Quantitative Microscopic evaluation of Melatonin treatment of Experimentally Atherosclerotic Rabbit Aorta T
he Egyptian Journal of Hospital Medicine Vol., 6 : 1- 14 March 2002

1687 - 2002

Quantitative Microscopic evaluation of Melatonin treatment of
Experimentally Atherosclerotic Rabbit Aorta

Mattar FE *; Ismael AA*; Kassem M.A **;Sharaf, H.A. and Ibrahim IK**

From the Department of Histology, Al-Azhar Faculty of Medicine in Cairo* , in Assiut
** and Pathology Dept. National Research Center, Dokki-Giza

Atherosclerosis is one of the most common diseases in Egypt. The exact etiology of the
disease is still vague. However, Oxidative modifications of low-density lipoprotein (LDL) have
been proposed to play a critical role in atherogenesis. Antioxidants have been proposed,
therefore, to have a possible role in protection and/or prevention of atherosclerosis. The pineal
gland hormone, melatonin, have been reported as an antioxidant. Recently melatonin has been
proposed to interfere with one of the chain of events in the onset of atherosclerosis, that is, the
formation of adhesion molecules. To test the role of melatonin in amelioration of atherogenesis,
this work was designed to study the atherosclerosis-.related microscopic changes in the aorta of
melatonin treated, high cholesterol fed rabbits.
Thirty adult male rabbits were used throughout this work. The animals were fed a standard
rabbit food free from cholesterol and antioxidants. The rabbits were divided into three groups
each of 10 animals: Control (A) group, was raised on the ordinary diet alone for 6 weeks.
Atherosclerotic induced, non treated (B) group was raised on the ordinary diet mixed with 1%
cholesterol powder for a period of 6 weeks. Atherosclerotic induced, melatonin treated (C)
was raised on the ordinary diet mixed with 1% cholesterol powder and drenched 4.2 mg
melatonin dissolved in saline solution daily at night for 6 weeks. The drug was given through
naso-gastric tube under light ether anesthesia
Paraffin sections of both control and experimental groups were prepared on the same slide.
The sections were stained by haematoxylin and eosin, Masson trichrome, orcein; or periodic
acid Schiff (PAS).
The endothelial thickness, relative collagen, elastic fibers, smooth muscle fibers and
glycosaminoglycan content in control and experimental groups were evaluated by computerized
image analyzer. Statistical analysis of the results was carried out using Microsoft Excel 2000.
The results revealed that the melatonin has antiatherosclerotic effects in experimental
atherosclerosis in the rabbit aorta. Melatonin significantly reduced specific atherosclerotic
lesions like fibrosis , smooth muscle atrophy, and decreased elastic fiber content in thoracic
aortic wall.
In conclusion, melatonin may be of value in the management of atherosclerosis by reducing its


Atherosclerosis is one of the common
plaque a white, pearly appearance (Walter
diseases among middle age and old age
& Israel, 1990) . These features result in the
Egyptians. The exact etiology of the disease
hardening of the arterial wall due to thicke -
is still indefinite. In atherosclerosis, fatty
ning and loss of elasticity (Cormack ,
material accumulates in the intima with
1997). Hemorrhage, thrombosis, ulceration
additional fibrosis . This material is covered
and calcification are advanced and late
by dense fibrous tissue, which gives the
complicated lesions of atherosclerosis



Full Paper (vol.6 paper# 1)

Mitochondrial NADH Dehydrogenase gene (mtND2) Phylogeny of Egyptian Tilapia species The Egyptian Journal of Hospital Medicine Vol., 6 : 15 - 26 March 2002

1687 - 2002

Mitochondrial NADH Dehydrogenase gene (mtND2) Phylogeny of
Egyptian Tilapia species

Mohammed H. Awwad

Department of Zoology, Faculty of Science, Zagazig University, Benha Branch

NADH dehydrogenase is a very important protein and is expressed by the mitochondrial
NADH dehydrogenase gene (mtND2). Dehydrogenase enzyme is used to remove hydrogen
from its substrate, which is used in the cytochrome (hydrogen carrier) system in respiration to
produce a net gain of ATP. Also, it reversibly catalyses the oxidation of NADH to NAD and
reduced acceptor. The size of mtDN2 of Tilapia species and their hybrids is ~1050 base pairs
and was detected by using the polymerase chain reaction technique. To identify the molecular
phylogeny and the physical characteristics of mtND2 gene of Tilapia species were done by
using the restriction fragment length polymorphisms (RFLPs) with some restriction
endonucleases(AccI, AvaII, AvaI, StyI, Bg1I and EaeI). The PCR-RFLPs of NADH dehydro-
genase gene of Tilapia species and their hybrids may prove that the gene is quite evolution
phylogenetic difference from one species to another. At the same time, This study investigated
the feasibility of mitochondrial DNA (mtDNA) based approaches in addressing problems of
identification of Tilapia species and their hybrids, isolated from the River Nile by using the
PCR-RFLPs analysis of mtND2 gene.

Key words; Tilapia spp., PCR-RFLP, mtND2, Polymorphism

Tilapias are a group of 40-50 species
AFLP markers (Kocher et al., 1998). The
of cichlid fish (genus Oreochromis,
second generation map is based on an F2
Sarotherodon and Tilapia) native to Africa,
cross between O. niloticus and O. aureus,
in particular Egypt. The significance of
and contains over 500 microsatellite
Tilapia for human culture predates their
markers (Danley and Kocher, 2001).
Tilapia fish have received wide
Oreochromis niloticus, a geographically
attention from evolutionary biologists for
widespread Tilapia, was already known to
more than 100 years because of their
the early Egyptian cultures and played a
extremely diverse morphology, behavior,
significant role in their lives, as indica -ted
and ecology (Fryer and Iles, 1972; Liem
by their presence in ancient Egyptian art
and Osse, 1975; Greenwood, 1978;
(Fryer and Iles, 1972). Tilapias continue to
Trewavas, 1983; Oliver, 1984; Meyer et al.,
have great econo -mic importance because
1990; Keenleyside, 1991; Meyer, 1993;
several species are an important source of
Meyer et al., 1994; Stiassny and Meyer,
protein in human diets, particularly in
Egypt, and increasingly, through aquacul-
Phylogenies for Tilapia genera were
ture, worldwide (Pullin and Lowe-Mc
inserted at positions suggested by Sodsuk
Connell, 1982 and Trewavas, 1982).
(1993) and Schliewen et al. (1994), and
Widespread exploitation and aquaculture of
Sturmbauer et al. (1994), respectively.
Tilapia has lead to their introduction in all
Previous research on Tilapia focused on all
tropical regions around the globe, often
aspects of their biology, including behavior,
(Keenleyside, 1991). The recently gathered
The first genetic linkage map of the
knowledge of Tilapia spp. phylogenetic
O. niloticus linked 162 microsatellite and
relation -ships (e.g., Oliver, 1984; Stiassny,

Full Paper (vol.6 paper# 2)

Protective Effects of Garlic Oil against Liver Damage Induced by Combined Administration of Ethanol-Carbon Tetrachloride in Ra The Egyptian Journal of Hospital Medicine Vol., 6 : 27 - 36 March 2002

1687 - 2002

Protective Effects of Garlic Oil against Liver Damage Induced by Combined
Administration of Ethanol and Carbon Tetrachloride in Rats

Ashraf B. Abdel-Naim a, Amani E. Khalifaa, Sherif H. Ahmed b

a Department of Pharmacology and Toxicology, Faculty of Pharmacy,
Ain Shams University, Cairo, Egypt.
b Department of Biochemistry, Faculty of Agriculture,
Cairo University, Cairo, Egypt.

Herbs are known to play a vital role in the management of various liver diseases. Garlic
oil (GO) contains numerous organosulfur compounds with potential hepatoprotective effects.
The present work was planned to evaluate the possible preventive role of GO on biochemical
and histopathological alterations induced by combined administration of ethanol (EOH) and
carbon tetrachloride (CCl4) in rat liver. Two dose levels of GO (5 or 10 mg/kg/day) were
administered orally to rats for 7 consecutive days with EOH + CCl4-induced liver damage.
Activity of GO against liver damage was compared with that of silymarin (25 mg/kg/day, p.o.
for 7 consecutive days). Biochemical parameters including serum aspartate aminotransferase
(AST), alanine aminotransferase (ALT), gamma glutamyl transpeptidase (­GT), alkaline
phophatase (ALP) and bilirubin were estimated to assess the liver function. In addition, the level
of total proteins, triglycerides, total cholesterol, glutathione (GSH), and thiobarbituric acid
reactive substances (TBARS), in liver tissues were estimated. Liver damage was evidenced by
an increase in the activity/level of AST, ALT, -GT, ALP and bilirubin in sera of rats after the
combined administration of EOH and CCl4 compared to normal animals. Pretreatment of rats
with GO reduced the EOH + CCl4-induced elevated levels of the above indices. Similarly, GO
significantly prevented the decline in total proteins and the increase in triglycerides and total
cholesterol resulted after EOH + CCl4 administration in rat liver homogenates. In addition, GO
pretreatment restored liver GSH levels decreased due to EOH + CCl4 administration. The
elevation in liver TBARS level due to EOH + CCl4 administration was also prevented by
pretreatment with both low and high doses of GO. Histopathological examination indicated that
GO exhibited an obvious preventive effect against the centrilobular necrosis and nodule
formation induced by EOH + CCl4 administration. In conclusion, GO exerts hepatoprotective
actions against EOH + CCl4-induced toxicity in rats.

Key Words:
garlic oil, alcohol, carbon tetrachloride, hepatotoxicity, rats

Liver diseases remain one of the
documented to exhibit hepatoprotective
serious health problems. The high
properties (Handa et al., 1986; Mehandale,
prevalence of bilharziasis and viral hepatitis
in Egypt resides behind the wide
For thousands of years, garlic extracts
occurrence of liver disorders especially
have been used to treat infectious diseases.
hepatic fibrosis and cirrhosis (el-Zayadi et
Contents of garlic oil, particularly E-ajoene,
al., 1992). Liver injury caused by toxic
have shown a broad activity against several
chemicals and certain drugs has been
DNA and RNA viruses (Sheen et al., 1999).
recognized as a toxicological problem. In
GO has been reported to reduce serum
the absence of reliable liver protective
drugs in medical practices, herbs play a role
(Fenwick and Hanley 1985). GO contains
in the management of various liver
other organosulfur compounds including
disorders. A number of plants have been
diallyl disulfide. These compounds are

Full Paper (vol.6 paper# 3)

ž’ The Egyptian Journal of Hospital Medicine Vol., 6 : 37 ­ 44 March 2002
I.S.S.N: 12084
Gastrocnemius ­ Soleus Mechanical Stretch in Relation to Gait in
Lower Limb Burn

Z. Mowafy Emam Mowafy* ; Adel A. Nossier*; Salah Abd El Ghany** and
Amal M. Abd El Baky***
* Department of P.T. For Surgery, Faculty of Physical Therapy, Cairo University, Egypt.
** Department of Surgery, Faculty of Medicine, Cairo University, Egypt.
*** M.Sc. Student.

This study has been conducted to evaluate the effect of the prolonged mechanical passive
stretch (PMS) in elongating the gastrocnemius ­ soleus muscles and the reflection of that on
some quantitative gait parameters and the range of motion (ROM) of both knee extension and
ankle dorsiflexion. Twenty male patients were participated in this study, who were selected
from Cairo University Hospitals with a second degree posterior leg burn and a total body
surface area of 15-20%. They were divided randomly into two groups of equal number, group
(A), 10 patients who were 20-30 years of age (X=25+3.4), that received the PMS plus the
traditional physical therapy program (positioning and exercises) and group (B), 10 patients who
were 20-30 years of age (X=24.4+3.75), that received only the same traditional physical therapy
program and acted as a control group. Measurements were recorded before treatment and at the
end of the first, second and third week of treatment (session for 30 minutes, twice/day, on 5
days / week bases for a total period of 3 weeks). A 16-meter walkway, tape measure and stop
watch were used to measure the quantitative gait parameters, while a standard gomiometer was
used to measure the knee extension and ankle dorsiflexion ROM. Results of this study revealed
that the PMS was effective in elongating, the gastrocnemius ­ soleus muscles as reflected by the
improved gait parameters as well as the ROM of both knee extension and ankle dorsiflexion.


Skin is the largest organ in the human
general, stretching exercises are the most
body, burn injury causes destruction and
important modality in elongating the
disturbance of the normal physiological
functions of the skin, 19.20.21.
structures and thereby increasing the range
of motion (ROM), 1.6.18.
rehabilitation always has been to return

patient back into society in as near to their
Material and Methods
normal function capacity as what existed

prior to the burn injury, 20.21. Patients with
lower limb burns, often spend significant
Twenty male patients ranging in age
time and effort attempting to regain their
from 20-30 years, participated in this study,
who were selected from Cairo University
gastrocnemius ­ soleus muscles result in
Hospitals, with a second degree posterior
temporary versus a permanent gait and
leg burn and a total body surface area of 15-
postural deformities depends on the
patient's compliance with an overall
Patients were randomly divided into
rehabilitation program established in
two groups of equal number, the PMS
consultation with the physical therapist,23.24.
group (study group), that received the
There are many approaches for
prolonged mechanical passive stretch plus
lengthening the shorted muscles as gait
the traditional physical therapy program
training, passive and active stretch, in
(positioning and exercises), and the control


Full Paper (vol.6 paper# 4)

Histological and Histochemical Studies on the Role of Caffeine on Radiation damage of the Liver The Egyptian Journal of Hospital Medicine Vol., 6 : 45 ­ 62 March 2002
I.S.S.N: 12084

Histological and Histochemical Studies on the Role of Caffeine on Radiation
damage of the Liver

* Omaima , S. Essa ; ** Abdel Mawgood A.Isamil and ** Ashraf, M. Moustafe

* National Center for Radiation and Technology.
**Histology Dept. Faculty of Medicine, AL-Azhar University Cairo, Egypt.

The present study was designed to evaluate effect of caffeine to gamma irradiated rats.
Forty eight female albino rats (120-150 gm) were divided into four groups (each of them
12 rats). The first group was considered as control. The animals in the second group were
injected I.P. with caffeine (80 mg/Kg b.w.), the third group exposed to (7Gy) -radiation while
the animals in the fourth group injected I.P with caffeine (80 mg/Kg. b. w.) 1 h prior to whole-
body dose of -rays (7 Gy).
Fresh liver specimens were obtained after 1 and 3 days of treatment. Paraffin sections
were prepared for histological study (Hx & E.) and frozen sections were prepared for
histochemical study for localization of both acid, alkaline phosphatases and succinic
dehgdrogenase enzymes.
The enzymatic activity was quantitatively evaluated and statistically analyzed. The
obtained results showed that whole body -irradiation of rats induced definite microscopic
changes in the liver represented by periportal fibrosis with some necrotic hepatic tissue.
Meanwhile caffeine alone showed histological pattern more or less near to normal. On the other
hand -rays combined with I.P. injection of caffeine revealed vaculated hepatocytes with dilated
blood sinusoids. Histochemical changes were also observed showing significant increase in
both acid phosphatase and succinic dehydrogenase enzymes activity after one and three days
post exposure while the alkaline phosphatase enzyme activity showed significant decrease after
one day and increase after three days of exposure.
Administration of caffeine (1, 3, 7-trimethylzanthine), a major component of coffee (80
mg/Kg body weight), 60 min prior to whole body lethal dose of gamma-irradiation (7 Gy)
resulted in improvement of the radiation induced changes in the activity of measured enzymes.


Exposure to high doses of ionizing
oxygen reactive sites (Kesaven et al.,
radiation is associated with utilization of
oxygen by cells resulting in production of
Protection from -rays, still nterest
highly reactive free radicals implicating
of many radiobiologists, trying to
tissue damage (Packer 1991) and appears in
achieve less toxic substance with
several forms of cellular injury (Boner et
maximum protecting action against
al., 1988).
radiation effects.
Caffeine, a major component of

coffee is an antioxidant radioprotector
Material and Methods:
against the oxic pathway of radiation
Forty eight female albino rats weighing
damage in a wide range of cells and
from 120-150 gm were used. Animals were
organisms (George et al., 1999). Moreover,
housed in cages under good ventilation
radioprotection could also be possibly due
condition and adequate stable standard diet.
to a mutually inhibitory reaction of caffeine
Irradiation was performed by gamma
molecules with the -radiation induced
cell 40 (cesium-137) source belonging to

Full Paper (vol.6 paper# 5)

I.S.S.N: 12084
Fluoxetine Hcl Induced Intrauterine Foetal Growth Retardation And
Skeletal Malformation In Pregnant Mice

Ali,M.O. ;Sharf-El Deen, U.A.. ; El Menshawy,O.M. and Bakry,S.A.

Zoology Dept. Fac. Sci., Al-Azhar Univ., Nasr city , Cairo, Egypt.

Fluoxetine is antidepressant drug which widely known as Prozac®, is a fluorinated methyl
phenoxy derivative of phenylpropylamine. Fluoxetine is a bicyclic antidepressant that differs
structurally and pharmacologically from other currently available antidepressant agents. 80
pregnant mice were administrated oral doses of Fluoxetine (0.052, 0.104 & 0.208 mg/mouse
/day) From 6th up to both 15th and 19th days of gestation.
The pregnant mice treated with Fluoxetine HCl showed states of instability, nervousness,
twitching of head, agitation, hazy movement and marked reduction in food intake as well as
reduction in the body weight. The results of uteri examination of pregnant mice groups on both
15th and 19th days of gestation showed remarkable reduction in their size, dismorphology, length
and number of implantation sites as well as large reduction in the number of still live embryos.
Increase in the number of dead and resorbed mouse embryos was dose dependant. Also,
the results showed reduction in both body weight and crown rump of mouse embryos. The
treated mouse foetuses showed several malformations as diminution in size, exenocephalia and
skeletal malformations.

Key words: Fluoxetine, Mice, Teratogenicity, Malformations, Dead and resorbed embryos.

The problem of psychotropic drug
and its metabolite (Norfluoxetine) secreted
teratogenicity has apparent in the very
in human breast milk. Limited data indicate
recent years and directed a great attention
that Fluoxetine and Norfluoxetine concentr-
of the authors toward this problem. Over
ations are (20-30%) concurrent maternal
10.000 cases of congenital anomalies in
plasma drug concentration. Fluoxetine was
not teratogenic in rat and rabbits at doses
equivalent to (9 and 11) fold the maximum
Tranquilizers) have been recorded between
daily human therapeutic dose respectively.
1960 and 1962. Sommi et al. (1987) and El-
A slight decrease in maternal survival was
Nahass, 1988 ) showed that when rats,
seen in rats exposed to (5 and 9) times the
administrated doses up to nine times of
maximum human therapeutic dose; this
Fluoxetine of those recommended for
may have been due to maternal toxicity
humans did not affect fertility but caused
(Goldstein, 1990). The exposure of mouse
reduced maternal food intake and weight
embryos in whole embryo culture to
gain and a slight reduction in neonatal
Sertarline (SSRI) at concentration, which
survival. No teratogenic effects have been
produced no evidence of general embry-
observed in vivo. Fluoxetine and its
otoxicity, caused by craniofacial malfor-
metabolite Norfluoxetine have been shown
mations consistent with direct action of 5-
to cross the placenta in the rats and are at
HT (5-hydroxytryptamine) uptake sites
highest levels in foetal brain and thymus
(Shuey et al., 1992).
(Pohland et al., 1989). The same results
Both Fluoxetine and Sertarline are inhibit
were reported by many investigators
the proliferation of cardiac mesenchyme
(Cooper, 1988; Hoyt,1989; Byrd &
endocardium. These effects were most
Markham ,1994 and Vorhees et al., 1994).
pronounced when exposure began at
Isenberg (1990) revealed that, Fluoxetine
embryo day 9th. These results suggested that

Full Paper (vol.6 paper# 6)

Comparative study on the effect of Gliclazide and two The Egyptian Journal of Hospital Medicine Vol., 6 : 80 ­ 98 March 2002
I.S.S.N: 12084

Comparative study on the effect of Gliclazide and two
Antidiabetic plants used in Folk Medicine on Albino Rat's fetuses

Ibrahim G. Ibrahim*, Boshra El-Salkh**, Nagwa Shawki**,
Shadia M.*** and Hala M. Abou El-Fotouh***

* Zoology department, Faculty of Science, Al ­ Azhar University for boys.
** Zoology department, Faculty of Science, Al ­ Azhar University for girls.
*** Faculty of Pharmacy Cairo University.


The present study was designed to assess the adverse effects of gliclazide and two
antidiabetic plants extracts on 15 &18 days fetuses of albino rat (Rattus norvegicus). The two
antidiabetic plants extracts were aquatic extract of Aremisia herba alba (shih-balady) and
alcoholic extract of Salix babylonica L. leaves (Om-Ashoor) which is known by salicin. The
doses used were 4 mg / kg (Katsumata, and kastumata 1990) for the gliclazide drug (diamicron),
450 mg/kg for Artemisia herba alba (Al-Waili, 1986) and 1.5 gm/kg for salicin. Pregnant rats
were given orally the applied does level every other day from the onset of gestation till the 15th
or to the 18th day. The pregnant rats were divided into control group (G1); diabetic pregnant
female rats (G2); Artemisia treated group (G3); salicin treated group (G4) and glicalzide treated
group (G5). Fetuses showed diminution of size. Crown-rump lentght while the number of
embryos increased in case of salicin treatment and decreased in case of gliclazide treatment due
to increased resorption rate.
The endoskeleton system of fetuses obtained from pregnant diabetic mothers (G2) showed
normal ossification in all fetuses with enlarged skeleton.
Some fetuses of Artemisia treated group showed major enoskeletal anomalies in the from of
incomplete ossification of the skull bones, or missing in some skull bones, and also missing of
caudal vertebrae.
Salicin treated group fetuses showed lack in ossification of the nasal, frontal and complete
absence of parietal bone. While gliclazide treated group fetuses showed moderate ossification of
nasal and frontal bones. On the other hand parietal bone exhibited complete missing.


Traditional antidiabetic plants might
Soler et al. (1976) stated that the
provide a useful source of new oral hypolg-
congenital malformations in infants of
lycemic compounds for the development as
diabetic mothers are three to four times
pharmaceutical entities or as simple dietary
higher than in the normal populations of
adjuncts for existing therapies (Marles, and
Birmingham. The malformations were fatal
Farnsworth 1995).
and ccounted for 26 percent of the perinatal
The present study concerned with the
mortality. Central nervous system, cardiov-
effect of two hypoglycemic plants used in
ascular and skeletal abnormalities were
folk medicine and gliclazide (diamicron) as
equally common and there was a high
a urea sulphony drug. The two antidiabetic
incidence of anencephalus, Spina bifida,
plant extracts were aquatic extract of
transposition of the great vessels and sacral
Artemisia herba alba sand alcoholic
extracts of Salix babylonica L. leaves (Om-
Temesio and poses, (1977) noticed
Ashoor) which known by salicin.
the congenital malformation in diabetic


Full Paper (vol.6 paper# 7)