Hery Wijayanto

A whole-arm translocation (WAT8/9) separating Sumatran and Bornean agile gibbons, and its evolutionary features.

Gibbons, like orangutans, are a group of threatened Asian apes, so that genetic monitoring of each species or subspecies is a pressing need for conservation programmes. We conducted a project to take, as far as possible, samples of known origin from wild-born animals from Sumatra and Borneo (Central Kalimantan) for genetic monitoring of agile gibbons. As a result, we found a whole arm translocation between chromosomes 8 and 9 (WAT8/9) specific to Sumatran agile gibbons. Furthermore, population surveys suggested that the form with the WAT8/9 seems to be incompatible with an ancestral form, suggesting that the former might have extinguished the latter from Sumatran populations by competition. In any case, this translocation is a useful chromosomal marker for identifying Sumatran agile gibbons. Population genetic analyses with DNA showed that the molecular genetic distance between Sumatran and Bornean agile gibbons is the smallest, although the chromosomal difference is the largest. Thus, it is postulated that WAT8/9 occurred and fixed in a small population of Sumatra after migration and geographical isolation at the last glacial period, and afterwards dispersed rapidly to other populations in Sumatra as a result of the bottleneck effect and a chromosomal isolating mechanism.

Human umbilical mesenchymal stem cells conditioned medium promote primary wound healing regeneration.

Aim: This research was conducted to clarify the capability of human umbilical mesenchymal stem cells conditioned medium (HU-MSCM) to promote regenerations of primary wound healing on the incision skin injury. Materials and Methods: In this study, two approaches in vitro and in vivo already done. On in vitro analysis, tube formation was performed using HU vein endothelial cells in the presence of HU-MSCM, in some experiments cells line was incubated prior the presence of lipopolysaccharide and HU-MSCM then apoptosis assay was performed. Furthermore, in vivo experiments 12 female rats (Rattus norvegicus) were used after rats anesthetized, 7 mm wound was made by incision on the left side of the body. The wound was treated with HU-MSCM containing cream, povidone iodine was run as a control. Wound healing regenerations on the skin samples were visualized by hematoxylin-eosin staining. Results: In vitro models elucidate HU-MSCM may decreasing inflammation at the beginning of wound healing, promote cell migration and angiogenesis. In addition in vivo models show that the incision length on the skin is decreasing and more smaller, HE staining describe decreasing of inflammation phase, increasing of angiogenesis, accelerate fibroplasia, and maturation phase. Conclusions: Taken together our observation indicates that HU-MSCM could promote the acceleration of skin tissue regenerations in primary wound healing process.

Study of Genetic Marker of Cuscuses (Marsupialia: Phalangeridae) from Maluku and Papua Based on Cytochrome b Gene Sequences

Cuscuses is marsupials animal (Phalangeridae) which has limited spread in eastern Indonesia (Sulawesi, Maluku, Papua and Timor islands), Australia and Papua New Guinea. The ex-situ and in-situ conservation of cuscuses under captivating condition is an alternative solution to protect from extinction. This study aimed to determine nucleotide sequences and genetic marker on cyt b gene with sequencing method of each species on two provinces. Whole genome DNA was extracted from 22 samples of cuscuses obtained from different habitats, Maluku (13 individuals) and Papua (8 individuals) according to the protocol of Qiamp DNA Blood Mini Kit (Qiagen) and then it was used as template for amplification of cyt b gene by using PCR method. The PCR product were then purified using column chromatography and were used as template for sequencing reaction. Results sequencing of cyt b gene were analyzed using MEGA program versions 6.0. The PCR product gives results nucleotides of 982 bp according to database GeneBank and sequencing product gives results nucleotides of 771 bp. Nucleotides alignment of Phalanger members was found 24 nucleotides distinguishing and Spilocuscus members was found 11 nucleotides distinguishing, which can be used as genetic marker between Phalanger and Spilocuscus members from Papua and Maluku.

Genetic Differentiation of Agile Gibbons Between Sumatra and Kalimantan in Indonesia

The gibbons (Hylobatidae) are a diverse group of small apes that have adapted to the rain forests of South and Southeast Asia and radiated into numerous (12–14) discrete species. Recently, the four subgenera of small apes [Hoolock, (previously Bunopithecus, Mootnick and Groves 2005), Hylobates, Symphalangus, and Nomascus] were all raised to the level of genera because the genetic distances between them indicated by mitochondrial DNA were larger than those between Homo andPan (Hayashi et al. 1995; Roos andGeissmann 2001). Some aspects of gibbon classification are still controversial, in particular the differentiation of subspecies (Groves 2001; Brandon-Jones et al. 2004; Mootnick 2006; Chatterjee this volume). Accurate determination of collection localities is critical for diagnosing subspecies in this group, which is morphologically very diverse in some physical features (e.g., pelage pattern, Marshall and Sugardjito 1986; Mootnick 2006). Thus it is important to use animals of known origin for genetic studies, which are crucial for planning the conservation of evolutionarily significant units (ESUs) (Crandall et al. 2000; Frankham et al. 2002). The taxonomy of agile gibbons is disputed (Chatterjee this volume), with some researchers recognizing a single species (H. agilis), and others recognizing one species (H. agilis) on the Asian mainland and Sumatra and a second species (H. albibarbis, or the whitebearded gibbon) on Borneo. In this chapter, we refer to all gibbons in this group as agile gibbons, but identify the Bornean taxon asH. albibarbis.We have aimed to collect samples from gibbons of known origin, and consequently have been able to demonstrate cytogenetic and molecular genetic differentiation of agile gibbon taxa between Sumatra and Kalimantan. These results provide important information on their biogeography and ESUs and a basis for future comprehensive evolutionary genetic investigations of small apes.We summarize the essential points of the data obtained so far, and give our point of view in this chapter.

KAJIAN DIVERSITI GENETIKA Tarsius sp. ASAL INDONESIA MENURUT URUTAN GEN NADH DEHIDROGENASE SUBUNIT 4 (ND4)

The aim of the study was to assess the genetic diversity ND4 coding genes in Tarsius bancanus, T. b. borneanus, T. dianae, and T. spectrum, and to reconfirm the taxonomy of Tarsius sp. Deoxyribonucleotides (DNA) specimens were isolated from tissue biopsy of each species of Tarsius using DNA extraction method then used for DNA template in the amplification process by means of polymerase chain reaction (PCR). Primers used in this study were designed specifically to amplify ND4 gene. The PCR result was then run for electrophoresis. PCR amplification results were then purified and used as template DNA for the reaction determination sequences of nucleotide. Sequences of ND4 nucleotide gene were blasted with other primates’ genes from Genbank using Clustal W. Furthermore, ND4 gene was analyzed based on amino acids sequence that were translated using Vertebrate mitochondrial translation code MEGA program software version 4.1 and phylogenetic tree were create using Neighbor Joining method. The results showed that 119 nucleotides sites out of 1378 were diverse. Genetic distance based on ND4 gene nucleotide that was calculated using Kimura two-parameter model shown the smallest value was 0.6%, while the largest value was 13%, and the average value is 6.1%. The phylogram based on the result of nucleotide sequence of ND4 genes using the Neighbor Joining method is useful for Tarsius sp. identification and distinguishing between species of Tarsius branch.