Parasites of Stray Cats in Iran: A Parasitological and Histopathological Study.

Many zoonotic parasitic diseases, including Toxocara cati, may be spread by stray cat populations. This study aimed to determine the prevalence of parasites by performing parasitological and histopathological examinations on stray cats in Shiraz, Iran. A total of 106 stray cats from different geographical areas of Shiraz, southern Iran, were examined for the presence of parasites. The overall prevalence was found to be 83.02% (88/106), and eight parasites were found. The parasites included three genera of cestodes [Joyeuxiella echinorhynoides (52.83%), Taenia taeniaeformis (21.70%), and Dipylidium caninum (1.89%)], three nematodes [Physaloptera praeputialis (23.59%), Toxocara cati (15.09%), and Rictularia sp. (1.89%)], one protozoa [Isospora spp. (6.60%)], and one arthropod [Ctenocephalides felis (5.66%)]. The prevalence did not significantly differ between males and females. It did appear, nevertheless, that the age of cats may be regarded as a risk factor for these parasitic infections. Histopathological examination revealed some parasite-induced lesions in the intestine and stomach, including hyperemia, hemorrhage, mucosal destruction and inflammation. The lung tissues showed some histopathological lesions such as hemorrhage, edema, emphysema and mild inflammation, and dormant larvae were found in one tongue sample. The results of the present study showed that parasitic infections and, more importantly, T. cati are relatively prevalent in stray cats, and the people living in this area are at serious risk of this zoonotic disease. The cats in this region need to be monitored, and specific preventive measures should be developed by public health officials.

A novel outlook in the delivery of artemisinin: production and efficacy in experimental visceral leishmaniasis.

The visceral form of leishmaniasis (VL), due to infection by Leishmania infantum, is a neglected tropical disease. The accessible therapeutic options are limited. Artemisinin is an efficient antileishmanial product with poor biological availability that requires high repetition of therapeutic doses in VL. Solid lipid nanoparticles (SLNs) provide targeted delivery, increase bioavailability and reduce toxicity of the traditional therapeutic strategy. The spherical shape artemisinin-loaded SLNs were prepared in a particle diameter of 222.0 ± 14.0 nm. The SLNs showed no particular toxic effect on the parasites, whereas the native artemisinin demonstrated a significant toxicity rate of 31% in viability of the promastigotes at the 250 µg/ml concentration. The therapeutic efficacy of the artemisinin-loaded SLNs was demonstrated in the experimental VL, using the L. infantum-infected BALB/c mice, in the present study. The 10 and 20 mg/kg doses of artemisinin-loaded SLNs showed higher level of antileishmanial efficacy compared with the free artemisinin. There was a significant diminishing of the parasite burden in liver (84.7 ± 4.9%) and spleen (85.0 ± 3.1%) and hepatosplenomegaly by the artemisinin-loaded SLNs treated at 20 mg/kg compared to the free artemisinin. Therefore, the present study supports the superior efficacy of artemisinin-loaded SLNs over the free artemisinin and could be considered as a new therapeutic strategy in the treatment of leishmaniasis.

Healing potential of curcumin nanomicelles in cutaneous burn wounds: an in vitro and in vivo study.

PURPOSE/AIM OF THE STUDY: Curcumin is the active substance of turmeric and has been shown to enhance the healing potential of burn wounds. However, its high hydrophobicity and rapid degradability are great challenges for its clinical applications. The development of new curcumin formulations may provide a potential solution to these issues. METHODS AND RESULTS: In this study, we investigated the use of curcumin nanomicelles for wound dressing and evaluated their effects on fibroblast migration and proliferation in vitro. We found that the application of curcumin nanomicelles to the wounds significantly improved wound contraction and increased the expression of transforming growth factor-1 and basic fibroblast growth factor at day 14 of the healing process. Furthermore, curcumin nanomicelles reduced the expression of interleukin-1 at days 7 and 14 post-wounding. Histopathological analysis revealed that the curcumin nanomicelles-treated burn wounds exhibited more organized granulation tissue, improved angiogenesis, and enhanced re-epithelialization. Additionally, the curcumin treatment led to increased hydroxyproline content and enhanced TGF-ß1 expression level in the wounds. The in vitro studies also demonstrated that the curcumin nanomicelles induced proliferation and migration of fibroblasts. CONCLUSION: Overall, our findings suggest that curcumin nanomicelles can be a promising candidate for the treatment of burn wounds.

Effects of isochronal induced feed restriction during the transition period on mRNA abundance of the hepatic genes related to lipid metabolism in fat-tailed ewes.

The process of fat mobilization during the transition period (TP) requires deep re-orchestration of the energy indices, and understanding its mechanism has generated considerable interest among the TP-related studies. The present study aims to validate the effect of feed restriction and TP on the mRNA abundance of hepatic genes related to fat metabolism in fat-tailed sheep. Twenty pregnant ewes with the age of 40.8 ± 6.2 (mean ± standard error) month were randomly assigned to control (n = 10) or restriction (n = 10), and investigated from week - 5 to 5 relative to parturition. Control animals received 100% DM during the trial. Restriction animals received 100% DM through weeks - 5, - 1, 1 and 5 and were fed with 50, 65, and 80% DM in the weeks - 4, - 3, - 2 and 2, 3, and 4, respectively. On the third week of experiment (65%) during both pre- and post-partum, the hepatic tissue was biopsied, and the mRNA load of the fatty acid synthase, acetyl-CoA carboxylase, carnitine palmitoyltransferase (CPT) 1, CPT2, and acyl-CoA synthase long-chain family member-1 genes was quantified by the TaqMan qPCR technique. Data were analyzed using the mixed model procedure of SAS. The mRNA abundance of the target genes was not influenced by feed restriction, during the pre- and post-partum periods. Parturition suppressed the mRNA abundance of target genes in both groups. It can be concluded that the fat-tailed sheep are well adapted to feed scarcity in the harsh environment and would have a higher capacity for the metabolism of fat mobilization during the negative energy balance.

Relationship between the Structure and Chaperone Activity of Human αA-Crystallin after Its Modification with Diabetes-Associated Oxidative Agents and Protective Role of Antioxidant Compounds.

The study was aimed to evaluate the impact of peroxynitrite (PON, oxidative stress agent in diabetes), methylglyoxal (MGO, diabetes-associated reactive carbonyl compound), and their simultaneous application on the structural and functional features of human αA-crystallin (αA-Cry) using various spectroscopy techniques. Additionally, the surface tension and oligomer size distribution of the treated and untreated protein were tested using tensiometric analysis and dynamic light scattering, respectively. Our results indicated that the reaction of PON and MGO with human αA-Cry leads to the formation of new chromophores, alterations in the secondary to quaternary protein structure, reduction in the size of protein oligomers, and significant enhancement in the chaperone activity of αA-Cry. To reverse the effects of the tested compounds, ascorbic acid and glutathione (main components of lens antioxidant defense system) were applied. As expected, the two antioxidant compounds significantly prevented formation of high molecular weight aggregates of αA-Cry (according to SDS-PAGE). Our results suggest that the lens antioxidant defense system, in particular, glutathione, may provide a strong protection against rapid incidence and progression of diabetic cataract by preventing the destructive reactions of highly reactive DM-associated metabolites.

Network Meta-Analysis of Chicken Microarray Data following Avian Influenza Challenge-A Comparison of Highly and Lowly Pathogenic Strains.

The current bioinformatics study was undertaken to analyze the transcriptome of chicken (Gallus gallus) after influenza A virus challenge. A meta-analysis was carried out to explore the host expression response after challenge with lowly pathogenic avian influenza (LPAI) (H1N1, H2N3, H5N2, H5N3 and H9N2) and with highly pathogenic avian influenza (HPAI) H5N1 strains. To do so, ten microarray datasets obtained from the Gene Expression Omnibus (GEO) database were normalized and meta-analyzed for the LPAI and HPAI host response individually. Different undirected networks were constructed and their metrics determined e.g., degree centrality, closeness centrality, harmonic centrality, subgraph centrality and eigenvector centrality. The results showed that, based on criteria of centrality, the CMTR1, EPSTI1, RNF213, HERC4L, IFIT5 and LY96 genes were the most significant during HPAI challenge, with PARD6G, HMG20A, PEX14, RNF151 and TLK1L having the lowest values. However, for LPAI challenge, ZDHHC9, IMMP2L, COX7C, RBM18, DCTN3, and NDUFB1 genes had the largest values for aforementioned criteria, with GTF3C5, DROSHA, ATRX, RFWD2, MED23 and SEC23B genes having the lowest values. The results of this study can be used as a basis for future development of treatments/preventions of the effects of avian influenza in chicken.

Use of a gyrB PCR-RFLP method to diagnose tuberculosis and identify the causative Mycobacterium sp. in cattle and humans.

GyrB PCR-restriction fragment length polymorphism (RFLP) could be applied to diagnose bovine and human tuberculosis and detect the causative agent. The lymph nodes and lungs from 50 cattle positive in tuberculin skin test were examined by histopathology and PCR-RFLP of a 1020-bp fragment of the gyrB gene. Swab smear samples from the nasal cavity, pleural, and abdominal cavities were also evaluated by cytological methods. Furthermore, the cultures of 50 sputum samples from the patients were assessed by PCR-RFLP using RsaI, TaqI, SacII enzymes. In histopathology, 39 cattle were positive and the acid-fast bacilli were seen in the Ziehl-Neelsen stained sections. Using gyrB PCR-RFLP, M. bovis was found as the etiological agent in 41 cattle. In terms of the human samples, the causative agent in 41 samples was M. tuberculosis, and M. bovis was isolated from two samples. It seems that gyrB PCR-RFLP could be applied as an accurate and reliable method for identifying the M. tuberculosis complex (MBTC) MBTC species. The isolation of M. bovis from the human specimens should be considered in the control strategies for tuberculosis.

Following the Trace of HVS II Mitochondrial Region Within the Nine Iranian Ethnic Groups Based on Genetic Population Analysis.

The Iranian gene pool is seen as an important human genetic resource for investigating the region connecting Mesopotamia and the Iranian plateau. The main objective of this study was to explore gene flow in nine Iranian ethnic/subpopulation groups (402 samples) by examining mtDNA HVS2 sequence variations. This then allowed us to detect mtDNA HVS2 sequence mutations in two independent thalassemia and cystic fibrosis patient sample groups. The patient groups did not explicitly belong to any of the aforementioned nine subpopulations. Across all subpopulations, the haplogroups B4a1c3a, H2a2a1, N10b, H2a2a2, and J1 were seen to be predominant. High haplogroup diversities along with admixture of the exotic groups were observed in this study. The Arab subpopulation was shown to be independent from the others. It was revealed that there is a far distant relationship between Arab and Azeri groups. The thalassemia patient group, represented an almost random sample of most Iranian ethnic groups, and revealed few significant differences (P < 0.05) in their HVS2 sequence. It turned out that the IVS II-I (G → A) mutation in the thalassemia ß-globin gene was highly significant. Since the thalassemia patients in the present study represent many unique haplotypes, we can begin to comprehend the importance of mtDNA with this disease and the necessity for more studies in this context.

Effectiveness of the adipose stem cells in burn wound healing: literature review.

Adipose- stem cells (ASCs) have received much attention in the recent years and several articles have investigated the role of these cells on burn wound healing. To understand the outcomes of the ASCs therapy on burn wound healing, a systematic review was performed. This study was conducted by searching in Pubmed, ISI, and Scopus until May 2021. Thirty-six animal studies were included in this study. The findings revealed that although treatment with ASCs somewhat enhanced the healing rate, cultured ASCs on scaffolds or its combination with hydrogels could significantly increase the viability of ASCs and promote rate of healing. However, clinical studies are necessary to gain a better understanding of the role of ASCs in burn wound healing.

Effects of Combination of BMP7, PFG, and Autograft on Healing of the Experimental Critical Radial Bone Defect by Induced Membrane (Masquelet) Technique in Rabbit.

BACKGROUND: Healing of large segmental bone defects can be challenging for orthopedic surgeons. This research was conducted to provide further insight into the effects of BMP7 in combination with autograft and platelet fibrin glue (PFG) on bone regeneration by Masquelet technique (MT). METHODS: Twenty five domestic male rabbits, more than 6 months old, weighing 2.00±0.25 kg were randomly divided into five equal groups as follows: MT-blank cavity (without any biological or synthetic materials) (1), blank cavity (2), MT-autograft (3), MT-autograft-BMP7 (4), and MT-BMP7-PFG (5). A 20 mm segmental defect was made in radial bone in both forelimbs. The Masquelet technique was done in all groups except group 2. The study was evaluated by radiology, biomechanics, histopathology and scanning electron microscopy. RESULTS: The results showed that Masquelet technique enhanced the healing process, as, the structural and functional criteria of the injured bone showed significantly improved bone healing (P<0.05). Treatment by PFG-BMP7, Autograft-BMP7, and autograft demonstrated beneficial effects on bone healing. However, Autograft-BMP7 was more effective than autograft in healing of the radial defect in rabbits. CONCLUSION: Our findings introduce the osteogenic materials in combination with Masquelet technique as an alternative for reconstruction of the big diaphyseal defects in the long bones in animal models. Our findings may be useful for clinical application in future.

Effectiveness of nuchal ligament autograft in the healing of an experimental superficial digital flexor tendon defect in equid.

This study aimed to investigate nuchal ligament (NL) autograft on experimental tendon defect healing in donkeys. Eight healthy donkeys were used. The left forelimb's superficial digital flexor (SDF) tendon was assigned as treatment, and the right forelimb was allocated as the control group (without surgical intervention). A 3×1.5 cm segment of the funicular part of the NL was excised. A full thickness defect created in the treatment tendon and was grafted with the excised NL. The following parameters were evaluated in 120 days postoperatively: clinical, ultrasonography, radiography, histopathology, biomechanical properties, and scanning and electronic transmission microscopy. There were no significant changes observed in the neck angle so that it was confirmed this treatment regimen preserved the head and neck situation without any considerable neck swelling. Weight-bearing in gait and trot was similar between both forelimbs at the end of the study. Mild to moderate adhesion was detected in the dorsal surface of the SDF tendon. There was no significant difference in the echogenicity and fiber alignment, respectively, on days 90 and 120 after surgery. Treatment significantly amplified the collagen diameter and enhanced the collagen fibril diameter and density considerably compared to the NL. The transplanted tissue was mostly in the remodeling or maturation phase, on day 120 postoperatively. It seems that the NL is biocompatible, almost biodegradable, and effective in tendon healing without metaplasia or tissue rejection.

Immunotherapy in treatment of leishmaniasis.

Leishmaniasis caused by various species of protozoan transmitted by sand fly vectors occurs as a spectrum of clinical features including cutaneous, mucocutaneous and visceral forms. It is a geographically distributed parasitic disease and a major public health problem in the world. The clinical syndromes are highly variable depending on the parasite species, host genetics, vectors and environment. To date, there is no effective vaccine and traditional treatments are toxic, expensive with long administration duration and many adverse side effects and/or drug resistance. Instead of treatments based on chemotherapy, certain strategies aim to recover leishmaniasis and reduce the parasitic burden. Immunotherapy has focused on the induction of effective immune response to rapidly control the disease. Recent studies have indicated that a single dose of a suitable therapeutic vaccine induces a quick and lasting recovery in patients. Immunotherapy reduces the toxicity of drug and the emergence of resistance dramatically. It could be an effective addition to chemotherapy with a safe and potent drug compared with monotherapy, resulting in a prophylactic and therapeutic cure of leishmaniasis. This review has focused on treatment of leishmaniasis with particular emphasis on immunotherapy as an alternative to conventional drug treatment.

Role of bone 1stem cell-seeded 3D polylactic acid/polycaprolactone/hydroxyapatite scaffold on a critical-sized radial bone defect in rat.

Osteoconductive biomaterials were used to find the most reliable materials in bone healing. Our focus was on the bone healing capacity of the stem cell-loaded and unloaded PLA/PCL/HA scaffolds. The 3D scaffold of PLA/PCL/HA was characterized by scanning electron microscopy (SEM), rheology, X-ray diffraction (XRD), and Fourier transform-infrared (FT-IR) spectroscopy. Bone marrow stem cells (BMSCs) have multipotential differentiation into osteoblasts. Forty Wistar male rats were used to organize four experimental groups: control, autograft, scaffold, and BMSCs-loaded scaffold groups. qRT-PCR showed that the BMSCs-loaded scaffold had a higher expression level of CD31 and osteogenic markers compared with the control group (P < 0.05). Radiology and computed tomography (CT) scan evaluations showed significant improvement in the BMSCs-loaded scaffold compared with the control group (P < 0.001). Biomechanical estimation demonstrated significantly higher stress (P < 0.01), stiffness (P < 0.001), and ultimate load (P < 0.01) in the autograft and BMSCs-loaded scaffold groups compared with the untreated group and higher strain was seen in the control group than the other groups (P < 0.01). Histomorphometric and immunohistochemical (IHC) investigations showed significantly improved regeneration scores in the autograft and BMSCs-loaded scaffold groups compared with the control group (P < 0.05). Also, there was a significant difference between the scaffold and control groups in all tests (P < 0.05). The results depicted that our novel approach will allow to develop PLA/PCL/HA 3D scaffold in bone healing via BMSC loading.

Effectiveness of a biodegradable 3D polylactic acid/poly(ɛ-caprolactone)/hydroxyapatite scaffold loaded by differentiated osteogenic cells in a critical-sized radius bone defect in rat.

The effects of a scaffold made of polylactic acid, poly (ɛ-caprolactone) and hydroxyapatite by indirect 3D printing method with and without differentiated bone cells was tested on the regeneration of a critical radial bone defect in rat. The scaffold characterization and mechanical performance were determined by the rheology, scanning electron microscopy, energy-dispersive X-ray spectroscopy, X-ray diffraction, and Fourier transform infrared spectrometry. The defects were created in forty Wistar rats which were randomly divided into the untreated, autograft, scaffold cell-free, and differentiated bone cell-seeded scaffold groups (n = 10 in each group). The expression level of angiogenic and osteogenic markers, analyzed by quantitative real time-polymerase chain reaction (in vitro), significantly improved (p < 0.05) in the scaffold group compared to the untreated one. Radiology and computed tomography scan demonstrated a significant improvement in the cell-seeded scaffold group compared to the untreated one (p < 0.001). Biomechanical, histopathological, histomorphometric, and immunohistochemical investigations showed significantly better regeneration scores in the cell-seeded scaffold and autograft groups compared to the untreated group (p < 0.05). The cell-seeded scaffold and autograft groups did show comparable results on the 80th day post-treatment (p > 0.05), however, most results in the scaffold group were significantly higher than the untreated group (p < 0.05). Differentiated bone cells can enhance bone regeneration potential of the scaffold.

Effects of bisphosphonates on osteoporosis: Focus on zoledronate.

Osteoporosis is a bone disease that mainly affects older people and postmenopausal women. Lack of proper treatment for this disease gives rise to many problems in patients and occasionally leads to death. Many drugs have been utilized to treat osteoporosis but the most effective one is the bisphosphonates (BPs) family. This family has several positive effects on bone tissue, including promoting bone healing, enhancing bone mineral density, reducing bone resorption, preventing pathologic fractures, suppressing bone turnover, and modulating bone remodeling. On the other hand, there have also been inconclusive reports that BPs might have a desirable or even adverse impact on osteoporotic patients. Therefore, we set out to examine the positive and negative effects of this family, with a focus on the most potent one that is zoledronate (Zol), in clinical usage. Zoledronate is an amino-BPs and nitrogen-containing drug which is the most powerful BPs on osteoporosis treatment or prevention. Many studies showed its effectiveness in the treatment of osteoporosis and bone healing. As Zol enjoys a considerable potential in treating and preventing osteoporosis, it can be used as one of the effective treatments in this field.

Comparison of botulinum toxin type A and aprotinin monotherapy with combination therapy in healing of burn wounds in an animal model.

Burns are one of the most common injuries that are complicated by many challenges including infection, severe inflammatory response, excessive expression of proteases, and scar formation. The aim of this study was to investigate the effect of botulinum toxin type A (BO) and aprotinin (AP) separately or in combination (BO-AP) in healing process. Four burn wounds were created in each rat and randomly filled with silver sulfadiazine (SSD), BO, AP and BO-AP. The rats were euthanized after 7, 14, and 28 days, and their harvested wound samples were evaluated by gross pathology, histopathology, gene expression, biochemical testing, and scanning electron microscopy. Both BO and AP significantly reduced expression of interleukin-1ß (IL-1ß) and transforming growth factor-ß1 (TGF-ß1) at the 7th post wounding day. Moreover, they inhibited scar formation by reducing the TGF-ß1 level and increasing basic fibroblast growth factor (bFGF) at the 28th day. AP by decreasing protease production showed more effective role than BO in wound regeneration. AP increased tissue organization and maturation and improved cosmetic appearance of wounds, at 28 days. The best results gained when combination of BO and AP were used in healing of burn wounds. Treatment by BO-AP significantly subsided inflammation compared to the BO, AP, and SSD treated wounds. Treatment with BO-AP also reduced collagen density and led to minimal scar formation. Combination of botulinum toxin type A and aprotinin considerably increased structural and functional properties of the healing wounds by reducing scar formation and decreasing production of proteases.

Hyaluronic acid hydrogel loaded by adipose stem cells enhances wound healing by modulating IL-1ß, TGF-ß1, and bFGF in burn wound model in rat.

Application of hydrogels can be an effective technique in transferring the adipose-derived stem cells (ASCs) to injured tissue and their protection from further complications. Besides, acellular dermal matrix (ADM) has successfully been used in treatment of wounds. In this study, a combination of hylauronic acid (HA) and ASCs (HA/ASCs) was applied on burn wounds and the injured area was then covered by an ADM dressing in a rat model (ADM-HA/ASCs). Wound healing was evaluated by histopathological, histomorphometrical, molecular, biochemical, and scanning electron microscopy assessments on days 7, 14, and 28 post-wounding. ADM-HA/ASCs stimulated healing significantly more than the ADM-HA and ADM treated wounds, as it led to reduced inflammation, and improved angiogenesis and enhanced granulation tissue formation. Expression of interleukin-1ß (IL-1ß) and transforming growth factor-ß1 (TGF-ß1) was lower in the ADM-HA/ASCs treated wounds than the ADM-HA and ADM groups, at the seventh post-wounding day. ADM-HA/ASCs also enhanced the expression level of TGF-ß1 mRNA at 14 day post-wounding that was parallel to the experimental data from histological and biochemical assessments and confirmed the positive role of ASCs in repair of burn wounds. Additionally, increase in basic fibroblast growth factor (bFGF) expression and decreased TGF-ß1 level on the 28th post-wounding day indicated the anti-scarring activity of ASCs. HA loaded by adipose stem cells can represent a promising strategy in accelerating burn wound healing.