Khadiga Gaafar

Abstract. The present study aimed to investigate the potential role of leptin in the progression of breast cancer and the associated cell proliferation signalling pathway(s). A total of 44 female patients diagnosed with breast cancer and 24 healthy donors from Ain Shams University Hospitals (Cairo, Egypt) were enrolled in the present study. The present study assessed leptin expression in breast cancer tissues at the gene and protein level using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and immunohistochemistry. The results demonstrate that the expression of leptin was significantly higher in tissue of breast cancer samples from obese patients than overweight and control samples (P<0.001). ELISA results indicated a significant increase (P<0.001) of leptin expression in obese patients. To investigate whether there is any difference in leptin expression between the peripheral and tumor microenvironment blood of patients with breast cancer, the concentration of leptin was assessed in plasma from both using ELISA assays. The results demonstrated a statistically significant increase in the level of leptin in plasma samples from the tumor microenvironment of obese patients with estrogen receptor positive (ER+) breast cancer, compared with peripheral plasma samples. Furthermore, the leptin gene was overexpressed in obese ER+ breast cancer tissue. RT‑qPCR was also performed to assess the expression of genes involved in proliferation pathways including leptin receptor (LEPR), aromatase, mitogen activated protein kinase (MAPK) and signal transducer and activator of transcription‑3 (STAT3). A positive association between leptin expression, LEPR, aromatase, MAPK and STAT3 was detected in tissue samples of patients with breast cancer. The current study concluded that leptin may enhance breast cancer progression by inducing the expression of JAK/STAT3, ERK1/2 and estrogen pathways in obese patients breast cancer.

Introduction: In Egypt, disorders of sex development (DSD) constitute a significant entity among the birth defect list. Previous studies have reported that end organ androgen unresponsiveness, i.e. Androgen resistance, was the most prevalent underlying mechanism among Egyptian 46,XY DSD cases. Based on cytogenetic and hormonal diagnostic criteria as well as few sporadic case reports, it was proposed that androgen receptor (AR) defects [i.e. Androgen insensitivity syndrome (AIS), OMIM#300068] might constitute a major etiology within this category. However, this has never been systematically ascertained through an AR molecular diagnostic approach. Aim of the Work: The current study aimed to assess the role of AR mutations as an underlying etiology among a sample of Egyptian 46,XY DSD pediatric patients presenting with androgen end organ unresponsiveness. Patients and Method: In the current study, 21 children [age<18years] with male undermasculinization due to androgen end organ unresponsiveness were selected from 46,XY DSD cases. The selection criteria included ambiguous genital phenotype or genitalia discordant to the genotypic sex; 46,XY Karyotype and normal testicular response to HCG stimulation in prepubertal patients or normal basal testosterone (T) levels in postpubertal subjects. Molecular studies of the AR entailed PCR amplification for screening of major deletions/ insertions, single stranded conformational polymorphism (SSCP) screening for point mutations in the AR 2-8 exons followed by sequencing of these exons for all cases. Results: The results showed that none had major deletions/insertions. Five exons out of 147 (3.4%) showed abnormal SSCP migrational patterns. Out of those 5, two mutations in two Egyptian patients were detected by sequencing. The first was R840G (Arginine 840 glycine), in exon 7 (The ligand binding domain). The other was A596T (Alanine 596 Threonine) in exon 3 (The DNA binding domain). Conclusion:, This study shows that AR mutation is an uncommon underlying etiology among Egyptian paediatric 46,XY cases.

Recently in Egypt, there was particular attention given to fish welfare. Many water bodies cover Egypt; the Nile River is the main bloodline, in addition to many fresh as well as salty lakes. Nearly 2.5 million fish species are being used in scientific research representing 94% of all research animals. The new 2014 Egyptian Constitution stated that fish resources are to be preserved and fish under the threat of extinction are to be protected. Although there are legislation for farmed animals and wildlife in both the Environmental and Agriculture laws, there are none concerning fish. Since the application of animal welfare guidelines has recently been implemented by the IACUC in Faculty of Science, Cairo University to gain its approval for scientific protocols, a need to investigate the extent of fish welfare awareness and international guidelines application in protocols using fish as research models was revealed.

Although animal research ethics committees (AREC) are
well established in Western
countries this field is
weakly developed and its concept is poorly understood
in the Middle East and North Africa region. Our
main objective was to establish the first animal ethics committee to introduce the concept and
requirements of ethical approaches in dealing with experimental animal models in Egypt.
To achieve this we established our standard oper
ating producers (SOP) according to international
guidelines. Our SOP constituted the AREC responsi
bilities, committee member
s selection, evaluation
procedures emphasizing the regional cultures, etc. T
he challenges encountered by the committee were
diverse such as the absence of laws that control the
use of animal models in scientific research, lack of
guidelines (protocols for animal subjects in research) and mandatory ethical approval for any
experimental animal research. To ov
ercome the challenges, high univer
sity authorities were approached
to approve the establishment of an AREC committee.
Moreover, workshops were instigated in different
universities to: 1) emphasize the legal requirement
of animal research ethics approval; 2) describe the
guidelines how ethical evaluation of scie
ntific proposal will be carried out; 3)
facilitate interaction with best
practices and research experts and 4) teach young rese
arches how to write protocol involving the use of
animal subjects in research. In three months, we were able to conduct three workshops after each
workshop questionnaire and evaluation was obtained to overcome weak points.

The aim of the present work was to clarify the involvement of free radicals, cytochrome P450 toxic metabolites, and deregulation of calcium homeostasis in the mechanism of diethyldithiocarbamate (DDC) hepatotoxicity. This was elucidated through the preadministration of ascorbic acid (a free radical scavenger), cimetidine (an inhibitor of cytochrome P450 enzymes), or nifedipine (a calcium-blocking agent) before DDC treatment to male albino rats. DDC was administered either as a single dose [800 mg/kg body weight (b.w.), subcutaneously, s.c.] or daily repeated doses for 30 days (400 mg/kg b.w., s.c.). Oxidative stress indicators [e.g., malondialdehyde (MDA), reduced glutathione (GSH), and superoxide dismutase enzyme (SOD)] showed that single or repeated DDC doses induce an increase in MDA level and a decrease in SOD activity in the liver, whereas it causes depletion in hepatic GSH after a single dose and an elevation in its value after repeated doses. Severe histopathological changes were also observed in the livers of rats treated with single or repeated DDC doses. Ascorbic acid, cimetidine, and nifedipine pretreatments were found to induce highly protective effects against the evinced DDC hepatotoxicity, manifesting that free radical, cytochrome P450, and calcium-dependent processes contribute to DDC liver toxicity. Finally, although multiple mechanisms may be involved in the hepatotoxic changes induced by DDC, calcium disarrangement and free radical formation play a more critical role than cytochrome P450 in metabolic events leading to toxic effects of DDC.

The reproductive activity of Damascus goats was studied in does that were not allowed to mate (non-lactating does) (Group C) and lactating does with different suckling treatments (Groups A and B). Time resumption of sexual activity was not significantly different between lactating and non-lactating does or between does’ groups of different suckling treatments. All does started to demonstrate estrous behavior during autumn (September). In Groups A, B and C the mean length of the estrous cycle was 19.0±0.6, 19.5±0.3 and 22.0±2.0 days, duration of the estrous period 29.6±3.0, 31.0±5.3 and 25.3±5.8h, mean corpus luteum (CL) counts 1.6±0.2, 1.8±0.2 and 1.8±0.3, while mean litter size was 1.1±0.1, 1.2±0.2 and 1.3±0.5, respectively.In the three groups an increase in the plasma progesterone concentrations started from summer (mid-June), indicating the occurrence of one or more ovulatory anestrus (silent ovulation). Plasma prolactin levels were higher (P

Objectives: To evaluate the levels of some parameters related to bone metabolism during pregnancy i.e. total calcium (CA), 1,25-dihydroxyvitamin D (Vit.D), parathormone hormone (PTH), phosphorous, osteocalcin, total acid phosphatase and its tartrate resistant isoform (TrACP), total alkaline phosphatase (TALP) and its isoenzmes activities, total protein, albumin, globulin in sera and haemoglobin in blood in a group of egyptian pregnant women who are living in cairo and under reasonable medical care.
Cases: The study was conducted on 33 healthy pregnant women (age range from 20 to 38 years) and 16 healthy non-pregnant women within the same age range to serve as controls. Twenty blood samples were collected from the pregnant women at the end of first trimester of pregnancy (Week-11), twenty four were collected at the end of third trimester (Week-38).
Results: Regarding the osteoblastic activity, the T.ALP was significantly increased due to placental T.ALP and to a lesser degree to bone ALP, while osteocalcin levels. Showed a non significant increment in the 1st and 2nd trimesters of pregnancy with significant increase in the 3rd trimester of pregnancy .concerning the osteoclastic activity, The TrACP isoform decreased significantly in the 2nd trimester of pregnancy, Then returned to the normal values of non-pregnant control levels. Women in the last trimester of pregnancy have higher estrogen level than any other time of life. Although, estrogens at physiologic concentration inhibit osteoclastic activity, hight estrogen concentration increases osteoclastic activity.
As regards the factors that responsible for calcium homeostasis, significant decrease in serum intact PTH was concurrent with significant increase in Vit.D. The total serum calcium level was significantly decreased through the pregnancy, while the ionized calcium did not change, and the serum phosphorus was highly significantly increased.
Conclusion: From these results it has been concluded that, the pregnancy imposes major changes in the mother's nutritional requirement and causes physiological alternation in maternal bone metabolism. Minimal changes affecting the bone turnover were demonstrated among egyptian women throughout pregnancy due to the nutritional statue of the women and thier supplementation with Ca and/or Vit.D. Also,the changes in the haemodynamic parameters of the mother may be due to heamodilution that occur during pregnancy. The obtained patterns showed that the egyptian women who are living in urban areas (e.g. cairo), and under reasonable medical care are in good health compared to other communities in which the nutritional status and the poor medical care are the main reasons for the dramatic changes in the bone metabolism.

The long- and short-term effects of chloroquine (CAS 54-05-7) on glucose metabolism in rats were assessed. The long-term chronic chloroquine administration (5 and 10 mg/kg b.w. 6 days a week for 6 months) caused a decrease in serum glucose, insulin, calcium, potassium and protein levels, while the glucagon level increased. The short-term acute effect of chloroquine administration (10 mg/kg b.w. 6 days for one week) caused an improvement in glucose tolerance as shown by the decrease in glucose and insulin curves after an oral glucose tolerance test. This was accompanied by an increase in insulin activity, corrected insulin response, and glucose tolerance parameter and a decrease in glucose and insulin areas. Lactate dehydrogenase and glucose-6-phosphate dehydrogenase activities were increased, too, indicating an increase which provides the needed energy for overcoming the injurious effect of chloroquine.

The protective effects of vitamins E and C against methanol-induced free radical production with its subsequent tissue injury in liver and retina of male albino rats were assessed. The rats were divided into four groups: (1) control, (2) antioxidant (Ao) control group receiving 5 mg of each vitamin, E and C, (3) daily ip-injected methanol (2 mL/kg b.wt.) group, killed after three and six doses, and (4) Ao/methanol group administered the vitamins 3 weeks prior to and along with methanol injection and killed as the latter group [Sharpe et al. Methanol Optic Neuropathy: A Histopathological Study. Neurology 1982, 32 (10), 1093–1100]. Methyl alcohol drastically altered the Ao defense system and energy status of rat liver, where highly significant depletion of glutathione levels and inhibition of superoxide dismutase activity, concurrent with significant increase in thiobarbituric acid reactive substances indicating marked elevation in lipid peroxidation. These effects were reversed when the vitamins were administered denoting their role in promoting the Ao defense system. Furthermore, they also increased the methanol-induced depletion in adenylate energy charge, phosphate potential, and ATP values. The amelioration in the energy status as a result of vitamins E and C supplementation suggests that their role as Aos is effective in relieving the impaired oxidative phosphorylation in order to increase the energy demand under physiologically stressful conditions. Histopathological and ultrastructural results of rat retina confirmed the protective effect of Ao vitamins. As compared to the methanol-intoxicated group, the protected group showed preservation of the membranous structures of the retinal cells, especially mitochondria that assumed their normal shape. This may be attributed to the inhibition of free radical production and lipid peroxidation and subsequently minimum degree of tissue damage. Amelioration of mitochondrial structure reflected the improvement of impaired oxidative phosphorylation of intoxicated animals with an approximately normal level of energy demand. This suggests that Ao vitamins may alter the retinotoxic effects of methanol by promoting the internal Ao defense system and preserving the energy status of the animal.

The effects of chronic chloroquine administration on serum proteins and free amino acids of rabbits and the accompanying ultrastructural changes in the retina were investigated.
Thirty pigmented rabbits were injected intramuscularly with chloroquine diphosphate (14 mg/Kg bw). Fifteen rabbits served as control. Blood samples drawn after 1, 2, 3, 4, 5, 6 and 12 months were assayed for serum proteins by Bio-analytic kits and horizontal electrophoretic technique. Free amino acids were determined by Beckman analyzer. Central retinal tissue was fixed in 4% glutaraldehyde, embedded in araldite CY212 and sectioned for ultrastructural examination.
Histopathologic changes were apparent within three months, initially affecting the pigment epithelium and photoreceptors and later involving the remaining neuroretinal layers. Hypoproteinemia gradually developed mainly due to a sharp drop in albumin and alpha 1 and 2 globulin fractions, inspite of an increase in beta and gamma globulin fractions. The concentration of non-essential amino acids varied considerably from a depletion of taurine, aspartine, glutamine, tryptophan and arginine to an increase of serine, alanine, GABA and ornithine. The most outstanding effect was the disappearance of tyrosine and its return to normal value at the end of the experiment.
The disturbances in protein pattern and free amino acid levels are an indication of chloroquine toxicity and may be implicated in the retinopathy.

capozide was administered in a dose of 1.012 mg/200g rat body weight once a day for 4 weeks to normal and diabetic rats. It had no effects on serum potassium urea or creatinine. No increase in lipid concentration to negate its beneficial hypotensive effect, was noted. Form oral glucose tolerance test, it was calculated that the drug improved glucose tolerance concomitant with the increased beta cell function and peripheral insulin response.
The interaction between the two components of capozide, captopril an angiotensin converting enzyme inhibitor, and hydrochlorothiazide, a diuretic, is discussed in regards to carbohydrate and lipid homeostasis.

Serum ferritin, Alpha-1 Acid Glycoprotein (α1AGP), c-reactive protein (CRP), and ceruloplasmin (CER) were studied in 83 children with an age range of three to thirty six months. Fifty nine cases suffered from protein energy Malnutrition (PEM), 29 marasmus and 30 kwashiorkor (KWO). Twenty four healthy children were included as controls. the malnourished group exhibited significant reduction in anthropometric measurement scores and in serum albumin and prealbumin levels than those of the control group. CER was significantly reduced in PEM cases. Evidence of infection by an elevated α1AGP, A positive CRP and the preence of diarrhea was also demonstrated in the malnourished group but not in the control one. Ferritin level was significantly elevated in the KWO group, While in the marasmus one the elevation was non-significant. At the same time 100% of the KWO and 89.7% of the marasmus cases had a hemoglobin level of less than 11 g/dL. Also 46.4% of the KWO and 48.2% of the marasmus cases had a serum iron of less than 50 µg/dL. In such circumstances the elevation in serum ferritin level, in accordance with other acute phase proteins, could reflect increased secretion and/or possible leakage from damaged tissues especially when evidence of increased body iron stores is lacking.

The effect of one month treatment with Hydrochlorothiazide (HCT); Captopril, angiotensin converting enzyme inhibitor (ACEI), Diclofenac, nonsteroidal anti-inflammatory drug (NSAID) and their different combinations was studied in DOCA-salt hypertensive rats (HR). It was observed that HCT causes a reduction of serum sodium, potassium and creatinine levels concomitant with an increase in serum aldosterone. Captopril also reduced serum sodium and creatinine but increased serum potassium and had no effect on aldosterone. The administration of HCT with Captopril produced a simple additive interaction, thus reducing serum sodium and creatinine, prevented hyperaldosteronism and reversed the hypokalemic effect induced by HCT. On the other hand, Diclofenac alone exerted a significant increase in serum sodium and creatinine while its biosynthesis of renal and/or extrarenal prostaglandins which are compounds known to modulate the vasoconstrictive effect of norepinephrine and to represent important mediators for the activation of the renin-angiotensin-aldosterone system (Brown et al., 1986). Moreover, Brown et al. (1986) reported that sodium retention is a characteristic effect of virtually all NSAIDs.

The effect of long-term treatment of chloroquine (CAS 54-05-7) (20 mg/kg body weight) on serum lipid components and its relation to to the retinotoxic effect was studied in albino rats. Chloroquine was found to form lamellar lysosome-like structures within the photoreceptive layer, as well as the pigment epithelium and neurooretinal layers. Biochemically, hypolipidemia in the serum was observed mainly due to the decrease in phospholipid portion. It was hypothesized that due to the inhibition of the degradation process in the defective lysosomes, the retinal cells were denied the re-use of their own phospholipids, and thereby resort to their uptake from the serum.

Captopril was effective in the long term reduction of serum sodium and aldosterone in normotensive rats, the addition of HCT produced a further decrease in serum sodium and was also useful in preventing hypokalemia produced by HCT. On the other hand, the concurrent therapy with Diclofenac attenuated the hypotensive effect of Captopril and HCT, it was observed that to combine Diclofenac with captopril was more beneficial as regards the metabolic parameters.

Glucose tolerance (GT), peripheral insulin activity (A) and beta cell function (CIR) were improved on prolonged chloroquine administration to normal male rats, while the acute effects comprised hyperinsulinaemia with unaltered blood glucose levels. This study explains some of the seemingly conflicting results due to chloroquine treatment and concludes that the drug is not damaging to glucose homeostasis in long term treatment.

The effect of methanol on some of the lipid components in serum was studied in rats. Methanol was administered by stomach tube in doses of 2 and 6 ml/kg b.wt daily for 21 and 6 days, respectively. Methanol was found to accumulate lipids; thus, cholesterol, phospholipids and triglycerides increased significantly. Concurrently, modification of the lipoid content of organs has been considered. It was concluded that methanol and not only formate, is toxic to rats, inspite of the alleged difference in routes of its metabolism in primates and rodents.

Glucose tolerance curve, glycogen and the calculated glucose tolerance (GT), peripheral insulin activity (A value), beta cell function (CIR) and glucose and insulin areas were studied in rats intoxicated with methanol. The study revealed that a sublethal dose of methanol (2 ml/kg b.wt) increases glucose consumption at the same time having no effect on insuline secretion, While the continued administration of methanol depletes glycogen due to its breakdown to restore glucose levels to normal values. The lethal dose (6 ml/kg b.wt) cause insuline resistance.

The effects of the non-selective beta-blocker propranolol (CAS 525-66-6) and the cardioselective drug atenolol (CAS 29122-68-7) on serum glucose, insulin levels and some serum lipid components, were compared in normal and diabetic rats receiving glibenclamide. The two beta-blockers, when administered concurrently with glibenclamide in normal rats, exerted a significant hypoglycemic effect (p < 0.01), while in diabetic rats the two drugs caused a more significant decrease (p < 0.001 and p < 0.01, resp.). Serum insulin levels were mainly unaffected by the two beta-blockers. Propranolol was found to exhibit a hypolipidemic effect in diabetic rats when administered alone or in combination with glibenclamide. In comparison atenolol, when used alone, exerted a significant increase in triglycerides and total lipid levels (p < 0.05) in normal rats. This effect was masked when atenolol was administered concurrently with glibenclamide, but a hypercholesterolemic effect (p < 0.01) was noticed. Paradoxically in diabetic rats, atenolol caused a significant decrease (p < 0.05) in triglycerides level, while its combined use with glibenclamide showed no effect.

The long term (6 Weeks) effect of both methanol (2 ml/kg) and ethanol (2 ml/kg) on GSH and SOD activities in rats was studied. Blood SOD and GSH activity declined sharply (1st week), then started to increase gradually. Liver GSH was seen to increase significantly (4 and 6 weeks post treatment). This was attributed to their swift usage in scavenging the free radical produced from the metabolism of the alcohols, and their protective response whereby an increase in their synthesis occurs.