Meloidogyne . This study characterized species of Meloidogyne in a total of 36 samples associated with vegetable crops by using isoenzyme electrophoresis, SCAR markers, and morphological markers, in addition to validation of SCAR markers for accurate identification of these species. The species M. incognita , M. javanica , M. hapla, M. morocciensis, and M. arenaria were identified, with the first two being the most frequent. The species M. arenaria parasitizing garden eggplant and M. morocciensis parasitizing pumpkin and cabbage were reported in Brazil for the first time. Esterase electrophoresis efficiently separated the species of Meloidogyne found in vegetable crops. However, SCAR markers were effective for the identification of only M. incognita , M. javanica, and M. hapla, as the primer pair Far/Rar did not yield any amplification to confirm the identity of M. arenaria . The species M. arenaria and M. morocciensis could not be distinguished by the female perineal patterns. Based on these results, new primers should be designed to identify M. arenaria and M. morocciensis .

Five marine teleost fishes inhabiting different marine depths namely Hippocampus hippocampus (Linnaeus, 1758), Sardina pilchardus (Walbaum, 1792), Gobius niger (Linnaeus, 1758), Mullus barbatus barbatus (Linnaeus, 1758) and Solea solea (Linnaeus, 1758) were used in the present study. Their retinae and lenses were subjected for histological, scanning electron microscopy, SDS-PAGE and isoenzyme electrophoresis of alkaline phosphatase, malate and glucose-6-phosphate dehydrogenase. The present findings showed variant histological structures with characteristic photoreceptors mainly of either rods for H. hippocampus , M. barbatus and S. solea or cones in S. pilchardus . Mixed photoreceptors are identified in G. niger . The fishes exhibited diversity in protein band expression coincides with change of pattern orientation in lens fibers arrangement and histological structures of retina. Isoenzyme electrophoresis of estimated isoenzyme showed differences between lens and retina of fishes especially H. hippocampus . It can be concluded that the retina and lens of the studied teleost fishes showed apparent varying structure reflecting the isoenzyme characteristic for preserving functional characteristic of vision according to the marine habitat depths.

Analysis of variation in the four enzymatic systems of three populations of Anthyllis vulneraria was made. Different polymorphism of enzyme .protein® in six terms of plant development was found by isoenzyme electrophoresis on polyacrylamide gel. Each population had a specific isoenzyme pattern and specific variability in the terms.

Background Macrocreatine kinase (MCK) type 1 is a high molecular weight form of CK that is non-pathological. Identification of MCK is beneficial in preventing unnecessary investigations that may follow a persistently elevated CK of unknown origin. Currently, gel filtration chromatography can be used as a confirmatory technique, but it is laborious, time-consuming and expensive. The aim of this work, carried out as part of a larger investigation into the prevalence of MCK, was to determine whether trypsin can be used as an alternative to confirm the presence of MCK on isoenzyme electrophoresis. Methods Five samples found to have bands running in the MCK region on isoenzyme electrophoresis were treated with trypsin. Electrophoresis was carried out using the Helena Biosciences Sas-1 Plus system. These samples were also analysed by the confirmatory technique of gel filtration chromatography. Results Of the five samples treated with trypsin, three were found to be MCK-positive and two MCK-negative. These results correlated with those obtained by the reference method of gel filtration chromatography. Conclusions There appears to be a potential use of trypsin in confirming the presence of MCK following isoenzyme electrophoresis. If these findings were verified, this would provide a less labour-intensive, less time-consuming and more cost-effective confirmatory technique. Further study is required, which needs to be expanded to include a larger number of patients, before this method can be adopted routinely.

A total of 50 potato (Solanum tuberosum L.) varieties were maintained in vitro or under field conditions at Keshan National Genebank (KNG) for 15 years. At the end of the conservation period, a two-year field trial was conducted to evaluate the effect of conservation methods on the genetic stability of accessions, using agronomic characters, isoenzyme markers, and DNA molecular markers. None of the 50 varieties maintained under field conditions had completely preserved agronomic characters. Plants kept in vitro demonstrated still lower level of stability than those conserved under field conditions. Differences in the isoenzyme electrophoresis patterns were found in ten of the varieties tested. The SSR (simple sequence repeat) analysis indicated genetic variation between each variety maintained in vitro and under field conditions and showed the genetic similarity coefficient ranged from 0.735 to 0.993. Three of the accessions maintained in vitro and under field conditions did not cluster together. The results suggest that the genetic stability of varieties maintained under field conditions was higher than those maintained in vitro. Therefore, in order to maintain the genetic integrity of potato germplasm resources, field conservation should be used for long-term conservation of potato germplasm collections, and the genetic stability should be checked regularly.

Isoenzyme electrophoresis was employed to examine the relationships of 21 individuals representing four populations of Astragalus gossypinus complex as well as 20 individuals representing five populations of Astragalus persicus complex. A total of 27 bands from three enzyme systems (superoxide dismutase, esterase, and peroxidase) were obtained. UPGMA clustering method resulted in two distinct clusters for different populations corresponding to the two species complexes analysed. In both species, populations distributed on Alborz mountain range in northern Iran form separated clusters from those distributed on Zagros mountain range in the West. The results also show that both species complexes exhibit a high diversity based on Shannon–Weaver diversity index ( H ′) compared with other species of Astragalus reported previously. The mean number of bands per each presumed isoenzyme ranges from 2.14 to 3.57. The value of Euclidean distance ranges from 1.251 to 3.152. Our data suggest that both species should be circumscribed wider than that treated by most taxonomists, and several taxonomic names should be reduced under synonymy of the corresponding species.

Background Anopheles funestus is a major vector of malaria in sub-Saharan Africa. In order to apply effective control measures against this vector, it is necessary to understand the underlying physiological factors that play a critical role in its development, reproduction, fertility and susceptibility to insecticides. One enzyme family involved in the above mentioned biological pathways is the esterases. The aim of this study was to analyse esterase activity levels at different ages during the life-span of adult Anopheles funestus Giles in order to better understand the complex biological processes in this species. Methods Isoenzyme electrophoresis (IEE) was used to examine the esterase activity in laboratory colonised An. funestus adults aged between 2 h (h) and 30 days post eclosion as well as in wild An. funestus adults aged between 2 h and 15 days post eclosion. Esterase activity was quantified by densitometry analysis of the IEE gels. Esterases were classified according to their activity inhibition by organic phosphates, eserine sulphate and sulphydryl reagents. Results Nine esterases IEE profiles were common to both the laboratory colonised and wild An. funestus adults. These esterases were further divided into acetylesterases, arylesterases, carboxylesterases and acetylcholinesterase. The activity level of certain specific esterases was primarily influenced by age and/or gender. Conclusions The information from this study contributes towards the general understanding of esterase enzyme activity variation in adults of a major malaria vector An. funestus. This variation likely carries physiological and adaptive significance and may influence specific characteristics, such as reproductive fitness and insecticide resistance that are epidemiologically important.

A parasitological study was conducted on 66 dogs seroreactive for Leishmania captured as a control measure of visceral leishmaniasis in the State of Rio de Janeiro, Brazil. Biological samples from different anatomical sites were collected during autopsy of the animals and cultured on biphasic medium (NNN/Schneider). The Leishmania isolates were characterized by isoenzyme electrophoresis. Leishmania was isolated from 80.3% of the animals: 12 animals with Leishmania (Viannia) braziliensis isolated exclusively from cutaneous lesions, 39 with L. (L.) chagasi isolated from different sites in the same animal, and 2 with simultaneous isolation of L. (V.) braziliensis from cutaneous lesions and L. (L.) chagasi from different sites. Isolation in culture revealed the absence of Leishmania parasites in 13 animals. The results obtained confirm the existence of mixed infections in dogs in Rio de Janeiro and indicate the need to complement the investigation of seroreactive dogs using methods for the parasitological diagnosis and identification of Leishmania species.

In the present work, isoenzyme electrophoresis was used to analyze the variability and phenetic relationships among seven American species of genus Prosopis belonging to three different sections: P. argentina (Monilicarpa), P. glandulosa, P. velutina, P. flexuosa, P. ruscifolia, P. kuntzei (Algarobia), and P. reptans (Strombocarpa). The genetic variability in P. argentina, P. reptans, and P. kuntzei was significantly lower than in the rest of the species analyzed. The species belonging to different sections are highly differentiated, but the relationships retrieved among species belonging to the section Algarobia suggested that the series of this section are not natural groups. P. kuntzei is as differentiated from the remaining species of Algarobia as from P. reptans or P. argentina, suggesting that this species might be included in a different section. The series within section Algarobia are not supported by the clusters retrieved in the phenogram based on isoenzymatic data. The results suggest that the two North American species (P. velutina and P. glandulosa) would have originated in different founder events.

Background: The presence of macroenzymes can cause significant diagnostic confusion and their detection can involve relatively cumbersome analytical procedures. Methods: Using a simplified polyethylene glycol precipitation technique and isoenzyme electrophoresis, this report describes the construction of reference ranges of precipitable activity for each of seven commonly measured enzymes in plasma. Results: The proposed reference ranges are reported. Since introducing the protocol, 12 cases of macroenzymaemia have been encountered. Three typical case histories are described in some detail. Conclusions: The polyethylene glycol precipitation method has thus far proved to be a simple and effective additional test for the detection of macroenzymes when the plasma enzyme activity is elevated.

INTRODUCTION: Visceral leishmaniasis presents urban behavior in some Brazilian cities, with domestic dogs as the main infection source. In Cuiabá, MT, canine visceral leishmaniasis was diagnosed and characterized as recommended by the Ministry of Health. METHODS: Biological samples from suspected canine carriers were analyzed by the isoenzyme electrophoresis technique. The 6PGDH enzyme and reference strain IOC/L0566 (MHOM/BR/1975/M2903) of Leishmania (Leishmania) infantum was used as one of the controls. RESULTS: Electrophoresis analysis revealed that the canine isolates belonged to the species L. (L.) infantum. CONCLUSIONS: The authors emphasize the importance of species characterization, particularly in areas of mixed infection like Cuiabá.

Objectives To determine the cause of discrepancies in the alkaline phosphatase (ALP) activities of quality control (QC) materials in two different analyzers using IFCC method. Design and methods ALP activities of patients' samples and QC materials (QC1 and QC2 from Bio-Rad) measured using TBA-200FR and Synchron LX-20 analyzers were compared and isoenzyme electrophoresis was done. Fractional mixing of bone or liver ALP with placental ALP was performed. ALP activities of QC materials were measured in TBA-200FR with pH-modified reagents. Results ALP activities of QC materials were significantly lower in TBA-200FR than in LX-20. Placental ALP comprised 57% of QC1 and 95% of QC2. Higher placental ALP proportion in the mixture with bone or liver ALP resulted in lower ALP activities in TBA-200FR. The discrepancy in ALPs of QC materials decreased when measured with pH-modified reagents. Conclusion Placental ALP in QC materials and differences in reagents' pH caused the discrepancy in ALP activities.

The analysis of sixteen Komagataea (Williopsis) pratensisisolates from Caucasian and Tien Shan soils by the PCR, blot hybridization, and isoenzyme electrophoresis techniques showed that fifteen of them do belong to the species K. pratensis. The isolates from the two geographic areas differed in some physiological characteristics and in the PCR product profiles obtained with the microsatellite primers (CAC)5and (GACA)4.

Este estudio evaluó los cambios en el perfil isoenzimático y antigénico de la cepa Munantá de Trypanosoma cruzidespués de dos pases consecutivos por ratón, mediante electrofóresis isoenzimática, electrofóresis en PAGE-SDS e immunoblot. Los resultados obtenidos muestran diferencias estadísticamente significativas entre la cepa antes y después de los pases en los animales, lo cual sugiere que el ratón no es un modelo recomendable para obtener las formas tripomastigotas de la cepa Munanta del parásito destinadas a estudios de la respuesta inmune.

A fortuitously discovered case of feline leishmaniosis is reported. The parasites were found in the skin and the bone marrow of a domestic female cat that spontaneously died after a few weeks of evolution. Serological tests for FeLV, FIV and PIF virus detection gave negative results. By using Western blot serology, a characteristic pattern of leishmaniosis was obtained and by performing an isoenzyme electrophoresis, a Leishmania infantum MON-1 strain was identified. The same zymodeme is implicated in most of the canine and human leishmaniosis in Southern Europe. A study on the prevalence of asymptomatic feline leismaniosis is foreseen.

Based on isoenzyme electrophoresis, 28 reputed cultivars of the date palm ( Phoenix dactylifera L.) previously characterized by morphological characters, were identified using the more variable enzyme systems in the date palm: esterases (EST), endopeptidases (ENP) and glutamate oxaloacetate transaminase (GOT), which showed, respectively, eight, six and seven electrophoretic phenotypes in leaf material. The phenotype combinations lead to an isoenzymebased key for date palm cultivar determination. Cultivar composition of 11 date palm groves, differing in their geographical origins, were compared. Unidentified material represented about 67% (±11%) of each date palm grove with the exception of populations at M’hamid and Rissani. This result confirmed the predominance of date palms derived from germinated seeds and called locally ‘Khalts’. The most frequent cultivars within the 11 date palm groves were Jihel and Bou-Feggouss, which exhibit good fruit qualities. Based on the cultivar composition of each population, the factorial component analysis showed that the Marrakech date palm grove situated at the northern side of High Atlas, was related to date palm groves of Skoura, Ait Atman, Ksar Jdid and Ksar Sifa. Date palm populations of Zagora, M’hamid and Rissani, each represented a unique type, were characterized by cultivars Bou-Sthammi Noire, Bou-Skri (and Iklane) and Bou-Feggouss, respectively. Cultivar compositions of date palms groves, determined separately on the basis of isoenzymes phenotypes and fruit qualities, were compared and discussed.

We analyzed the clinical and epidemiological characteristics of 90 patients with Cutaneous Leishmaniasis (CL) in Rio Preto da Eva, Amazonas state, Brazil, between 2005 and 2012. All patients had skin lesions, whose isolates were characterized by isoenzyme electrophoresis and identified as Leishmania (Viannia) guyanensis (n=80), Leishmania (V.) naiffi (n=3) and Leishmania (Leishmania) amazonensis (n=7). Most patients were male (70%) and the age group with the highest number of cases was 11 to 20 years old (31.7%). In terms of occupation, most were farmers or home caretakers (37.7%). Around 95% of cases of CL were from rural areas of endemic foci and the settlement known as Ipora, located at KM 127 of the AM 010 road, had the highest number of cases (33.3%). We conclude that American Tegumentary Leishmaniasis is a parasitic disease endemic in Rio Preto da Eva (Amazonas State), occurring mainly along the 010 AM Highway, and that infection was mainly acquired through contact with the sylvatic transmission cycle, although the possibility of disease transmission peridomiciliary was also analysed.

International audience; AIM: Three forms of cutaneous leishmaniasis (CL) are endemic in Tunisia. The identification of the causative species is useful to complete epidemiological data and to manage the cases. The aim of this study is to assess PCR-RFLP technique in the identification of Leishmania species responsible of CL in Tunisia and to compare the results of this technique to those of isoenzyme analysis. PATIENTS AND METHODS: Sixty-one CL lesions were sampled. Dermal samples were tested by culture on NNN medium and analyzed by PCR-RFLP assay targeting the ITS1 region of ribosomal DNA. Species identification was performed by both iso-enzymatic typing for positive cultures and analysis of restriction profiles after enzymatic digestion by HaeIII of the obtained amplicons. RESULTS: Thirty-eight (62%) samples were positive by culture. The iso-enzymatic typing of 32 isolates identified 3 L. infantum, 23 L. major MON-25 and 6 L. tropica MON-8. Sixty samples were positive by PCR. The PCR-RFLP digestion profiles of the 56 PCR products identified 12 L. infantum, 38 L. major and 6 L. tropica. The results of both techniques were concordant in the 32 strains identified by both techniques. Species identification correlated with the geographical distribution of CL forms endemic in Tunisia. CONCLUSION: Results of PCR-RFLP revealed highly concordant with those of isoenzyme electrophoresis. Thanks to its simplicity, rapidity and ability to be performed directly on biological samples, this technique appears as an interesting alternative for the identification of Leishmania strains responsible of CL in Tunisia.

International audience; BACKGROUND: Leishmaniasis is endemic in Iran. Different species of Leishmania (L.) parasites are causative agents of this disease. Correct identification of Leishmania species is important for clinical studies, prevention, and control of the diseases. Mix up of Leishmania isolates is possible in the laboratory, so there is need for verification of species for isolates of uncertain identity. Different methods may be used for this purpose including isoenzyme electrophoresis and molecular methods. The isoenzyme electrophoresis, due to its drawbacks, is feasible only in specialized laboratories while molecular methods may be more feasible. The aim of this research was to study the application of the internal transcribed spacer 1 (ITS1) sequencing method, in comparison to isoenzyme electrophoresis method, for verification of Leishmania species. METHODS: Six Leishmania isolates were received from different research institutions in Iran. The species of these isolates were known by donating institution according to their isoenzyme profile. The species of these isolates were re-identified in Pasteur Institute of Iran by PCR amplification of ITS1 followed by sequencing and comparison of these sequences with Leishmania sequences in GenBank. Isoenzyme electrophoresis was performed for confirmation of the results of ITS1. RESULTS: ITS1 sequence showed that some isolates were mixed up or contaminated with Crithidia. Isoenzyme electrophoresis confirmed the results of ITS1 sequences. CONCLUSION: ITS1 sequencing is relatively more feasible than the traditional isoenzyme electrophoresis method and is suggested for verification of Leishmania species.

OBJECTIVE To discuss the problem about the origin of Oviduetus Ranae in the Chinese Pharmacopoeia according to historical documents, the researches reported recently and the author research. METHOD Through comprehensive analysis of the documents and materials reported, the original animal sources of Oviduetus Ranae was discussed in terms of historical records, morphology, karyotype, Ag-Belt and isoenzyme electrophoresis, gene levels and so on. RESULT AND CONCLUSION The original animal sources of Oviduetus Ranae is Rana dybowskii,its order element is an effective species in China. In order to avoid the problem of species confusion about the origin of Oviduetus Ranae, author suggests that R. dybowskii should be the original animal of Oviduetus Ranae.

Fourteen Giardia duodenalis isolates were examined using the RAPD (random amplified polymorphic deoxyribonucleic acid) technique. Simple reproducible polymorphisms were generated using 3 different RAPD primers. The results generated by each primer were very similar and were significantly correlated with each other. These data were then compared to existing isoenzyme electrophoresis data on the same isolates. The RAPD data divided the isolates into 10 groupings or rapdemes while the isoenzyme data divided them into 10 similar zymodemes. Both methods grouped 4 isolates (BAH42, BAH44c9, BAH12c9 and BAH39c7), which comprised a phenotypically heterogeneous assemblage with respect to growth rate and metabolism, into similar groupings. The 2 methods were significantly correlated (P

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Isoenzyme electrophoresis using 13 enzyme systems was applied to 31 Australian and 7 Swiss isolates of Giardia of human, cat, cattle, dog, sheep and rat origin. The Portland (ATCC No. 30888) reference strain was also included. The 39 isolates were divided into 22 different zymodemes. These consisted of 19 zymodemes containing the P1 and Australian isolates and three zymodemes containing Swiss isolates only. Differences in enzyme profiles between zymodemes was measured by euclidean distance and it was found that Australian isolates of Giardia exhibited more variation than the Swiss isolates. Relationships between zymodemes determined by clustering analysis are discussed with particular reference to the zoonotic potential of Giardia.

Summary: The accurate identification of the causative organisms of disease is fundamental to the study of epidemiology. Hence molecular tools are now widely used to detect and distinguish pathogens, and have greatly improved our understanding of epidemiology. David Godfrey pioneered the use of molecular markers in the epidemiology of African trypanosomiasis, thus enabling the light of reliable evidence to shine on this previously problematic and controversial subject area. From the early 1970’s David’s group employed first isoenzyme electrophoresis and subsequently DNAbased characterization methods to aid identification of trypanosomes collected from a range of endemic countries across Africa. These investigations had a major impact on our understanding of the zoonotic nature of human trypanosomiasis in Africa and of the genetic diversity of African trypanosomes.

The limited usefulness of traditional taxonomic methods combined with the discovery of a Strongyloides parasitic in man in Papua New Guinea (PNG) that resembles S. fuelleborni, a parasite of man and other primates in tropical Africa, has precipitated the need to apply new methods to the taxonomy of the genus. In this study isoenzyme electrophoresis has been used on Strongyloides isolates of many different origins. Cluster analysis of the data suggested that existence of three main groups within the material considered, consisting of (1) isolates of S. stercoralis, (2) isolates from PNG domestic animals, and (3) isolates from PNG man and African non-human primates. The taxonomic implications of these groups are considered.

Com o objetivo de avaliar a variabilidade genética e as relações de afinidade entre dezenove populações de oito raças de milho (Zea mays L.) - as comerciais antigas Cateto Sulino, Cateto Sulino Grosso, Cateto Nortista e Canario de Ocho, e as raças indígenas Moroti, Lenha, Entrelaçado e Caingang - analisaram-se os seguintes sistemas enzimáticos: glutamato oxalacetato transaminase (GOT), esterase (EST) e malato desidrogenase (MDH). Observou-se maior semelhança entre as raças pertencentes a um mesmo grupo, mas as populações analisadas não se agruparam de acordo com as raças, classificadas anteriormente segundo caracteres morfológicos. Os sistemas enzimáticos utilizados não permitiram a caracterização individual de cada uma das raças analisadas. As indígenas apresentaram maior variabilidade do que as comerciais antigas quanto ao número de alelos por loco e à porcentagem de locos polimórficos.The objective of this study was to evaluate the genetic variability and affinity relationships among eight races of maize (Zea mays L.): four ancient varieties (Cateto Sulino, Cateto Sulino Grosso, Cateto Nortista and Canario de Ocho) and four indigenous (Moroti, Lenha, Entrelaçado and Caingang), through isoenzymatic polymorphisms. The following isoenzymatic systems were evaluated: Glutamate Oxaloacetate Transaminase (GOT), Esterase (EST) and Malate Dehydrogenase (MDH). It was observed a higher identity among races belonging to the same racial group; however, populations within each race were not grouped according, to previous morphological classification. Indigenous races showed higher average number of alleles per loci and percentage of polymorphic loci than the ancient varieties. This fact might be due to the selection the ancient varieties had been gone through.