Leukemia cell microvesicles promote survival in umbilical cord blood hematopoietic stem cells

Microvesicles can transfer their contents, proteins and RNA, to target cells and thereby transform them. This may induce apoptosis or survival depending on cell origin and the target cell. In this study, we investigate the effect of leukemic cell microvesicles on umbilical cord blood hematopoietic stem cells to seek evidence of apoptosis or cell survival. Microvesicles were isolated from both healthy donor bone marrow samples and Jurkat cells by ultra-centrifugation and were added to hematopoietic stem cells sorted from umbilical cord blood samples by magnetic associated cell sorting (MACS) technique. After 7 days, cell count, cell viability, flow cytometry analysis for hematopoietic stem cell markers and qPCR for P53 gene expression were performed. The results showed higher cell number, higher cell viability rate and lower P53 gene expression in leukemia group in comparison with normal and control groups. Also, CD34 expression as the most important hematopoietic stem cell marker, did not change during the treatment and lineage differentiation was not observed. In conclusion, this study showed anti-apoptotic effect of leukemia cell derived microvesicles on umbilical cord blood hematopoietic stem cells

Busulfan Treatment Effects on Testicular Tissue and Serum Levels of Anti- Mullerian Hormone and Testosterone in Adult Mice

BACKGROUND: Busulfan, a chemotherapeutic drug, is an alkylating antineoplastic agent that belongs to the class of alkyl sulfonates and has some side effects on fertility. This research was aimed to investigate the effects of busulfan on testicular tissue and serum levels of anti-Mullerian hormone (AMH) and testosterone in adult mice.METHODS: Eighteen adult male Balb/C mice were collected randomly and were assigned in three groups including; control, azoospermia and spontaneous recovery. The groups, except for control group, received two injections of busulfan (10 mg/kg) intraperitoneally with 21-days interval in order to induce azoospermia. Thirty-five and 140 days after the last injection, the effects of busulfan on testicular tissue were evaluated by histologic, histomorphometric and hormonal changes. AMH and testosterone were measured by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) kits, respectively.RESULTS: Hormonal analyses showed no significant differences in AMH and testosterone serum levels. Histologic and histomorphometric evaluation showed disrupted spermatogenesis in azoospermia group, and restoration of spermatogenesis spontaneously in spontaneous recovery group.CONCLUSION: Busulfan at doses was used had no effect on AMH and testosterone serum levels. Busulfan can also induce azoospermia on a temporary basis however, in long term, spontaneous recovery can occur. The results indicated that some side effects are reversible and may depend on the dose applied.KEYWORDS: Busulfan, anti-Mullerian hormone, testosterone, mouse, testi

Growth kinetics and characterization of human dental pulp stem cells: comparison between third molar and first premolar teeth

Dental pulp stem cells (DPSCs) play an important role in tissue regeneration. This study compares the growth kinetics and characterization of third molar and first premolar human DPSCs. Dental pulp tissues were isolated from human first premolar and third molar teeth and were digested by treating them with collagenase type I. Single-cell suspensions from each dental pulp were seeded in T25 culture flasks and the media were replaced every 3 days until 70% confluence. The cells were enumerated to determine the population doubling time (PDT). Cells were characterized using flow cytometry, RT-PCR and osteogenic medium for differentiation of DPSCs. Karyotyping assay was also performed till passage 7th. The DPSCs had spindle-shaped morphology. There was an increase in PDT in third molar DPSCs when compared to first premolar teeth. Positive expression of CD44, CD73, and CD90 and negative expression of CD34 and CD45 were illustrated. A normal karyotype was visible for all seven passages. The Alizarin red staining was positive for osteogenic induction of DPSCs. When DPSCs are needed, third molar teeth can be a good and convenient candidate for cell transplantation, yielding high number of cells with mesenchymal characteristics. They can be a source for further investigations in vitro and work on tissue engineering protocols

Učestalost i jačina invazije oblićem Toxocara cati u mačaka lutalica u Shirazu, Iran.

A cross-sectional survey was undertaken to study the prevalence and intensity of infestation with Toxocara cati in 108 stray cats, including 44 males and 64 females, from December 1996 to May 1997 in Shiraz city, southern Iran. A total of 57 cats (52.8%) were infected with T. cati. There was no significant difference between the rate of infection between males and females, young and adult cats, or the prevalence of infection in different geographical areas. The number of recovered worms per cat or intensity of infection ranged from one to a maximum of 50 worms per animal, with an average of 6.53 T. cati in each cat.Istraživanje je provedeno na 108 mačaka lutalica i to 44 mužjaka i 64 ženke u razdoblju od prosinca 1996. do svibnja 1997. na području grada Shiraza u Iranu. Potvrđeno je da je čak 52,8% pretraženih životinja bilo invadirano oblićem Toxocara cati. Nije utvrđena značajna razlika u učestalosti invazije s obzirom na spol i dob te mjesto istraživanja. Broj oblića kretao se od 1 do 50, u prosjeku 6,53 oblića po mački

Active immunization using exotoxin A confers protection against Pseudomonas aeruginosa infection in a mouse burn model

<p>Abstract</p> <p>Background</p> <p><it>Pseudomonas aeruginosa </it>is an important cause of nosocomial infection and may lead to septicemia and death. We evaluated the immunogenicity of semi-purified exotoxin A from the bacterium in a mouse burn model.</p> <p>Methods</p> <p>The toxoid was prepared from exotoxin A taken from toxigenic strains of <it>P. aeruginosa </it>(PA 103). 50 mice were immunized with the toxoid, burned with hot metal and infected with 1 × 10⁸CFU of toxigenic strains of <it>P. aeruginosa </it>(experimental group); 25 non-immunized mice were also burned and infected (control group). The mortality rate and presence of any exotoxin and <it>P. aeruginosa </it>in the sera, liver and spleen were determined.</p> <p>Results</p> <p>In the experimental group, 2 mice died before the burns were administered and were excluded from the study. The remainder (48 mice) were challenged with a lethal dose of <it>P. aeruginosa </it>and followed for 70 days. 3 of these mice died. Neither <it>P. aeruginosa </it>nor exotoxin A was not detected in the liver, spleen or sera of the surviving mice. The protective efficacy of toxoid vaccination was therefore 93.8%. In the control group, all mice died from bacteremia and septicemia, most (80%) within 6 days, and <it>P. aeruginosa </it>and exotoxin A were isolated from sera, spleen and liver.</p> <p>Conclusion</p> <p>Active immunization of mice using a semi-purified exotoxin A derived from <it>P. aeruginosa </it>was 93.8% effective at protecting mice from subsequent <it>P. aeruginosa </it>infections in a mouse burn model.</p

Caprine Endometrial Mesenchymal Stromal Stem Cell: Multilineage Potential, Characterization, and Growth Kinetics in Breeding and Anestrous Stages

The endometrial layer of the uterus contains a population of cells with similar characteristics of mesenchymal stem cells (MSCs). In the present study, caprine endometrial mesenchymal stromal stem cells (En-MSCs) characters and differentiation potential to chondrogenic, osteogenic, and adipogenic cell lines as well as their growth kinetics in breeding and anestrous stages were evaluated. En-MSCs were enzymatically isolated from endometrial layer of the uterus of adult goats and were cultured and subcultured until passage 4. The growth kinetics and population doubling time (PDT) of caprine En-MSCs in breeding and anestrous stages were determined. En-MSCs in passage 4 were used for the karyotyping and differentiation into chondrocytes, osteocytes, and adipocytes. The PDT in anestrus phase was 40.6 h and in cyclic goats was 53 h. En-MSCs were fibroblast-like in all passages. The number of chromosomes was normal (2n=60) with no chromosomal instability. Chondrogenic, osteogenic, and adipogenic differentiation of En-MSCs was confirmed by staining with Alcian blue, Alizarin red, and Oil Red O, respectively. Caprine En-MSCs demonstrated to be an alternative source of MSCs for cell therapy purposes in regenerative medicine

Cutaneous leishmaniasis in Iran: A review of epidemiological aspects, with emphasis on molecular findings

Leishmania parasites can cause zoonotic cutaneous leishmaniasis (CL) by circulating between humans, rodents, and sandflies in Iran. In this study, published data were collected from scientific sources such as Web of Science, Scopus, PubMed, Springer, ResearchGate, Wiley Online, Ovid, Ebsco, Cochrane Library, Google scholar, and SID. Keywords searched in the articles, theses, and abstracts from 1983 to 2021 were cutaneous leishmaniasis, epidemiology, reservoir, vector, climatic factors, identification, and Iran. This review revealed that CL was prevalent in the west of Iran, while the center and south of Iran were also involved in recent years. The lack of facilities in suburban regions was an aggravating factor in the human community. Some parts of southern Iran were prominent foci of CL due the presence of potential rodent hosts in these regions. Rhombomys opimus, Meriones lybicus, and Tatera indica were well-documented species for hosting the Leishmania species in Iran. Moreover, R. opimus has been found with a coinfection of Leishmania major and L. turanica from the northeast and center of Iran. Mashhad, Kerman, Yazd, and sometimes Shiraz and Tehran foci were distinct areas for L. tropica. Molecular identifications using genomic diagnosis of kDNA and ITS1 fragments of the parasite indicated that there is heterogeneity in leishmaniasis in different parts of the country. Although cutaneous leishmaniasis has been a predicament for the health system, it is relatively under control in Iran

Povezanost između radiografskih nalaza, magnetske rezonancije i histopatoloških nalaza kod pokusne rupture prednje križne sveze u kunića.

Experimental osteoarthritis (OA) was induced in the knee joints of rabbits and the trend of changes were compared by radiography, Magnetic Resonance Imaging (MRI) and histopathology. Twenty rabbits were randomly divided into two equal groups based on short (30 days) and long-term (180 days) follow ups. In half of the animals in each group (n = 5) OA was induced by sectioning the cranial cruciate ligament and in the other half, only arthrotomy was performed as a sham operation. Radiography and MRI were carried out on days 0 and 30 in the group of short term studies, and on days 0, 90 and 180 in the other group. Histopathological examinations were performed on day 30 in the short-term group after the animals had been sacrificed and in the other group on day 180. The slope of changes over the course of the study between all 3 methods and the grade of changes, were both highest in histopathology, and then in MRI and radiology respectively. The slope of changes was 0.01 for histopathology, 0.009 for MRI and 0.004 for radiology. The ratios of slopes, when compare to each other, were as follows: His./MRI = 1.1, His./Rad. = 2.5, MRI/Rad. = 2.2. Comparison of MRI with radiology revealed that radiology would not show signs of OA when the MRI grade is less than a grade of 0.27. Comparing both imaging techniques with histopathology showed that whenever the histopathological grade was below 0.22, radiology would not show signs of OA involvement, while MRI was capable of showing signs of OA involvement whenever it was more than 0.018 on histopathological grade.Nakon što je u kunića pokusno izazvan osteoartritis koljenog zgloba, trend promjena promatran je pomoću radiografije, magnetske rezonancije (MRI) i patohistoloških nalaza. Dvadeset kunića bilo je metodom slučajnog odabira podijeljeno u dvije jednake skupine od kojih je jedna promatrana tijekom kratkotrajnog (30 dana), a druga tijekom dugotrajnog (180 dana) razdoblja. U polovice životinja iz svake skupine (n = 5) osteoartroza je bila uzrokovana sekcijom prednje križne sveze, a kod druge polovice primijenjena je samo artrotomija kao lažna operacija. Radiografija i magnetska rezonancija obavljene su 0-ti i 30. dan tijekom kratkotrajnog promatranja te 0-ti, 90. i 180. dan kod dugotrajnog promatranja. Histopatološke pretrage obavljene su 30. dan kod kratkotrajnog praćenja nakon što su životinje bile usmrćene, a kod dugotrajnog 180. dan. Slabljenje promjena tijekom promatranja svima trima metodama kao i stupanj promjena pokazali su se najvećima kod histopatoloških nalaza, zatim kod magnetske rezonancije te nakon toga kod radioloških snimki. Slabljenje promjena bilo je 0,01 za histopatološke nalaze, 0,009 za magnetsku rezonanciju i 0,004 za radiološke snimke. Kad se uzmu u obzir međusobni odnosi slabljenja su bila sljedeća: His./MRI = 1,1, His./Rad. = 2,5, MRI/Rad. = 2,2. Usporedba magnetske rezonancije i radiologije potvrdila je da radiologija neće pokazati znakove osteoartritisa kada je stupanj magnetske rezonance niži od 0,27. Usporedbom obje tehnike snimanja s histopatološkim nalazima utvrđeno je da radiologija neće pokazati znakove osteoartritisa kad je stupanj histopatoloških nalaza ispod 0,22. Primjenom magnentske rezonancije moguće je utvrditi znakove osteoartritisa svaki put kada je njezin stupanj veći od 0,018 u odnosu na stupanj histopatoloških nalaza

Indukcija spermatogeneze nakon terapije azoospermične zamorčadi matičnim stanicama

Bone marrow-derived mesenchymal stem cells (BM-MSCs) provide a large quantitative alternative source for regenerative medicine. This study was undertaken to determine the effect of BM-MSCs in the treatment of busulfan-induced azoospermia in guinea pigs. BM-MSCs were isolated from the femur bones of 6 adult guinea pigs as the donor group, and characterized by morphology, MSC markers and osteogenic and adipogenic differentiation. A dose of 40 mg/kg of busulfan was administered at a 21 day interval to induce azoospermia in 6 guinea pigs. Thirty-five days after the second injection of busulfan, transplantation of 1×106 BM-MSCs was performed into the seminiferous tubules of the left testes. The right testis was considered as the positive busulfan treated control. The testes of the donor group were applied as an intact normal control. Then, 60 days after cell therapy, histopathological and histomorphometric evaluations of the testes were performed. The seminiferous tubules treated with BM-MSCs, similar to the intact group, showed a normal appearance of spermatogenesis in comparison to the busulfan-induced azoospermic testes. In conclusion, BM-MSCs were effective in the treatment of azoospermia in a guinea pig model where they restore the fertility of busulfan-induced azoospermic animals after transplantation of BM-MSCs. Therefore, this report could open a window in future to the possibility of BM-MSCs transplantation in the treatment of azoospermia in humans, but more studies should be undertaken for further verification.Mezenhimne matične stanice podrijetlom iz koštane srži (MMSKS) pružaju velike mogućnosti alternativnog izvora u regenerativnoj medicini. Ovo je istraživanje provedeno radi određivanja učinka MMSKS u svrhu liječenja azoospermije u zamorčadi izazvane busulfanom. MMSKS su bile izdvojene iz srži bedrene kosti od šestero odraslih zamorčića kao donorske skupine te im je određena morfologija, markeri te osteogenična i adipogenična diferencijacija. Doza od 40 mg/kg busulfana bila je zamorčićima primijenjena u razmaku od 21 dana. Tridesetipet dana nakon posljednje primjene busulfana provedena je transplantacija 1×106 MMSKS u sjemenonosne tubule lijevih sjemenika. Desni sjemenik uzet je kao pozitivna busulfan kontrola. Sjemenici donorske skupine uzeti su kao netaknuta normalna skupina. Nakon stanične terapije u trajanju od 60 dana provedena je histopatološka i histomorfometrijska prosudba sjemenika. Sjemenonosni tubuli obrađeni MMSKS, slično kao i neobrađena kontrolna skupina, pokazivali su normalnu spermatogenezu u usporedbi s busulfanom induciranim azoospermičnim sjemenicima. Zaključno se može reći da su MMSKS bile učinkovite u liječenju azoospermije na modelu zamorčadi te da se plodnost u azoospermičnih zamorčića vratila nakon transplantacije MMSKS. Stoga ovo istraživanje može ubuduće otvoriti prozor za mogućnost transplantacije MMSKS u svrhu liječenja azoospermije u čovjeka, premda su za potvrdu toga potrebna daljnja istraživanja