Native Smear Research Articles

LABORATORY TESTING RESULTS ANALYSIS OF GIARDIASIS PREVALENCE AMONG DIFFERENT POPULATION GROUPS IN THE NENETS AUTONOMOUS DISTRICT

The Nenets Autonomous District (NAD) refers to the territories of the Russia’s Far North. The country is the only region of Russia, where there are still no roads. The extreme weather conditions, as well as residential patterns in the NAD reduce the availability of specialized medical care, as indigenous people and other nationalities living in the district, which requires the use of health care management specific forms. The article presents data on giardiasis studies amongst different categories of the NAD population, obtained as a result of the population surveys in remote regions of the Russian Federation Arctic zone. Samples studied were serum and faecal emulsion. The study used the following laboratory methods: for serum — Giardia lamblia antibody screening method using ELISA kit, emulsion stool — native smear microscopy analysis, the method of enrichment using disposable concentrators «Parasep» immunological diagnostic methods based on the detection of specific lamblia antigen in faeces using ELISA kits. For statistics was used descriptive statistics analysis (mean values, the percentage error of the mean, the construction of the trend line) with program SPSS 20.00, Excel 2010, the method of analysis of official statistical reporting hospitals survey on giardiasis, data of Rospotrebnadzor (Russian Sanitary service) for NAD and data from own research from 2002 to 2013. Just in the process of research carried out research on the 10 356 giardiasis, registered diseases by 3470 cases of giardiasis, accounting for 99.14% of the total number of protozoonosises registered and 37.4% of all registered in the NAD parasitosis. The comparison was made among the population of the county towns, home to both the indigenous people, nomadic, and representatives of other ethnic groups, sedentary with urban population of the city of Naryan-Mar. The article analyzes the incidence rates for giardiasis according to district official statistical reporting compared with the incidence in the Russian Federation for the same period of time. Statistical analysis was performed using the software package Excel 2010. The study found an association of giardiasis prevalence in the population of the NAD on criteria such as location, gender, occupation and age structure examined.

Effects of infection intensity with Strongyloides papillosus and albendazole treatment on development of oxidative/nitrosative stress in sheep

The objective of this study was to estimate and evaluate oxidative/nitrosative stress parameters in sheep infected with Strongyloides papillosus and after antihelminthic treatment with albendazole (ABZ). This parasite, especially during development stages can seriously damage parenchaematous organs during migration within the host. The presence of parasites leads to increased productions of reactive oxygen species (ROS) and reactive nitrogen species (RNS). It is also well known that certain drugs can be very harmful for the delicate oxidant–antioxidant equilibrium, provoking oxidative stress during their biotransformation. ABZ is a broad spectrum antihelminthic drug, frequently used in veterinary medicine for therapy of parasitic infections. The current research was performed on female Württemberg sheep (n=48). The distribution of parasites in sheep was evaluated using the native smear coprological technique, by sedimentation and flotation methods, revealing the presence of S. papillosus. The degree of infection intensity per sheep was quantitatively established by the method of McMaster, the animals having been divided into three groups according to the intensity of infection; mild, moderate and high. The control group consisted of sheep negative to the parasites. After determining the type of parasite infection, the sheep were treated with ABZ, per orally, in single doses of 5mg/kg per body weight. Sampling of feces for parasitological and blood for biochemical assaying was performed on the 0 and 21st day after treatment with ABZ. The oxidative stress parameters were measured for catalase activity (CAT), the red cell membrane damage by level of malondialdehyde (MDA), while carbonyl and thiol plasma protein group concentrations were used as indicators of the degree of protein oxidative modification. The activity of Cu,Zn-superoxide dismutase (SOD) and relative distribution of lactate dehydrogenase (LDH1–LDH5) activity were determined electrophoretically. The distribution of LDH isoenzymes in sheep moderately and highly infected with S. papillosus revealed that the parasite induced damage to the myocardial (LDH2), lung (LDH3) and liver cells (LDH5) in infected animals, while ABZ treatment only damaged liver cells (LDH5). The MDA concentration revealed that lipid peroxidation increased both in the presence of parasites and the antihelminthic formulation tested (p<0.001) when compared to the control sheep, while the increase of carbonyl concentration (p<0.001), as well as the observed decrease of thiol concentration (p<0.001) indicated significant oxidative damage of plasma proteins in experimental sheep, when compared to the control animals. Our results indicate that S. papillosus induces oxidative/nitrosative stress in sheep. The antihelminthic treatment with ABZ further promotes the disbalance of oxidative–antioxidative equilibrium in all tested sheep.

IFISH analysis of native smears from bone marrow and blood for the monitoring of chimerism and clonal markers after stem cell transplantation in children

After stem cell transplantation (SCT) close follow-up of chimerism and/or clonal disease markers is essential for early treatment of graft failure or relapse. We wanted to assess the sensitivity, clinical reliability and practicability of inter-phase FISH on untreated, native smears of BM or PB for this purpose. We investigated 23 children after SCT with sex mismatch (MM) and/or clone specific markers (monosomy 7, trisomy 8, MLL rearrangement, bcr-abl, TEL-AML-1). Diagnoses were ALL (8), AML (6), MDS (2), CML (2), large cell anaplastic lymphoma (1) and SAA (4). Eighteen children were transplanted from sex-mismatched donors, seven among them had shown a clonal marker at diagnosis. The remaining five patients with sex matched donors also had a clonal marker. For FISH, we used commercial probes on fresh or stored unmanipulated smears of PB or BM. Cut-off levels for clonal markers were established on control probands without hematologic disease, for host sex on probands of the opposite sex, respectively (mean +3 SD). The presence of host cells and/or clonal markers established at diagnosis by conventional karyotyping was followed up after SCT at regular intervals by FISH. Nineteen of the 23 patients studied achieved and maintained complete continuous hematologic remission with corresponding absence of host and/or disease markers. In one of them, a fatal extramedullary relapse occurred. The associated mixed chimerism was confirmed by FISH. In all four cases with hematological relapse, the respective marker (MLL, bcr-abl, Mo 7) reappeared and was successfully monitored during DLI and repeat SCT in two as well as parallelled by simultaneous demonstration of host cells in the two sex mismatched cases among them. We demonstrate the usefulness of FISH on native smears for clinical routine follow-up of SCT patients. FISH allowed identification of cell origin in non-hematologic material (spinal fluid, pericardial effusion). Chimerism analysis in BM was slightly more sensitive than in PB. FISH was feasible on frozen stored smears as well.

Punktatdiagnostik von Gicht und Pseudogicht

Introduction Radiological and morphological findings in advanced arthritis urica and pyrophosphate arthropathy are well known. In contrast, the early changes of synovial membrane in these disturbances of metabolism pose diagnostic problems. With the assistance of various cytological techniques and polarizing microscopical as well as electron microscopical investigation it was examined to what extent needle biopsies can be helpful in the differential diagnosis of gout and pseudogout. Material and Methods In 8 patients with gout and 11 patients with pseudogout synovial fluid and small tissue specimens could be obtained with the aid of the Parker-Pearson needle. Both fluid and tissue specimens were investigated light and electron microscopically. Cell counts were evaluated in a Rosenthal chamber. The differentiation of the cells in stained smears was done by counting 200–600 cells per case. Crystals were identified by polarizing microscopy in wet preparations of freshly aspirated synovial fluid. Results Polarizing microscopy of synovial fluid detected intra- as well as extracellular urate and pyrophosphate crystals. The wedge-shaped urate crystals and the larger partly polygonal pyrophosphate crystals showed different polarizing microscopical properties and a negative birefringence. The absolute cell counts in gout were higher than those in pseudogout. The relative cell counts of the different cell types in synovial fluid showed more variation in gout than in pseudogout. Cases with acute gout developed a relative leukocytosis in contrast to a relative lymphocytosis in chronic gout. A relative leukocytosis was constant in all patients with pseudogout. Sclerosed areas with scarce and plump villi as well as sometimes hyperplastic and polymorphous synovial cell layers could be demonstrated histologically in the tissue specimens of the needle biopsies in cases with gout. Urate crystals were less frequent in specimens fixed in formalin. The histological alterations in pseudogout were uniform, 2–4 rows of slightly pleomorphic synovial cells lined the inner surface of the joint capsule, sclerosing alterations were less frequent. Pyrophosphate crystals and calcified particles were seen within the synovial lining cells, the connective tissue and the endothelial cells of the blood vessels in pseudogout specimens. Intra- as well as extracellular crystals could also be demonstrated with the aid of scanning electron microscopy in sediments of synovial fluid in gout and pseudogout. Transmission electron microscopical investigations of synovial tissue specimens detected proliferated and pleomorphic synovial lining cells in gout in contrast to a more monomorphic appearance of these cells in pseudogout. The crystals were washed out during the preparation techniques for transmission electron microscopy so that needle-like empty spaces resulted within cytoplasm of the phagocytic cells. These clefts were surrounded by phagosomal structures and densified cytoplasmic ground substance; sometimes they were also lined by membranes. Histochemical examinations of the cells with crystalline needles stowed an increase of acid phosphatase around the needle-like clefts. Discussion The used investigational methods are helpful in differential diagnosis of gout and pseudogout. Polarizing microscopy of native smears allows to differentiate the crystals in gout and pseudogout; in addition, a non-crystalline arthritis can be excluded by this test. Comparative electron microscopical investigations elucidated the phagocytosis of crystals by synovial, leukocytic and monocytic cells of synovial fluid. The cellular reaction with formation of phosphatase activity around the crystalline needles was interpreted as a delimitation phenomenon of cells phagocytosing larger needles incompletely. The demonstrated histo- and cytotoxicity of the crystals can explain the variability of the cytological appearance of cellular sediments in gouty and pseudogouty articular effusions.