Targeted therapies in HER2-positive breast cancer with receptor-redirected Arazyme-linker-Herceptin as a novel fusion protein.

BACKGROUND: Targeted treatment of different types of cancers through highly expressed cancer cell surface receptors by fusion proteins is an efficient method for cancer therapy. The HER2 receptor is a member of the tyrosine kinase receptors family, which plays a notable role in breast cancer tumor development. About 25-30% of breast cancers overexpress human epidermal growth factor receptor 2 (HER2). METHODS: In this study, we evaluated the particulars of a designed recombinant protein formed by HER2-specific Mab Herceptin linked with Arazyme on a HER2-overexpressing breast cancer cell line (SKBR3). Arazyme, a metalloprotease produced by Serratia proteamaculans was fused to the variable area of light and heavy chains of the Herceptin. The cytotoxic assay of the Arazyme-linker-Herceptin in the SKBR3 and MDA-MB-468 cells was evaluated by the MTT and flow cytometry techniques. The Caspase­3 activity determination and adhesion assay were performed to evaluate the antitumor activity of the Arazyme-linker-Herceptin against SKBR3 cells. Furthermore, RT-PCR was used to measure the expression levels of the Bcl-2, Bax, MMP2, MMP9, and RIP3 genes. RESULTS: The Arazyme-linker-Herceptin showed higher cytotoxicity in SKBR3 cells compared to MDA-MB-468 cells. In addition, flow cytometry results revealed that the Arazyme-linker-Herceptin can significantly induce apoptosis in the HER2-overexpressing breast cancer cell line (SKBR3), which was confirmed by Bax upregulation and the decrease in adhesion of tumor cells and MMP2/MMP9. CONCLUSION: The findings of this study demonstrated that the Arazyme-linker-Herceptin induced apoptosis and decreased metastatic genes in SKBR3 cells; however, further research is required to confirm the effectiveness of the fusion protein.

Aqueous extract-mediated green synthesis of CuO nanoparticles: Potential anti-tuberculosis agents.

The emergence of drug-resistant strains in tuberculosis treatment underscores the urgency for novel therapeutic approaches. This study investigates the anti-tuberculosis activity of green-synthesized copper oxide (CuO) nanoparticles (NPs) using garlic and astragalus extracts. The physicochemical characterization of the nanoparticles confirms successful synthesis, followed by assessment of their antibacterial properties and safety profile. Rats infected with Mycobacterium tuberculosis are treated with nanocomposites derived from garlic extract at doses of 50 mg/kg and 100 mg/kg body weight. Evaluation includes the analysis of Early secreted antigenic target of 6 kDa (ESAT-6) expression and confirmation of antibodies through molecular assays. Administration of garlic and nanocomposites demonstrates significant inhibitory effects on tuberculosis progression in rats, validated by safety assessments and antibacterial efficacy. Notably, the 100 mg/kg dosage exhibits pronounced mitigation of tuberculosis-induced oxidative stress and lung damage. In conclusion, the combined administration of garlic extracts and green-synthesized nanocomposites shows promising efficacy in reducing tuberculosis infection, highlighting a potential avenue for anti-tuberculosis interventions.

An In Vivo Study of Lactobacillus rhamnosus (PTCC 1637) as a New Therapeutic Candidate in Esophageal Cancer.

Objective: This study is aimed at investigating the effect of probiotic Lactobacillus rhamnosus on esophageal cancer in vivo and in vitro. Methods and Results: In this study, the cytotoxicity effects of L. rhamnosus supernatant and whole-cell culture on a cancer cell line (Kyse30) compared to 5fu were evaluated by the MTT assay. The real-time PCR method was used to analyse the L. rhamnosus supernatant effect on the expression of Wnt signaling pathway genes. An in vivo investigation in nude mice was done to assess the anti-tumor activity of L. rhamnosus supernatant and whole-cell culture. Both supernatant and whole-cell culture of L. rhamnosus reduced cell survival (Kyse30) P < 0.001. The supernatant of this bacterium significantly reduced the expression of Wnt signaling pathway genes. Administration of supernatant and whole-cell culture of L. rhamnosus expressively reduced tumor growth compared to the control group. The effects of this bacterium on tumor necrosis were quite evident, pathologically P < 0.01. Conclusion: This study is the first report that assessed the potential impact of L. rhamnosus, especially its supernatant on esophageal cancer and Wnt signaling pathway genes. Therefore, this bacterium can be a harmless candidate for esophageal cancer therapy.

Interference of Lactiplantibacillus plantarum With Pseudomonas aeruginosa on the Infected Burns in Wistar Rats.

Burns are the most prevalent type of trauma in the world, and they have a high fatality rate. For cutaneous wound healing, modern and natural therapies, particularly probiotic supplements, have lately been considered. The goal of this study was to see how Lactiplantibacillus plantarum affected wound healing as well as the antibacterial activity of probiotic lactobacilli against Pseudomonas aeruginosa. The glass slide method was used to assess anti-adhesion activity, and the high-performance liquid chromatography method was used to quantify anti-adhesion chemicals in cell-free supernatant. Multidrug-resistant (MDR) P. aeruginosa was administered subcutaneously directly on the burn after induction of second-degree wounds. Three groups of animals were created. Every day, the supernatants were sprayed for therapy, and the wound healing was monitored. Lactobacilli bacteria had good anti-adhesion effects on P. aeruginosa, according to our findings, and high-performance liquid chromatography research revealed that their inhibitory effect could be attributable to four main organic acids: lactic acid, acetic acid, citric acid, and succinic acid. When the effect of treatments on fibroblastic cells was examined, it was discovered that the group treated with L. plantarum supernatants had the most fibroblastic cells when compared to the nontreated group. Furthermore, the bacteria increased the number of fibroblastic cells, re-epithelialization in the wound area, and the thickness of the epidermis and dermis layers. Lactobacilli bacteria's antimicrobial activity against MDR P. aeruginosa was determined by prevents infection. These findings revealed that L. plantarum can treat a P. aeruginosa infection in a second-degree burn and can significantly reduce inflammation.

Interference of Lactobacillus casei with Pseudomonas aeruginosa in the treatment of infected burns in Wistar rats.

OBJECTIVES: Burns are the most common type of trauma with a high mortality rate worldwide. The use of modern and natural medicines, especially probiotic products, has been recently considered for cutaneous wound healing. The present study was designed to investigate the effect of Lactobacillus casei on wound healing caused by Pseudomonas aeruginosa. MATERIALS AND METHODS: In this study, the anti-adhesion activity of L. casei was examined by the glass slide method, and inhibitory substances in the cell-free supernatant (CFS) were quantified by high-performance liquid chromatography (HPLC). Following the induction of second-degree wounds, multidrug-resistant (MDR) P. aeruginosa was injected subcutaneously and directly on the burn. The animals were divided into four groups. The supernatant of L. casei was sprayed for treatment every day and wound healing was examined. RESULTS: Based on our findings, the supernatant of L. casei showed considerable anti-adhesion effects on P. aeruginosa. HPLC analysis indicated that the inhibitory effect of this supernatant can be due to four main organic acids including lactic acid, acetic acid, citric acid, and succinic acid. The effect of treatment on fibroblastic cells showed that the treated group by supernatant of L. casei had more fibroblastic cells compared with the non-treated group. Moreover, this supernatant increased the rate of fibroblastic cells, re-epithelialization in the wound area, and the largest thickness of the epidermis and dermis layers. CONCLUSION: The present findings showed that L. casei supernatant significantly reduced inflammation and could be used to treat P. aeruginosa infection in second-degree burns.

Biofilm formation, antimicrobial susceptibility, serogroups and virulence genes of uropathogenic E. coli isolated from clinical samples in Iran.

BACKGROUND: Uropathogenic Escherichia coli O- Serogroups with their virulence factors are the most prevalent causes of UTIs. The present research performed to track common uropathogenic E.coli serogroups, antibiotic resistance pattern of strains and prevalence of virulence genes in isolations having the ability to constitute biofilm. METHODS: In this research 130 E.coli isolation from patients having UTI symptoms were collected and antimicrobial resistance pattern was performed by Kirby-Bauer method. Polymerase chain reaction was done using primer pairs to identify common serogroups of uropathogenic E.coli and studying virulence genes in isolations creating biofilm. RESULTS: Among 130 E.coli isolates, 80 (61.53 %) were able to make biofilm that 15 isolates (18.75 %) indicated strong reaction, 20 (25 %) of medium and 45 (56.25 %) of weak biofilm reaction. Among isolations creating biofilm, the highest resistance reported to Ampicillin (87.5 %) and the lowest to Nitrofurantoin (3.75 %). The frequency of fimH, pap, sfa and afa genes in isolations having the ability to create strong biofilm reported 93.33 %, 86.66 %, 86.66 % and 66.66 %, respectively. CONCLUSIONS: The findings indicated the importance of virulence genes in serogroups producing uropathogenic E.coli biofilm. It is recommended that strains producing biofilm before antibiotic use should be studied.