ABSTRACT

BACKGROUND: on the last decade in Libya, the oncology center in Misurata become the most

important cancer diagnosis and follow up center on the western region of Libya. The Libyan

society was worried about the increase of the percentage of tumor incidences among the Libyans

on the last few years. This study focuses on cancer prevalence in city of Misurata and the

surrounding area in central Libya from 2012 to 2016.

METHODS: A hospital-based registry of cancer patients was formed using records from oncology

center in Misurata, focusing on patients who were diagnosed between 2012 and 2016.

RESULTS: the incidence of tumor on females was higher than males from 2014 up to higher

deference on 2016, the most common malignancies in men were cancers of the lung on years from

2012 up to 2015 even though it decrease to 18% of males patients on 2016, followed by colon, its

prevalence rate ranged from 14% to 18% of recorded male patients (For women, they were found

to be cancers of the breast ranged from 29.84% on 2012 and raised to 43.08%. Additionally, agestandardized

rates (ASR) were determined for recording patients who leaved on the city of

Misurata and compared with published record from the other countries. The incidence rates given

for the city of Misurata can be considered with the counties with low malignancy rate

CONCLUSION: Proper surveillance programs need to be in place and healthcare policy should be

adjusted to consider the more prevalent and pressing cancers in society.

Key words : Misurata cancer center, breast cancer, lung cancer, colon cancer, Libya

ABSTRACT

Fat cadherins comprise the largest of all known members of cadherin superfamily. They are present

in all multicellular organisms and retain a high degree of structural conservation. In Drosophila

there are two Fat genes: Fat and Fat-like, whilst in vertebrates there are four members called Fat1,

Fat2, Fat3 and Fat4. Our lab group focused on the use of various biochemical methods to analyse

expression of the FAT1 protein. Because of the high molecular size of FAT1 protein our laboratory

has used affinity purification to prepare bespoke rabbit anti-FAT1 antibodies using FAT1 protein

prepared as GST fusions. five overlapping segments of the FAT1 cytoplasmic tail designated A, B,

C, D and E as GST-fusion proteins contained within pGEX plasmids were assigned in our lab.

According to their sequences and hosted rabbit name antibodies were named N34B, N35B and

N34E. To test the efficacy of each antibody preparation, two different techniques were undertaken,

first Western blotting and second immunofluorescent staining. By this analysis the

immunoreactivity of N34B, N35B and N34E compare favourably with the CTD-pAb. N34E

produced less than satisfactory results in this test with low signal to noise and these results are

omitted here. This staining pattern was largely consistent between N34B and N34E, but the signal

to background was a considerably high. According to that the use of the new B and E antibodies

was restricted to Western blotting where these reagents fulfilled the functional requirements.

Key words: Fat1, Fat1 antibodies, cadherins, western blot. Immunofluorescence staining

Abstract

The loss of cognitive function accompanying healthy aging is not associated with extensive or characteristic patterns of cell death, suggesting it is caused by more subtle changes in synaptic properties. In the hippocampal CA1 region, long-term potentiation requires stronger stimulation for induction in aged rats and mice and long-term depression becomes more prevalent. An age-dependent impairment of postsynaptic calcium homeostasis may underpin these effects. We have examined changes in presynaptic calcium signalling in aged mice using a transgenic mouse line (SyG37) that expresses a genetically encoded calcium sensor in presynaptic terminals. SyG37 mice showed an age-dependent decline in cognitive abilities in behavioural tasks that require hippocampal processing including the Barnes maze, T-maze and object location but not recognition tests. The incidence of LTP was significantly impaired in animals over 18 months of age. These effects of aging were accompanied by a persistent increase in resting presynaptic calcium, an increase in the presynaptic calcium signal following Schaffer collateral fibre stimulation, an increase in postsynaptic fEPSP slope and a reduction in paired-pulse facilitation. These effects were not caused by synapse proliferation and were of presynaptic origin since they were evident in single presynaptic boutons. Aged synapses behaved like younger ones when the extracellular calcium concentration was reduced. Raising extracellular calcium had little effect on aged synapses but altered the properties of young synapses into those of their aged counterparts. These effects can be readily explained by an age-dependent change in the properties or numbers of presynaptic calcium channels.

Neurodegenerative disorders results in inflammatory processes, including inflammatory cytokine secretion and concomitant superoxide production. Acetylcholinesterase inhibitors (AChEIs) influence neuro degeneration through anti-inflammatory effects. Donepezil as an AChEIs also provide neuro protection. This study aimed to evaluate the effect of donepezil (DZ) as a potent acetyl-cholinesterase (AChE) inhibitor, L-NG-Nitroarginine methyl ester (L.NAME) as non-selective nitric oxide synthase inhibitor and 7-nitroindazole (7-NI) as a selective neuronal nitric oxide synthase (NOS) inhibitor against Amyloid beta-peptide (1-42) (Aβ(1-42)) induced neurological disorder. Rats were divided into six groups including control, Aβ(1-42) ,Aβ(1-42) +L-arginine, Aβ(1-42) +L-NAME ,Aβ(1-42) +7-NI, and Aβ(1-42) +DZ. Brain AChE, malondialdehyde, nitric oxide, super oxidedismutase, catalase, reduced glutathione, interleukin 1-beta (IL-1β) and tumor necrosis factor alpha (TNF-α) were measured. Also, brain fatty acids fractions were estimated by high performance liquid chromatography (HPLC). Aβ(1-42) significantly alter levels of brain antioxidant, inflammatory markers and fatty acids content compared to the control group. Treatment with L.NAME, 7-NI and especially DZ improve these parameters. Administration of acetylcholinesterase inhibitors such as DZ attenuates neurodegenerative disorders through decreasing oxidation and inflammation.

Keywords: Neurotoxicity, Aβ (1-42), Donepezil, Nitric oxide, 7-nitroindazole, Fatty acids, HPLC

Genetically encoded calcium indicators (GECIs) have gained widespread use for measurement of neuronal activity but their low expression levels in transgenic mice tend to limit sensitivity. We have developed a transgenic mouse line (SyG37) that expresses a ratiometric calcium sensor, SyGCaMP2-mCherry, that is expressed throughout the brain but targeted to presynaptic terminals. Within the CA1 and CA3 regions of hippocampus of male and female mice, SyGaMP2 fluorescence responds linearly up to 10 electrical stimuli at frequencies up to 100 Hz and it can detect responses to a single stimulus. Responses in single boutons can be measured using multiphoton microscopy. The ensemble amplitude of SyGCaMP2 responses is a function of the number of stimuli applied and the number of contributing boutons. The peak responses and initial rates of calcium influx in single boutons in CA1 and CA3 were similar but the rate of calcium clearance from CA3 boutons after stimulation was significantly faster. In CA1, DNQX reduced SyGCaMP2 responses to Schaffer collateral stimulation to 86% of baseline indicating that 14% of the total response originated from presynaptic terminals of neurones synaptically driven via AMPA receptors. Theta burst stimulation induced long-term potentiation (LTP) of both SyGCaMP2 and fEPSP responses in both young and 18-month-old mice. The proportion of postsynaptically connected terminals increased significantly to 76% of the total after LTP induction. The SyG37 mouse allows stable optical detection of synaptic activation and connectivity at the single bouton level and can be used to characterize the contributions of presynaptic calcium to synaptic transmission and plasticity.

Alzheimer’s disease (AD) is a progressive age-related neurodegenerative disorder characterized by progressive impairment of memory and cognitive functions. Oxidative stress, neuroinflammation, and neurotransmitters disturbance may play an important role. This study aimed to evaluate the effect of donepezil alone and in combination with nitric oxide synthase (NOS) substrate L-arginine, and nitric oxide synthase inhibitor (NOSi) Ng-nitro-l-arginine methyl ester hydrochloride (L-NAME) and 7-nitroindazole against aluminum chloride (AlCl3) induced neurotoxicity. A total of 36 male albino rats were divided to six groups: control normal saline, AlCl3. ,Donepezil, Donepezil+ L-arginine, Donepezil+ L-NAME, and Donepezil+7-nitroindazole. Neurotoxicity was induced by AlCl3 in a dose of 10 mg/Kg subcutaneously for 30 days. After the experimental period, brain was removed for determination of acetylcholinesterase activity (AChE), levels of fatty acids fractions by high performance liquid chromatography (HPLC), interleukin 1β (IL-1β) and tumor necrosis factor alpha (TNF-α). AlCl3 significantly increase brain levels of AChE, IL-1β, and TNF-α, and arachidonic acid (AA) and linoleic acid (LA). Treatment with Donepezil alone and in combination with L.NAME and 7-nitroindazole significantly attenuates these changes. In conclusion, regulation of nitric oxide (NO) level is the key for stability and prevention of neurodegenerative diseases. Combination of Donepezil with nitric oxide synthase inhibitors especially 7-nitroindazole attenuates the oxidant and inflammatory induced by AlCl3.

Keywords: AlCl3, L. NAME, Donpezil, fatty acids, HPLC

In this project we used a combination of electrophysiology and fluorescent imaging to monitor synaptic transmission and calcium signalling in synaptic terminals. The study as a whole intended to examine how presynaptic calcium contributes to normal synaptic transmission within different hippocampal neuronal pathways. To this end, we used a transgenic mouse strain known as SyG37 that stably expresses a calcium sensor, SyGCaMP2-mCherry that is expressed in subsets of CNS neurones under the control of the Thy1 promoter. Our findings indicate that this new ratiometric sensor, in the SyG37 mouse strain, provides an excellent tool for detecting neural activity in acute brain slices. First, we showed that evoked calcium transients can be detected in acute brain slices prepared from SyG37 mice where electrical activation of Schaffer collaterals or mossy fibres elicited large calcium transients in area CA1 and CA3, respectively. Using immunohistochemical techniques, SyGCaMP2-mCherry co-localised with presynaptic proteins such as Bassoon, VGLUT1 and VGAT, confirming that it is expressed presynaptically in both excitatory and inhibitory terminals. Blocking fast glutamatergic and GABA/Glycinerergic transmission reduced the size of calcium transients in CA1 and CA3 by only 25 and 20% respectively indicating that the majority of the signals originated from first order presynaptic terminals. Pharmacologically, manipulating the adenosine receptor signalling pathway showed that the actions of adenosine, via the A1 receptor subtype, were different in the CA3 region compared to those in CA1. Forskolin also caused a small, concentration dependent effect on SyGCaMP2 fluorescence in response to electrical stimulation within both CA1 and CA3 regions with pronounced effects on field potential recordings. Together, with this SyG37 strain of transgenic mouse, it is possible to detect neuronal activity with fast temporal and high spatial resolution without the need for pre-incubation with organic calcium dyes or invasive viral transduction procedures.

Alzheimer's disease is an age-associated, irreversible, progressive neurodegenerative disease that is characterized by severe memory loss, unusual behavior, personality changes and a decline in memory function. It is the most common form of dementia and affects an estimated 10 million people worldwide. Alzheimer’s disease demolishes the vital brain cells, causing trouble with memory, thinking and behavior, brutal enough to affect work, lifelong hobbies and social life. Recognized factors in Alzheimer's disease include acetylcholine deficiency, free radicals and inflammation of the brain tissue. There is no cure for Alzheimer's disease, but drugs designed to slow down the disease progression are available. Some herbs may help to improve brain function, but scientific evidence to prove that they can treat Alzheimer's disease. Medicinal plants have been the single most productive source of leads for the development of drugs, and over a hundred new products are already in clinical development. Indeed, several scientific studies have described the use of various medicinal plants and their constituents for treatment of Alzheimer's disease. This review gathers research on various medicinal plants that have shown promise in reversing the Alzheimer's disease pathology. The report summarizes information concerning applications of these various plants in order to provide sufficient baseline information that could be used in drug discovery campaigns and development process, thereby providing new functional leads for Alzheimer's disease.

Since last two decades the work on oleogels is being exploited in pharmaceutical, cosmetics, and nutraceutical industries for their desired rheological, physical, and chemical stabilities in semisolid formulations. Recently, we had developed a

stable and efficacious oleogel containing diclofenac diethylamine for topical application.The present review article deals with the literature of oleogels including its application in various fields from last few decades till date. The literature reveals that the oleogels have simplicity in manufacturing, high physical, chemical, and mechanical stability and better invivo efficacy,which make them appropriate to employ as bases for topical formulations.

Tiagabine (TGB) is an antiepileptic agent enhancing the activity of GABA at neuronal and glial region. It has recently been shown that enhancement of TGB chemical stability was improved by complexation with 2-hydroxypropyl-beta-cyclodextrin (2-HPβCD). The aim of this project is to explain the improvement of the chemical stability of complexed TGB by studying the inclusion properties and factors affecting the complexation selectivity between 2-HPβCD and TGB. Analysis of the interaction between 2-HPβCD and TGB and the effect of 2-HPβCD on TGB solubility was performed by phase solubility method described by Higuchi and Connors; the complexation was followed by characterization using DSC, FTIR and NMR spectroscopy. In aqueous media, the analysis of NMR proton shift change continuous variation method (Job’s plot) and the NMR diffusion-ordered spectroscopy (DOSY) measurements clearly show that TGB form 1:1 inclusion complex with 2-HPβCD with an association constant (K a) of 3396 M⁻¹. More detailed information about the inclusion mode and the geometry of the complex was obtained by the analysis of 2D NMR NOESY experiment and molecular modelling calculations. The inclusion process indicates that A-ring, C10–C11 double bond and the half of the B-ring of TGB molecule were located inside the cavity while the nipecotic acid part of TGB (Ring C) was exposed towards the outside of the 2-HPβCD cavity. These results suggest that the inclusion of the C10–C11 double bond in the 2-HPβCD cavity may possibly the reason of improvement of TGB chemical stability.