Antibacterial effects of Eucalyptus globulus leaf extract on pathogenic bacteria isolated from specimens of patients with respiratory tract disorders

ABSTRACTThe antibacterial activity of Eucalyptus globulus leaf extract was determined for 56 isolates of Staphylococcus aureus, 25 isolates of Streptococcus pyogenes, 12 isolates of Streptococcus pneumoniae and seven isolates of Haemophilus influenzae obtained from 200 clinical specimens of patients with respiratory tract disorders. MIC50s for these species were 64, 32, 16 and 16 mg/L, respectively; MIC90s were 128, 64, 32 and 32 mg/L, respectively; and MBCs were 512, 128, 64 and 64 mg/L, respectively. These results suggest that further studies to clarify the possible therapeutic role of E. globulus leaf extract in the treatment of respiratory tract infection are warranted

The clinical and environmental spread and diversity of toxigenic Clostridium difficile diarrhea in the region of the Middle East.

Stool samples of 1822 hospitalized patients with nosocomial diarrhea and 100 environmental samples were collected at three teaching hospitals and PCR amplification of rRNA intergenic spacer regions (ISR) was conducted. Bacterial cytotoxicity was assayed by conducting three assays namely toxigenic culture on vero cells, stool cytotoxin, and enzyme immunoassay. ISR was carried out using two universal primers complementary to conserved regions in the 16S and 23S rRNA genes. It was found that the toxigenic culture, stool cytotoxin and enzyme immunoassay showed close rates of detection of toxigenic C. difficile, 124, 121, and 122 /1822 (6.8, 6.64., and 6.7%) respectively. In addition, 32 different ribotypes for toxigenic C. difficile were detected, 28 in clinical and 6 in environmental isolates. The predominant ribotypes from the clinical isolates were 13-15, 35.6%, of isolates. Ribotypes were associated with age, location of isolation, and severity of symptoms of clostridial diarrhea (P<0.05). Ribotypes 6-9 affected children only. The most common ribotype of C. difficile , no. 13, as well as ribotypes 16, 20, and 4 covered almost the whole range of severity of symptoms. Ribotypes 21-27, 1, 3, 6, 7, 9, 11, 14, and 19 caused mild-moderate CDAD symptoms while ribotypes 5, 10 8, 12, 15, 17, and 28 were dominantly of severe symptoms (P<0.05). Environmental isolates showed 17% toxigenic strains composed of 4 different ribotypes while ribotypes 5 was shared with clinical isolates. These findings showed that C. difficile associated with diarrhea were genetically diverse and linked to environmental strains

Optimization and efficient purification in production of Brucella melitensis recombinant HSP and TF proteins with low endotoxin contents

Background: The development of an effective subunit vaccine against brucellosis is a research area of intense. But optimization of recombinant proteins production in Escherichia coli and content of endotoxins associated with final recombinant proteins are very important. Objectives: In the present study, expression and purification of Brucella melitensis rHSP and rTF were optimized to reduce endotoxin contaminants. Materials and Methods: pDEST-tf and pDEST-hsp were transformed into E. coli BL21 (DE3), and then B. melitensis recombinant HSPA and TF proteins were overexpressed. Purification of these proteins was optimized to remove most of endotoxin contaminants from the end product using 0.1 Triton X-114 in washing buffers. Results: An endotoxin reduction of less than 0.05 EUmg/1 was achieved with protein recovery close to an 80 yield. Conclusions: As this new protocol requires only one step to simultaneously purify tagged proteins and eliminate endotoxins, it represents a substantial advantage in time, effort, and expense. © 2013, Ahvaz Jundishapur University of Medical Sciences; Licensee Kowsar Ltd

Can concurrent lower gastrointestinal manifestations help the timely diagnosis of small intestinal bacterial overgrowth in CVID patients?

Summary Introduction and objective. Gastrointestinal complications are considered as one of the most common manifestations in patients with Common Variable Immunodeficiency (CVID). These complications can result from Small Intestinal Bacterial Overgrowth (SIBO). Hydrogen breath test is extensively used to diagnose SIBO. The objective of this study was to evaluate the prevalence of SIBO using the Hydrogen Breath Test (HBT) in patients with CVID. Materials and methods. Twenty-seven patients with CVID entered this cross-sectional study. Demographic and lower gastrointestinal symptoms were recorded in a check list. Hemoglobin level was measured in all patients. The concentration of IgA and IgG was assessed using nephelometry. Moreover, SIBO was detected by means of Glucose hydrogen breath test. Results. The mean (± SD) age of the patients was 35.25 (± 11.69) years. Twenty patients (74.1) manifested at least one lower gastrointestinal symptom. The most frequent lower gastrointestinal manifestations were bloating (66.7) and chronic diarrhea (40.7), respectively. IgA level less than 10 mg/dl and IgG level less than 600 mg/dl were determined in 77.8 and 25.9 of patients, respectively. Positive HBT was detected in 40.7 (n = 11) of the patients. In the positive HBT group, bloating, chronic diarrhea and abdominal pain were the most common lower GI manifestations. There was no significant difference in terms of age, BMI, IgA level, and duration of CVID between the positive and negative HBT groups. The significant association of co-occurrence of anemia and abdominal pain with positive HBT (positive predictive value: 100) might be considered as a clue to SIBO diagnosis. Conclusions. Regarding the high prevalence and non-specific manifestation of SIBO, it is suggested to consider concurrent symptoms in patients with CVID to manage the timely and precise diagnosis of SIBO. © 2021 Associazione Allergologi Immunologi Italiani Territoriali e Ospedalieri-AAIITO. Published by EDRA SpA. All rights reserved

The global burden of cancer attributable to risk factors, 2010–19: a systematic analysis for the Global Burden of Disease Study 2019

BACKGROUND: Understanding the magnitude of cancer burden attributable to potentially modifiable risk factors is crucial for development of effective prevention and mitigation strategies. We analysed results from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2019 to inform cancer control planning efforts globally. METHODS: The GBD 2019 comparative risk assessment framework was used to estimate cancer burden attributable to behavioural, environmental and occupational, and metabolic risk factors. A total of 82 risk–outcome pairs were included on the basis of the World Cancer Research Fund criteria. Estimated cancer deaths and disability-adjusted life-years (DALYs) in 2019 and change in these measures between 2010 and 2019 are presented. FINDINGS: Globally, in 2019, the risk factors included in this analysis accounted for 4·45 million (95% uncertainty interval 4·01–4·94) deaths and 105 million (95·0–116) DALYs for both sexes combined, representing 44·4% (41·3–48·4) of all cancer deaths and 42·0% (39·1–45·6) of all DALYs. There were 2·88 million (2·60–3·18) risk-attributable cancer deaths in males (50·6% [47·8–54·1] of all male cancer deaths) and 1·58 million (1·36–1·84) risk-attributable cancer deaths in females (36·3% [32·5–41·3] of all female cancer deaths). The leading risk factors at the most detailed level globally for risk-attributable cancer deaths and DALYs in 2019 for both sexes combined were smoking, followed by alcohol use and high BMI. Risk-attributable cancer burden varied by world region and Socio-demographic Index (SDI), with smoking, unsafe sex, and alcohol use being the three leading risk factors for risk-attributable cancer DALYs in low SDI locations in 2019, whereas DALYs in high SDI locations mirrored the top three global risk factor rankings. From 2010 to 2019, global risk-attributable cancer deaths increased by 20·4% (12·6–28·4) and DALYs by 16·8% (8·8–25·0), with the greatest percentage increase in metabolic risks (34·7% [27·9–42·8] and 33·3% [25·8–42·0]). INTERPRETATION: The leading risk factors contributing to global cancer burden in 2019 were behavioural, whereas metabolic risk factors saw the largest increases between 2010 and 2019. Reducing exposure to these modifiable risk factors would decrease cancer mortality and DALY rates worldwide, and policies should be tailored appropriately to local cancer risk factor burden

The Effect of Moisture Content and Temperature on the Specific Heat Capacity of Nut and Kernel of Two Iranian Pistachio Varieties

Pistachio has a special ranking among Iranian agricultural products. Iran is known as the largest producer and exporter of pistachio in the world. Agricultural products are imposed under different thermal treatments during storage and processing. Designing all these processes requires thermal parameters of the products such as specific heat capacity. Regarding the importance of pistachio processing as an exportable product, in this study the specific heat capacity of nut and kernel of two varieties of Iranian pistachio (Kalle-Ghochi and Badami) were investigated at four levels of moisture content (initial moisture content (5%), 15%, 25% and 40% w.b.) and three levels of temperature (40, 50 and 60°C). In both varieties, the differences between the data were significant at the 1% of probability; however, the effect of moisture content was greater than that of temperature. The results indicated that the specific heat capacity of both nuts and kernels increase logarithmically with increase of moisture content and also increase linearly with increase of temperature. This parameter has altered for nut and kernel of Kalle-Ghochi and Badami varieties within the range of 1.039-2.936 kJ kg-1 K-1, 1.236-3.320 kJ kg-1 K-1, 0.887-2.773 kJ kg-1 K-1 and 0.811-2.914 kJ kg-1 K-1, respectively. Moreover, for any given level of temperature, the specific heat capacity of kernels was higher than that of nuts. Finally, regression models with high R2 values were developed to predict the specific heat capacity of pistachio varieties as a function of moisture content and temperatur

Immunization of mice with a novel recombinant molecular chaperon confers protection against <em>Brucella melitensis</em> infection.

Brucella spp. are zoonotic Gram-negative intracellular pathogens with the ability to survive and replicate in phagocytes. It has been shown that bacterial proteins expressed abundantly in this niche are stress-related proteins capable of triggering effective immune responses. BMEI1549 is a molecular chaperone designated DnaK that is expressed under stress conditions and helps to prevent formation of protein aggregates. In order to study the potential of DnaK as a prospective Brucella subunit vaccine, immunogenicity and protective efficacy of recombinant DnaK from Brucella melitensis was evaluated in BALB/c mice. The dnak gene was cloned, expressed in Escherichia coli, and the resulting recombinant protein used as subunit vaccine. DnaK-immunized mice showed a strong lymphocyte proliferative response to in vitro antigen stimulation. Although comparable levels of antigen-specific IgG2a and IgG1 were observed in immunized mice, high amounts of IFN-&gamma;, IL-12 and IL-6, no detectable level of IL-4 and very low levels of IL-10 and IL-5 were produced by splenocytes of vaccinated mice suggesting induction of a Th1 dominant immune response by DnaK. Compared to control animals, mice vaccinated with DnaK exhibited a significant degree of protection against subsequent Brucella infection (p&lt;0.001), albeit this protection was less than the protection conferred by Rev.1 (p&lt;0.05). A further increase in protection was observed, when DnaK was combined with recombinant Omp31. Notably, this combination, as opposed to each component alone, induced statistically similar level of protection as induced by Rev.1 suggesting that DnaK could be viewed as a promising candidate for the development of a subunit vaccine against brucellosis

Gold Nanoparticles and Polyethylene Glycol Alleviate Clinical Symptoms and Alter Cytokine Secretion in a Mouse Model of Experimental Autoimmune Encephalomyelitis

Gold nanoparticles (GNPs) are attractive nanoparticles with unique electronic and optical properties in the nanotechnology field and are widely used in various biomedical fields. Studies have shown that these particles also exhibit antioxidant and anti-inflammatory properties. On the other hand, polyethylene glycol (PEG) that used to stabilize GNPs also exhibits antioxidant and anti-inflammatory properties due to their membrane resealing properties. The aim of this study was to evaluate the ameliorative effect of GNPs and PEG in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). EAE was induced in female C57BL/6 mice with injection of an emulsion of myelin oligodendrocyte glycoprotein (MOG35-55) peptide and Freund's adjuvant. GNPs measuring 25 nm were prepared, and their size was determined using transmission electron microscopy (TEM), then intraperitoneal injection of GNPs and PEG (MW 1500; 30 w/v) was initiated after immunization and continued until the day 27 postimmunization (13 injections in total). The EAE clinical scores and body weights were evaluated. We analyzed cental nervous system's cell infiltration and demyelinated lesions using hematoxylin and eosin and luxol fast blue staining, respectively. Also, interleukin-23 and interleukin-27 were examined using the ELISA technique. The severity of MS symptoms was significantly decreased in the treated groups with GNPs and PEG. Histological examination of the spinal cord showed that the number and severity of cells� infiltration and demyelinated lesions decreased significantly, and also the cytokine levels of IL-23 and IL-27 altered in treated groups. These results show that GNPs and PEG ameliorate the clinical course of EAE in mice. Our findings demonstrate proof of principle for potential of GNPs and PEG as novel agents for therapeutic approaches in the alleviated clinical symptoms of MS. © 2019 IUBMB Life, 1�9, 2019. © 2019 International Union of Biochemistry and Molecular Biolog

Simultaneous immunization of mice with Omp31 and TF provides protection against <em>Brucella melitensis</em> infection.

Brucella vaccines consisting of live attenuated Brucella strains are currently used in livestock, but safety concerns preclude their application in humans. Subunit vaccines have recently emerged as safe and efficacious alternatives in both humans and animals. In this study, subunit vaccines were developed that consisted of a recombinant outer membrane protein (rOmp31) and the trigger factor chaperone protein (rTF) of Brucella melitensis, either alone or in combination. BALB/c mice that were immunized with rOmp31+rTF showed comparable but slightly higher TF-specific IgG1 and IgG2a antibodies as compared to mice with rTF alone. Indeed, mice given this combination had titers of rOmp31-specific antibodies similar to those immunized with rOmp31 alone. In lymphocyte reactivation experiments, the splenocytes of immunized mice, whether given either of these antigens alone or as a cocktail, exhibited a strong antigen-specific recall proliferative response and expressed high amounts of IFN-&gamma;, IL-12, IL-10 and IL-6. Both rTF and rTF+rOmp31 vaccinated mice exhibited significantly higher CD4 and CD8 levels compared to the PBS group. The combination of rOmp31 and rTF provided protection against B. melitensis infection comparable to that of vaccine strain Rev.1. In comparison to rTF alone, combination of rTF and rOmp31 caused only a slight increase in protection level. Although combination of rTF and rOmp31 caused a non-significant increase in IFN-&gamma; induction, antibody level, proliferation index and CD4 and CD8 frequencies compared to rTF alone, its cumulative effects on aforesaid parameters may be viewed as a better efficacy

Glibenclamide mitigates cognitive impairment and hippocampal neuroinflammation in rats with type 2 diabetes and sporadic Alzheimer-like disease

A growing body of evidence suggests that type 2 diabetes (T2D) is a risk factor for cognitive impairment and dementia. Both preclinical and clinical studies have provided evidence that brain insulin resistance is associated with cognitive decline in patients with T2D and sporadic Alzheimer disease (AD). Accordingly, antidiabetic medications have been suggested as potential drugs for the treatment of cognitive impairments in patients with sporadic AD. This study set out to determine whether glibenclamide (GBC), an antidiabetic agent, can ameliorate cognitive impairments in rats with T2D and sporadic AD. Both animal models were treated with GBC for 23 consecutive days. To assess working and spatial memory, animals were subjected to the Y-maze and Morris water-maze tests. We measured glucose and insulin levels in the blood, and inflammatory cytokines such as tumor necrosis factor (TNF)-α and interleukin (IL)-6 in the hippocampus of animals. Our findings indicated that T2D and sporadic AD impaired memory and elevated TNF-α and IL-6 in the hippocampus. We found increased glucose and insulin levels in the blood of T2D-induced rats but not of sporadic AD rats. In contrast, GBC treatment improved memory impairment, increased insulin, and reduced glucose and hippocampal inflammation in rats with T2D and sporadic AD. This study suggests that GBC could be considered as a potential treatment for cognitive deficits in patients with T2D and sporadic AD. Taken together, this study highlights the need for further studies in humans to test whether GBC treatment is associated with cognitive improvement in sporadic AD patients. © 2019 Elsevier B.V