Infected Rats Research Articles

Chemotherapeutic potentials of β-ionone against Trypanosoma congolense infection: Inhibitionofparasite proliferation, anemia development, trans-sialidase (TconTS3 and TconTS4) gene expressions, and phospholipase A2.

Trypanosoma congolense is a pathogenic African animal trypanosome species causing devastating conditions leading to death of an infected host. The drawbacks of the existing trypanocidal drugs have led to the search for new drug candidates. In this study, β-ionone at 15 and 30mg/kg body weight (BW) was orally administered to T.congolense infected rats for 14days followed by an assessment of anemia, organ damages, and the expression of T.congolense trans-sialidase gene variants. A significant decrease in parasitemia (p<.05) was observed in the animals treated with 15mg/kg BW β-ionone besides increased animal survival rate. A trypanosome-induced decrease in packed cell volume (PCV) and histopathological changes across tissues was significantly (p<.05) ameliorated following treatment with both doses of β-ionone. This is in addition to reversing the parasite-induced upsurge in free serum sialic acid (FSA) and expression of T.congolense trans-sialidase gene variants (TconTS1, TconTS3, and TconTS4). Correlation analysis revealed a positive correlation (p>.05) between FSA with the TconTS gene expressions. In addition, the compound inhibited partially purified T.congolense sialidase and phospholipase A2 via mixed inhibition pattern with inhibition binding constants of 25.325 and 4.550µM, respectively, while molecular docking predicted binding energies of -5.6kcal/mol for both enzymes. In conclusion, treatment with β-ionone suppressed T.congolense proliferation and protected the animals against some of the parasite-induced pathologies whilst the effect on anemia development might be due to inhibition of sialidase and PLA2 activities as well as the expression levels of TconTS3 and TconTS4.

The therapeutic potential of skin mucus from Asian swamp eel (Monopterus albus): In vivo evaluation and histological evidence

ObjectivesThe Asian swamp eel (Monopterus albus), is commonly distributed in Asian countries. However, its therapeutic potential has not been thoroughly investigated yet. The current study aimed to evaluate the in-vivo therapeutic properties of the skin mucus of this fish. MethodsThe eel mucus was collected fleshly and topical gel with carbopol 934 was formulated to study the antibacterial activity on the infected skin of the rats. Sprague Dawley rats were used in the study and divided into 4 groups negative, positive, normal control, and treated groups. ResultsIntracutaneous injections of pathogenic bacteria (Streptococcus pyogenes, Staphylococcus aureus) and fungi (Microsporum gypseum, Candida albicans) were injected into the rats. The development of tinea capitis, impetigo, and cutaneous candidiasis in the animal model was confirmed based on clinical and histopathological observations. To treat the infected rats, a formulated gel of eel skin mucus was applied on the infected rat’s skins topically. The histological analysis confirms a complete recovery in the skin tissues similar to commercial antifungal and antibacterial agents used in the positive control groups. ConclusionThe present novel eel skin mucus is an efficient therapeutic candidate in treating skin infections associated with pathogenic microbes.

Alloyed nanostructures integrated metal-phenolic nanoplatform for synergistic wound disinfection and revascularization

New materials for combating bacteria-caused infection and promoting the formation of microvascular networks during wound healing are of vital importance. Although antibiotics can be used to prevent infection, treatments that can disinfect and accelerate wound healing are scarce. Herein, we engineer a coating that is both highly compatible with current wound dressing substrates and capable of simultaneously disinfecting and revascularizing wounds using a metal-phenolic nanoplatform containing an alloyed nanostructured architecture (Ag@Cu-MPNNC). The alloyed nanostructure is formed by the spontaneous co-reduction and catalytic disproportionation reaction of multiple metal ions on a foundation metal-phenolic supramolecular layer. This synergistic presence of metals greatly improves the antibacterial activity against both Gram-negative and Gram-positive pathogenic bacteria, while demonstrating negligible cytotoxicity to normal tissue. In infected rat models, the Ag@Cu-MPNNC could kill bacteria efficiently, promoting revascularization and accelerate wound closure with no adverse side effects in infected in vivo models. In other words, this material acts as a combination therapy by inhibiting bacterial invasion and modulating bio-nano interactions in the wound.

Infection with intestinal helminth (Hymenolepis diminuta) impacts exploratory behavior and cognitive processes in rats by changing the central level of neurotransmitters.

Parasites may significantly affect the functioning of the host organism including immune response and gut-brain-axis ultimately leading to alteration of the host behavior. The impact of intestinal worms on the host central nervous system (CNS) remains unexplored. The aim of this study was to evaluate the effect of intestinal infection by the tapeworm Hymenolepis diminuta on behavior and functions of the CNS in rats. The 3 months old animals were infected, and the effects on anxiety, exploration, sensorimotor skills and learning processes were assessed at 18 months in Open Field (OF), Novel Object Recognition (NOR) and the Water Maze (WM) tests. After completing the behavioral studies, both infected and non-infected rats were sacrificed, and the collected tissues were subjected to biochemical analysis. The levels of neurotransmitters, their metabolites and amino acids in selected structures of the CNS were determined by HPLC. In addition, the gene expression profile of the pro- and anti-inflammatory cytokines (TNF-α, IL-1β, IL-6 and IL-10) was evaluated by Real-Time PCR to determine the immune response within the CNS to the tapeworm infection. The parasites caused significant changes in exploratory behavior, most notably, a reduction of velocity and total distance moved in the OF test; the infected rats exhibited decreased frequency in the central zone, which may indicate a higher level of anxiety. Additionally, parasite infestation improved spatial memory, assessed in the WM test, and recognition of new objects. These changes are related to the identified reduction in noradrenaline level in the CNS structures and less pronounced changes in striatal serotonergic neurotransmission. H. diminuta infestation was also found to cause a significant reduction of hippocampal expression of IL-6. Our results provide new data for further research on brain function during parasitic infections especially in relation to helminths and diseases in which noradrenergic system may play an important role.

Immunopotentiating properties of chimeric OprF-OprI-PopB protein against Pseudomonas aeruginosa PAO1 in the infected burned rat model.

Objective(s): Pseudomonas aeruginosa, as an opportunistic pathogen, is known to cause nosocomial infections among patients suffering from burn injuries and also cystic fibrosis patients. The objective of our research was to develop a novel vaccine against P. aeruginosa. Materials and Methods:A recombinant P. aeruginosa subunit vaccine based on the outer membrane proteins, including the OprF-OprI region and its major protein in the type III secretion system, PopB (called FIB protein) was formulated. To induce a robust immune response, our preferred regions were conjugated to a carrier protein, GMCSF (Granulocyte-macrophage colony-stimulating factor). FIB protein’s immunogenicity with and without adjuvant was evaluated in vaccinated rats and also burned rat models, which were subcutaneously challenged by the PAO1 strain of P. aeruginosa.Results:Antibody levels were increased in sera of rats in this study. Assessment of the resident memory CD4+ T cells in splenocytes from vaccinated rats demonstrated that the FIB conjugated with GMCSF could cause higher responses in comparison with other groups. Moreover, immunization with the FIB plus adjuvant protein could improve interferon-gamma (IFN-γ) production, interleukin 17A (IL-17A), and IL-4, contributing to elicit humoral and cellular immune responses and decreased post-infection bacterial loads after PA challenge, pathology, and inflammatory cell infiltration. Conclusion:These observations demonstrated that FIB conjugated with GMCSF can be a putative vaccine candidate against P. aeruginosa in burnt rat models.

A targeted nanozyme based on multiple porphyrins for enhanced photodynamic antibacterial application

• The targeted nanozyme has the catalase- and peroxidase-like activities. • This phenylboronic acid-functionalized nanozyme can capture the bacteria. • The nanozyme can decompose the overexpressed H 2 O 2 to O 2 and generate ROS in situ. • The nanozyme can alleviate hypoxia and enhance PDT for bacterial infection ulcers. High H 2 O 2 content at the bacterial infection site causes irreversible damage to DNA, RNA and proteins, and influences intracellular signaling through essential signaling proteins that control cell proliferation. Catalase can decompose H 2 O 2 to produce O 2 in situ, which is an ideal strategy to reduce H 2 O 2 in the infectious tissue. In this work, a novel targeted nanozyme Co Ⅱ TBPP(bpy) is precisely synthesized by utilizing supramolecular self-assembly. The prepared nanozyme exhibits excellent ability to capture bacteria due to its phenylboronic acid groups and has broad-spectrum antibacterial efficiency through the synergy of PDT, the change of membrane permeability and the peroxidase-like • OH production. In addition, Co Ⅱ TBPP(bpy) has significant effect on anti-inflammatory and promoting wound healing, which is proved in E. coli -infected rat ulcers model in vivo and shows very low cytotoxicity to normal mammalian cells. With further development and optimization, as a new type of antibacterial nanozyme, Co Ⅱ TBPP(bpy) has a great potential for the treatment of wound infections.

Treatment of Infection-Related Non-Unions with Bioactive Glass-A Promising Approach or Just Another Method of Dead Space Management?

The treatment of infected and non-infected non-unions remains a major challenge in trauma surgery. Due to the limited availability of autologous bone grafts and the need for local anti-infective treatment, bone substitutes have been the focus of tissue engineering for years. In this context, bioactive glasses are promising, especially regarding their anti-infective potential, which could reduce the need for local and systemic treatment with conventional antibiotics. The aim of this study was to investigate the osteoinductive and osteoconductive effects, as well as the anti-infectious potential, of S53P4 using a standardized non-union model, which had not been investigated previously. Using an already established sequential animal model in infected and non-infected rat femora, we were able to investigate bioactive glass S53P4 under realistic non-union conditions regarding its osteoinductive, osteoconductive and anti-infective potential with the use of µCT scans, biomechanical testing and histological, as well as microbiological, analysis. Although S53P4 did not lead to a stable union in the non-infected or the infected setting, µCT analysis revealed an osteoinductive effect of S53P4 under non-infected conditions, which was diminished under infected conditions. The osteoconductive effect of S53P4 remained almost negligible in histological analysis, even 8 weeks after treatment. Additionally, the expected anti-infective effect could not be demonstrated. Our data suggested that S53P4 should not be used in infected non-unions, especially in those with large bone defects.

Evaluation of immune response and haematological parameters in infected male albino rats by giardiasis.

The present work aimed to study the effects of Giardia lamblia infection on immunological and haematological studies and to evaluate immunoglobulins and some cytokines. Fifty male albino rats were divided into six groups. The control group includes 20 rats and the infected group includes 30 rats. All the estimations were checked all over five checkpoints (CP) (7, 14, 21, 28 and 35days post-infection). Serum levels of IgA, IgG, IgM and IgE. Cytokines INF-γ, TNF-alpha, IL-4, IL-10 and haematological parameters were determined. Cyst and trophozoite were counted. A considerable increase in the level of immunoglobulins and cytokines in all infected groups compared with the control group was documented. Furthermore, a significant decrease in red blood corpuscles, haemoglobin and mean corpuscular haemoglobin concentration levels was observed, whereas substantial increases in mean corpuscular volume, mean corpuscular haemoglobin and platelets were observed. Moreover, infected rats had a substantial rise in WBCs, lymphocytes and eosinophil counts compared with the control group, whereas neutrophils and monocytes had a significant decrease. The number of trophozoites and cysts was significantly increased in infected groups before diminishing after day 28. The current results showed that Th1 and Th2 immune responses, which are characterized by the production of TNF-α, IFN-γ, IL-4 and IL-10, are important for protection against Giardia infections and also verified the balance between these cytokines and the timing of their production was crucial in G.lamblia immune response.

​​Pathogenicity of Drug-resistant Canine Isolate of Trypanosoma brucei brucei in Rats

Background: Trypanosomosis is a neglected tropical disease. Drug resistance in trypanosomosis has been on the increase in recent years. It is presumed that drug resistant trypanosomes are less pathogenic and may be very suitable for rodent models of human African trypanosomosis (HAT). Methods: The T. b. brucei used was locally isolated from a dog and characterized by PCR. Drug resistance trait of the isolate was evaluated using diminazine aceturate and isometamidium chloride. Healthy rats were inoculated with the isolate, IP. Some haemato-biochemical parameters were evaluated. Three rats were euthanized on day 70 PI and necropsied. Data generated from the study were analyzed by student’s t-test and significance was accepted at probability, p less than 0.05. Result: The isolate was confirmed to be drug resistant T. b. brucei and caused no mortalities in the infected rats. The low pathogenicity in rats was discussed and the typical CNS lesions of chronic-stage HAT produced by the isolate makes it a suitable candidate for rodent models of HAT.

Pharmacological Mechanisms of Shangke Huangshui against Skin and Soft Tissue Infection.

Background Skin and soft tissue infections (SSTIs) are a group of common diseases, usually caused by bacteria. Shangke Huangshui (SK) has been widely used to treat various SSTIs diseases for many years, but its mechanism is unclear. Therefore, this study was designed to investigate the anti-infective effect of SK on different skin and soft tissue infection diseases and to explore its underlying mechanism. Methods The subcutaneous abscess mouse model, the dermal ulcer rat model, and the infectious soft tissue injury rat model were established by injection of Staphylococcus aureus to evaluate the anti-inflammatory and antibacterial effects of SK. Abscess volume, local appearance score and histological changes, bacterial contents, and hydroxyproline concentration in the skin or soft tissue were analyzed. The levels of NO, TNF-α, IL-1β, and IL-8 in the serum and tissue were determined by ELISA method. The mRNA expression levels of TLR2, MyD88, TAK1, NF-κB, AP-1, and other genes were measured with qRT-PCR method, and the protein expression of TLR2, MyD88, TAK1, NF-κB, and AP-1 was detected by western blot method. Results SK had an obvious therapeutic effect on skin and soft tissue infections. It reduced the volume of abscess and promoted the healing of skin ulcer, improved pathological phenomena, such as inflammatory infiltration of skin and soft tissue, reduced the levels of white blood cells and NO in the blood, decreased bacteria contents in the skin and soft tissue. Furthermore, SK decreased the mRNA expression of TLR2, MyD88, TAK1, NF-κB and AP-1, and other related genes and also downregulated the protein expression of TLR2, MyD88, TAK1, NF-κB, and AP-1. Conclusion The experiments provide evidence that SK can treat skin and soft tissue infection diseases based on its comprehensive antibacterial and anti-inflammatory effects. The therapeutic mechanism may be associated with the inhibition of TLR2/MyD88/NF-κB signaling pathway.

Evaluation of the Efficacy of Vancomycin-Soaked Autograft to Eliminate Staphylococcus aureus Contamination After Anterior Cruciate Ligament Reconstruction: Based on an Infected Rat Model

Background: Vancomycin-soaked autograft application in anterior cruciate ligament reconstruction (ACLR) significantly reduces postoperative infection rates. However, the optimal vancomycin concentrations and time of vancomycin presoaking of autografts for preventing infection are still unknown. Purpose: To evaluate the efficacy of vancomycin-soaked autografts in preventing infection in rats with ACLR. Study Design: Controlled laboratory study. Methods: A total of 102 tendons of Wistar rats were harvested under sterile conditions from fresh cadaveric legs. Contamination with 2.0 × 104 colony forming units per milliliter of Staphylococcus aureus and soaking in different vancomycin concentrations for different soaking times was performed in vitro. In vivo, after being contaminated with S. aureus and soaked with optimal vancomycin solution treatment and sterile saline, the grafts were implanted in rat knees to finish ACLR surgery. At 2, 4, and 12 weeks after surgery, samples were harvested to observe signs of infection and tendon-bone incorporation via general postoperative conditions, serum inflammatory markers, microbiological counting, knee radiographs, micro–computed tomography, histologic staining, scanning electron microscopy, and biomechanical testing. Results: Bacterial contamination was eliminated when at least 5 or 10 mg/mL of vancomycin was applied for 30 minutes in vitro. Rats in the vancomycin-soaked graft group (5 mg/mL of vancomycin for 30 minutes) showed no significant signs of infection and fewer positive cultures than did those without presoaking. The vancomycin-soaked graft group had reduced serum inflammatory markers, tissue scores, inflammatory reactions in the joint tissue, and radiographic evidence of periarticular osseous destruction compared with the control group. At postoperative week 12, the vancomycin-soaked graft group showed good outcomes in tendon-bone incorporation via micro–computed tomography, histologic staining, and biomechanical testing. Conclusion: In a rat model of infection after ACLR, presoaking grafts in a 5-mg/mL vancomycin solution for 30 minutes could effectively prevent S. aureus contamination without affecting tendon-bone incorporation and knee function. Clinical Relevance: The present study could provide a specific solution for the use of vancomycin in the prevention of infection after ACLR clinically.

Assessment ofthe efficacy of thymol against Toxocara vitulorum in experimentally infected rats.

The effect of thymol and ivermectin on the development and embryonation of Toxocara vitulorum (T. vitulorum) eggs, as well as their migration in albino rats was investigated both in vitro and in vivo. A total of forty male albino rats were divided into four groups for an in vivo experiment. The first group was uninfected; the second group was infected but left untreated; the third group was infected and received thymol at a dose of 40mg/kg; and the fourth group was infected and received ivermectin (0.2mg/kg). In vitro, thymol inhibited the development of Toxocara larvae within the eggs. However, ivermectin, produced inconsistent results. The in vivo results indicated that the recovery rates of Toxocara larvae from the liver and lungs on day 7 post-infection were significantly lower in the thymol or ivermectin-treated groups than in the infected untreated control. Albumin levels were significantly increased in the thymol-treated group as compared to thepositive control and ivermectin groups.Nitric oxide, IL-4, and IFN- levels in the serum of the thymol or ivermectin-treated groups were significantly lower than that of thepositive control group. Histopathological examination demonstrated that thymol and ivermectin were effective in reducing larval load, reducing the number and size of granulomas in the absence of larvae, and improving tissuearchitecture. The current study concluded that thymol possessed anti-Toxocara activity in a rat model. Additionally, thymol possessed ovicidal properties and may be used as a disinfectant.

Surface modification of titanium implants by pH-Responsive coating designed for Self-Adaptive antibacterial and promoted osseointegration

The development of responsive antibacterial implants is of high significance for the treatment of implant-associated infections. In this study, a self-adaptive antibacterial Ti-based implant with pH-sensitive coating was flexibly designed and constructed, with the dual functions of antibacterial and promoted osseointegration. The pH-sensitive antibacterial copolymer containing cationic quaternary ammonium salts (QAs) and carboxyl groups was firstly synthesized. Then, the prepared copolymer was coated on the surface of Ti metal by a simple layer-by-layer (LBL) self-assembly method to construct an antibacterial coating. The surface charge reversal was caused by the acidic microenvironment induced by the infected bacteria. The positively charged QAs possessed excellent antibacterial property against E. coli and S. aureus in vitro. The negatively charged carboxyl groups promoted the adhesion, proliferation and osteogenic differentiation of MC3T3-E1 cells in vitro. And the results of a S. aureus infection rat model further confirmed that the coated screws possessed good abilities of antibacterial and anti-inflammatory of implantation, and showed good osteointegration performance compared with the control ones. This strategy of self-adaptive antibacterial coating will provide a potential application for the prevention of Ti implant-associated bacterial infections.

The ameliorative efficacy of Thymus vulgaris essential oil against Escherichia coli O157:H7-induced hematological alterations, hepatorenal dysfunction and immune-inflammatory disturbances in experimentally infected rats.

The purpose of the present study was to evaluate the possible ameliorative role of Thymus vulgaris (T. vulgaris) essential oil against Escherichia coli O157:H7 (E. coli O157:H7) deleterious effects in both blood and different tissues of rats by assessing the hematological, biochemical and immune-inflammatory parameters besides the histopathological alterations in the different organs. Forty male rats were randomly divided into four equal groups as follows: group I served as control, group II orally inoculated with E. coli O157:H7 at a dose of 1.0 × 109cfu/ml, group III orally received 250mg/kg BW T. vulgaris oil daily for 7days and group IV orally inoculated with E. coli O157:H7 as the same dose of group II and orally received T. vulgaris oil as the same dose and duration of group III. Bacterial challenge in groups II and IV was once at the beginning of experiment and administration of oil began after 72h from bacterial inoculation. At the end of the study, blood was sampled and complete blood picture, liver and kidney function alongside immunoglobulins and cytokines concentrations were estimated and tissues of large intestine (colon), liver and kidneys were collected for histopathological examinations. The results revealed that there was an increase of red blood cells count, hematocrit value and hemoglobin concentration besides white blood cells and thrombocytes counts and substantial increment of serum markers of hepatorenal damage such as the activities of transaminases and concentrations of bilirubin (total, direct and indirect), total proteins, albumin, creatinine and urea in E. coli O157:H7-challenged group. Also, there was a considerable increase in serum immunoglobulins M and G, interleukin 6 and 8 and tumor necrosis factor alpha as well as decreased serum alkaline phosphatase activity. Moreover, T. vulgaris oil could partially improve the hematological, biochemical and histopathological alterations induced by E. coli O157:H7 without any significant alterations in all measured parameters when used alone. The study concluded that the T. vulgaris oil relatively diminished the alterations in hematological parameters, hepatic and renal function markers and immune-inflammatory variables alongside the histopathological changes in different organs induced by E. coli O157:H7. The ameliorative effects of T. vulgaris oil are mediated through its anti-inflammatory, antioxidant and immunomodulatory activities.

Angiostrongylus cantonensis an Atypical Presenilin: Epitope Mapping, Characterization, and Development of an ELISA Peptide Assay for Specific Diagnostic of Angiostrongyliasis.

Background: Angiostrongyliasis, the leading cause universal of eosinophilic meningitis, is an emergent disease due to Angiostrongylus cantonensis (rat lungworm) larvae, transmitted accidentally to humans. The diagnosis of human angiostrongyliasis is based on epidemiologic characteristics, clinical symptoms, medical history, and laboratory findings, particularly hypereosinophilia in blood and cerebrospinal fluid. Thus, the diagnosis is difficult and often confused with those produced by other parasitic diseases. Therefore, the development of a fast and specific diagnostic test for angiostrongyliasis is a challenge mainly due to the lack of specificity of the described tests, and therefore, the characterization of a new target is required. Material and Methods: Using bioinformatics tools, the putative presenilin (PS) protein C7BVX5-1 was characterized structurally and phylogenetically. A peptide microarray approach was employed to identify single and specific epitopes, and tetrameric epitope peptides were synthesized to evaluate their performance in an ELISA-peptide assay. Results: The data showed that the A. cantonensis PS protein presents nine transmembrane domains, the catalytic aspartyl domain [(XD (aa 241) and GLGD (aa 332–335)], between TM6 and TM7 and the absence of the PALP and other characteristics domains of the class A22 and homologous presenilin (PSH). These individualities make it an atypical sub-branch of the PS family, located in a separate subgroup along with the enzyme Haemogonchus contournus and separated from other worm subclasses. Twelve B-linear epitopes were identified by microarray of peptides and validated by ELISA using infected rat sera. In addition, their diagnostic performance was demonstrated by an ELISA-MAP4 peptide. Conclusions: Our data show that the putative AgPS is an atypical multi-pass transmembrane protein and indicate that the protein is an excellent immunological target with two (PsAg3 and PsAg9) A. costarisencis cross-reactive epitopes and eight (PsAg1, PsAg2, PsAg6, PsAg7, PsAg8, PsAg10, PsAg11, PsAg12) apparent unique A. cantonensis epitopes. These epitopes could be used in engineered receptacle proteins to develop a specific immunological diagnostic assay for angiostrongyliasis caused by A. cantonensis.

Investigating essential factors in the spread of lassa fever dynamics through sensitivity analysis

Lassa fever is a zoonotic acute viral illness caused by Lassa virus. Since there is no vaccine yet to protect against contracting the virus, it continues to spread in West Africa. In this paper, a mathematical model of lassa transmission that considers two classes of rats: house rat and bush rat, is proposed. Theoretically, global stability of the model disease-free and endemic equilibria are established by constructing a global Lyapunov function. Sensitivity indices of the basic reproduction number are derived using the normalised forward approach to evaluate the effectiveness of control measures. The disease-free equilibrium is globally asymptotically stable when the basic reproduction number R0 1. Results from sensitivity analysis reveals that rat biting rate for infectious house rats RFH and infectious bush rats R_FB, transmission probability per contact with infectious house and bush rats (R_FH and R_FB), human recruitment rate and transmission probability per contact with infectious human hosts are highly significant in determining the severity of lassa infection. On the other hand, natural death rate of rats, natural death rate of human hosts, recovery and hospitalization rates of human hosts are critical for lassa transmission reduction. Plans that target the contact rate between house and bush rats (i.e use of indoor residual spray, fumigation of environment with pesticide) and those that target recovery rate of human hosts (i.e treatment of infectious human hosts) are recommended to control the disease.

Nanoparticle-mediated delivery system alleviates the formation of infection stones by activating TRPV5.

Infection stones constitute a small but intractable group of diseases of urinary system. In this study, we explored the potential therapeutic effect of a small activation RNA, ds-320, encapsulated in chitosan (320-chitosan). Western blot analysis verified the downregulation of TRPV5 in patients and rat model of infection stones, as well as the stimulation of ds-320 on TRPV5 expression. MTT assay showed that chitosan-mediated delivery was less cytotoxic to ds-320 compared with liposome delivery. Further a modified invasion assay revealed an inhibitory effect of 320-chitosan on bacterial invasion into normal rat kidney epithelial NRK-52E cells. The establishment of infection stone model was performed by intravesical injection of 1×108 CFU of Proteus mirabilis. In animal experiments, no visible stones were obtained. The number of live bacteria and white blood cells in urine showed no difference among all infected rats at the time of sacrifice. However, we observed a decline in urine calcium and pH, suggesting the effect of acidification. Overall, our study provides evidence for the protective effect of 320-chitosan, for its ability to down-regulate urinary calcium, acidify urine, and inhibit bacteria from invading renal epithelial cells. Thus, it can be served as an important complementary therapy for infection stones.

Codelivery of Doxycycline and Hydroxychloroquine to Treatment of Brucellosis: An Animal Study

Evaluation of various biochemical and immunological parameters in infectious diseases is one of the best indicators for a diagnosis and treatment process. The main goal of this project is to determine the effect of hydroxychloroquine and doxycycline loading into solid‐lipid‐nanoparticles (DOX‐HCQ‐SLN) on the both acute and chronic phases of brucellosis. In addition, evaluate some biochemical factors, trace elements, and inflammatory elements. Blood serum levels of Zn, Fe, Na, and K and hepatic biochemical parameters (AST, ALT, ALP, and TBil) were remarkably different between infected and healthy rats. Vitamin D was decreased, and CRP was increased in chronic and acute brucellosis. Quantitative evaluation of these mentioned parameters can be useful to diagnose brucellosis in advance. Due to the good effect of the synchronized use of hydroxychloroquine and doxycycline in the form of nanoparticles, the manipulation of these nanoparticles can help for better treatment and also reduction in brucellosis reinfection.

Efficacy of Solanum torvum Sw. as a Potential Antifungal Medicinal Plant against Candida albicans in In Vitro and In Vivo Experimental Models

Solanum torvum Sw. is a native plant of India found abundantly in North East. It has many medicinal properties and its berries are used to treat fungal infections by the natives of the study area. The efficacy of Solanum torvum Sw. was evaluated using both in vitro antimicrobial methods and in vivo model of vaginal Candida albicans infection. Two in vitro models, disc diffusion assay and micro dilution broth assay was used and in in vivo study laboratory female rats were used. In the disc diffusion assay highest concentration i.e. 200 mg/ml Solanum torvum Sw. methanolic extract dose shows larger inhibition zone of fungal growth than the other concentrations of extract. In the microdilution broth assay, 150 mg/ml concentration of extract shows highest fungal growth inhibition. Histological study was done by loading fungal conidia on vaginas of in vivo model (rat) where 600 mg/kg extract treated group showed complete reduction of fungal growth over vaginal epithelium and vaginal cavity. In result, this plant proves its efficacy in reducing the fungal load from vaginas of infected rat as well as showing good minimum inhibitory concentration and zone of inhibition at in vitro study. From the above study, it was clear that Solanum torvum Sw. showed promising positive result against Candida albicans.

The effect of ethanolic extract of mangosteen peel to malondialdehyde level, colony number, and pulmonary damage level score of tuberculosis model rats

Mycobacterium tuberculosis is an intracellular pathogen that can replicate in host macrophages. This research examined the medication mechanism of mangosteen peel extract as anti-tuberculosis towards rats infected by Mycobacterium tuberculosis H37Rv. The mangosteen peel has chemical antituberculosis properties, such as xanthone, flavonoid, and tanin. Rats were invected by M. tuberculosis 105 (100 μl) by intratracheal for 4 weeks, then divided into 6 experimental groups: K1 (infected rats were treated by standard feed); K2 (infected rats were treated by INH); P1 (infected rats were treated by INH and extract 100mg/kg BW/day); P2 (infected rats were treated by INH and extract 200mg/kg BW/day); P3 (infected rats were treated by INH and extract 300mg/kg BW/day); and P4 (infected rats were treated by INH and extract 400mg/kg BW/day). INH and extract were administered orally for 4 weeks. On the 29th rats were euthanized to collect the left lung for MDA measurement and germs planting, and the right lung was collected for Hematoxyline eosin preparation. The data was analyzed by SPSS. The mangosteen peel ethanolic extract showed significant differences between treatments towards Tuberculotic rats treated by INH. The MDA level decreased with a value of p=0,000 (p&lt;0,005) r= 82,2% ; the decrease of colony count with a value of p=0,000(p&lt;0,005) r= 93,3%; the decrease of pulmonary damage level score with p=0,001(p&lt;0,005) r= 50%. Mangosteen peel ethanolic extract reduced MDA levels, colony counts, and lung damage levels at a dose of 200mg/kg BW/day of tuberculosis-model rats.