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The Sumatran Rhinoceros Dicerorhinus sumatrensis is on the edge of extinction. The decline of this species was initially attributed to poaching and habitat loss, but evidence presented here indicates that reproductive failure has also been a significant cause of loss, and continues to affect wild populations. Indonesia’s remaining populations of Sumatran Rhino are small and scattered, with limited access to breeding opportunities with unrelated mates. This leaves them subject to inbreeding and isolation-induced infertility, linked to fertility problems analyzed here. Sumatran Rhino females in captivity showed high rates (>70%) of reproductive pathology and/or problems with conception, which has significantly hindered the breeding program. Technological advances enabling examination immediately after capture revealed similarly high rates and types of reproductive problems in individuals from wild populations. The last seven Sumatran Rhino females captured were from areas with small declining populations, and six had reproductive problems. Going forward, capturing similarly compromised animals will take up valuable space and resources needed for fertile animals. The high risk of infertility and difficulty of treating underlying conditions, coupled with the decreasing number of remaining animals, means that the success of efforts to build a viable captive population will depend upon utilizing fertile animals and applying assisted reproductive techniques. Decades of exhaustive in situ surveys have not provided information relevant to population management or to ascertaining the fertility status of individual animals. Thus the first priority should be the capture of individuals as new founders from areas with the highest likelihood of containing fertile rhinos, indicated by recent camera trap photos of mothers with offspring. In Sumatra these areas include Way Kambas and parts of the Leuser ecosystem.

An adult female Sumatran rhinoceros was observed with a swelling in the left infraorbital region in March 2017. The swelling rapidly grew into a mass. A radiograph revealed a cystic radiolucent area in the left maxilla. In June 2017, the rhinoceros was euthanized. At necropsy, the infraorbital mass measured 21 cm × 30 cm. Samples of the infraorbital mass, left parotid gland, and left masseter muscle were collected for histopathology (Hematoxylin & Eosin, Von Kossa, Masson’s trichrome, cytokeratin AE1/AE3, EMA, p53, and S-100). Numerous neoplastic epithelial cells showing pleomorphism and infiltration were observed. Islands of dentinoid material containing ghost cells and keratin pearls were observed with the aid of the two special histochemistry stains. Mitotic figures were rarely observed. All the neoplastic odontogenic cells and keratin pearls showed an intense positive stain for cytokeratin AE1/AE3, while some keratin pearls showed mild positive stains for S-100. All samples were negative for p53 and S-100 immunodetection. The mass was diagnosed as a dentinogenic ghost cell tumor.

Farahnaz Barahuie,1 Mohd Zobir Hussein,1 Shafinaz Abd Gani,2,3 Sharida Fakurazi,3,4 Zulkarnain Zainal11Materials Synthesis and Characterization Laboratory, Institute of Advanced Technology (ITMA), Universiti Putra Malaysia, 2Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 3Laboratory of Vaccines and Immunotherapeutics, Institute of Bioscience, Universiti Putra Malaysia, 4Department of Human Anatomy, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, MalaysiaBackground: We characterize a novel nanocomposite that acts as an efficient anticancer agent. Methods: This nanocomposite consists of zinc layered hydroxide intercalated with protocatechuate (an anionic form of protocatechuic acid), that has been synthesized using a direct method with zinc oxide and protocatechuic acid as precursors.Results: The resulting protocatechuic acid nanocomposite (PAN) showed a basal spacing of 12.7 Å, indicating that protocatechuate was intercalated in a monolayer arrangement, with an angle of 54° from the Z-axis between the interlayers of the zinc layered hydroxide, and an estimated drug loading of about 35.7%. PAN exhibited the properties of a mesoporous type material, with greatly enhanced thermal stability of protocatechuate as compared to its free counterpart. The presence of protocatechuate in the interlayers of the zinc layered hydroxide was further supported by Fourier transform infrared spectroscopy. Protocatechuate was released from PAN in a slow and sustained manner. This mechanism of release was well represented by a pseudo-second order kinetics model. PAN has shown increased cytotoxicity compared to the free form of protocatechuic acid in all cancer cell lines tested. Tumor growth suppression was extensive, particularly in HepG2 and HT29 cell lines. Conclusion: PAN is suitable for use as a controlled release formulation, and our in vitro evidence indicates that PAN is an effective anticancer agent. PAN may have potential as a chemotherapeutic drug for human cancer.Keywords: protocatechuic acid, nanocomposite, zinc layered hydroxide, zinc oxide, 3T3, HepG2, HeLa, HT29

Mothanna Sadiq Al-Qubaisi,1 Abdullah Rasedee,1,2 Moayad Husein Flaifel,3 Sahrim Hj Ahmad,3 Samer Hussein-Al-Ali,1 Mohd Zobir Hussein,4 Zulkarnain Zainal,4 Fatah H Alhassan,4 Yun H Taufiq-Yap,4 Eltayeb EM Eid,5 Ismail Adam Arbab,1 Bandar A Al-Asbahi,3 Thomas J Webster,6,7 Mohamed Ezzat El Zowalaty1,8,9 1Institute of Bioscience, 2Faculty of Veterinary Medicine, Universiti Putra Malaysia, 3Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 4Faculty of Science, Universiti Putra Malaysia, Selangor, Malaysia; 5College of Pharmacy, Qassim University, Buraidah, Saudi Arabia; 6Department of Chemical Engineering and Program in Bioengineering, Northeastern University, Boston, MA, USA; 7Center of Excellence for Advanced Materials Research, King Abdulaziz University, Jeddah, Saudi Arabia; 8Faculty of Pharmacy, Zagazig University, Zagazig, Egypt; 9Faculty of Public Health and Tropical Medicine, Jazan University, Jazan, Saudi Arabia Abstract: The long-term objective of the present study was to determine the ability of NiZn ferrite nanoparticles to kill cancer cells. NiZn ferrite nanoparticle suspensions were found to have an average hydrodynamic diameter, polydispersity index, and zeta potential of 254.2 ± 29.8 nm, 0.524 ± 0.013, and -60 ± 14 mV, respectively. We showed that NiZn ferrite nanoparticles had selective toxicity towards MCF-7, HepG2, and HT29 cells, with a lesser effect on normal MCF 10A cells. The quantity of Bcl-2, Bax, p53, and cytochrome C in the cell lines mentioned above was determined by colorimetric methods in order to clarify the mechanism of action of NiZn ferrite nanoparticles in the killing of cancer cells. Our results indicate that NiZn ferrite nanoparticles promote apoptosis in cancer cells via caspase-3 and caspase-9, downregulation of Bcl-2, and upregulation of Bax and p53, with cytochrome C translocation. There was a concomitant collapse of the mitochondrial membrane potential in these cancer cells when treated with NiZn ferrite nanoparticles. This study shows that NiZn ferrite nanoparticles induce glutathione depletion in cancer cells, which results in increased production of reactive oxygen species and eventually, death of cancer cells. Keywords: NiZn ferrite nanoparticles, cancer cells, reactive oxygen species, cytochrome C, mitochondrial membrane potential, p53

Mothanna Sadiq Al-Qubaisi,1 Abdullah Rasedee,1,2 Moayad Husein Flaifel,5 Sahrim HJ Ahmad,5 Samer Hussein-Al-Ali,1 Mohd Zobir Hussein,3 Eltayeb EM Eid,6 Zulkarnain Zainal,3 Mohd Saeed,1 Muna Ilowefah,4 Sharida Fakurazi,1 Norhaszalina Mohd Isa,1 Mohamed Ezzat El Zowalaty1,7 1Institute of Bioscience, 2Faculty of Veterinary Medicine, 3Department of Chemistry, Faculty of Science, 4Faculty of Food Science and Technology, Universiti Putra Malaysia, Selangor, Malaysia; 5School of Applied Physics, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Selangor, Malaysia; 6College of Pharmacy, Qassim University, Buraidah, Saudi Arabia; 7Faculty of Pharmacy, Zagazig University, Ash Sharqiyah, Egypt Abstract: In this study, in vitro cytotoxicity of nickel zinc (NiZn) ferrite nanoparticles against human colon cancer HT29, breast cancer MCF7, and liver cancer HepG2 cells was examined. The morphology, homogeneity, and elemental composition of NiZn ferrite nanoparticles were investigated by scanning electron microscopy, transmission electron microscopy, and energy dispersive X-ray spectroscopy, respectively. The exposure of cancer cells to NiZn ferrite nanoparticles (15.6–1,000 µg/mL; 72 hours) has resulted in a dose-dependent inhibition of cell growth determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The quantification of caspase-3 and -9 activities and DNA fragmentation to assess the cell death pathway of the treated cells showed that both were stimulated when exposed to NiZn ferrite nanoparticles. Light microscopy examination of the cells exposed to NiZn ferrite nanoparticles demonstrated significant changes in cellular morphology. The HepG2 cells were most prone to apoptosis among the three cells lines examined, as the result of treatment with NiZn nanoparticles. In conclusion, NiZn ferrite nanoparticles are suggested to have potential cytotoxicity against cancer cells. Keywords: NiZn ferrite nanoparticles, cancer cells lines, anticancer, apoptosis

Farahnaz Barahuie,1 Mohd Zobir Hussein,1 Samer Hasan Hussein-Al-Ali,2 Palanisamy Arulselvan,3 Sharida Fakurazi,3,4 Zulkarnain Zainal11Materials Synthesis and Characterization Laboratory, Institute of Advanced Technology (ITMA), 2Laboratory of Molecular Biomedicine, Institute of Bioscience, 3Laboratory of Vaccines and Immunotherapeutics, Institute of Bioscience, 4Department of Human Anatomy, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, MalaysiaAbstract: In the study reported here, magnesium/aluminum (Mg/Al)-layered double hydroxide (LDH) was intercalated with an anticancer drug, protocatechuic acid, using ion-exchange and direct coprecipitation methods, with the resultant products labeled according to the method used to produce them: “PANE” (ie, protocatechuic acid-Mg/Al nanocomposite synthesized using the ion-exchange method) and “PAND” (ie, protocatechuic acid-Mg/Al nanocomposite synthesized using the direct method), respectively. Powder X-ray diffraction and Fourier transform infrared spectroscopy confirmed the intercalation of protocatechuic acid into the inter-galleries of Mg/Al-LDH. The protocatechuic acid between the interlayers of PANE and PAND was found to be a monolayer, with an angle from the z-axis of 8° for PANE and 15° for PAND. Thermogravimetric and differential thermogravimetric analysis results revealed that the thermal stability of protocatechuic acid was markedly enhanced upon intercalation. The loading of protocatechuic acid in PANE and PAND was estimated to be about 24.5% and 27.5% (w/w), respectively. The in vitro release study of protocatechuic acid from PANE and PAND in phosphate-buffered saline at pH 7.4, 5.3, and 4.8 revealed that the nanocomposites had a sustained release property. After 72 hours incubation of PANE and PAND with MCF-7 human breast cancer and HeLa human cervical cancer cell lines, it was found that the nanocomposites had suppressed the growth of these cancer cells, with a half maximal inhibitory concentration of 35.6 µg/mL for PANE and 36.0 µg/mL for PAND for MCF-7 cells, and 19.8 µg/mL for PANE and 30.3 µg/mL for PAND for HeLa cells. No half maximal inhibitory concentration for either nanocomposite was found for 3T3 cells.Keywords: Mg/Al-LDH, direct coprecipitation method, ion-exchange method, MCF-7, HeLa, 3T3