A huge progress has been achieved in mammalian in vitro technique. Instead, of the many trials to develop marine invertebrate cell cultures, only a few have obtained them and only from few tissues. Since in vitro cell culture of invertebrates could be very useful for many aspects of basic and applied science, in this work we investigate and describe the development of a technique for the establishment of cell cultures from gill, mantle and gonadic tissue of the Manila clam (Ruditapes philippinarum). We maintained viable cultures for up to 25 days. Culture viability and proliferation were tested with 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) and with trypan blue, while an antibody against the ATP-dependent RNA helicase VASA, a protein expressed in the germline, and in multipotent stem cells of some animals, was used to verify the presence of these cell types. Following the described protocol: 1) explant resulted the better source to obtain cell cultures, when compared to enzymatic dissociation; 2) cultures of suspended cells were viable for longer period than adherent cells; 3) cell cultures obtained from tissues sampled in September-October performed better compared to other periods of the year, regarding maintenance and growth; 4) the tissue from which we obtained longer-lived cell cultures was gonadic tissue, especially form samples that show more undifferentiated germ cells and more VASA-stained cells. This study describes the challenges concerning the development of in vitro culture techniques for aquatic invertebrates.

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Aim: To evaluate the potential therapeutic role of Annona muricata (graviola) fruit and bee venom (BV) against N-methylnitrosourea (MNU)-induced breast cancer in pregnant female rats and complications in the ovaries. Methods: A total of 24 female rats were induced with a single dose of MNU (50 mg/kg body weight). After confirmation of positive tumor marker, female rats were placed with the males for mating. The pregnant rats were randomly divided into four groups (n=6): MNUinduced only (group 1), MNU-induced rats and supplemented with A. muricata 200 mg/kg diet (group 2), MNU-induced and treated with two doses of BV 75 μg/kg (group 3), and MNU-induced and treated with both A. muricata and BV (group 4). Results: In group 1, the breast tissue of mothers revealed pronounced cellular hyperplasia and histopathological signs. Also, the ovarian tissue of mothers and their offspring displayed deleterious histological changes. In groups 2 and 4, histopathological signs and cellular hyperplasia markedly disappeared in breast tissue. However, the histopathological signs induced by MNU in the ovarian tissue reversed to normal in groups 2-4. Also in groups 2-4, levels of serum MMP1, NFκB, and TNFα significantly decreased, and serum caspase 3 significantly increased either in mother rats or their offspring compared to the MNU-alone group. Levels of serum MDA significantly decreased; however, levels of serum antioxidants (CAT and SOD) significantly increased in all groups 2-4 compared to MNU-alone group. Conclusion:A. muricata has a more powerful therapeutic role than BV against MNU-induced breast cancer in rats; however, both have a powerful ameliorative role against ovarian histopathological alterations induced by MNU. Such ameliorative effects of A. muricata and BV are mainly attributed to their antioxidant, anti-inflammatory, and antiproliferative constituents.

Diabetes mellitus (DM) is a chronic metabolic disease. Olive leaves consists of phenolic compounds, flavonoids and volatile oil. The aim of this study was to evaluate the anti-diabetic and the curative effect of olive leaves extract on streptozotocin-induced diabetic rats. Forty male rats were divided into 4 groups, Group I; Animals were served as control. Group II; Animals were received oral Olive leaves extract (OLE) 0.5 mg/kg of body weight/day. Group III; Animals were injected intraperitoneally with a single dose of streptozotocin (STZ) (45mg /kg of body weight) to induce diabetes. Group IV, Animals were received a single dose of streptozotocin and after 7 days received Olive leaves extract for four weeks. The results indicatet that injection of STZ provoked a significant increase (P<0.005) in serum ALT, AST and lipids. Moreover, Serum malondialdehyde was increased and the antioxidant enzymes SOD and CAT decreased. Histopathologically, the OLE group didn't show any histopathological changes. Diabetic animals showed many histopathological changes in liver e.g. destruction of liver architecture, congestion of blood vessels, Leucocytic infiltration, and cytoplasmic vacuolization of hepatocytes. The pancreas showed severe damages in the pancreas architecture and atrophy of β-cells. When animals treated with OLE, an improvement was observed in the biochemical parameters and liver and pancreas histology of these animals. It is concluded that OLE exhibited a pronounced hypoglycemic, hypolipidemic and ameliorative effects in diabetic rats and this is may be attributed to the presence of its phenolic compounds.

Background: Cisplatin (CP) is one of the chemotherapeutic drugs widely utilized in the treatment of several malignancies. However, recently; its use has been limited because of its hazardous health drawbacks. Previous researches confirmed that CP has severe deleterious side effects on pregnant mothers and their fetuses. Garlic (Allium sativum) extract has been claimed to exhibit potent antioxidative and free radical scavenging abilities. Aim: This work is mainly designed to evaluate the potential therapeutic role of garlic extract against CP-induced nephrotoxicity in pregnant rats and their offspring. Methods: 24 pregnant rats were used in the current study. They were randomly allocated into four groups (n=6):  control, garlic, CP, and CP + garlic group. At the end of the weaning period, the mothers and the offsprings of all groups were sacrificed, the kidneys were immediately excised, and processed for histological and biochemical investigations. Also, blood samples were withdrawn and processed for estimation of the assigned biochemical parameters.   Results: The renal histological sections from CP-treated mother rats displayed pronounced histopathological lesions however, their offspring showed mild renal histopathological lesions if compared with those of their mothers. The levels of renal tissue Superoxide dismutase, catalase, and glutathione peroxidase enzymes were significantly decreased. On the contrary, the levels of malondialdehyde, serum urea, and creatinine were significantly increased in CP-treated mother rats and their offspring as compared with control. The percentage value of caspase 3 activity was markedly elevated in the renal tissues of CP-treated mother rats and their offspring compared to the control group. Supplementation of garlic extract to the CP treated rats; the overall histological lesions, as well as biochemical parameters, were restored nearly to the control ones. It is concluded that garlic (Allium sativum) extract has a powerful ameliorative role against CP-induced nephrotoxicity in pregnant rats and their offspring.

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Silver nanoparticles (AgNPs) have wide applications. Production of AgNPs can be occurred through different method chemical, physical, and green methods. The most popular methods are chemical approaches. Marine organisms exhibit a wide range of bioactivity. The present study was designed to establish the biosynthesis of silver nanoparticles from marine crustacean extract of the hard and soft parts of male and female E. massavensis. The microstructure, morphology and optical absorption properties of the nanoparticles were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and UV-visible spectroscopy. The formation of silver nanoparticles was confirmed by Uv-Vis absorption and the spectra were observed plasmon bands between 441.79-462.74 nm. XRD results show that the nanoparticles are crystalline in nature. SEM images detected the quasi-spherical AgNPs morphological shape. Silver nanoparticles from marine crustacean extract of the hard part of male E. massavensis (HM4) showed the best results in morphology and particle size. Evaluation of the cytotoxicity of AgNPs (HM4) on different cancer cell lines antiviral, anti-microbial, anti-diabetic, anti-arthritic, anti-aging and anti-inflammatory properties were assessed. AgNPs characterization may be introduced a promising applications in medical aspects.

Background: Cisplatin (CP) is one of the most powerful chemotherapeutic drugs that widely utilized in the treatment of several malignancies .However, in the late era; its use has been limited because of its hazardous health drawbacks. Previous researches confirmed that CP has severe deleterious side effects on pregnant mothers and their fetuses .Garlic (Allium sativum) extract has been claimed to exhibit potent antioxidative and free radical scavenging abilities in various disease conditions.Aim: The goal of the current investigation was to throw light on the CP-induced nephrotoxic effects in pregnant albino rats and their offspring and to clarify the possible ameliorating role of garlic (Allium sativum) extract on such alterations.Material & Methods: Thirty two adult Wistar albino rats (24 females and 8 males) were used in the current study. After mating and confirmation of pregnancy, the pregnant rats were randomly allocated into four equal groups (n=6).Group1: served as control, group 2: garlic extract supplemented, group 3: CP-treated and eventually group 4: received both CP and garlic extract. At the end of the weaning period, the mothers and the offsprings of all groups were sacrificed, the kidneys were immediately excised, weighed and processed for histological and biochemical investigations. Also, blood samples were withdrawn by direct cardiac puncture and processed for estimation of the assigned biochemical parameters. Results: The renal histological sections from CP-treated mother rats displayed pronounced histopathological lesions including little and atrophied glomeruli, damaged and dilated renal tubules. However, the offsprings suffered from mild renal histopatological lesions if compared with those of their mothers. Furthermore, the levels of renal tissue superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) enzymes were significantly decreased .On the contrary, the levels of malondialdehyde (MDA), serum urea and creatinine were significantly increased in CP-treated mother rats and their offsprings as compared with the corresponding values in the control group. On the other hand, the percentage value of caspase 3 activity was markedly elevated in the renal tissues of CP-treated mother rats and their offsprings compared to the control group. Following concomitant administration of both CP and garlic extract to the rats of group 4, the over-all histological scene of the kidney documented that there was an evident improvement so that it, more or less, resembled that of the control animals. In parallel with this, a noticeable improvement in the estimated levels of anti-oxidants, MDA, urea and creatinine as well as caspase 3activity, more or less, simulated the control levels.Conclusion: Based on the previous data, it is concluded that garlic (Allium sativum) extract has a lessening effect on the CP induced renal toxicity in pregnant rats and their offspring.