IN VITRO SUSCEPTIBILITY OF ISOLATED SHIGELLA FLEXNERI AND SHIGELLA DYSENTERIAE TO THE ETHANOLIC EXTRACTS OF TRACHYSPERMUM AMMI AND PEGANUM HARMALA

Objective: This study represents the first attempt to investigate the antimicrobial activity of Peganum harmala, and Trachyspermum ammi seeds extract against the isolated bacillary dysentery-causing microorganisms. Methods: T. ammi and P. harmala were extracted by 96% ethanol using Soxhlet apparatus. The extracts were screened for their phytochemical constituents. Their antimicrobial activity against the isolated dysentery-causing microorganisms was evaluated using the agar diffusion method. Results: The antimicrobial activity result showed that, the two isolated bacteria, Shigella flexneri, and Shigella dysenteriae were found to be sensitive to the extract of T. ammi seed with inhibition zones up to 25 mm, compared to the inhibition zone of 20 mm produced by Gentamycin standard drug, this is mainly due to the presence of the different phytochemical in the extract such as tannin, flavonoids, terpenoids which are well known for their antimicrobial effects. The two isolated bacteria were found to be insensitive (zero mm) to P. harmala extract, Amoxicillin, and Amoclan (Amoxicillin+clavulanic acid) standard drugs, this is due to the fact that, the phytochemicals constituents of P. harmala possess the antagonistic effect to each other’s. Addition to; these bacteria became resistant to both Amoxicillin and Amoclan. Conclusion: From the results it concludes, T. ammi seeds extract had a considerable level of antimicrobial activity against bacillary dysentery-causing microorganisms resistant to Amoxicillin and Amoxicillin+clavulanic acid drugs

Antitumor Effect of Zerumbone Isolated from Lempoyang (Zingiber Zerumbet) on Human Cervical Cancer Cells and Mouse Cervical Intraepithelial Neoplasia

Malaysia as a tropical country is a rich source of biologically active phytochemicals, which could be useful as an alternative to the current unsafe regimens of cancer treatment. This includes the use of cisplatin (CIS), the current chemotherapeutic drug to treat cervical cancer, the second most lethal cancer affecting women in Malaysia. Therefore, anti-tumor activities of zerumbone (ZER) were investigated in both in vitro and in vivo cervical cancer models. This natural compound was isolated from the edible plant Zingiber zerumbet, locally known as Lempoyang, through column chromatography and hydrodistillation methods. The chemical structure of ZER was confirmed using NMR. The cytotoxic effects of ZER were tested in human cervical cancer cell lines (HeLa) using MTT assay and compared concurrently to cisplatin. Zerumbone’s induction of HeLa cancer cell deaths were quantified using AO/PI double staining and flow cytometry. Transmission and scanning electron microscopic analyses were done to evaluate ultra-morphological changes. The effect of ZER on caspase-3 and caspase-9 was evaluated colorimetrically in HeLa cells. The in vivo model of cervical intraepithelial neoplasia (CIN) was induced in pregnant female Balb/c mice using Diethylstilboestrol (DES). Cervical tissues were stained with hematoxylin and eosin (H&E) and viewed under light microscopy and the in vivo antiproliferative properties of ZER was confirmed by the immunohistochemical staining of proliferating cellular nuclear antigen (PCNA) as a proliferation marker and the PCNA labeling index was obtained. Apoptosis (Bcl-2 & Bax) and G2/M-cell cycle arrest (cdc25B, cyclinB1 and Chk2) associated proteins were investigated using immunohistochemistry. Moreover, RT-PCR was used to amplify mRNA of Bcl-2, Bax, c-myc and β-actin genes. The genetic material was obtained by laser capture microdissection microscopy (LCMM). No previous toxicological investigations have been carried out on this compound. Hence, acute, sub-acute and sub-chronic toxicity studies and ZER was evaluated for its behavioural, biochemical and histo-pathological effects. Findings of NMR coincide to the previously published data. However, ZER was able to exert an antiproliferative effect towards HeLa when isolated by both hydrodistillation and column chromatography, with an IC50 of 20.30±1.1 μM and 20.41±0.9 μM (p>0.05, student t-test, n=3), respectively. AO/PI-stained HeLa cells showed that ZER induced apoptosis in a time-dependent manner with insignificant statistical (p>0.05) difference in necrosis between various doses of this compound. Moreover, flow cytometric evaluation of the effect of ZER on DNA content by cell cycle phase distribution revealed that the cell populations at G0 and G2/M phases were significantly different (p0.05) serum concentrations of AST, ALP, ALT and GGT. No histopathological changes were observed in the hepatic, renal, cardiac and gastrointestinal tissues. These histomorphological findings were supported by the insignificant differences (p>0.05) between the mean lesion scores of hepatic and renal tissues. Collectively, results presented in this study demonstrated that ZER causes metaphasal blockage in HeLa cells, leading to growth inhibition and apoptosis, which was later confirmed to be through mitochondrial pathways. As ZER exhibits similar pharmacological activity to CIS, it possesses the potential to be developed as an antiproliferative agent for cervical cancer but producing less side effects, as the compound was shown to have no toxicological signs compared to the clinical complications of CIS

Societal Marketing Orientation And Marketing Performance: The Mediating Role Of Key Market-Based Capabilities And The Moderating Effect Of Institutional Context

Societal issues appear to be increasingly important in today’s business climate. Response to these issues by adopting societal marketing orientation is expected to enable firms to gain better competitive advantage. However, literature search reveals that there is a lack of research offering valid measurement and a systematic framework that demonstrates the underlying mechanisms and contingencies through which societal marketing orientation can help firms to achieve that end. Using a priori approach the study intended to propose a valid measure of societal marketing orientation and to investigate the extent of this orientation among large manufacturing firms in Malaysia

Performance, genetic variation and interrelationships in different traits of some safflower (Carthamus tinctorius L.) genotypes

   The success of safflower improvement and production activities can be enhanced with scientific information generated from the study of genotypes, environments and genotype by environment interactions (GEI). In this study, 15 safflower genotypes were evaluated during two consecutive winter seasons (2013/14 and 2014/15) at three locations viz Gezira, El- Suki and Hudeiba Research Station farms of the Agricultural Research Corporation, Sudan. The objectives of this study were to evaluate seed yield potential, the extent of genetic variability and interrelationships in 15 safflower genotypes under different environments in Sudan. Data were collected on days to 50% flowering, days to maturity, plant height (cm), number of branches per plant, number of capitula per plant, number of seeds per capitula, 100 seed weight (g), seed yield (kg/ha), and oil content. A wide range of genetic variability was observed among the genotypes for most of the studied traits. The results also indicated that the tallest genotype (Saff 1) was the latest maturing, and the shortest genotype (Saff 13) was the earliest maturing. Simple linear correlation and path coefficient analyses indicated that days to 50% flowering, plant height, number of branches/plant and number of capitula per plant could be used as potential selection criteria in breeding programs for developing high yielding safflower genotypes. Genotypes Saff 1, Saff 12 and Saff 14 were high yielders (582.4, 507.9 and 572.8, kg/ha, respectively). Also, Saff 14 showed the highest oil content (37.5 %) followed by Saff 12 (31.0 %) and Saff 1 (29.1 %), therefore, they could be grown successfully under irrigated winter condition of central and northern Sudan.       القرطم من المحاصيل الزيتية المهمة في العالم، وفي السودان يزرع المحصول فقط في الولاية الشمالية في المساحة الواقعة علي طول النيل. إن نجاح تحسينِ محصول القرطمِ ونشاطاتِ إنتاجِه يُمْكِنُ أَنْ يُحَسّنا بالمعلومات العلمية التي تنتج منْ دراسة التراكيب الوراثية. قُيّمَت في هذه الدراسةِ خمسة عشرَ تركيباً وراثياً من القرطمَ علي مدي فصلي شتاء متتاليينِ (2013/14-2014/15) في ثلاثة مواقعِ هي محطات بحوث الجزيرة و السوكي والحديبة التابعة لهيئة البحوث الزراعية بالسودان. هدفت الدراسةِ إلي تُقييم هذه التراكيب الوراثية من حيث الانتاجية  ومكوّناتِها و دراسة العلاقات المترابطة بين الانتاجية والصفات المرتبطة بها بإستعمال الإرتباطَ الخطي البسيط و معامل تحليل المسار وتمييز التراكيب الأعلى إنتاجية. كانت الصفات المدروسة هي عدد الأيام حتي 50%  إزهار و عدد الأيام حتي النضج و طول النبات و عدد الفروع بالنبات و عدد الكبسولات بالنبات و عدد البذور بالكبسولة و وزن المائة حبة بالجرام و الإنتاجية بالكيلوجرام للهكتار ونسبة الزيت بالبذور. أوضحت النتائج وجود مدي واسع من التباين الوراثي بين التراكيب الوراثية لمعظم الصفات المدروسة. أوضحت النَتائِج إلي أنّ التركيب الوراثي (Saff 1) هو الاطول من بين التراكيب الوراثية الاخري و متأخر النُضُج، بينما التركيب الوراثي (Saff 13) مبكر النُضُج وهوأقصرالتراكيب الوراثية. من ناحية أخري، باستخدام الارتباط الخطي البسيط ومعامل تحليل المسار، أوضحت الدراسة  أن صفات عدد أيام 50% إزهار و طول النبات و عدد الفروع في النبات و عدد الرؤوس بالنبات يمكن استعمالهما كصفات إنتخاب غير مباشر في برامج التربية لاستنباط أصناف عالية الإنتاجية من القرطم. خلصت الدراسة إلي أن التراكيب الوراثية Saff 1 و Saff 12 و Saff 14 كانت عالية الإنتاجيةً ( 582.4 ، 507.9 و 572.8 :جم/ ه علي التوالي)، كما وأن التركيب الوراثي  Saff 14 له أعلي محتوي من نسبة الزيت  (37.5 %)  يعقبه في ذلك  (31.0 %) Saff 12    و(29.1 %) Saff 1   لذلك يمكن زراعتها بنجاح  تحت ظروف الري الشتوي بأواسط وشمال بالسودان

Assessment of genotype x environment interactions and stability for seed yield of selected safflower (Carthamus tinctorius L.) genotypes in central and northern Sudan

   Safflower is an important oilseed crop worldwide. In Sudan, safflower is only cultivated in the Northern State along the River Nile. The success of safflower improvement and production activities can be enhanced with scientific information generated from the study of genotypes, environments and genotype by environment interactions (GEI). In this study, 15 safflower genotypes were evaluated during two consecutive winter seasons (2013/14 and 2014/15) at three locations viz Gezira, El- Suki and Hudeiba Research Station Farms of the Agricultural Research Corporation, Sudan. The objectives of this study were to estimate G x E interactions and identify the highest yielding and stable genotypes under different environments. A wide range of genetic variability was observed among the genotypes for most of the studied traits. Combined analyses of variance revealed highly significant environment, genotype and genotype x environment (GE) component of interaction and indicated wide differences among the environments and differential genotypic behavior to the tested environments. Additive main effects and multiplicative interactions (AMMI) analyses have higher efficiency in partitioning and analyzing stability studies compared to regression analysis. First and second principal component axis (PCA 1 and PCA 2) in AMMI explained 64.6% and 25.3% of the interaction sum of squares, respectively, and together they accounted for 89.9 % of the G X E sum of squares, while the regression model accounted only for 13.8% of the G X E sum of squares. Hence, AMMI analysis was superior to regression techniques in accounting for the large part of the G X E sum of squares and more effective in partitioning the interaction sum of squares. In conclusion, genotypes Saff 1, Saff 12 and Saff 14 were high yielding (582.4, 507.9 and 572.8, kg/ha, respectively) and stable under all environments. Therefore, they are recommended to be grown under winter irrigated conditions of central and northern Sudan.     يعتبر القرطم من المحاصيل الزيتية المهمة في العالم و يزرع في السودان فقط في الولاية الشمالية في المساحة الواقعة علي طول النيل. إن نجاح تحسينِ محصول القرطمِ ونشاطاتِ إنتاجِه يُمْكِنُ أَنْ يُحَسّنا بالمعلومات العلمية التي تنتج منْ دراسة التراكيب الوراثية وتفاعلاتِ البيئة والتفاعل بين التراكيب الوراثية والبيئة. قُيّمَت في هذه الدراسةِ خمسة عشرَ تركيباً وراثياً من القرطمَ علي مدي فصلي شتاء متتاليينِ (2013/14-2014/15) في ثلاثة مواقعِ هي محطات بحوث الجزيرة و السوكي والحديبة التابعة لهيئة البحوث الزراعية بالسودان. هدفت الدراسةِ إلي تُقييم هذه التراكيب الوراثية من حيث الانتاجية  ومكوّناتِها و دراسة التفاعل بين البيئة و التراكيب الوراثية المختلفة وتمييز التراكيب الوراثية الأعلى إنتاجية و المستقرةّ في البيئاتِ المختلفة. أوضحت النتائج وجود مدي واسع من التباين الوراثي بين التراكيب الوراثية لمعظم الصفات المدروسة. أيضاً أظهرت النتائج وجود فروقات معنوية كبيرة بين التراكيب الوراثية و البيئة والتفاعل بينهما. أوضحت الدراسة أنه في طريقة الأثر التجميعي الرئيسي والتفاعل المتراكم لتحليل التباين  (AMMI)الناتج عن التفاعل بين التركيب الوراثي والبيئة أن محور المكون الأول والثاني قد  فسرا حوالي 64.6 % و25.3 %، علي التوالي من هذا التباين، بينما وجد في طريقة معامل الارتدادEberhart and Russell’s (1966)  أنه قد فُّسَر حوالي 13.8 % منه. عليه فإن طريقة الأثر التجميعي الرئيسي والتفاعل المتراكم أعلي كفاءةً من طريقة معامل الارتداد الخطي في تقسيم التباين الناتج عن التفاعل بين التراكيب الوراثية والبيئة. خلصت الدراسة إلي أن التراكيب الوراثية Saff 1 و Saff 12و Saff 14 كانت عالية الإنتاجيةً ولها ثباتاً وراثياً في كل بيئات التقييم  لذلك يوصى  بزراعتها بنجاح  تحت ظروف الري الشتوي بأواسط وشمال السودا

Effects of sowing date and intercropping on yield, yield components and oil content of sunflower and groundnut in a desert environment in northern Sudan

    The summer season in the Northern State, Sudan, is characterized by its limited field activities. The present and future expansion of agricultural production is targeted in desert plain soils away from the River Nile bank. This study was conducted during seasons 2014/15 and 2015/16 with the objectives of determining the effects of sowing date and intercropping system on yield, yield components and oil content of sunflower and groundnut. Split-plot design with randomized complete arrangement with four replicates was used. The main plots were assigned to the three sowing date namely: 1st June, 1st July and 1st August. The sub-plots were assigned to intercropping system: viz: sole cropped and intercropping (two rows of groundnut alternate with two rows of sunflower). The interaction between sowing date and intercropping had a significant (p ≤ 0.05) effect on yield component, total yield and oil content of both crops in both seasons. The highest seed yield was obtained by intercropped sunflower and sole groundnut for 1st June and 1st August, respectively. The highest oil content was attained by intercropped groundnut and sole sunflower, respectively, for 1st August.  First June sowing date resulted in the best land equivalent ratio in both seasons. Based on these results, to obtain high seed yield of high oil content, it could be recommended that 1st of June and 1st of August were optimum sowing dates for sunflower and groundnut, respectively in the desert environment of north Sudan. It was also recommended to intercrop sunflower with groundnut and sow them on the first of June for the highest LER.   النشاط الزراعي محدود جدا في الموسم الصيفي بالولاية الشمالية في السودان.  وأن التوسع الحالي والمستقبلي في الرقعة الزراعية يستهدف أراضي السهل الصحراوي والتروس العليا البعيدة عن حوض نهر النيل. نفذت هذه التجربة في المناخ الجاف علي تربة السهل الصحراوي في الولاية الشمالية في الموسم الصيفي 2014/15 و2015/16 بهدف تحديد أثر تاريخ الزراعة ونظام الزراعة البينية علي الإنتاجية ومكوناتها وكمية الزيت لمحصولي زهرة الشمس والفول السوداني. نظمت المعاملات إحصائيا باستخدام تصميم القطع المنشقة مع القطاعات العشوائية الكاملة بأربعة مكررات. حيث وضع في القطع الرئيسية ثلاثة تواريخ زراعة (1/6 و1/ 7 و1/8) وفي القطع الثانوية نظام الزراعة البينية: حيث زرع المحصول الواحد بمفرده والمحصولين مع بعض في زراعة بينية (صفان من الفول السوداني يَتناوبانِ مع صفين من زهرة الشمس). أظهرت النتائج أن التداخل بين تاريخ الزراعة والزراعة البينية كان له تأثير معنوي على مكونات الإنتاج والإنتاجية وكمية الزيت لكلا المحصولين في كلا الموسمين. أعلى إنتاجية بذور تحققت للزراعة البينية لزهرة الشمس ولمحصول الفول السوداني منفردا عند زراعتهما في تواريخ الأول من يونيو والأول من أغسطس على التوالي. أعلى نسبة زيت تحققت لمحصولي زهرة الشمس منفردا وللزراعة البينية للفول السوداني عند ما نفذت الزراعة بتاريخ الأول من أغسطس. قيمة مؤشر نسبة الأرض المكافئ أعلي من الرقم واحد في كل تواريخ الزراعة، وتاريخ أول يونيو أعطي أفضل مكافئ في الموسمين علي التوالي. للحصول على أعلى إنتاجية بذور ونسبة زيت لمحصولي زهرة الشمس والفول السوداني توصي الدراسة بأن الأول من يونيو  والأول  من أغسطس هي الأمثل لزراعة محصولي زهرة الشمس والفول السوداني علي التوالي في البيئة الصحراوية بشمال السودان, كما توصي أيضا بأن الزراعة البينية لزهرة الشمس والفول السوداني في أول يونيو هي الأفضل في استغلال الأرض. &nbsp

Screening the Efficacy of Some Traditional Herbal Drugs for Treatment of Hymenolepis diminuta Infection in Rats

Background: Hymenolepis nana (human infecting tapeworm) and H. diminuta (rodent infecting tapeworm) are currently incriminated to be the cause of non-specific bowel disturbances. They are in most instances resistant to the available anticestodal compounds due to misuse of drugs and probably adaptation of the parasites to the commercially available drugsObjective: Our objective is to study the toxicity and curative efficacy of different medicinal plants that are candidate for the treatment of tapeworm infections in man.Methods: Four medicinal plants were tested for their ability to treat Hymenolepis diminuta tapeworm infection in rats. These plants are Amaranthus viridis, Cucurbita maxima, Hagenia abyssinica and Balanites aegyptiaca. Selection of these plants was based on ethnobotanicalinformation. The evaluation of the efficiency of these medicinal plants was based on the “controlled test design”, modified from Moskey and Harwood10: Following pre-infection screening, and life cycle establishment rats were grouped to six experimental groups for each plant. Stool specimens were collected from all groups, the mean of eggs counts per gram of faeces were counted. The reduction percentage of eggs per gram (EPG) was calculated and time to clear eggs was compared with that of Niclosamide. Niclosamide drug was used in this study as a control treatment14.Results: There were no signs of toxic effect on the rats due to administration of any of the tested medicinal plants. Amaranthus viridis leavs exhibited a very weak efficacy. It did not reduce eggs ineither water or food significantly as compared to the untreated control group (p> 0.05). The deparasitization activity of this plant (35%) was not significant. Similarly, Balanites aegyptiaca seeds were not effective in treatment of the infection in rats. Egg counts and deparasitization infood and water, were not significantly (p> 0.05) different from those of the untreated control group. On the other hand, Cucurbita maxima and Hagenia abyssinica seeds were very effective in the treatment of Hymenolepis diminuta infection in rats. Egg reduction (100%) was highly significant (p< 0.01) in food and water as compared to that of the untreated control group of rats (zero%). C. maxima seeds in food deparasitized 80% of the worms, while Hagenia abyssinica deparasitized 100%.Conclusion: Our conclusion was that Hagenia abyssinica was the most active plant of this group in the treatment of Hymenolepis diminuta infection in rats

Tamoxifen drug loading solid lipid nanoparticles prepared by hot high pressure homogenization techniques.

As drug delivery systems Nanoparticulate widely investigated because of many advantages such as smaller size, controlled drug release potential, targeting ability, enhancement of therapeutic efficacy and reduction of toxicity. So, Solid Lipid Nanoparticles have recently received considerable attention as alternative drug delivery carrier. In this study Solid Lipid Nanoparticles (SLNs) containing tamoxifen, nonsteroidal antiestrogens have been loaded and to be used as breast cancer therapy, were prepared by hot High Pressure Homogenization techniques. Tamoxifen loaded SLNs seem to have dimensional properties useful for parenteral administration. Preliminary study of antiproliferative activity in vitro, carried out on MCF-7 cell line (human breast cancer cells), demonstrated that SLNs, containing tamoxifen showed an antitumoral activity comparable to free drug. Tamoxifen loaded SLNs seem to have dimensional properties useful for parenteral administration. SLN was characterized by Differential Scanning Calorimetery (DSC), Transmission Electron Microscopy (TEM), Zeta Potential and Particle Size. The results of characterization studies strongly support the potential application of Tamoxifen-loaded SLNs as a carrier system. The SLN presented here are well suited for certain drug delivery applications, particularly breast cancer therapy

Potential chemoprevention of diethylnitrosamine-initiated and 2-acetylaminofluorene-promoted hepatocarcinogenesis by zerumbone from the rhizomes of the subtropical ginger (Zingiber zerumbet)

Zerumbone (ZER), a monosesquiterpene found in the subtropical ginger (Zingiber zerumbet Smith), possesses antiproliferative properties to several cancer cells lines, including the cervical, skin and colon cancers. In this study, the antitumourigenic effects of ZER were assessed in rats induced to develop liver cancer with a single intraperitoneal injection of diethylnitrosamine (DEN, 200 mg/kg) and dietary 2-acetylaminofluorene (AAF) (0.02%). The rats also received intraperitoneal ZER injections at 15, 30 or 60 mg/kg body wt. twice a week for 11 weeks, beginning week four post-DEN injection. The hepatocytes of positive control (DEN/AAF) rats were smaller with larger hyperchromatic nuclei than normal, showing cytoplasmic granulation and intracytoplasmic violaceous material, which were characteristics of hepatocarcinogenesis. Histopathological evaluations showed that ZER protects the rat liver from the carcinogenic effects of DEN and AAF. Serum alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (AP) and alpha-fetoprotein (AFP) were significantly lower (P < 0.05) in ZER-treated than untreated rats with liver cancer. The liver malondialdehyde (MDA) concentrations significantly (P < 0.05) increased in the untreated DEN/AAF rats indicating hepatic lipid peroxidation. There was also significant (P < 0.05) reduction in the hepatic tissue glutathione (GSH) concentrations. The liver sections of untreated DEN/AAF rats also showed abundant proliferating cell nuclear antigen (PCNA), while in ZER-treated rats the expression of this antigen was significantly (P < 0.05) lowered. By the TUNEL assay, there were significantly (P < 0.05) higher numbers of apoptotic cells in DEN/AAF rats treated with ZER than those untreated. Zerumbone treatment had also increased Bax and decreased Bcl-2 protein expression in the livers of DEN/AAF rats, which suggested increased apoptosis. Even after 11 weeks of ZER treatment, there was no evidence of abnormality in the liver of normal rats. This study suggests that ZER reduces oxidative stress, inhibits proliferation, induces mitochondria-regulated apoptosis, thus minimising DEN/AAF-induced carcinogenesis in rat liver. Therefore, ZER has great potential in the treatment of liver cancers

In Vitro Ultramorphological Assessment of Apoptosis Induced by Zerumbone on (HeLa)

Zerumbone (ZER), a potential anticancer compound, isolated from the fresh rhizomes of Zingiber zerumbet. In this investigation, the cytotoxic properties of ZER were evaluated, on cancer cells of human cervix (HeLa), breast and ovary, and normal cells of Chinese Hamster ovary, using MTT assay. Apoptogenic effects of ZER on HeLa were studied using fluorescence microscopy (AO/PI double staining), scanning and transmission electron microscopy (SEM and TEM), and colorimetric assay of the apoptosis promoter enzyme, caspase-3. The results of MTT assay showed that ZER has less effect on normal cells compared to cancer cells. The lowest IC50 of ZER was observed on HeLa cells. Cytological observations showed nuclear and chromatin condensation, cell shrinkage, multinucleation, abnormalities of mitochondrial cristae, membrane blebbing, holes, cytoplasmic extrusions and formation of apoptotic bodies as confirmed collectively by double staining of AO/PI, SEM and TEM. Statistical analysis (two-tailed t-test) of differential counting of 200 cells under fluorescence microscope revealed significant difference in apoptotic cells populations between treated and untreated HeLa cells. In addition, ZER has increased the cellular level of caspase-3 on the treated HeLa cells. It could be concluded that ZER was able to produce distinctive morphological features of cell death that corresponds to apoptosis