Effects of garlic (Allium sativum L) and Citrullus colocynthis (L.) Schrad individually and in combination on male reproductive damage due to diabetes: suppression of the AGEs/RAGE/Nox-4 signaling pathway.

BACKGROUND: Diabetes Mellitus is associated with disturbances in male reproductive function and fertility. Studies have shown that oxidative stress with the subsequent inflammation and apoptosis cause these complications in diabetes. Garlic (G) (Allium sativum L) and Citrullus colocynthis (L.) Schrad (C) both have antidiabetic and antioxidant properties. Recently, we demonstrated their synergistic effects in alleviating reproductive complications when administered concomitantly. However, as even medicinal plants in long term usage may lead to some unwanted side effects of their own, we examined whether with half the original doses of these two medicinal plants we could achieve the desired results. METHODS: Thirty-five male Wistar rats were divided into five groups (n = 7/group): Control, Diabetic, Diabetic + G (0.5 ml/100 g BW), Diabetic + C (5 mg/kg BW) and Diabetic + GC (0.5 ml/100 g BW of garlic and 5 mg/kg BW of C. colocynthis) groups. The experimental period was 30 days. RESULTS: Oxidative stress, advanced glycation end products (AGEs), immunoexpression of caspase-3, and expression of mRNAs for receptor for advanced glycation end products (RAGE), NADPH oxidase-4 (NOX-4) and nuclear factor kappa B increased in testis of diabetic rats. Treatment with garlic and C. colocynthis alone showed some beneficial effects, but in the combination form the effectiveness was more profound. CONCLUSIONS: We conclude that the combination therapy of diabetic rats with lower doses is still as efficient as higher doses; therefore, the way forward for reducing complications in long term consumption.

Autophagy in combination therapy of temozolomide and IFN-γ in C6-induced glioblastoma: role of non-coding RNAs.

Aim: We predicted the modulation of autophagy and apoptosis in response to temozolomide (TMZ) and IFN-γ based on changes in the expression of non-coding RNAs in C6-induced glioblastoma (GBM). Materials & methods: Each rat received an intraperitoneal injection of TMZ (7.5 mg/kg) and/or IFN-γ (50,000 IU). Results: The reduced expression of H19 and colorectal neoplasia differentially expressed (CRNDE) was associated with a reduction in autophagy in response to TMZ, IFN-γ and TMZ + IFN-γ therapy, whereas the decreased level of miR-29a (proapoptotic miRNA) was associated with an increase in apoptosis. Conclusion: It appears that H19 promotes switching from autophagy to apoptosis in response to combination therapy of TMZ and IFN-γ through the miR-29a/autophagy-related protein 9A (ATG9A) pathway in C6-induced GBM.

Temozolomide (TMZ) is a drug for people with brain cancer. It can make it hard for patients to learn and think, and it can also make the drug stop working, which lets the tumor keep growing. Researchers are looking for other drugs or things that can be taken with TMZ to stop this from happening. In this study, we used a protein called interferon (IFN), which helps fight cancer. We gave mice with brain cancer both TMZ and IFN, and saw that the tumor cells died and the tumor got smaller. We also looked at how IFN and TMZ changed the genetic material of the mouse brain, called RNA. But we need to test this on people to be sure it works.

Nasopharyngeal myiasis due to Cephalopina titillator in Southeastern Iran: a prevalence, histopathological, and molecular assessment.

The Cephalopina titillator is one of the most important causative agents of nasal myiasis in camels. This study aimed to explore the prevalence, histopathological effects, and molecular identification of C. titillator infestation in camels of Kerman province, South-Eastern Iran, between 2019 and 2021. The larvae were placed in 10% formalin for histopathological evaluation and species identification. Pieces of larval abdominal segments of C. titillator were selected for extraction of DNA. Partial mitochondrial CO1 genes were sequenced for final analysis. Out of the 870 camels examined, 339 (38.9%) were infested with larval stages of C. titillator. There was a significant difference between age and infection rate (P = 0.001), while no association between males and females (P = 0.074) was found. The infection rate was significantly higher in the winter (P < 0.001) than in the other seasons. In this study, different lesions depending on duration, locations, and the depth of larval adhesion notably degeneration changes, necrosis, and ulceration were observed. Also, in chronic cases, granulation tissue reactions were organized. Cephalopina titillator was confirmed by PCR sequencing analysis using mitochondrial CO1 region. A 582 bp nucleotide sequence was deposited in GenBank under the MW136151 accession number. Phylogenetic analysis of CO1 produced a single uniform sister clade to MZ209004 and MW167083 records from China and Iraq, respectively. The high prevalence of C. titillator in camels in this region and other areas of Iran declares that the country is in an endemic status and displays the existence of the potential risk for camels.

Adverse effects of methamphetamine on vital organs of male rats: Histopathological and immunohistochemical investigations.

Objectives: Methamphetamine (named crystal, ice, and crank), is a strong psychostimulant drug with addictive and neurotoxic properties. It is absorbed by various organs and induces tissue damage in abusers. Most METH studies have focused on the central nervous system and its effects on other organs have been neglected. Experimental investigations of animal models are used to provide significant additional information. We have studied the histopathological effects of methamphetamine in the brains, hearts, livers, testes, and kidneys of rats. Materials and Methods: Methamphetamine (0.5 mg/kg) was administered subcutaneously for 21 days. Immunohistochemistry was carried out with markers including glial fibrillary acidic protein (GFAP) for reactive astrocytes, vimentin as an intermediate filament in different cells, and CD45 marker for the detection of reactive microglia in the brain. Also, some samples were taken from livers, kidneys, hearts, and testes. Results: Degenerative changes and necrosis were the most common histopathological effects in the liver, kidneys, heart, testes, and brains of rats treated with methamphetamine. Immunohistochemical analyses by vimentin and GFAP markers revealed reactive microglia and astrocytes with the appearance of swollen cell bodies and also short, thickened, and irregular processes. Moreover, the number of CD45-positive cells was higher in this group. Reactive cells were more noticeable in the peduncles and subcortical white matter of the cerebellum. Conclusion: Our results showed the toxic effects of methamphetamine on the vital organs and induction of neurotoxicity, cardiomyopathy, renal damage, and infertility in male rats. We could not attribute observed hepatic changes to METH and further evaluation is needed.

Evaluation the efficacy of oral immunization of broiler chickens with a recombinant Lactobacillus casei vaccine vector expressing the Carboxy-terminal fragment of α-toxin from Clostridium perfringens.

BACKGROUND: Clostridium perfringens (C. perfringens) is a serious anaerobic enteric pathogen causing necrotic enteritis (NE) in broiler chickens. Following the ban on antibiotics as growth promoters in animal feedstuffs, there has been a remarkable rise in occurrence of NE which resulted in considering alternative approaches, particularly vaccination. The objective of this work was to evaluate the recombinant Lactobacillus casei (L. casei) expressing the C-terminal domain of α-toxin from C. perfringens as a potential probiotic-based vaccine candidate to immunize the broiler chickens against NE. RESULTS: The broiler chickens immunized orally with recombinant vaccine strain were significantly protected against experimental NE challenge, and developed specific serum anti-α antibodies. Additionally, the immunized birds showed higher body weight gains compared with control groups during the challenge experiment. CONCLUSIONS: The current study showed that oral immunization of broiler chickens with a safe probiotic-based vector vaccine expressing α-toxin from C. perfringens could provide protective immunity against NE in birds.

Combination therapy with interferon-gamma as a potential therapeutic medicine in rat's glioblastoma: A multi-mechanism evaluation.

BACKGROUND: This study assessed the effects of single or combined administration of temozolomide (TMZ) and interferon-gamma (IFN-ᵞ) on anxiety-like behaviors, balance disorders, learning and memory, TNF-α, IL-10, some oxidant and antioxidants factors with investigating the toll-like receptor-4 (TLR4) and p-CREB signaling pathway in C6-induced glioblastoma of rats. METHODS: 40 male Sprague-Dawley rats bearing intra-caudate nucleus (CN) culture medium or C6 inoculation were randomly divided into five groups as follows: Sham, Tumor, TMZ, IFN-ᵞ and a TMZ + IFN-ᵞ combination. The open-field test (OFT), elevated plus maze (EPM), rotarod, and passive avoidance test (PAT) were done on days 14-17. On day 17 after tumor implantation, brain tissues were extracted for histopathological evaluation. TNF-α, IL-10, SOD, GPX, TAC, MDA, the protein level of TLR4 and p-CREB was measured. RESULTS: Combination therapy inhibited the growth of the tumor. Treatment groups alleviated tumor-induced anxiety-like behaviors and improved imbalance and memory impairment. SOD, GPX, and TAC decreased in the tumor group. The combination group augmented GPX and TAC. MDA decreased in treatment groups. TMZ, IFN-ᵞ reduced tumor-increased TNF-α and IL-10 level. The combination group declined TNF-α level in serum and IL-10 level in serum and brain. Glioblastoma induced significant upregulation of TLR4 and p-CREB in the brain which inhibited by IFN-ᵞ and TMZ+ IFN-ᵞ. CONCLUSION: The beneficial effects of TMZ, IFN-ᵞ, and TMZ+ IFN-ᵞ on neurocognitive functioning of rats with C6-induced glioblastoma may be mediated via modulating oxidative stress, reduced cytokines, and the downregulation of expression of TLR4 and p-CREB. Combination treatment appears to be more effective than single treatment.

The effects of methionine administration during the beginning postnatal days on the ovarian structures in adult rats.

OBJECTIVES: Methionine is known as an essential amino acid in mammals. Consuming excessive amounts of methionine has toxic effects. This study aimed at evaluating the histomorphometric and histopathologic changes of ovaries after methionine administration during follicle formation. MATERIAL AND METHODS: A total of 60 newborn female rats born under similar conditions were selected and randomly assigned into three groups including control, recipients of 50 and 200 mg/kg body weight of methionine for 5 days. On day 120, all 60 female rats were euthanized and the whole left ovary of each animal was taken in order to count the number of primordial, primary, secondary, antral, atretic follicles, as well as corpora lutea and also to conduct histopathologic study. RESULTS: According to the results, the 50 mg/kg methionine did not significantly change the number of primordial follicles compared to the control group but the 200 mg/kg dose significantly decreased the number of primordial follicles. There were no significant differences between the groups in the number of other types of follicles and also in the number of corpora lutea. There was no histopathological lesion in the groups. CONCLUSIONS: It seems that the high dose of methionine could exacerbate apoptosis of the primordial ovarian follicle during the follicle assembly process. However, the remaining were enough to form later stages of follicles after puberty.

Histopathologic and histomorphometric evaluation of Dipetalonema evansi infection in camel testicular tissue.

Camels are important sources of milk, meat, wool and leather, and are widely used in transportation in arid and semi-arid areas. But their illnesses, especially parasitic diseases, have not been taken into consideration. The Dipetalonema evansi microfilariae are in the blood. Adult nematode is only dedicated to camels and disrupts spermatic arteries, lung arteries, right atrium, and testicles. This study was carried out on testicular samples of camels infected with D. evansi referred from slaughterhouse. In each of the control and contaminated groups, 5 samples were examined. In this study, in addition to the qualitative description of parasite histopathologic lesions, the spermatogenesis process was evaluated quantitatively including spermatogenesis process, diameter of the seminiferous tubules and Johnsen ranking and compared with the control group. Histopathological examination of infected testis with D. evansi showed lumen obstruction of testicular blood vessels by parasites, hypertrophy of blood vessels, degenerative and necrosis changes in the tubules, decreased spermatogenetic activity, increased interstitial space tubules, destruction of the spermatogenic cells. Also, there was a significant difference in the control and contaminated groups in the parameters of spermatogenesis, diameter of the seminiferous tubules and Johnsen score.

In vitro and in vivo therapeutic potentials of 6-gingerol in combination with amphotericin B for treatment of Leishmania major infection: Powerful synergistic and multifunctional effects.

The ongoing conventional drugs for leishmaniasis treatment are insufficient. The present study aimed to assess 6-gingerol alone and in combination with amphotericin B on Leishmania major stages using experimental and in vivo murine models. Here, arrays of experimental approaches were designed to monitor and evaluate the 6-gingerol potential therapeutic outcomes. The binding affinity of 6-gingerol and IFN-γ was the basis for docking conformations. 6-Gingerol combined with amphotericin B represented a safe mixture, extremely leishmanicidal, a potent antioxidant, induced a remarkable apoptotic index, significantly increased the expression of the Th1-related cytokines (IL-12p40, IFN-γ, and TNF- α), iNOS, and transcription factors (STAT1, c-Fos, and Elk-1). In contrast, the expression of the Th2-related cytokines was significantly downregulated (p < 0.001). This combination was also potent when the lesion appearance was evaluated following three weeks of treatment. The histopathological and immunohistochemical patterns of the murine model represented clusters of CD4+ and CD8+ T lymphocytes which compressed and deteriorated the macrophages harboring Leishman bodies. The primary mode of action of 6-gingerol and amphotericin B involved broad mechanistic insights providing a coherent basis for further clinical study as a potential drug candidate for CL. In conclusion, 6-gingerol with amphotericin B synergistically exerted anti-leishmanial activity in vitro and in vivo and potentiated macrophages' leishmanicidal activity, modulated Th1- and Th2-related phenotypes improved the histopathological changes in the BALB/c mice infected with L. major. They elevated the leukocyte infiltration into the lesions. Therefore, this combination should be considered for treating volunteer patients with CL in clinical studies.

Major risk factors and histopathological profile of treatment failure, relapse and chronic patients with anthroponotic cutaneous leishmaniasis: A prospective case-control study on treatment outcome and their medical importance.

Over the last years, there has been a remarkable increase in the number of unresponsive patients with anthroponotic cutaneous leishmaniasis (ACL) reported worldwide. The primary objective of this study was to explore the role of demographic, clinical and environmental risk related-factors in the development of treatment failure, relapse and chronic cases compared to responsive patients with ACL. Moreover, molecular, histopathological and immunohistochemical (IHC) findings between these forms were explored. This work was undertaken as a prospective and case-control study in southeastern Iran. Culture media and nested PCR were used to identify the causative agent. Univariate multinomial and multiple multinomial logistic regression models and the backward elimination stepwise method were applied to analyze the data. A P<0.05 was defined as significant. Also, for different groups, skin punch biopsies were used to study the histopathological and immunohistochemical (IHC) profile. All samples showed that L. tropica was the only etiological agent in all unresponsive and responsive patients with ACL. Data analysis represented that 8 major risk factors including nationality, age groups, occupation, marital status, history of chronic diseases, duration of the lesion, the lesion on face and presence of domestic animals in the house were significantly associated with the induction of unresponsive forms. The histopathological and immunohistochemical findings were different from one form to another. The present findings clearly demonstrated a positive relation between ACL and distinct demographic, clinical and environmental risk determinants. Knowledge of the main risk factors for ACL infection is crucial in improving clinical and public health strategies and monitor such perplexing factors.

Partridge and embryonated partridge egg as new preclinical models for candidiasis.

Candida albicans (C. albicans) is the most common cause of candidiasis in humans and animals. This study was established to a new experimental infection model for systemic candidiasis using partridge and embryonated partridge egg. First, we tested the induction of systemic candidiasis in partridge and embryonated partridge egg. Finally, interaction between virulence factors of C. albicans and Bcl-2 family members was predicted. We observed that embryonic infection causes a decrease in survival time and at later embryonic days (11-12th), embryos showed lesions. Morphometric analysis of the extra-embryonic membrane (EEM) vasculature showed that vascular apoptotic effect of C. albicans was revealed by a significant reduction in capillary area. In immunohistochemistry assay, low expression of Bcl-2 and increased expression of Bax confirmed apoptosis. The gene expression of Bax and Bcl-2 was also altered in fungi-exposed EEM. Ourin silico simulation has shown an accurate interaction between aspartic proteinase, polyamine oxidase, Bcl-2 and BAX. We observed that the disease was associated with adverse consequences, which were similar to human candidiasis. Acquired results support the idea that partridge and embryonated partridge egg can be utilized as appropriate preclinical models to investigate the pathological effects of candidiasis.

Linguatula serrata in cattle in southeastern Iran: Epidemiological, histopathological and phylogenetic profile and its zoonotic importance.

This study aimed to determine the prevalence, histopathological observations, and phylogenetic analysis of L. serrata in cattle and its potential zoonotic and public health implications in southeastern Iran.The cross-sectional study was performed in cattle in southeastern Iran. Lymph nodes were collected from each cattle and examined by parasitological and histopathological techniques. A binary logistic regression and chi-square tests were implemented to analyze the data. Genomic DNA was randomly extracted from the nymphal stages of Linguatula isolates. Further characterization and phylogenetic relationships were done using two primers for amplification of partial DNA fragments of 18 s rRNA and cytochrome C oxidase subunit 1 (cox1), respectively.The results showed that 64 cattle of the total 404 were infected with L. serrata. There was no significant difference between linguatulosis infection and gender, while age was significantly different (P < 0.05). The cyst-like spaces containing the longitudinal and transverse sections of the L. serrata nymphs were surrounded by granulomatous reactions. The higher nucleotide variation in the cox1 region was supported by estimating the evolutionary divergence between L. serrata isolates and other Linguatula records of ruminants in Iran. The phylogenetic tree confirmed the close evolutionary relationships among all reported records of L. serrata in Iran.The high prevalence of linguatulosis caused by L. serrata declares the existence of a potential risk of FBPs for humans in southeastern Iran. This condition can advance more serious public health problems and requires a comprehensive control program and treatment strategies to prevent the disease.

Allogenic Bone Graft Enriched by Periosteal Stem Cell and Growth Factors for Osteogenesis in Critical Size Bone Defect in Rabbit Model: Histopathological and Radiological Evaluation.

BACKGROUND & OBJECTIVE: This study aimed to investigate the effect of decellularized allogeneic bone graft enriched by periosteal stem cells (PSCs) and growth factors on the bone repair process in a rabbit model, which could be used in many orthopedic procedures. METHODS: In this experimental study, a critical size defect (CSD) (10 mm) was created in the radial diaphysis of 40 rabbits. In group A, the defect was left intact with no medical intervention. In group B, the defect was filled by a decellularized bone graft. In group C, the defect was implanted by a decellularized bone graft enriched with platelet growth factors. In group D, the defect was treated by a decellularized bone graft seeded by periosteal mesenchymal stem cells (MSCs). Also, in group E, the defect was filled by a decellularized bone graft enriched with platelet growth factors and periosteal MSCs. Radiological evaluation was done on the first day and then in the second, fourth, and eighth weeks after the operation. The specimens were harvested on the 28th and 56th postoperative days and evaluated for histopathological criteria. RESULTS: The radiologic and microscopic analysis of the healing process in bone defects of the treated groups (C, D, and E) revealed more advanced repair criteria than those of groups A and B significantly (P<0.05). CONCLUSION: Based on this study, it appears that implantation of concentrated PSCs in combination with growth factors and allogeneic cortical bone graft is an effective therapy for the repair of large bone defects.

Experimentally Induced Cerebral Cystic Echinococcosis in Rats: A Suitable Animal Model for Cerebral Echinococcosis.

BACKGROUND: Echinococcus granulosus is a worldwide zoonotic cestode that lives mainly in the intestine of dog as definitive host. Its larval stage infects intermediate hosts and forms hydatid cysts mainly in the liver and lungs tissues and less other organs such as brain, eye and bone. In the experimental models, inoculation of protoscoleces into the peritoneum, thoracic cavity, subcutaneous and cerebrum produces hydatid cysts. Experimental echinococcosis in the animal models provides a good opportunity for study of the parasite-host relationship, different transmission ways of infection in the intermediate hosts and effect of new drugs. METHODS: The present study was conducted in the Veterinary School, Shahid Bahonar University of Kerman, Kerman, Iran in 2018. In this study, cerebral hydatidosis was investigated in 6 female Wistar rats weighing (200±20 gr). For this purpose, protoscoleces were collected from hydatid cysts of infected sheep liver. Overall, 300 protoscolices were injected directly in the lateral ventricle by an insulin syringe through the implanted cannula. RESULTS: After 4 months of inoculation, multiple thin-walled, transparent hydatid cysts were observed in the rat skull. All cysts were infertile. The cysts were localized prominently on the cerebral cortex and lesser in the parenchyma and ventricles. The cyst walls consisted of three layers consist of the outer layer (fibrous capsule), two parasitic layers and the endocyst layer (germinal layer). The cyst was surrounded by the inflammatory cells consist of lymphocytes, plasma cells and macrophages. CONCLUSION: To the best of our knowledge, this research is the first experimental cerebral hydatidosis arisen from larval stage of Echinococcus granulosus in the animal model.

Scanning electron microscopy of Onchocerca fasciata (Filarioidea: Onchocercidae) adults, microfilariae and eggs with notes on histopathological findings in camels.

BACKGROUND: Onchocerca fasciata is a prevalent filarial species in camelids of Asia and Africa forming nodules in the skin of dromedary and Bactrian camels. In spite of recent advances in the biology and epidemiology of this nematode species, a relatively scant number of studies have focussed on the morphology of this parasite. The main objective of this study was to describe morphological characteristics of adults, microfilariae and eggs of O. fasciata by scanning electron microscopy (SEM), staining and histology. METHODS: From April 2016 to March 2017 dromedary camels (n = 456) were inspected for infection with O. fasciata in a slaughterhouse in Kerman (south of Iran). Adult worms in nodules were isolated by digestion of nodules in collagenase and used for SEM. Skin nodules were also fixed, sectioned and stained with hematoxylin and eosin for histopathology. Skin microfilariae that were isolated from tissues surrounding the nodules were confirmed as O. fasciata by sequencing of the cytochrome c oxidase subunit 1 (cox1) and 12S rRNA genes and used for SEM and Giemsa staining. RESULTS: Single or multiple O. fasciata nodules (1.2-2.2 cm in diameter and 507-845 mg in weight) were found in 30.3% of the examined camels. SEM analysis helped identify 18 papillae in the caudal region of the male. Discontinuous longitudinal cuticular crests were observed in the posterior region of the male. In female nematodes, the ridges had a rounded shape with a height/width ratio of 7/16 in longitudinal sections. Unsheathed skin microfilariae with a rounded anterior extremity measured 210.7 × 2.5 µm on average. Developed eggs containing microfilariae measured 35.9 × 31.0 µm and their smooth shell surface had characteristic tongue-like appendages. In addition to inflammatory reactions surrounding the parasites, accumulation of intracellular ceroid pigment, golden-yellow to brown in colour, was observed within macrophages upon histopathological examination. CONCLUSIONS: We found longitudinal crests on the surface of the posterior region of the male nematode. Measurements of the main morphological features of microfilariae and eggs, and the shape index of ridges (height/width) in female nematodes are described for the first time.

Topical effect of allogenous serum rich in growth factors (SRGF) on diabetic skin wound in rat.

Diabetic foot ulcer is a major complication of diabetes mellitus with negative effects on the life quality. Management of diabetic foot ulcers is a big challenge with poor and low sufficient outcome management. Therefore, achievement to effective treatments may treat these ulcers. Nowdays, platelet products are used as an effective and safe agent for promotion of healing proposes in regenerative medicine. Serum rich in growth factors (SRGF) is a source of released growth factors from the platelets. In the present study, effect of allogenous SRGF was investigated on the streptozotocin (STZ)-induced diabetic wounds in rats. STZ (50 mg/kh, SC) caused significant increase in blood glucose and weight loss in rats. Full thickness cutaneous wounds (8 mm diameter) were created bilaterally on the dorsal of the diabetic rats. SRGF was injected at the edges of the wounds of one side only on the first day, and the contrary sides were considered as the control group. The percentage of wound contraction was demonstrared on day 3, 7, 11 after surgery. Tissue specimens were collected for microscopic study on days 7 and 14. Results showed a significant higher rate of wound contraction on days 5 and 7 in the treated group. Histopathologic findings displayed acceleration of re-epithelialization, increased angiogenesis and dense collagen fibers with better organization in the treated group. Current study suggests that SRGF was efficient because facilate wound healing and showed rapid re-epithelialization and increased angiogenesis.

Experimental study on healing of long bone defects treated with fibrin membrane enriched with platelet growth factors and periosteal mesenchymal stem cells in rabbit: Radiographical and histopathological evaluations.

The present study was designed to evaluate the effects of platelet growth factors and periosteal mesenchymal stem cells on bone healing process, radiographically. Forty male White New Zealand rabbits in five equal groups were used in this study. A 2 mm full thickness bone defect was made in left radial bone of each animal. In group A (control) the defect was left with no medical intervention. In group B the defect was covered by a fibrin membrane. In group C the defect was covered by a fibrin membrane plus platelet growth factors. In group D the defect was covered by a fibrin membrane plus periosteal mesenchymal stem cells, and in group E the defect was covered by a fibrin membrane enriched with platelet growth factors and periosteal mesenchymal stem cells. Radiological evaluation was done immediately after surgery (week 0) and then at the 1st, 2nd, 4th, 6th and 8th weeks after operation. At the end of the eighth week, bone samples were taken to evaluate the histopathology. The radiological and histopathological observations showed a superior bone healing in the groups D and E, after eight weeks in comparison with the groups A, B and C. According to this study, it could be concluded that the platelet growth factors and periosteal mesenchymal stem cells could promote bone regeneration in long bone defects in a rabbit model.

Chemical disbudding of goat kids with subcutaneous administration of synthetic eugenol: Histopathology and morphometry.

This study was conducted to evaluate the effect of subcutaneous administration of synthetic eugenol (EG) for disbudding of goat kids, as a new chemical method. Thirty apparently healthy Raieni (Cashmere) goat kids (five-day-old) were divided randomly into six groups (n = 5). In the pathology (P) groups (P3, P8 and P60 according to the sample collection day after injection) an amount of 0.10 mL of EG was subcutaneously administrated in both horn buds. In the disbudding 1 and 2 (DB1, DB2) groups, 0.10 mL of EG, and in the control (C) group 0.10 mL normal saline was subcutaneously injected in the right horn buds, respectively. Eugenol injection in DB2 group was done in twelve-day-old goat kids. The left horn buds of DB1, DB2 and C groups were considered as control of horn outgrowing. The horn buds, kidneys, liver, lung, brain and heart, tissue specimens were collected from P3 and P8 groups, and bud skin samples were collected from P60 group. The results showed that the EG was able to stop the horn growth in the first week of goat life. There was no significant difference between left and right horn size in the C group. Histopathological study revealed complete necrosis of bud tissue in dermal and epidermal layers, in P3 animals. Healing and re-epithelialization were seen in the samples taken from P8 group. Subcutaneous injection of the synthetic EG can be considered as a new method for goat kids disbudding.

Histomorphometric evaluation of mice testicular tissue following short- and long-term effects of lipopolysaccharide-induced endotoxemia.

OBJECTIVES: Lipopolysaccharide (LPS)-induced endotoxemia is known to cause male infertility. This study was designed to explore the effects of bacterial LPS on histomorphometric changes of mice testicular tissues. MATERIALS AND METHODS: In experiment 1, a pilot dose responsive study was performed with mice that were divided into five groups, receiving 36000, 18000, 9000, and 6750 µg/kg body weight (B.W) of LPS or only saline (control). White blood cells (WBC) were observed for 3 days after LPS inoculation. In experiment 2, two groups of mice were treated with 6750 µg/kg B.W of LPS or only saline (control). Five cases from each experimental group were sacrificed at 3, 30, and 60 days after LPS inoculation. Left testes were fixed in Bouin's solution, and stained for morphometrical assays. RESULTS: Time-course changes of WBC obtained from different doses of LPS-treated mice showed that inoculation of 6750 µg/kg B.W produced a reversible endotoxemia that lasts for 72 hr and so it was used in the second experiment. In experiment 2, during the first 3 days, no significant changes were observed in the evaluated parameters instead of seminiferous tubules diameter. Spermatogenesis, Johnsen's score, meiotic index, and epithelial height were significantly affected at 30th day. However, complete recovery was only observed for the spermatogenesis at day 60. Interestingly, deleterious effects of LPS on spermatogonia were only seen at 60th day (P<0.05). CONCLUSION: Endotoxemia induced by LPS has long-term detrimental effects on spermatogonia and later stage germ cells, which are reversible at the next spermatogenic cycle.

Low susceptibility of domestic cats to experimental Leishmania infantum infection.

BACKGROUND AND OBJECTIVES: : The dogs are considered the main reservoir of visceral leishmaniasis (VL), but lately the disease incidence has been reported in cats also. In this study, the susceptibility of domestic cats to experimental Leishmania infantum infection was assessed by different diagnostic methods. METHODS: : A total of 12 healthy adult male cats were captured by double door live trap cages containing baits. Of them eight cats were intraperitoneally inoculated with 107 L. infantum promastigotes (stationary phase), and four cats were used as controls. Whole blood and serum samples were collected at weekly intervals for 16 wk after inoculation for testing by polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) methods. Aspirates of prescapular lymph nodes and bone marrow were obtained at monthly intervals. Clinical examination was performed twice weekly and histopathological evaluation was done on necropsy samples at the termination of the study. Results: One week after inoculation, blood nested PCR was able to detect the L. infantum infection and it remained positive until 16 wk. ELISA test remained negative during the study. Amastigote phase of parasite was not observed in bone marrow aspiration and necropsy samples. INTERPRETATION AND CONCLUSION: : The feline model described in this work would be useful in further understanding of L. infantum immunopathogenensis in cats. The results of this preliminary study suggest that cats might be resistant to VL as the inoculation dose which induces pathognomonic clinical features in dogs, just creates asymptomatic parasitaemia in cats. Though, due to long-lasting parasitaemia, cats may act as appropriate reservoir for transmission of VL to human population. Further studies are needed to describe the possible role of cats in the epidemiology of VL in endemic areas.