Circulating microRNAs as novel biomarkers for measuring the potency of ginger extract against cyclophosphamide toxicity in rat renal tissues: molecular and histopathological study.

OBJECTIVE: This study aims to explore underlying molecular variations in the expression of miRNAs in kidney tissues of ginger-treated and non-treated cyclophosphamide (CP)-intoxicated rats. MATERIALS AND METHODS:   A total of 40 adult male Wistar rats were randomly divided into four groups of 10 each: Group I (control: received normal food and water), Group II (received ginger at a dose of 300 mg/kg), Group III (received CP 75 mg/kg, i.p.), and Group IV (received the same dose of CP and ginger extract).  Rats received a single injection of 75 mg/kg CP on days 3, 4, 5, 19, 20, and 21. In CP-intoxicated rats, the treatment with ginger extract at a dose of 300 mg/kg was received by oral gavage starting seven days before CP and continuing throughout the duration of the experiment for four weeks. Molecular variations in the expression of miRNAs, apoptotic genes, histological kidney damage, and abnormal kidney function in control, ginger, and CP-intoxicated rats were identified by using real-time RT-PCR Analysis, immunohistochemical, and colorimetric assays. In addition, HPLC analysis and liquid chromatography spectrophotometry analysis using Diphenyl-1-picrylhydrazyl (DPPH) radical, and Β-Carotene-linoleic acid reagents were applied respectively for in-vitro screening of phytoconstituents and antioxidant activity for ginger extract. RESULTS: The kidney tissues of CP-intoxicated rats displayed an increase in lipid peroxidation marker malonaldehyde (MDA), DNA damage, and fibrosis markers like hyaluronic acid (HA) and hydroxyproline Hypx) with a decrease in the superoxide dismutase (SOD) and total antioxidant capacity (TAC). In addition, molecular expressions of mRNA fibrotic genes such as collagen, type 1, alpha 1 (COL1A1), and α-smooth muscle actin (αSMA). Molecular expressions of levels of B-cell lymphoma 2 (BCl-2) mRNA gene were down-regulated, and the expression of mRNA apoptotic; BCL2 associated X gene (Bax), caspase-3, Bax/BCl-2 ratio genes were significantly up-regulated respectively. Moreover, cellular oxidative genes, erythroid 2-related factor (Nrf2), and heme oxygenase-1 (HO-1) were down-regulated, respectively. The miR-155-5p, miR-34a-5p, miR-21-5p significantly increased while the miR-193b-3p, miR-455-3p, and miR-342-3p significantly decreased. Ginger also increased the expression of Nrf2, HO-1, and BCl-2 genes in the kidneys of rats induced with CP. In addition, active phytoconstituents, particularly 6]]-shogaol and 6]]-gingerol, were significantly identified in ginger extract using HPLC analysis. Antioxidant activity of these active metabolites were shown to be higher against in vitro free radicals (DPPH and Β-Carotene-linoleic acid), suggesting the potential antioxidant and antiapoptotic properties of ginger against CP-toxicity. CONCLUSIONS: Treatment with ginger in rats induced with CP resulted in significant improvement in the expression of certain molecular miRNAs. The kidney tissues of these rats showed a marked decrease in the expression of miR-155-5p, miR-34a-5p, and miR-21-5p, while the levels of miR-193b-3p, miR-455-3p, and miR-342-3p were observed to increase significantly. In conclusion, ginger can protect rats from CP-induced nephrotoxicity.

Back to nature: henna extracts from nanotech to environmental biotechnology - a review.

The Lythraceae family includes henna (Lawsonia inermis), which thrives in subtropical and tropical climates. One of its many and long-standing uses is in cosmetics as a pigment to color hair and nails. Additionally, it serves as a disinfectant against microbiological infections and has traditional applications in the textile industry, specifically for coloring wool and nylon. The dried leaves of henna contain a significant amount of lawsone, an active substance bestowing them with staining abilities. Environmental biotechnology, a subfield of biotechnology, engages in the production of biomass or renewable energy sources and the elimination of pollutants, utilizing either entire organisms or their by-products. Recent research indicates that henna, owing to its sustainability, abundant production, simplicity of preparation, low cost, and reputation for being safe and ecologically benign, is exceptionally well-suited to participate in the realm of environmental biotechnology. This review navigates through the most recent studies exploring the use of henna and its extracts for related purposes. These encompass a spectrum of applications, including but not limited to nanobiotechnology, fabric dyeing, corrosion resistance, colored solar cells, carbon dots, and new renewable energy exemplified by biofuel and biohydrogen. Furthermore, henna extracts have been deployed to function as antimicrobials and ward off dangerous insects.

Identifying 3rd larval stages of common strongylid and non-strongylid nematodes (class: Nematoda) infecting Egyptian equines based on morphometric analysis.

Strongylid and non-strongylid nematodes are one of the most important parasites infecting equines. The traditional method to identify these nematodes is through coproscopy and fecal culture. Because of the scarcity of data published in Egypt discussing the morphometric features of infective 3rd larvae of these nematodes, this study aims to provide a morphometric key for L3 of common strongylid and non-strongylid nematodes infecting Egyptian equines. For this reason, we cultured fecal samples containing GINs eggs and 3rd larval stages were identified based on their morphology (i.e., shape and number of intestinal cells (IC), shape of the esophagus, and shape of the tail sheath) in addition to computing their dimensions (i.e., length of larvae with sheath, length of the esophagus, length of intestinal cells, and body breadth). We identified 3rd larval stages of four strongylid nematodes (Cyathostomum sensu lato, Strongylus vulgaris, Strongylus equinus, and Strongylus edentatus) as well as two non-strongylid nematodes (Strongyloides westeri, and Trichostrongylus axei). Statistically, our results revealed significant differences in terms of total length, body width, esophagus length, and gut length among 3rd larvae identified in the current study. The combination of both morphological and metric keys will allow the better identification of common strongylid and non-strongylid nematodes infecting equines.

Immunological and histopathological evaluation of Eimeria tenella oocysts Egyptian local isolate vaccine and its comparative efficacy with a commercial live vaccine.

Coccidiosis is the most important protozoan disease in broilers all over the world. Controlling of broilers coccidiosis via vaccination rather than chemicals is a new trend with promising results. Thus, the present work describes an evaluation of Eimeria tenella Lab-made vaccine of local Egyptian strain and its comparative efficacy with a commercial live vaccine "Fortegra®". Eighty broiler chickens one day old were used; they were divided in to 4 equal groups; 20 chicks each. Group 1 (G1) kept as control negative, G2 administrated orally with lab-made sporulated oocysts vaccine at 5 days old, the birds of G3 vaccinated orally with Fortegra® at day 6 of age, and G4 served as control positive. All birds were challenge by 50,000 sporulated oocysts of E. tenella at day 21. For testing the efficacy and comparison; OPG (oocyst per gram), serum Interleukin4 (IL4) levels, Immunoglobulin A (IgA) levels in both serum and ceca, cecal lesion score, as well as histopathological changes in ceca of tested groups were evaluated. The results demonstrated significantly elevated IL4 level in serum and IgA level in serum and cecum of G2 than G3. IgA in cecum significantly elevated in G2 than G3. OPG significantly decreased in both vaccinated groups (G2 and G3), and have lower lesion score than nonimmunized group. Cecal tissues of vaccinated groups had mild pathological changes. Conclusively, good immunization by the currently tested vaccine, against experimental E. tenella infection was observed.

Immunological and histopathological evaluation of Eimeria tenella oocysts Egyptian local isolate vaccine and its comparative efficacy with a commercial live vaccine

@#Coccidiosis is the most important protozoan disease in broilers all over the world. Controlling of broilers coccidiosis via vaccination rather than chemicals is a new trend with promising results. Thus, the present work describes an evaluation of Eimeria tenella Lab-made vaccine of local Egyptian strain and its comparative efficacy with a commercial live vaccine “Fortegra®”. Eighty broiler chickens one day old were used; they were divided in to 4 equal groups; 20 chicks each. Group 1 (G1) kept as control negative, G2 administrated orally with lab-made sporulated oocysts vaccine at 5 days old, the birds of G3 vaccinated orally with Fortegra® at day 6 of age, and G4 served as control positive. All birds were challenge by 50,000 sporulated oocysts of E. tenella at day 21. For testing the efficacy and comparison; OPG (oocyst per gram), serum Interleukin4 (IL4) levels, Immunoglobulin A (IgA) levels in both serum and ceca, cecal lesion score, as well as histopathological changes in ceca of tested groups were evaluated. The results demonstrated significantly elevated IL4 level in serum and IgA level in serum and cecum of G2 than G3. IgA in cecum significantly elevated in G2 than G3. OPG significantly decreased in both vaccinated groups (G2 and G3), and have lower lesion score than nonimmunized group. Cecal tissues of vaccinated groups had mild pathological changes. Conclusively, good immunization by the currently tested vaccine, against experimental E. tenella infection was observed.

In vitro assessment of the bioactivities of sericin protein extracted from a bacterial silk-like biopolymer.

Sericin is one of the main components of silk proteins, which has numerous biomedical applications because of its antioxidant, anticancer and antimicrobial properties. We recently isolated and characterized a novel silk-like protein named BNES. It is of non-animal origin and is like a bacterial polymeric silk. Sericin is a very popular protein compound that is effective in treating cancerous tumors. The process of extracting it from natural silk produced by silkworms or spiders is both complex and expensive. From the published scientific literature, it has been shown that sericin has not been previously extracted from a bacterial source. In the present study, sericin was extracted from bacteria capable of producing a biopolymer named BNES whose chemical composition is like that of natural silk and its bio-therapeutic effects were evaluated for the first time. The antioxidant activity of BNES measured by DPPH and ABTS assays showed IC50 values of 0.38 and 0.41 mg mL-1, respectively. BNES displayed satisfactory cytotoxic effect against four cancer cell lines, including Huh-7, Caco-2, MCF-7 and A549 cells, with IC50 values in the ranges of ca. 0.62 ± 0.17, 0.72 ± 0.27, 0.76 ± 0.36 and 0.83 ± 0.31 mg mL-1, respectively, after 24 h of treatment and 0.51 ± 0.22, 0.49 ± 0.19, 0.41 ± 0.25 and 0.55 ± 0.38, respectively, after 48 h of treatment, without affecting normal cells (WI38 cells). The antitumor activity of BNES was established to be an apoptosis-dependent mechanism determined via cellular morphology alterations, cell cycle arrest in the sub-G1 phase and nuclear staining with highly fluorescent fragments. The antimicrobial effects of BNES were examined with yeast and Gram-negative and Gram-positive bacteria. The results confirmed its antimicrobial activity against all tested organisms at concentrations of up to 1.33 mg mL-1. The competitive advantage of the bacterial sericin BNES over sericin extracted from spider or silkworm sources is that it can be produced in very large quantities through large-scale bio-fermenters, which reduces the expected cost of production, in addition to having sustainable bacterial production source.

Disinfection of water and wastewater by biosynthesized magnetite and zerovalent iron nanoparticles via NAP-NAR enzymes of Proteus mirabilis 10B.

Disinfection of water and wastewater strongly contributes to solving the problem of water shortage in arid/semi-arid areas; cheap and ecofriendly approaches have to be used to meet water quality standards. In the present study, a green synthesis of iron nanoparticles (INPs) under aerobic and anaerobic conditions via nitrate reductases (NAP/NAR) enzymes produced by Proteus mirabilis strain 10B were employed for this target. The biosynthesized INPs were characterized; UV-Vis spectroscopy revealed surface plasmon resonance at 410 (aerobic) and 265 nm (anaerobic). XRD indicated crystalline magnetite ((MNPs) aerobically synthesized) and zerovalent INPs (ZVINPs anaerobically synthesized). EDX demonstrated strong iron signal with atomic percentages 73.3% (MNPs) and 61.7% (ZVINPs). TEM micrographs illustrated tiny, spherical, periplasmic MNPs (1.44-1.92 nm) and cytoplasmic ZVINPs with 11.7-60.8 nm. Zeta potential recorded - 31.8 mV (ZVINPs) and - 66.4 mV (MNPs) affirming colloidal stability. Moreover, the disinfection power of INPs was evaluated for standards organisms and real water (fresh, sea and salt mine) and wastewater (municipal, agricultural and industrial) samples. The results reported that INPs displayed higher antagonistic effect than iron precursor, 700 and 850 µg/mL of MNPs and ZVINPs, respectively, was sufficient to show a drastic algicidal effect on algal growth. Both types of INPs demonstrated obvious dose-dependent antibiofilm efficiency. Due to their smaller size, MNPs were more efficient than ZVINPs at the suppression of microbial growth in all examined water samples. Overall, MNPs showed superior antagonistic activity, which promotes their exploitation in enhancing water/wastewater quality.

Detection of infectious laryngotracheitis virus (Gallid herpesvirus-1) from clinically infected chickens in Egypt by different diagnostic methods.

Infectious laryngotracheitis (ILT) disease is an acute highly contagious viral disease leading to massive economic losses to the national poultry industry. This study aimed to identify the most accurate and rapid diagnostic methods to rescue layer poultry farms from intense outbreaks in Egypt. Fifty pathological specimens were collected and subjected to virus isolation (VI), histopathology, direct fluorescent antibody technique (FAT) and polymerase chain reaction (PCR). Egg inoculation revealed stunted growth and white pock lesions on chorioallantoic membranes (CAM) in 23 samples. Isolation and propagation of infectious laryngotracheitis virus (ILTV) in cell culture showed syncytia formation 5 days post infection in 20 inoculated samples. PCR resulted in successful amplification of a 647 bp fragment of the thymidine kinase (TK) gene in 25 field samples. Histopathological examination of inoculated CAM showed intranuclear inclusion bodies with infiltration of inflammatory cells. Direct FAT showed intra-cytoplasmic apple green reactions in 18 examined tracheal tissues. PCR has been shown to be more sensitive, accurate and rapid than VI, FAT and histopathological examination.

Sarcocystis chloropusae (protozoa: Sarcocystidae) n. sp. from the common moorhen (Gallinula chloropus) from Egypt.

A new name Sarcocystis chloropusae is proposed for a parasite previously found in two of 25 common moorhen (Gallinula chloropus) from Brolos Lake, Egypt. Sarcocysts were microscopic, up to 650 µm long, the cyst wall was up to 4.5 µm thick, and contained villar protrusions that were up to 4 µm long and up to 2 µm wide. The villar protrusions were crowded, contained vesicles but lacked microtubules. The ground substance layer was smooth. The bradyzoites were up to 12 µm long and up to 2 µm wide. Molecular characterization and phylogenetic analysis of the (ITS-1) supported the conclusion that the Sarcocystis in G. chloropus is a distinct species.

Effects of curcumin on Cryptosporidium parvum in vitro.

Cryptosporidium parvum is a zoonotic protozoan parasite having peculiarities among the apicomplexa that could be responsible for its resistance to some drugs and disinfectants against coccidia. The awareness of Cryptosporidium as a health problem in man and animal is increasing and potent drugs are urgently needed. Curcumin, a natural polyphenolic compound, has been found to be active against a variety of diseases including anticarcinogenic, antimicrobial, and antiprotozoal effects. We investigated the effects of curcumin on infectivity and development of C. parvum in a recently established in vitro system combining infection of human ileocecal adenocarcinoma cell cultures with quantification of intracellular parasites by quantitative polymerase chain reaction. Curcumin was found to be effective (>95% inhibition of parasite growth) at 50 microM for 24 h when infected cultures were exposed for more than 12 h. Withdrawal of curcumin after 24 h of exposure did not result in a significant resumption of C. parvum growth. The invasion of host cells by sporozoites (infectivity) was found to be inhibited at least 65% in the presence of 200 microM curcumin. No significant reduction of viability of C. parvum oocysts after incubation with curcumin was recorded. Altogether, curcumin showed promising anticryptosporidial effects under in vitro conditions and deserves further exploration.

Biosynthesis of polyhydroxyalkanotes in wild type yeasts.

Biosynthesis of biodegradable polymers polyhydroxyalkanotes (PHAs) have been studied extensively in wild type and genetically modified prokaryotic cells, however the content and structure of PHAs in wild type yeasts is not well documented. The purpose of this study was to screen yeast isolates collected from different ecosystems for their ability to accumulate PHAs. Identification of the isolates and characterization of PHAs produced by the positive isolates were investigated. One positive isolate (strain Y4) was identified by both API20C system and 18S rDNA sequencing. The data revealed that isolate Y4 belongs to the yeast genus Rhodotorula and exhibits 18S rDNA similarity value >99% to the species Rhodotorula minuta. Quantification of PHAs yield of strain Y4 in glucose, oleic acid and tween 60 containing medium for over a growth period of 96 h gave 2% of PHAs in biomass. The nature of produced PHAs was analyzed by infrared spectroscopy and nuclear magnetic resonance (1H and 13C NMR) and found to contain polyhydroxybutyrate and polyhydroxyvalerate (PHBV).

Isolation and characterization of extracellular bioflocculants produced by bacteria isolated from Qatari ecosystems.

Compared with conventional synthetic flocculants, bioflocculants has special advantages such as safety, strong effect, biodegradable and harmlessness to humans and the environment, so they may potentially be applied in drinking and wastewater treatment, downstream processing, and fermentation processes. To utilize bioflocculants widely in industrial fields, it is desirable to find various microorganisms with high bioflocculant-producing ability and improve the flocculating efficiency of the bioflocculant. In the present study, screening of new flocculant-producing microorganisms was carried out using samples collected from different Qatari ecosystems. The flocculating activity of the novel bioflocculants produced by isolated microorganisms was investigated. A total of 5 g/l Kaolin suspension was used to measure the flocculating activity. Isolated bioflocculant-producing bacteria were identified by 16S rDNA analysis, using PCR with universal primers. Comparative analysis of the 16S rDNA sequence (approximately 550 bp) in the GenBank database revealed that these bacteria are related to the genus Bacillus. FT-IR spectrometry analysis of the extracted bioflocculants indicated the presence of carboxyl, hydroxyl and amino groups preferred for the flocculation process. Influences of pH and bioflocculant dosage on the flocculation were also examined. The maximum flocculating rates were observed at pH 7, 7 and 3 of the bioflocculants derived from strains QUST2, QUST6 and QUST9, respectively. However, 20.0 mg/l was the dose that gave the highest flocculating rate with all examined bioflocculants. The elemental analysis of examined bioflocculants revealed the mass proportion of C, H, N and S. Carbon and nitrogen contents of examined bioflocculants were in the range of 42-48% and 11-12%, respectively.

Leishmania donovani: expression and characterization of Escherichia coli-expressed recombinant chitinase LdCHT1.

Leishmania parasites produce chitinase activity (EC. 3.2.1.14) thought to be important in parasite-sandfly interactions and transmission of the parasite to the vertebrate host. Previous observations have suggested that parasite chitinases are involved in degradation of the sandfly peritrophic matrix and the chitinous layer of the cardiac valve cuticle. This chitinase activity is thought to produce an incompetent pharyngeal valve sphincter and a route of egress that allow Leishmania promastigotes to be regurgitated into the site of blood feeding. In the studies reported here, enzymatically active L. donovani chitinase LdCHT1 was expressed as a thioredoxin fusion protein in Escherichia coli strain AD494 (DE3). Recombinant LdCHT1 had a predominantly endochitinase activity, in contrast to previous reports of both exo- and endochitinase activities in axenic culture supernatants of diverse Leishmania spp. promastigotes. The predominant endochitinase activity of recombinant LdCHT1 is consistent with the presumed function of the enzyme in disrupting chitinous structures in the sandfly digestive system to allow transmission.

Uptake and utilization of n-octacosane and n-nonacosane by Arthrobacter nicotianae KCC B35.

Arthrobacter nicotianae KCC B35 isolated from blue-green mats densely covering oil sediments along the Arabian Gulf coast grew well on C10 to C40 n-alkanes as sole sources of carbon and energy. Growth on C20 to C40 alkanes was even better than on C10 to C18 alkanes. Biomass samples incubated for 6 h with n-octacosane (C28) or n-nonacosane (C29) accumulated these compounds as the predominant constituent alkanes of the cell hydrocarbon fractions. The even chain hexadecane C16 and the odd chain pentadecane C15 were the second dominant constituent alkanes in C28 and C29 incubated cells, respectively. n-Hexadecane-incubated cells accumulated in their lipids higher proportions of C16-fatty acids than control cells not incubated with hydrocarbons. On the other hand, C28 and C29-incubated cells did not contain any fatty acids with the equivalent chain lengths, but the fatty acid patterns of the cell lipids suggest that there should have been mid-chain oxidation of these very long chain alkanes. This activity qualifies A. nicotianae KCC B35 to be used in cocktails for bioremediating environments polluted with heavy oil sediments.

Hepatitis-B virus and schistosomiasis infections in childhood proteinuria.

Hepatitis-B surface antigen (HBsAg), circulating anti-schistosomal IgG (CSAb) and circulating specific schistosomal immune complexes (CIC) were detected, using ELISA, in sera of 40 active nephrotic children, 40 active S. mansoni infected cases and 20 apparently normal age-matched controls. The presence of HBsAg cases was significantly higher among nephrotic cases (20%), active S. mansoni cases (17.5%) than controls. Moreover, HBsAg cases were significantly higher in positive CIC S. mansoni cases than negative CIC ones. The mean O.D. readings of CSAb was significantly higher in positive HBsAg nephrotic cases than negatives. At the same time, the anti-schistosomal antibodies were higher in S. mansoni cases with proteinuria than those without. Specific CIC level was significantly higher among nephrotic and schistosomiasis cases than controls. The CIC were significantly higher in schistosomiasis cases with positive HBsAg than those with negative HBsAg and were detected in 80% of cases with proteinuria compared to 37% of cases without proteinuria with a statistically significant difference. On the other hand, CIC level was not influenced, in nephrotic cases, by the presence or absence of HBsAg. It was concluded that the presence of proteinuria was considered as a good monitor of the kidney affection either with schistosomiasis or the nephrotic syndrome or the HBsAg. The detection of CIC can be used as a good monitor too and could be included in methods of early diagnosis and/or following the disease prognosis.