Flavonoids of Chromolaena odorata (L.) R.M.King & H.Rob. as potential leads for treatment against tuberculosis.

Tuberculosis (TB) is currently rated as the 13th leading cause of mortality and the second leading cause of death after COVID-19, and above AIDS. Existing challenges relating to the development of multidrug-resistant strains and dangerous side effects of currently used drugs add impetus to the search for additional TB treatments. Hence, interest has grown in the use of medicinal plants as a source of bioactive preparations with efficacy against TB-causing organisms, and also with the ability to ameliorate the negative effects of TB drugs. This study aimed to evaluate the antimycobacterial and hepatoprotective potentials of extracts and isolated flavonoid compounds from invasive Chromolaena odorata. Test organisms used were pathogenic Mycobacterium bovis and M. tuberculosis H37RV, and the fast-growing M. aurum, M. fortuitum and M. smegmatis. The selectivity index (SI) values of the test substances were determined through cytotoxicity assays to promote these extracts and compounds as leads for the development of effective and safe anti-tubercular drugs. The antimycobacterial activity was evaluated using a serial microdilution method, and the SI was calculated from the 50% lethal concentrations calculated from cytotoxicity tests. Hepatoprotective activity was determined using HepG2 liver cells treated with rifampicin as a toxin. The extracts and compounds had a range of antimycobacterial activity with minimum inhibitory concentration (MIC) values ranging from 0.031 to 2.5 mg/mL. Two flavonoid compounds, 5,7,4'-trimethoxy flavanone and 5­hydroxy-3,7,4'-trimethoxyflavone showed promising antimycobacterial potential, and minimal toxicity was observed, as most SI values were higher than 1. The flavonoid compound 5,7,4'-trimethoxy flavanone had the highest SI (6.452), which was against M. tuberculosis H37RV. The HepG2 cells were reduced to 65% due to toxicity by rifampicin, however, the flavonoid compounds were able to improve cell viability to between 81 and 89% at different concentrations tested. Results obtained indicate that C. odorata may serve as a lead for the development of safe and effective antimycobacterial and hepatoprotective drugs.

Anatomical features in the kidney involved in water conservation through urine concentration in dromedaries (Camelus dromedarius).

The aim of this study was to report some of the morphological characteristics of the kidney involved in urine concentration and hence water conservation in the dromedaries. A total of 20 fresh kidneys of 10 apparently healthy camels were used in this study. The architecture of the renal pelvis was revealed by dissection and polyvinyl chloride corrosion casts. Samples were also processed for histology and for enzyme histochemistry. The camel kidney is bean shaped, smooth, multilobar, unipapillary, in which the fusion of renal papillae is complete forming a common renal papilla or crest, which channel urine into a central renal pelvis. It is more or less similar to equine, caprine, ovine and canine kidney. Under certain anatomical requisites the renal pelvis is known to play a role in urine concentration through recycling of urea to increase the medullary osmotic concentration which favors the counter-current mechanism. One of these requisites is an elaborate renal pelvis which is closely associated with the renal medulla. The renal pelvis of the camel has a main crescentic cavity following the long axis and curvature of the kidney. A thick extensive renal crest projects into the cavity of the pelvis. The thick renal crest contains large numbers of long loops of Henle and vasa recta which are important for urine concentration. The renal crest is formed by convergence of the medullary pyramids before it projects into the cavity of the renal pelvis. The crescentic main cavity of the pelvis forms 20-24 three dimensional radiating collateral recesses which contain the medullary pyramids. This close association of the renal pelvis and medulla provide a large surface area for the recycling of urea and hence urine concentration. This large pelvic-medullary interface is lined by simple low cuboidal epithelium which enhances the recycling of urea and water from the pelvic urine into the medulla and directly contributes to urine concentration. The rest of the wall of the renal pelvis and its recesses facing away from the renal crest and medullary pyramids is lined by impermeable transitional epithelium. Another feature is the intense activity of alkaline phosphatase demonstrated in the proximal convoluted tubules which indicates increased membrane transport. It is concluded that the kidney in dromedaries has the anatomical and histochemical requisites for the production of concentrated urine. These requisites enable the kidney to adequately contribute to the ability of the camel to conserve water and withstand the aridity of its habitat.

Sublethal effects of buprofezin on development and reproduction in the white-backed planthopper, Sogatella furcifera (Hemiptera: Delphacidae).

In the present study, the effects of sublethal concentrations of buprofezin on life-table traits of S. furcifera were evaluated for two consecutive generations (F0 and F1). Our results exhibited that the fecundity, life span (longevity) and hatchability of the F0 and F1 generations were significantly decreased at LC30 compared to the control. However, copulation was not significantly affected for the F0 or F1 generations at sublethal concentrations. The female life span was affected negatively at both treatments in F0 and at LC30 in F1, compared to the control. Furthermore, significant effects of the sublethal concentrations were found on the developmental rate of all instars except the 3rd instar of F1. However, the pre-adult period, total pre-oviposition period (TPOP) and adult pre-oviposition period (APOP) significantly increased in F1 individuals at LC30 and LC10 compared to the control. Our findings revealed that demographic characters (survival rate, intrinsic rate of increase (ri), finite rate of increase (λ), net reproductive rate (R 0), and gross reproductive rate (GRR)) of the F1 generation (from F0 parents) significantly decreased compared to the untreated group; however, the generation time (T) increased at LC10. Therefore, the results suggested that buprofezin could adversely affect individuals in the successive generation.

Cloning, expression, and functional analysis of two acetylcholinesterase genes in Spodoptera litura (Lepidoptera: Noctuidae).

Two acetylcholinesterase genes (SlAce1 and SlAce2) were cloned from Spodoptera litura, which is an important pest that causes widespread economic damage to vegetables and ornamental plants. We analyzed their expression patterns and compared their biological functions by using RNA interference. Our results showed that SlAce1 and SlAce2 cDNA contains 2085bp and 1917bp nucleotides and encoding proteins of 694 and 638 amino acid residues, respectively. Phylogenic analysis indicated that the lineage of SlAce genes and SlAce1 was completely different from SlAce2. Although both genes were expressed in all developmental stages and majorly in the brain. The expression levels of the both genes were suppressed by inserting their related dsRNA in the 6th instar larvae, which led to 47.3% (SlAce1) and 37.9% (SlAce2) mortality. Interestingly, the suppression of the SlAce2 transcripts also led to significant reductions in the fecundity, hatching, and offspring in the parental generation of S. litura. It is concluded that SlAce2 is responsible for the hydrolysis of acetylcholine and also plays role in female breeding, embryo progress, and the development of progeny. Considerable larval mortality was observed after both AChE genes (i.e. Ace1 and Ace2) were silenced in S. litura confirms its insecticidal effectiveness, which provided a molecular basis in biological pest control approach.

The use of dynamic external fixation in the treatment of dorsal fracture subluxations and pilon fractures of finger proximal interphalangeal joints.

The aim of this study was to review the outcome of the treatment of finger proximal interphalangeal joint dorsal fracture subluxations and pilon fractures with a modified external fixator. We treated 36 patients (36 fingers). We assessed the ranges of motion and patient satisfaction. At final follow-up, 23 patients had no pain; 11 had pain in the cold; and two also had mild pain. None had moderate or severe pain. The mean range of proximal interphalangeal joint motion was 86° (60°-100°). The mean total active range of finger motion was 244° (range 200°-265°). This system is simple, cheap and relatively easily applied. It gives stable fixation that allows early mobilization. LEVEL OF EVIDENCE: IV.

Genetic diversity of donkey populations from the putative centers of domestication.

Donkey domestication drastically changed ancient transport systems in Africa and Asia, enabling overland circulation of people and goods and influencing the organization of early cities and pastoral societies. Genetic studies based on mtDNA have pointed to the African wild ass as the most probable ancestor of the domestic donkey, but questions regarding its center of origin remain unanswered. Endeavoring to pinpoint the geographical origin of domestic donkey, we assessed levels and patterns of genetic diversity at 15 microsatellite loci from eight populations, representing its three hypothesized centers of origin: northeast Africa, the Near East and the Arabian Peninsula. Additionally, we compared the donkey genotypes with those from their wild relative, the African wild ass (Equus africanus somaliensis) to visualize patterns of differentiation among wild and domestic individuals. Obtained results revealed limited variation in levels of unbiased expected heterozygosity across populations in studied geographic regions (ranging from 0.637 in northeast Africa to 0.679 in the Near East). Both allelic richness (Ar) and private allelic richness presented considerably higher values in northeast Africa and in the Arabian Peninsula. By looking at variation at the country level, for each region, we were able to identify Sudan and Yemen as the countries possessing higher allelic richness and, cumulatively, Yemen also presented higher values for private allelic richness. Our results support previously proposed northeast Africa as a putative center of origin, but the high levels of unique diversity in Yemen opens the possibility of considering this region as yet another center of origin for this species.

Prenatal diagnosis of beta-thalassemia in Egypt: implementing accurate high-tech methods did not reflect much on the outcome.

The clinical severity of thalassemia major makes it a priority genetic disease for prevention programs through prenatal diagnosis for carrier couples. Incorporation of automated DNA sequencing that enables the characterization of mutations not detected by other mutation specific detection procedures was a prime goal of this work. Automated DNA sequencing was offered on fetal tissues in 30 pregnancies during the year 2005. The pregnancies were at high risk for homozygosity or compound heterozygosity for beta-thalassemia based on mutation analysis of both parents before prenatal diagnosis. Both parents have beta-thalassemia trait. Fetal samples were collected by chorionic villus sampling (CVS) in the first trimester and by amniocentesis in the second trimester. The point mutations were characterized by PCR (ARMS). The absence of the expected fragment with all the mutant ARMS primers insinuated an uncharacterized DNA segment that was further subjected to direct automated fluorescent DNA sequencing in an attempt to know if the fetus was affected by parents' mutations. If no mutation was detected using the PCR ARMS, the sample was further analyzed using direct automated fluorescent DNA sequencing. The mean gestation when carrying out the invasive procedure was 14 (10 -18) weeks. All mothers had a previous affected pregnancy, and 13 had two or more previous affected pregnancies. Pregnancies were: 8 carrier fetuses (trait) and 22 affected fetuses in which 2 were homozygous and 20 double heterozygous. Fourteen parents of affected fetuses preferred to continue pregnancy and the babies were born as diagnosed. The other 8 parents decided on termination and DNA of the abortuses proved to be as previously diagnosed by DNA sequencing. The use of PCR amplification and direct sequencing have permitted the accurate characterization of unidentified alleles and successfully solved 100% of the examined samples. However, it has resulted in minor changes of the outcome as the majority of couples preferred continuation of pregnancy.

Nephrotoxicity of low-dose methotrexate in guinea pigs: an ultrastructural study.

The nephrotoxicity of low-dose methotrexate (MTX) and the rescue effect of leucovorin were studied by electron-microscopic examination of the kidney of guinea pigs. One group received MTX as a single weekly dose of 10 mg/kg, i.p.; a second group received a similar dose divided into three equal fractions. The third group received MTX rescued with an equal dose of leucovorin. The distal convoluted tubule showed cell swelling, fragmentation of the endoplasmic reticulum and loss of mitochondrial cristae and matrix. Leucovorin minimized these changes. The proximal tubule and the glomerulus were not affected. Low-dose MTX induced a nephrotoxicity in the distal convoluted tubule which could be minimized by leucovorin. Mild myelosuppression was also observed.

The poll glands of the dromedary (Camelus dromedarius): ultrastructural characteristics.

The poll glands of the camel are tubuloalveolar glands. They consist of lobules separated by interlobular connective tissue. Adrenergic axons and blood vessels including fenestrated capillaries are present in the intralobular connective tissue in close proximity to the secretory cells. The prominent features of the secretory cell cytoplasm are many mitochondria, smooth endoplasmic reticulum, and many vesicles in various secretory stages. It is concluded that the secretory cells have apocrine as well as merocrine modes of secretion. The glands may have the ability to uptake hormones and concentrate them. Additional work is needed before considering these glands as steroid-secreting glands.

Cellular and humoral immunological changes in chronic renal failure associated with schistosomiasis.

The changes that occur in cell mediated and humoral immunity in patients suffering from chronic renal failure associated with schistosomiasis were studied. There was a decrease in T3%, T4% and T4/T8 ratio. IgG, IgM and IgE levels were found to be increased in such cases. It was concluded that there was a reciprocal relationship between cell mediated immunity and IgE levels in cases with chronic renal failure associated with bilharzial hepatosplenomegaly.

Characterization of serum lysosomal enzymatic activities. III. Effect of infectious influenza in Egyptian equines.

An outbreak of infectious influenza was recognized in Menofeia governorate in October 1989. Eight naturally influenza infected as well as 8 healthy control horses, mules and donkeys were selected for collection of blood and sera separation to estimate four lysosomal enzymatic activities and to describe the clinical findings, which were fever, congested nasal, conjunctival membranes and cough. Bronchopneumonia followed later with bilateral purulent nasal discharge as a complication in 2 donkeys. Thereafter laboured breathing occurred. Therefore a therapeutic penicillin-streptomycin dose was injected to safeguard against the secondary bacterial invasion. All lysosomal enzyme levels in serum of diseased equines were subjected to consistent regression except N-acetyl-beta-glucosaminidase -beta-NAG) that behaved very highly significant activity, probably due to the acid pH resulting from bronchopneumonia and respiratory acidosis which override the inhibitory action of the antibiotics. Influenza virus may inhibit the synthesis of the pulmonary surfactant in alveolar and bronchial epithelium, thereafter, the animals suffered from respiratory distress and bronchospasm with resultant decreased acid phosphatase (ACP) value which was histochemically located in both sites. The disappearance of alpha- and beta-galactosidases (alpha-GAL & beta-GAL) from the serum of diseased animals can be attributed to the antibiotic dose.

Characterization of serum lysosomal enzymatic activities. II. Effect of lumpy skin disease in Egyptian cows.

Clinical findings and lysosomal enzymes (LYE) in eight lumpy skin diseases (LSD) cows and same number of healthy ones were reported in Tal-El Baker village and Tal Alkabir centre, Ismailia province, Egypt. LSD began with fever, anorexia, skin lesions in form of nodules all over the body, which disappeared spontaneously or gathered to form large lumps. It was complicated with respiratory manifestation, corneal opacity, mastitis, dehydration and later on recumbency. It is noteworthy that the level of 3 LYE showed the same trend of significant reduction in acute stage of the disease (5 days after occurrence of LSD) probably due to injection of animals with a therapeutics dose of terramycin. Acid-phosphatase (ACP) enzyme is the sole that behaved very high significant increase in the serum in acute stage of LSD due to the damaged tissues caused by the virus. It underwent insignificant decrease in late stage of the disease (20 days after its occurrence) to restore the normal LYE level in control cows indicating recovery. Alpha-galactosidase (alpha-GAL) decreased perpetually by the progression of LSD because of the decreased bactericidal index which ist in concomitance with the secondary bacterial invader. N-acetyl-beta-glucosaminidase (beta-NAG) and beta-galactosidase (beta-GAL) in LYE had the same fluctuating manner. The activities showed very highly significant decrease in acute stage, followed by highly significant and significant increases (late LSD stage) respectively. The appreciable significant increase of beta-GAL may declare the effect of anorexia on LSD. In view of these findings, it can be postulated that LSD may be diagnosed and prognosed through LYE changes in the serum.

Characterization of serum lysosomal enzymatic activities. I. Effect of dietary urea in dairy cows.

The experiment was carried out on ten Holstein dairy cattle fed on conventional concentrate as base diet for 15 days, then turned on unconventional concentrate diet supplemented with 3% urea (CM-U) for 45 days to study the effect of four lysosomal enzymes in both states. The N-acetyl-beta-glucosaminidase enzyme level reflected the highest value among other enzymes inspite of its significant drop in the serum after feeding cows on the unconventional ureated diet. Beta-galactosidase was the sole enzyme to rise significantly in the serum of cows during the unconventional ureated diet feeding. Acid phosphatase and alpha-galactosidase showed highly significant decreases due to the effect of the tested diet.

Light and electron microscopic observations on the cerebellum of guinea pigs following low-dose methotrexate.

The neurotoxicity of single and divided low-dose methotrexate (MTX) and the rescue effect of leucovorin (folic acid) were investigated by light and electron microscopic examination of the cerebellum of Guinea pigs. One group of animals received a single weekly intraperitoneal MTX in a dose of 10 mg/kg, a second received a similar dose divided into three equal fractions, and the third group received MTX rescued with an equal dose of leucovorin. The results showed degeneration of Purkinje cells indicated by shrinkage and distortion of perikarya, dissolution of mitochondrial cristae, and nuclear and nucleolar changes in all groups which received MTX. There was also astrocytic proliferation and perivascular edema. Lesser changes were found in the group which was rescued with leucovorin. The degenerative changes in the cerebellum particularly those of the Purkinje cells explain the neurological signs reported in cancer patients treated with MTX.

Studies on gangrenous mastitis in goats.

One hundred and fifty goats were diagnosed as having proven gangrenous mastitis. The disease was categorized into early, intermediate and late stages. Gangrenous mastitis in goats is typified by a sudden onset, dark hyperemia, and edema with progressive discoloration of the distal part of the udder. The disease affected lactating goats but not the dry ones. Coagulase positive Staphylococcus aureus was isolated from 60% of composite and half milk samples obtained from the diseased goats. The histopathological changes mainly comprised proliferation of connective tissue, thrombosis and necrosis involving a group of lobules. Treatment of the early and intermediate stages of the disease was successful through the administration of systemic and intramammary terramycin together with diuretics and topical antiseptic cream. The late stage of the disease was successfully treated only through surgery.

Ultrastructural morphometric observations on the extramural aortico-pulmonary bodies of the domestic fowl.

The extramural aortico-pulmonary bodies of the domestic fowl were analysed in electron micrographs by point-counting morphometry. The Type I cell constituted about 34% of the total volume of the parenchyma, whereas the supporting cells (Type II cells, pericytes and Schwann cells) constituted about 26%. The blood capillaries occupied the lowest volume density of about 5%. The mitochondria and the dense-cored vesicles occupied about the same volume (8%) of the Type I cell cytoplasm. The results obtained in the present study are compared with other similar studies on the aortic and carotid bodies of mammals.

Morphometric and histological studies on the adrenal glands of the camel Camelus dromedarius.

The adrenal gland of the camel consists of an outer cortex and an inner medulla. The general disposition of the cortex and medulla, however, differs occasionally from that of other mammals. Extensions of medulla could reach as far as the periphery of the cortex. Islet of medullary tissue may be found in sections of the cortex and cortical tissue consisting of all zones of the cortex may occur around arteries or nerves in the medulla. The medulla may be separated from the cortex by connective tissue especially in old camels. The arrangement of noradrenaline-secreting cells is different from that in other ruminants; they are found in groups scattered between the adrenaline-secreting cells. Bundles of smooth muscle occur in venules at the corticomedullary interface. Accessory adrenal glands are found embedded in the renal fat. They are similar in structure to the adrenal gland. The adrenal cortex forms 74% of the volume of the gland and the ratio of the cortex to medulla is 4:1. The zona glomerulosa, fasciculata and reticularis constitute about 13%, 53%, and 29% by volume of the cortex, respectively.