Use of lemongrass essential oil as a feed additive in quail's nutrition: its effect on growth, carcass, blood biochemistry, antioxidant and immunological indices, digestive enzymes and intestinal microbiota.

The present study was designed to assess the impact of dietary supplementation of lemongrass essential oil (LGEO) on growth performance, carcass traits, liver and kidney function, immunity, antioxidant indices and caecal microbiota of growing quail. A total of 200 Japanese quails at 1-week-old were haphazardly allotted to 5 groups of 40 chicks in five replicates (8 per replicate). The first group was the control group, while LGEO was added at levels of 150, 300, 450, and 600 mg/kg diet in the 2nd, 3rd, 4th and 5th groups, respectively. Dietary supplementation of LGEO (150, 300 and 450 mg/ kg diet) increased body weight at 3 and 5 wk of age, and increased body weight gain during all periods compared with the control group (P < 0.05). All levels of LGEO improved feed conversion ratio during the periods from 1 to 3 and 1 to 5 wk of age. During 3 to 5 wk, feed conversion ratio was improved in quails fed LGEO (300 and 450 mg/kg diet) compared with the control and other treatments. Carcass traits, plasma globulin, alanine aminotransferase, and urea values did not differ among the treatments (P > 0.05), but the activity of aspartate aminotransferase in the plasma was significantly decreased (P < 0.05) in LGEO-treated groups. The total protein and albumin values were significantly increased (P < 0.05) in quails fed levels of LGEO (except 600 mg/kg diet) compared with the control. The inclusion of LGEO in quail diets improved (P < 0.05) plasma lipid profile. The dietary supplementation of LGEO increased (linear and quadratic, P < 0.05) plasma immunoglobulins (IgM, IgG, and IgA) levels, lysozyme values and activities of superoxide dismutase, total antioxidant capacity, reduced glutathione and catalase compared with the control group. The caecal Coliform, E. coli and Salmonella were lowered (P < 0.0001) in the quails treated with all LGEO levels, but the total bacterial count and Lactobacillus count were increased with dietary supplementation of LGEO levels (300 and 450 mg/kg) compared with those in the control group. The activities of digestive enzymes were significantly higher in birds fed the diet supplemented with LGEO levels than those fed the control diet. In conclusion, dietary supplementation of LGEO can improve the performance, lipid profile, immunity and antioxidant indices and decline intestinal pathogens and thus boost the health status of growing quail.

DOWN REGULATION OF CLASSICAL MONOCYTES SUBSET IN PATIENTS WITH HCV RELATED LIVER FIBROSIS.

Chronic liver disease is a worldwide common pathology characterized by inflammatory and fibrotic processes that may lead to progressive evolution from chronic hepatitis to cirrhosis. Peripheral blood monocytes may play an important role in the pathogenesis and resolution of liver fibrosis. These cells may offer new approaches for better understanding the pathogenesisof liver fibrosis. This work defined the proportion of circulating classical monocyte subset with hematopoietic origin in peripheral blood to establish the possible potential role of this subset as non-invasive biomarker of liver fibrosis in patients with HCV related chronic liver disease. Forty patients with HCV induced chronic liver disease were classified according to the stage of liver fibrosis after METAVIR score into 4 groups, patients with stages F1, F2, F3 & F4 liver fibrosis (10 patients each) and 10 healthy subjects served as normal controls. Flowcytometric analysis for immunophenotypic characterization for identification of levels of circulating peripheral blood classical monocytes subset in different groups studied was carried out using monoclonal antibodies anti-CD45, anti-CD 14 and anti-CD 16. THE RESULTS: data demonstrated a significant down regulation (p< 0.01) in the proportion of classical monocytes subset (CD45+ve, CD 14+ve and CD 16-ve) in patients with chronic hepatitis C related liver disease compared to healthy subjects. Data also demonstrate that down regulation of the expression of classical monocyte subset paralleled the worsening severity of liver disease and the progression of liver fibrosis.

Role of fibrogenic markers in chronic hepatitis C and associated hepatocellular carcinoma.

Detection and follow up of fibrogenesis in chronic hepatitis C (CHC) is mandatory for early treatment and risk stratification. The current study included 120 patients with CHC, of whom 30 had liver cirrhosis (LC) and 30 had hepatocellular carcinoma (HCC). 15 wedge liver biopsies, taken during laparoscopic cholecystectomy, were included as normal controls. Cases were subjected to laboratory investigations, serologic markers for viral hepatitis and assessment of circulating levels of hyaluronic acid (HA) and platelet-derived growth factor (PDGF). Immunohistochemical expression of connective tissue growth factor (CTGF), PDGF and transforming growth factor-ß1 (TGF-ß1) was also carried out. A significant increase (p < 0.01) in serum HA was noticed in CHC, LC and HCC compared to controls. Although, a significant decrease in serum PDGF was detected in CHC and LC compared to controls, HCC values were comparable. A significant up-regulation of CTGF was detected in CHC, LC and HCC (p < 0.01) in contrast to its limited mild expression in normal livers. Intense PDGF positive staining was noticed in CHC, LC and HCC compared to scattered faint expression in controls. The significant expression and marked intensity of PDGF staining matched the progress to tumorigenesis. A positive TGF-ß1 immunostaining was also noticed in CHC, LC and HCC. An intense and extensive cytoplasmic expression of TGF-ß1 was encountered in patients with LC revealing that CTGF, PDGF and TGF-ß1 act synergistically in LC. Data revealed that HA and CTGF may be implicated as important diagnostic parameters for assessment of hepatic fibrosis and PDGF for monitoring malignant transformation in CHC.

Environmental exposure to asbestos and the exposure-response relationship with mesothelioma.

An epidemiological and environmental study was carried out in Shubra El-Kheima city, greater Cairo, of the exposure-response relationship between asbestos and malignant pleural mesothelioma. Radiological screening was done for 487 people occupationally exposed to asbestos, 2913 environmentally exposed to asbestos and a control group of 979 with no history of exposure. Pleural biopsy was done for suspicious cases. The airborne asbestos fibre concentrations were determined in all areas. There were 88 cases of mesothelioma diagnosed, 87 in the exposed group. The risk of mesothelioma was higher in the environmentally exposed group than other groups, and higher in females than males. The prevalence of mesothelioma increased with increased cumulative exposure to asbestos.

Environmental exposure to asbestos and the exposure-response relationship with mesothelioma

An epidemiological and environmental study was carried out in Shubra El-Kheima city, greater Cairo, of the exposure-response relationship between asbestos and malignant pleural mesothelioma. Radiological screening was done for 487 people occupationally exposed to asbestos, 2913 environmentally exposed to asbestos and a control group of 979 with no history of exposure. Pleural biopsy was done for suspicious cases. The airborne asbestos fibre concentrations were determined in all areas. There were 88 cases of mesothelioma diagnosed, 87 in the exposed group. The risk of mesothelioma was higher in the environmentally exposed group than other groups, and higher in females than males. The prevalence of mesothelioma increased with increased cumulative exposure to asbestos

Field and storage testing Bt potatoes for resistance to potato tuberworm (Lepidoptera: Gelichiidae).

Potato tuberworm, Phthorimaea operculella (Zeller), is the most serious insect pest of potatoes worldwide. The introduction of the Bacillus thuringiensis (Bt) toxin gene through genetic engineering offers host plant resistance for the management of potato tuberworm. We report on the field and storage studies to evaluate Bt-cry5 potato lines for resistance to potato tuberworm in Egypt under natural infestations and their agronomic performance in both Egypt and Michigan. From 1997 to 2001, field experiments were conducted at the International Potato Center (CIP) Research Station, Kafr El-Zyat, Egypt, and/or Agricultural Genetic Engineering Institute (AGERI), Giza, Egypt, to evaluate resistance to tuberworm. A total of 27 Bt-transgenic potato lines from six different Bt constructs were evaluated over a 5-yr period. After harvest and evaluation of the agronomic trials, storage evaluation of potato tuberworm damage was done at the CIP Research Station. The 1997 field trial was the first field test of genetically engineered crops in Egypt. Field tests to assess potato tuberworm resistance in Egypt were able to differentiate between the Bt-transgenic lines and the nontransgenic lines/cultivars in 1999, 2000, and 2001. The Bt-cry5-Spunta lines (Spunta-G2, Spunta-G3, and Spunta-6a3) were the most resistant lines in field with 99-100% of tubers free of damage. In the 2001 storage study, these lines were also over 90% free of tuberworm damage after 3 mo. NYL235-4.13, which combines glandular trichomes with the Bt-cry5/gus fusion construct, also had a high percentage of clean tubers in the field studies. In agronomic field trials in Michigan from 1997 to 2001, the Bt-transgenic lines in most instances performed similar to the nontransgenic line in the agronomic trials; however, in Egypt (1998-1999), the yields were less than one-half of those in Michigan. Expression of the Bt-cry5 gene in the potato tuber and foliage will provide the seed producer and grower a tool in which to reduce potato tuberworm damage to the tuber crop in the field and storage.

Wolbachia infection and the expression of cytoplasmic incompatibility in sandflies (Diptera: Psychodidae) from Egypt.

A PCR-based method was used to screen four laboratory colonies of sandflies for Wolbachia infection. The colonies - one of Phlebotomus langeroni, one of P. bergeroti and two of P. papatasi - were all derived from sandflies collected in Egypt. Only one of the colonies, derived from P. papatasi collected in Sinai, was found infected. The sequence of the PCR product for this colony was identical to that previously reported for the Wolbachia in P. papatasi from Israel. The induction with tetracycline of cytoplasmic incompatibility (CI) in flies from the P. papatasi (Sinai) colony was then investigated, through reciprocal crosses between treated and untreated P. papatasi siblings. Partial CI expression was attained in the crosses involving antibiotic-treated (i.e. uninfected) females, whether the males used were infected with Wolbachia or had also been cleared of Wolbachia by antibiotic treatment. Most (75%) of the eggs oviposited by uninfected females that had been crossed with infected males, and most (58%) of those laid by uninfected females that had been crossed with uninfected males, failed to hatch. These results provide the first published evidence showing that Wolbachia infection in sandflies is advantageous to the insects. The failure to detect Wolbachia in one of the colonies derived from Egyptian P. papatasi or in the colonies derived from Egyptian P. bergeroti and P. langeroni may indicate that the inter- and intra-specific spread of Wolbachia is discontinuous, even within one country.

An association between acquired epiphora and the signs and symptoms of chronic rhinosinusitis: a prospective case-control study.

BACKGROUND: It has been suggested that chronic rhinosinusitis may lead to epiphora because of inflammatory edema at the nasal end of the nasolacrimal duct and that treatment of the underlying nasal disease may obviate the need for dacryocystorhinostomy. The aim of this study was to establish whether or not there is an association between the signs and symptoms of chronic rhinosinusitis and a complaint of epiphora in a blinded, prospective case-control study. METHODS: A consecutive series of 15 adult patients presenting to the ophthalmology department with acquired epiphora were compared with 29 patients presenting with chronic open-angle glaucoma over the same study period. Nasal symptoms and the findings on nasal endoscopy were recorded by a single otolaryngologist blinded to the diagnosis and using a standardized staging system RESULTS: The age and sex distributions of the two groups were similar. Scores for headache and altered smell were significantly higher in the epiphora cases than in controls (p = 0.05 and 0.03, respectively). Trends for higher scores for congestion and discharge were not statistically significant (p = 0.06 and 0.07, respectively). Mucosal edema and discharge were significantly more common on endoscopy in the epiphora cases than in the control cases (p < 0.02). CONCLUSION: We have shown an association between rhinosinusitis and acquired epiphora, and this would be consistent with chronic rhinosinusitis being the cause of the epiphora in some cases.

Cloning and expression of a Ralstonia eutropha HF39 gene mediating indigo formation in Escherichia coli.

On complex medium Escherichia coli strains carrying hybrid plasmid pBEC/EE:11.0, pSKBEC/BE:9.0, pSKBEC/PP:3.3, or pSKBEC/PP:2.4 harboring genomic DNA of Ralstonia eutropha HF39 produced a blue pigment characterized as indigo by several chemical and spectroscopic methods. A 1,251-bp open reading frame (bec) was cloned and sequenced. The deduced amino acid sequence of bec showed only weak similarities to short-chain acyl-coenzyme A dehydrogenases, and the gene product catalyzed formation of indoxyl, a reactive preliminary stage for production of indigo.

Sensitive detection of the Egyptian species of sugarcane streak virus by PCR-probe capture hybridization (PCR-ELISA) and its complete nucleotide sequence.

A rapid and sensitive assay for the specific detection of Sugarcane streak virus (SSV) using PCR-probe capture hybridization (PCR-ELISA) was developed. Nucleic acids suitable for PCR were extracted from SSV-infected tissue using organic solvents or Fast DNA kit. SSV cDNA was amplified using viral specific primers and the amplified SSV cDNA (amplicon) was DIG-labelled during the amplification process. The amplicon was then detected in a colorimetric hybridization system by a microtiter plate using a biotinylated cDNA (22 nt), cDNA (789 nt) or cRNA (789 nt) capture probe. This system combines the specificity of molecular hybridization, the ease of the colorimetric protocol, and is 10-100 fold more sensitive than agarose gel electrophoretic analysis in detecting the amplified product. Long cDNA or cRNA capture probe was 2-7 fold more sensitive than the oligo cDNA probe for the detection. Complete nucleotide sequence of SSV from Naga Hammady, Egypt, revealed that SSV-EG is a new species of SSV that shares 66% nucleotide identity with the virus species from Natal, South Africa.

Evaluation of potato tuber moth (Lepidoptera: Gelechiidae) resistance in tubers of Bt-cry5 transgenic potato lines.

The potato tuber moth, Phthorimaea operculella (Zeller), in tropical and subtropical countries, is the most destructive pest of potato, Solanum tuberosum L. The larvae attack foliage and tubers in the field and in storage. The purpose of this study was to evaluate the efficacy of a Bt-cry5 transgene to control the potato tuber moth in tuber tissues. Tuber bioassays using stored (11-12 mo old) and newly harvested tubers of Bt-cry5-Lemhi Russet and Bt-cry5-Atlantic potato lines showed up to 100% mortality of 1st instars. Mortality was lowest in the newly harvested tubers of Bt-cry5-Atlantic lines (47.1-67.6%). Potato tuber moth mortality was 100% in the Bt-cry5-Spunta lines that were transformed with Bt-cry5 gene controlled by the CaMV 35S promoter (pBIML5 vector) and in 2 of 3 lines transformed with Bt-cry5 gene controlled by the Gelvin super promoter (pBIML1 vector). The transgenic Spunta lines expressing Bt-cry5 controlled by the patatin promoter (pBMIL2 vector) showed the lowest tuber moth mortality (25.6 and 31.1%). The Bt-cry5 transgenic lines with high tuber expression of B. thuringiensis have value in an integrated pest management system to control potato tuber moth.

Structure of the Na+-driven flagellum from the homoacetogenic bacterium Acetobacterium woodii.

The Na+-dependent flagellum of Acetobacterium woodii was characterised. Flagellin and whole flagella were purified and analysed by SDS-PAGE and electron microscopy. The structure and dimensions of the filament and the hook-basal body, as revealed by electron microscopy, resemble those of H+-dependent flagella from gram-positive bacteria. Intramembrane particle rings were present at the cell pole in freeze-fractured A. woodii cells, which might correspond to the mot complex.

Analysis of the PHA granule-associated proteins GA20 and GA11 in Methylobacterium extorquens and Methylobacterium rhodesianum.

Electrophoretic analysis of the proteins bound to poly(3-hydroxybutyric acid), PHB-, granules in Methylobacterium extorquens, M. rhodesianum as well as the PHB-leaky mutants Mu 1 and Mu 11, which were isolated from the latter, resulted in two dominant low-molecular weight proteins, which were referred to as GA11 and GA20. After purification of these proteins antibodies against the GA11 and GA20 protein of M. extorquens were obtained. Both proteins bound to the surface of PHB granules as revealed by immunoelectron microscopy of whole cells of M. extorquens and M. rhodesianum. With cells of the PHB-leaky mutants Mu 1 and Mu 11 no specific labeling was observed. The N-terminal amino acid sequences of the GA11 and the GA20 protein were determined. We found significant homologies between the sequences of the investigated strains. The use of oligonucleotide probes based on the N-terminal sequences of the GA20 protein from M. rhodesianum to identify the corresponding structural genes in various genomic libraries failed.

Reproductive hazards of lead exposure among urban Egyptian men.

Fifty-five urban Egyptian males, aged 20-40, were assigned to two main groups to study the effects of their exposure to lead (Pb). Group I, infertile men (INF, n = 30), was divided into environmentally exposed (INF-E, n = 15) and environmentally and occupationally exposed (INF-EO, n = 15). A matching group (II) of fertile men (F, n = 25) was divided into fertile, environmentally exposed (F-E, n = 10), which was the control group, and fertile, environmentally and occupationally exposed (F-EO, n = 15). Semen parameters (i.e., count, morphology, motility, and volume), blood and semen Pb levels, and reproductive hormonal indices (i.e., serum testosterone, FSH, and LH) were measured in all subjects. Lead levels were always higher in blood than semen. Semen lead levels were significantly higher in all groups vs. the control (F-E) group. While no changes were observed in testosterone levels across groups, variable effects on LH and FSH levels were observed. Infertile-EO subjects showed a definite pattern of impaired semen parameters in comparison with infertile-E. No abnormalities were detected in hematologic, hepatic or renal function.

Identification of the region of a 14-kilodalton protein of Rhodococcus ruber that is responsible for the binding of this phasin to polyhydroxyalkanoic acid granules.

The function of the polyhydroxyalkanoic acid (PHA) granule-associated GA14 protein of Rhodococcus ruber was investigated in Escherichia coli XL1-Blue, which coexpressed this protein with the polyhydroxybutyric acid (PHB) biosynthesis operon of Alcaligenes eutrophus. The GA14 protein had no influence on the biosynthesis rate of PHB in E. coli XL1-Blue(pSKCO7), but this recombinant E. coli strain formed smaller PHB granules than were formed by an E. coli strain that expressed only the PHB operon. Immunoelectron microscopy with GA14-specific antibodies demonstrated the binding of GA14 protein to these mini granules. In a previous study, two hydrophobic domains close to the C terminus of the GA14 protein were analyzed, and a working hypothesis that suggested an anchoring of the GA14 protein in the phospholipid monolayer surrounding the PHA granule core by these hydrophobic domains was developed (U. Pieper-Fürst, M. H. Madkour, F. Mayer, and A. Steinbüchel, J. Bacteriol. 176:4328-4337, 1994). This hypothesis was confirmed by the construction of C-terminally truncated variants of the GA14 protein lacking the second or both hydrophobic domains and by the demonstration of their inability to bind to PHB granules. Further confirmation of the hypothesis was obtained by the construction of a fusion protein composed of the acetaldehyde dehydrogenase II of A. eutrophus and the C terminus of the GA14 protein containing both hydrophobic domains and by its affinity to native and artificial PHB granules.

Post-heatstroke parkinsonism and cerebellar dysfunction.

A case is presented of a 45-year-old woman who developed parkinsonism, cerebellar dysfunction, aphasia and flaccid paralysis secondary to heat stroke. Magnetic resonance imaging revealed irregular patchy areas of increased signal intensity in the white matter of both cerebral hemispheres and the left striatum, remaining unchanged over a follow-up period of two years. Cerebellar atrophy, while not seen on initial examination, was present on the follow-up study done one year later and became more marked after two years.

Considerations on the structure and biochemistry of bacterial polyhydroxyalkanoic acid inclusions.

Some mathematical calculations were done that provided information about the structure and biochemistry of polyhydroxyalkanoic acid (PHA) granules and about the amounts of the different constituents that contribute to the PHA granules. The data obtained from these calculations are compared with data from the literature, which show that PHA granules consist not only of the polyester but also of phospholipids and proteins. The latter are referred to as granule-associated proteins, and they are always located at the surface of the PHA granules. A concept is proposed that distinguishes four classes of structurally and functionally different granule-associated proteins: (i) class I comprises the PHA synthases, which catalyze the formation of ester linkages between the constituents; (ii) class II comprises the PHA depolymerases, which are responsible for the intracellular degradation of PHA, (iii) class III comprises a new type of protein, which is referred to as phasins and which has most probably a function analogous to that of oleosins in oilseed plants, and (iv) class IV comprises all other proteins, which have been found to be associated with the granules but do not belong to classes I-III. Particular emphasis is placed on the phasins, which constitute a significant fraction of the total cellular protein. Phasins are assumed to form a close protein layer at the surface of the granules, providing the interface between the hydrophilic cytoplasm and the much more hydrophobic core of the PHA inclusion.

Purification and characterization of the poly(hydroxyalkanoic acid) synthase from Chromatium vinosum and localization of the enzyme at the surface of poly(hydroxyalkanoic acid) granules.

A recombinant strain of Escherichia coli, which overexpressed phaC and phaE from Chromatium vinosum, was used to isolate poly(3-hydroxyalkanoic acid) synthase. The isolation was performed by a two-step procedure including chromatography on DEAE-Sephacel and Procion Blue H-ERD. The poly(3-hydroxyalkanoic acid) synthase consisted of two different kinds of subunit (PhaC, M(r) 39,500 and PhaE, M(r) 40.500). PhaC was separated from the poly(3-hydroxyalkanoic acid) synthase complex by chromatography on phenyl-Sepharose: PhaE was enriched by solubilization of protein inclusion bodies. The stoichiometry of PhaC and PhaE in the enzyme complex was not determined. The poly(3-hydroxyalkanoic acid) synthase (PhaEC) exhibited a native relative molecular mass of M(r) 400,000 and most probably consists of ten subunits. The Km value of the enzyme for D(-)-3-hydroxybutyryl-CoA was 0.063 mM. The enzyme synthesized poly(3-hydroxybutyric acid) in vitro from D(-)-3-hydroxybutyryl-CoA or, together with propionyl-CoA transferase in a coupled enzyme reaction, synthesized the same product from acetyl-CoA plus D(-)-3-hydroxybutyric acid. Antibodies were raised against both subunits of the poly(3-hydroxyalkanoic acid) synthase. By immunoelectron microscopy, the poly(3-hydroxyalkanoic acid) synthase was localized within the cytoplasm in cells of C. vinosum grown under non-storage conditions. In cells grown under poly(3-hydroxybutyric acid) storage conditions, the enzyme was observed to be located at the surface of the poly(3-hydroxybutyric acid) granules. Immunoblots with anti-PhaC, anti-PhaE IgG and crude extract proteins indicated that poly(3-hydroxyalkanoic acid) synthases with partial sequence similarities are widespread among purple sulphur bacteria.

Purification and characterization of a 14-kilodalton protein that is bound to the surface of polyhydroxyalkanoic acid granules in Rhodococcus ruber.

The N-terminal amino acid sequence of the polyhydroxyalkanoic acid (PHA) granule-associated M(r)-15,500 protein of Rhodococcus ruber (the GA14 protein) was analyzed. The sequence revealed that the corresponding structural gene is represented by open reading frame 3, encoding a protein with a calculated M(r) of 14,175 which was recently localized downstream of the PHA synthase gene (U. Pieper and A. Steinbüchel, FEMS Microbiol. Lett. 96:73-80, 1992). A recombinant strain of Escherichia coli XL1-Blue carrying the hybrid plasmid (pSKXA10*) with open reading frame 3 overexpressed the GA14 protein. The GA14 protein was subsequently purified in a three-step procedure including chromatography on DEAE-Sephacel, phenyl-Sepharose CL-4B, and Superose 12. Determination of the molecular weight by gel filtration as well as electron microscopic studies indicates that a tetrameric structure of the recombinant, native GA14 protein is most likely. Immunoelectron microscopy demonstrated a localization of the GA14 protein at the periphery of PHA granules as well as close to the cell membrane in R. ruber. Investigations of PHA-leaky and PHA-negative mutants of R. ruber indicated that expression of the GA14 protein depended strongly on PHA synthesis.