Herbal-based compounds: A review on treatments of cryptosporidiosis.

Cryptosporidium, a monoxenous apicomplexan coccidia, is a prevalent diarrhetic and an opportunistic agent, mainly in immunocompromised individuals. As there are few chemotherapeutic compounds that have limited efficacy, we need to identify new compounds or specific parasite targets for designing more potent drugs to treat cryptosporidiosis. Herbal products with low toxicity, environmental compatibility, wide therapeutic potential, and abundant resources can be considered alternatives for treatment. The current review tried to summarize the studies on plants or herbal bioactive constituents with anti-cryptosporidial activities. Based on constituents, plants act via different mechanisms, and further investigations are needed to clarify the exact mechanisms by which they act on the developmental stages of the parasite or host-parasite relationships.

Immuno-reactivity evaluation of Mce-truncated subunit candidate vaccine against Mycobacterium avium subspecies paratuberculosis challenge in the goat models.

BACKGROUND: Detection of an appropriate antigen with high immunogenicity can be a big step in the production of an effective vaccine for control of Johne's disease (JD). The aim of this study was to evaluate the efficacy of Mce-truncated protein as a subunit vaccine candidate for the control of JD in experimentally challenged goats. MATERIALS AND METHODS: Six healthy goat kids were immunized with Mce-truncated protein, and two goats were kept as controls. All kids were twice challenged orally with live Mycobacterium avium subspecies paratuberculosis(MAP) strain and half the goats from both the categories were sacrificed at 7 and 10 months after start of challenge study. Culture of MAP was performed from all the necropsied tissues to determine the true JD infection status. RESULTS: Mce-truncated protein only reacted with pooled vaccinated goat sera in western-blot. A significant increase in humoral immune response against Mce protein was also observed in vaccinated goats. Compared to the control group, vaccinated goats gained higher body weights and none of them shed MAP or showed histopatological lesions or colonization of MAP in their necropsy tissues. CONCLUSIONS: The new Mce protein based vaccine provided significant immunity in goats as they could meet the challenge with live MAP bacilli. Although the vaccine used in this study showed the high potential as a new effective vaccine for the control of JD, further validation study is still required to successfully implement the vaccine for JD control program.

Evaluation of delaying effects of different short-term dosage regimens of topical ciprofloxacin on corneal ulcer healing in an avian model.

BACKGROUND: Knowledge on possible delaying effects of topical ciprofloxacin on corneal ulcer healing is scarce in avian patients. OBJECTIVES: The study evaluates effects of different dosage regimens of topical ciprofloxacin on healing of corneal ulcer in an avian model. METHODS: One hundred and fifty adult layers were randomly allocated into two equal categories each consisted of 5 groups (n = 15): 1, negative control (NC, normal cornea); 2, positive control (PC) (birds with experimental corneal ulcer); and 3, 4 and 5, birds with corneal injury that received ciprofloxacin 0.3% topically q6h, q8h and q12h, respectively for 3 (category 1) or 5 days (category 2). Corneas were excised for histopathological evaluation and determination of MMP-9 expression. RESULTS: While no significant difference was observed in daily-measured fluorescein-stained ulcer size among ciprofloxacin-treated birds and PC group in category 1, birds in PC group of category 2 had significantly smaller ulcers as compared to antibiotic-treated birds at the end of experiment (p < 0.01 for all cases). Histopathological evaluations at the end of the experiment showed no significant difference among PC and ciprofloxacin-treated birds of both categories for almost all of the assayed parameters. Over expression of MMP-9 mRNA was observed in PC group after 3 and 5 days of ulcer induction compared to NC groups. Its expression in ciprofloxacin-treated birds of both categories remained close to PC groups. CONCLUSIONS: While ciprofloxacin administration for 3 days does not affect ulcer healing, it delays healing process at the end of 5 days of treatments in an avian model of corneal ulcer injury. This delaying effect is not associated with a drastic change in MMP-9 expression.

In vivo assessment of simultaneous G1 cyclins silencing by a tumor-specific bidirectional promoter on the mammary tumor in nude mice.

Dysregulation of G1 cyclins (cyclins D1 A and E) expression contributes to the loss of standard cell cycle control during tumorigenesis. This study aims to evaluate the inhibitory effect of G1 cyclins in nude mice. The human breast cancer MDA-MB-231 cells were subcutaneously transplanted into the supra-femoral right side of female Balb/c-nude mice. The dual shRNA vector harboring G1 cyclins shRNAs (bipSUR) was intratumorally injected by the in vivo jetPEI transfection reagent for 2 weeks. We have evaluated tumor growth and tumor weight as parameters of tumor progression. Finally, necropsy, histopathological analysis, and immunodetection of G1 cyclins were assessed. Also, apoptosis induction in tumor tissues was evaluated by TUNEL assay. No toxicity and metastasis was observed in the tumor-bearing mice treated by the bipSUR. Tumor weight and volume were significantly lower in the bipSUR treated mice than untreated tumor-bearing mice and control. Histopathological observations revealed more apoptotic foci and lower mitotic cells in tumor sections in the treated mice than in control groups. A significant reduction of G1 cyclins at the protein level was indicated in the bipSUR treated mice than in other groups. Apoptosis in tumor tissues was remarkably induced in response to the bipSUR (42.53%). The bipSUR reduced the protein expression of G1 cyclins and exhibited an inhibitory effect on MDA-MB-231 xenograft mice through apoptosis induction. Further research is demanded to identify the protein partners of G1 cyclins involved in the cancer pathways. These may offer new insight into the biomedical function of G1 cyclins in breast cancer progression.

The effect of Urtica dioica extract on oxidative stress, heat shock proteins, and brain histopathology in multiple sclerosis model.

Multiple sclerosis (MS) results from the destruction of myelin and focal inflammation. The study aimed to evaluate the effect of hydroalcoholic extract of Urtica dioica on oxidative stress, heat shock proteins, and brain histopathology in multiple sclerosis model. Sixty male C57BL/6 mice were divided into six groups of 10. Groups included positive control, negative control, and treatment groups with U. dioica extract at a dose of 50, 100, 200, and 400 mg/kg for 21 days (three times a week). The MS model was developed by a diet containing 0.2% cuprizone for 6 weeks. A section of brains was evaluated with Luxol Fast Blue staining and the other part evaluated with heat shock protein (HSP) kits 60 and 70, total antioxidant capacity (TAC), and malondialdehyde (MDA). In sections of corpus callosum, the highest amount of myelin was observed in the negative controls, while the use of cuprizone in the positive controls caused the destruction and reduction of myelin. The use of U. dioica extract in therapeutic groups except at a dose of 50 mg/kg could reduce myelin degradation to some extent and lead to remyelination. However, myelin levels in treatment groups were not significantly different from any of the negative and positive controls. Although HSP60 decreased in the treatment groups, there was no significant difference between the positive and negative controls. Treatment with this extract significantly reduced the amount of HSP70 compared with the positive controls. The decreased TAC and increased MDA in positive controls indicated oxidative stress, respectively. Furthermore, the extract led to an increase and decrease of TAC and MDA in the treatment groups, respectively. However, only the MDA level was significantly different from that of the positive controls. Therefore, the antioxidant effects of U. dioica extract could decrease cuprizone-induced oxidative stress and be effective in improving demyelination.

Comparison of the Efficacy of HIV-1 Nef-Tat-Gp160-p24 Polyepitope Vaccine Candidate with Nef Protein in Different Immunization Strategies.

OBJECTIVES: One of the promising strategies for effective HIV-1 vaccine design involves finding the polyepitope immunogens using T cell epitopes. METHODS: Herein, an HIV-1 polyepitope construct (i.e., Nef-Tat-Gp160-P24) comprising of several epitopes from Nef, Tat, Gp160, and P24 proteins was designed. To improve its immunogenicity in BALB/c mice, cell-penetrating peptides (HR9 and MPG for DNA delivery, and LDP-NLS and Cy- LoP-1 for protein transfer), Montanide adjuvant, and heterologous DNA prime/polypeptide boost strategy were used. To compare the immunogenicity, Nef was utilized as a vaccine candidate. The levels of total IgG and its subclasses, cytokines, and Granzyme B were assessed using ELISA. RESULTS: Immunological studies showed that heterologous prime-boost regimens for both antigens could considerably augment the levels of IgG2a, IgG2b, IFN-γ, and Granzyme B directed toward Th1 and CTL immune responses in comparison with homologous prime-boost strategies. The levels of IFN-γ, IL-10, total IgG, IgG1, and IgG2b were drastically higher in groups immunized with Nef-Tat-Gp160-P24 in heterologous prime-boost regimens than those in groups immunized with Nef. CONCLUSION: The use of the Nef-Tat-Gp160-P24 polyepitope immunogen in heterologous primeboost strategy could generate the mixture of Th1 and Th2 responses directed further toward Th1 response as a hopeful method for improvement of HIV-1 vaccine.

Treatment effects of Shilajit on aspirin-induced gastric lesions in rats.

The present study investigated the effects of Shilajit extract on aspirin-induced gastric lesions in rats. We evaluated macroscopic and histopathological lesions in the stomach, measured the activity of oxidative stress enzymes in gastric tissue homogenates, and assessed serum electrolytes and parameters of kidney and liver function. Forty-five male rats were allocated to five groups: Normal control, positive control, omeprazole treatment, Shilajit treatment, and Shilajit control. The treatment period lasted for four consecutive days. The size and number of gastric lesions were significantly reduced in the Shilajit and omeprazole groups compared to the positive control group, indicating a reduction in mucosal damage and the severity of edema and leukocyte infiltration in tissue sections. A significant increase was observed in the levels of all oxidative stress parameters, except malondialdehyde, in rats treated with Shilajit and omeprazole compared to those in the positive control group. The effect of the aqueous extract of Shilajit was comparable to that of omeprazole. These results indicated the protective effects of Shilajit against aspirin-induced gastric lesions.

Protective effect of black mulberry (Morus nigra L) fruit hydroalcoholic extract against testosterone-induced benign prostatic hyperplasia in rats / Efecto protector del extracto hidroalcohólico de las moras (Morus nigra L) en la hiperplasia prostática benigna no inducida por testosterona, en ratas

BACKGROUND: Finding new agents for prevention and/or treatment of benign prostatic hyperplasia (BPH) especially from natural sources is a demanding field. OBJECTIVES: This study aimed to evaluate the effect of black mulberry (BM) (Morus nigra) fruit hydroalcoholic extract on the establishment of BPH in rats. MATERIALS AND METHODS: Forty-nine adult male rats were randomly assigned into 7 equal groups: I: Sham control (SC), a sham surgery was performed. II: positive control (PC), rats were castrated and received testosterone propionate, at 10mg/kg/day S.C. for BPH induction. III: comparative control (CC), BPH was induced and the rats received finasteride at 5mg/kg/day P.O. IV-VII: (T1-T4): BPH was induced and the rats received BM extract at 25, 50, 100 and 200mg/kg/day P.O. for 4 consecutive weeks. RESULTS: Finasteride and/or BM extract especially at the two higher dosages, significantly affected prostate weight, prostatic index, percent of inhibition, serum and prostatic levels of dihydrotestosterone (DHT), serum prostate-specific antigen (PSA), antioxidant parameters of prostatic tissue as well as histopathological and histomorphometric parameters (epithelial thickness and acinar area) of prostate. CONCLUSIONS: BM extract has protective effects against experimentally-induced BPH in rats with regard to histopathological and biochemical parameters which may be related to its antioxidant as well as DHT reducing properties in prostatic tissue

ANTECEDENTES: El hallazgo de nuevos agentes para prevenir y/o tratar la hiperplasia prostática benigna (HBP), procedentes especialmente de fuentes naturales, es un campo exigente. OBJETIVOS: El objetivo de este estudio fue evaluar el efecto del extracto hidroalcohólico de las moras (Morus nigra) sobre el establecimiento de HBP en ratas. MATERIALES Y MÉTODOS: Se asignaron aleatoriamente 49 ratas adultas macho en 7 grupos iguales: I: grupo control (GC), en el que se practicó cirugía de control; II: control positivo (CP), en el que se castró a las ratas y se les administró propionato de testosterona, a una dosis de 10mg/kg/día sc para inducción de BPH. III: control comparativo (CC), en el que se indujo BPH y se administró a las ratas finasterida a una dosis de 5mg/kg/día po; IV-VII: (T1-T4), en el que se indujo BPH y se administró a las ratas extracto de moras a una dosis de 25, 50, 100 y 200mg/kg/día po durante 4 días consecutivos. RESULTADOS: La finasterida y/o el extracto de moras, especialmente en 2 dosis elevadas, afectaron significativamente al peso prostático, al índice prostático, al porcentaje de inhibición, a los niveles séricos y prostáticos de dihidrotestosterona (DHT), al antígeno prostático específico sérico (PSA), a los parámetros antioxidantes del tejido prostático, así como a los parámetros histopatológicos e histomorfométricos (espesor epitelial y área acinar) de la próstata. CONCLUSIONES: El extracto de mora tiene efectos protectores frente a HPB experimentalmente inducido en ratas, con respecto a sus parámetros histopatológicos y bioquímicos, y que pueden guardar relación con sus propiedades antioxidantes y reductoras de DHT en el tejido prostático

Lumpy skin disease, an emerging transboundary viral disease: A review.

Lumpy skin disease is an emerging bovine viral disease, which is endemic in most African countries and some Middle East ones, and the elevated risk of the spread of disease into the rest of Asia and Europe should be considered. The recent rapid spread of disease in currently disease-free countries indicates the importance of understanding the limitations and routes of distribution. The causative agent, Capripoxvirus, can also induce sheeppox and goatpox. The economic significance of these diseases is of great concern, given that they threaten international trade and could be used as economic bioterrorism agents. The distribution of capripoxviruses seems to be expanding due to limited access to effective vaccines and poverty within farming communities. This is largely due to the economic effects of the Covid-19 pandemic and the imposition of crippling sanctions in endemic regions, as well as an increase in the legal and illegal trade of live animals and animal products, and also global climate change. The present review is designed to provide existing information on the various aspects of the disease such as its clinicopathology, transmission, epidemiology, diagnosis, prevention and control measures, and the potential role of wildlife in the further spread of disease.

In vivo delivery of a multiepitope peptide and Nef protein using novel cell-penetrating peptides for development of HIV-1 vaccine candidate.

OBJECTIVES: A potent HIV vaccine should overcome some limitations such as polymorphism of human HLA, the diversity of HIV-1 virus, and the lack of an effective delivery system. In this study, a DNA construct encoding Nef60-84, Nef126-144, Vpr34-47, Vpr60-75, Gp16030-53, Gp160308-323, and P248-151 epitopes was designed using bioinformatics tools. The pcDNA3.1-nef-vpr-gp160-p24 and pcDNA3.1-nef constructs were prepared in large scale as endotoxin-free form. Moreover, the recombinant Nef-Vpr-Gp160-p24 polypeptide and Nef protein were generated inE. coli. These constructs were delivered using cell penetrating peptides (CPPs) in vivo, and immune responses were assessed for different modalities in BALB/c mice. RESULTS: The recombinant DNA constructs were confirmed as the ~ 867 bp and ~ 648 bp bands related tonef-vpr-gp160-p24 andnef genes on agarose gel. Moreover, the purified Nef-Vpr-Gp160-p24 polypeptide and Nef protein showed the ~ 32 kDa and ~ 30 kDa bands on SDS-PAGE, respectively. The results of immune responses indicated that the heterologous prime/boost regimens using both Nef-Vpr-Gp160-P24 and Nef antigens induced significantly the secretion of IgG2a, IgG2b, IFN-γ and Granzyme B compared to other groups. The levels of Granzyme B in mice immunized with Nef antigen were higher than those immunized with Nef-Vpr-Gp160-P24 antigen. The CPPs showed the same potency with Montanide adjuvant for eliciting immune responses. CONCLUSIONS: The heterologous prime/boost regimens for both antigens could significantly direct immune responses toward Th1 and CTL activity compared to other regimens. Comparing the efficiency of Nef-Vpr-Gp160-P24 and Nef constructs, the Nef-Vpr-Gp160-P24 constructs delivered by CPPs showed promising results as an HIV vaccine candidate.

Protective effect of black mulberry (Morus nigra L.) fruit hydroalcoholic extract against testosterone-induced benign prostatic hyperplasia in rats.

BACKGROUND: Finding new agents for prevention and/or treatment of benign prostatic hyperplasia (BPH) especially from natural sources is a demanding field. OBJECTIVES: This study aimed to evaluate the effect of black mulberry (BM) (Morus nigra) fruit hydroalcoholic extract on the establishment of BPH in rats. MATERIALS AND METHODS: Forty-nine adult male rats were randomly assigned into 7 equal groups: I: Sham control (SC), a sham surgery was performed. II: positive control (PC), rats were castrated and received testosterone propionate, at 10mg/kg/day S.C. for BPH induction. III: comparative control (CC), BPH was induced and the rats received finasteride at 5mg/kg/day P.O. IV-VII: (T1-T4): BPH was induced and the rats received BM extract at 25, 50, 100 and 200mg/kg/day P.O. for 4 consecutive weeks. RESULTS: Finasteride and/or BM extract especially at the two higher dosages, significantly affected prostate weight, prostatic index, percent of inhibition, serum and prostatic levels of dihydrotestosterone (DHT), serum prostate-specific antigen (PSA), antioxidant parameters of prostatic tissue as well as histopathological and histomorphometric parameters (epithelial thickness and acinar area) of prostate. CONCLUSIONS: BM extract has protective effects against experimentally-induced BPH in rats with regard to histopathological and biochemical parameters which may be related to its antioxidant as well as DHT reducing properties in prostatic tissue.

Detection of Mycoplasma capricolum subsp. capripneumoniae and Mannheimia haemolytica as causative agents of pleuropneumonia in goats / Detección de Mycoplasma capricolum subsp. capripneumoniae y Mannheimia haemolytica como agentes causales de pleuro-neumonia en cabras

Abstract Objective: The aim of the study was detection of two major causative agents of pleuropneumonia, Mycoplasma capricolum subsp. capripneumoniae (Mccp) and Mannheimia haemolytica, in goats. To the best of our knowledge, this study is the first investigation of Mccp in Iran. Methods: 50 grossly suspected lungs to pleuropneumonia and 10 healthy samples were collected from Shiraz abattoir. Results: Histopathological evaluation of tissue samples showed various diagnosed pneumonias including 40% bronchointerstitial pneumonia (20 samples), 34% interstitial pneumonia (17 samples), 10% fibrinopurulent bronchopneumonia (5 samples), 12% purulent bronchopneumonia (6 samples) and 4% chronic pneumonia (2 samples). In molecular study, all 50 suspected samples and 10 healthy ones by PCR showed no Mccp positive sample, but the detection rate of M. haemolytica in suspected samples was 14% and in healthy lungs was zero. Conclusions: It may be concluded that goats referred to Shiraz abattoir is free of Mccp. Further sampling and molecular testing at the level of suspected herds to CCPP can be useful.

Resumen Objetivo: El objetivo del estudio fue la detección de dos agentes causantes principales de pleuroneumonía, Mycoplasma capricolum subsp. Capripneumoniae (Mccp) y Mannheimia haemolytica, en cabras. Hasta donde sabemos, este estudio es la primera investigación de Mccp en Irán. Métodos: 50 pulmones muy sospechosos de pleuroneumonía y 10 muestras sanas se obtuvieron del matadero de Shiraz. Resultados: La evaluación histopatológica de muestras de tejido mostró varias neumonías diagnosticadas, incluyendo 40% de neumonía broncointersticial (20 muestras), 34% de neumonía intersticial (17 muestras), 10% de bronconeumonía fibrinopurulenta (5 muestras), 12% de bronconeumonía purulenta (6 muestras) y 4% neumonía crónica (2 muestras). En un estudio molecular, las 50 muestras sospechosas y 10 sanas por PCR no mostraron una muestra positiva de Mccp, pero la tasa de detección de M. haemolytica en muestras sospechosas fue del 14% y en pulmones sanos fue cero. Conclusiones: se puede concluir que las cabras referidas al matadero Shiraz están libres de Mccp. La realización de muestreo adicional y pruebas moleculares a nivel de rebaños sospechosos para CCPP puede ser útil.

Histopathological and immunohistochemical characterization of testicular T-cell lymphoma in a dog.

Canine lymphoma is the most common neoplasm of the hematopoietic system with the most frequently diagnosed malignancy. Also, dogs affected by T-cell lymphoma displayed a lower rate of complete chemotherapy response and have a high risk of death in the early stage of the disease. A 5-year-old male mixed dog was presented to the Small Animal Hospital, School of Veterinary Medicine, Shiraz University, Shiraz, Iran with a history of left testicle swelling for four months. The testicle had approximately 13.00×10.00 cm size with a greyish-white cut surface. Histopathologically, the testicular tissue was infiltrated by neoplastic cells as only a few degenerated seminiferous tubules have remained. The neoplastic cells were round to oval with pleomorphic nuclei and single or multiple prominent nucleoli. Immunohistochemical analysis revealed positive and negative immunoreactivity for CD3 and CD20, respectively. According to histopathological and immunohistochemical features, the tumor was concluded as a testicular T-cell lymphoma. It seems that recognition of lymphoma type could be helpful for clinicians therapeutic protocols.

Regenerative effect of mesenteric fat stem cells on ccl4-induced liver cirrhosis, an experimental study.

BACKGROUND: Liver cirrhosis is a chronic disease in which normal liver tissue is replaced by fibrous tissue, leads to liver malfunction. Although transplantation is the most certain cure, stem cell therapies are shedding light on efforts to regenerate cirrhotic liver. The purpose of this study was to evaluate the regenerative potential of mesenteric fat stem cells in CCL4-induced liver cirrhosis in an animal model. METHODS: Thirty rats were treated with the mixture of CCL4 and olive oil intraperitoneally by a dose of 0.2 ml (0.1 ml CCL4 and 0.1 ml olive oil) every other day for 16 weeks till cirrhosis signs appeared. Fifteen rats were randomly selected as control group. Others treated by mesenteric fat derived mesenchymal stem cells transferred into the liver parenchyma. RESULTS: After 5 weeks, rats received stem cells had improved clinically by increased movements, appetite, improvement in overall behavior and decreased abdomen size. Histopathologically, liver cells showed state of regeneration and forming new colonies. CONCLUSION: Liver cirrhosis was induced. The mesenchymal stem cells derived from mesenteric adipose tissue could improve hepatic status of the rats, as cirrhotic livers were regenerated back into normal appearing parenchyma. Rats' clinical behavior also reached healthy status.

First Cases of SARS-CoV-2 in Iran, 2020: Case Series Report.

In Jan 2020, the outbreak of the 2019 novel coronavirus (SARS-CoV-2) in Wuhan, Hubei Province of China spread increasingly to other countries worldwide which WHO declared it as a public health emergency of international concern. Iran was included in the affected countries. Throat swab specimens were collected and tested by using real-time reverse transcription PCR (RT-PCR) kit targeting the E region for screening and RNA dependent RNA polymerase for confirmation. Conventional RT-PCR was conducted for the N region and the PCR products were sequenced by Sanger sequencing. The first seven cases of SARS-CoV-2 infections were identified in Qom, Iran. This report describes the clinical and epidemiological features of the first cases of SARS-CoV-2 confirmed in Iran. Future research should focus on finding the routes of transmission for this virus, including the possibility of transmission from foreign tourists to identify the possible origin of SARS-CoV-2 outbreak in Iran.

In vitro and in vivo activity of cinnamaldehyde against Eimeria kofoidi in chukar partridge (Alectoris chukar).

One hundred and twenty one-day-old chukar partridges were randomly divided into eight groups which received diets with different supplementations. There were four unchallenged groups. One group received salinomycin (50 ppm), two groups received cinnamaldehyde (CINN) (100 and 200 mg/kg of diet), and another one received only the basal diet from the 1st to the 31st day. There were also four corresponding groups orally challenged by 3 × 105Eimeria kofoidi sporulated oocysts at the 21st day. Three samplings were done at the 24th, 26th, and 31st days of rearing for pathological and biochemical assessments. Fecal samples were daily taken to check the pattern of oocyst shedding from the 26th to 31st day. The body weight of birds was measured at 21st and 31st days. Along with the in vivo experiment, an in vitro sporulation inhibition test was carried out. The in vitro results showed that CINN decreased sporulation rate at 1 and 0.5 mg/ml. In vivo, it was found that CINN did not prevent the oocyst shedding. Furthermore, the histopathological findings revealed that CINN and salinomycin had no effect on infection establishment. However, our findings showed that CINN (200 mg/kg of diet) could enhance the body weight and improve antioxidant status. Although our results did not support the in vivo anticoccidial activity of CINN, it had a promising potential to improve antioxidant status and body weight in the chukar partridge.

Delivery of HIV-1 Polyepitope Constructs Using Cationic and Amphipathic Cell Penetrating Peptides into Mammalian Cells.

BACKGROUND: An effective vaccine against human immunodeficiency virus 1 (HIV-1) is an important global health priority. Despite many efforts in the development of the HIV-1 vaccine, no effective vaccine has been approved yet. Recently, polyepitope vaccines including several immunogenic and conserved epitopes of HIV-1 proteins have received special attention. METHODS: In this study, HIV-1 Nef, Tat, Gp160 and P24 proteins were considered for selection of immunodominant and conserved epitopes due to their critical roles in the viral life cycle and pathogenesis. At first, the Nef60-84-Nef126-144-Tat29-49-Gp16030-53-Gp160308-323-P248-151 DNA construct was designed using in silico studies. Then, the DNA construct was subcloned in pEGFP-N1 and pET- 24a (+) expression vectors and the rNef-Tat-Gp160-P24 polyepitope peptide was generated in E.coli expression system for in vitro delivery using novel cell-penetrating peptides (CPPs), LDP-NLS and CyLoP-1, in a non-covalent manner. Also, the HR9 and MPG CPPs were used to transfer the DNA construct. RESULTS: Our results showed that the recombinant polyepitope peptide generated in Rosetta strain migrated as a clear band of ~31 kDa in SDS-PAGE. The SEM data confirmed the formation of stable nanoparticles with a size below 250 nm. MTT assay revealed that the complexes did not represent any considerable cytotoxic effect compared to untreated cells. The results of fluorescence microscopy, flow cytometry and western blotting indicated that these CPPs successfully delivered polyepitope constructs into HEK-293T cell line. CONCLUSION: These data suggested that these CPPs can be used as a promising approach for the development of the HIV-1 vaccine.

Immunomodulatory effects of Calcitriol in acute spinal cord injury in rats.

Pharmacological therapy options for spinal cord injury (SCI) in acute phase have so far been limited, thus we focused on Calcitriol, FDA-approved biologically active form of vitamin D whose neuroprotective effects are increasingly recognized, to ameliorating damage following acute SCI in rats. Calcitriol (1 µg/kg) treatment for 7 consecutive days after SCI was compared SCI control and Sham control rat groups. Calcitriol-treated group had significantly improved outcome in standard functional recovery evaluation test (BBB) 12 weeks after SCI compared to SCI control, which was confirmed by increased ventral horn motor neurons in Calcitriol-treated group. In addition, proliferation test performed on lymphocytes from spleen and lymph nodes one week after SCI showed that calcitriol injection has a significant regulatory effect on Division Index (DI) in response to MBP stimulation compared to control SCI groups, which was associated with significant reduction in IFN-γ and IL-17A secretion and leukocyte infiltration into injury site. Along with confirmation of immunoregulatory aspects of Calcitriol treatment against myelin antigens in SCI, this study has shown that reducing the extent of progressive tissue loss by Calcitriol therapy in acute phase, could result in better recovery after SCI.

A Laboratory Strain of Leishmania major: Protective Effects on Experimental Leishmaniasis.

PURPOSE: Leishmaniasis, as one of the most important vector-borne and zoonotic diseases, can be seen in different forms and is more prevalent in developing countries worldwide. Due to the absence of effective strategies in its prevention, treatment, and control, investigation of effective control strategies against the disease is necessary. In this research, we evaluated the immunogenicity of a cold-adapted laboratory strain of Leishmania major (LMC) in the mouse model. METHODS: Twenty BALB/c mice were divided into two groups. LMC group received 4 × 106 of LMC strain in 0.5 ml DMEM, and VLM group, as the control group, received 0.5 ml Dulbecco's modified Eagle's medium. Both groups were challenged with virulent L. major 3 weeks after inoculation. RESULTS: The data obtained from the analysis of immune responses and histopathological changes interestingly revealed protection against L. major in immunized mice. Compared with the VLM group, the mice immunized with LMC strain of L. major in the LMC group showed a significant increase in IFN-γ and IgG2a levels (P < 0.05) which are important indexes for Th1-related immune responses. Additionally, significant differences in concentration of IgG1 and IgG total before and after the challenge was observed in LMC group (P < 0.05). Furthermore, the immunized mice showed a significant reduction in mean sizes of skin lesion and liver damage compared to the VLM group. CONCLUSION: Based on the present findings on immunogenicity of LMC strain, it seems this strain is able to induce both humoral and cellular immunity and a significant protection against L. major in the mouse model.

Curcumin: A promising treatment for Cryptosporidium parvum infection in immunosuppressed BALB/c mice.

Members of the genus Cryptosporidium are frequent protozoan pathogens in humans and a wide range of animals. There is no consistently effective treatment against cryptosporidiosis, especially in immunodeficient patients. The present study was carried out to study the therapeutic effects of curcumin against cryptosporidiosis in immunosuppressed BALB/c mice. Mice were divided into five groups and immunosuppressed by dexamethasone. Three groups were inoculated with C. parvum oocysts, administered with curcumin, paromomycin, and without treatment. The reminders were regarded as controls. The oocysts in the fecal smear were counted daily. At days 0, 3, 7, and 11 post-treatment, the mice were sacrificed, and the efficacy of drugs was evaluated by comparing the histopathological alterations in jejunum and ileum, measuring the total antioxidant capacity, and malondialdehyde in the affected tissues. The infection was completely eliminated in the curcumin-treated group, and oocyst shedding stopped with no recurrence after drug withdrawal. On the contrary, paromomycin was unable to eliminate C. parvum infection completely, and oocyst shedding continued even 10 days after the drug withdrawal. Based on these findings, curcumin can be a trustworthy compound for the elimination of infection in immunosuppressed hosts. Further evaluation to find its accurate mechanism of action should be considered.