Development of a DNA marker by minisatellite associated sequence amplification (MASA) from the endangered Indian rhino (Rhinoceros unicornis)

Rhinoceroses are highly endangered species and their protection warrants immediate remedial measures. Development of DNA markers is envisaged to complement global efforts of the conservation of these extant animals. Minisatellite associated sequence amplification (MASA) of DNA from Indian rhinoceros (Rhinoceros unicornis) and three sub-species of South African black rhinoceros (Diceros bicornis) was carried out using a primer based on consensus sequence of the minisatellite repeat locus 33.15. Several bands in the range of 3.0 kilobases (kb) to 650 base pairs (bp) were identified that were useful for successful differentiation of R. unicornis from D. bicornis. Of these fragments, a 688 bp one, unique to R. unicornis was cloned and sequenced (Accession No. AF-296689). The band patterns uncovered by MASA and the species-specific hybridisation of pSG5 may be utilised as a tool for differentiating the R. unicornis genome from that of D. bicornis. This approach may also be adopted for the development of DNA-based genetic marker(s) useful for identification of other endangered species

CXCR4 is a response gene for parathyroid hormone which affects osteoblast and osteoclast function in vitro

AimsTo investigate the role of CXCR4 in response to teriparatide (TPTD) treatment in osteoblasts and osteoclasts.MethodsPrimary murine and human osteoblasts and osteoclasts, MC3T3 cell lines, and hMSC-TERT4 cell lines were treated with TPTD and/or AMD3100, a pharmacological inhibitor of CXCR4. Changes in gene expression, osteoblast viability, mobility, mineralization capacity, and alkaline phosphatase activity were investigated. Osteoclastogenesis and cell fusion were also assessed in response to both treatments.ResultsTPTD increased messenger RNA levels of CXCR4 in all stages of both murine and human osteoblast differentiation. Mineralization analysis showed that CXCR4 was involved in bone matrix formation in response to TPTD. Alkaline phosphatase activity was also impaired by CXCR4 inhibition at early stages of osteoblast differentiation, while it was promoted at late stages, suggesting that CXCR4 could produce a delay in osteoblast maturation. Moreover, we also found a direct activation of osteoclastogenesis after TPTD treatment in murine and human osteoclasts. This process seems to involve CXCR4 activity, since AMD3100-induced CXCR4 inhibition led to a reduction in both murine and human osteoclastogenesis. This process, however, could not be prevented by TPTD treatment.ConclusionOur results suggest that CXCR4 is a responsive gene to TPTD treatment, involved in the regulation of osteoblast and osteoclast generation and function. Further in vivo studies are required to confirm this role, and to determine whether pharmacological strategies targeting CXCR4 could potentially improve the treatment outcome for osteoporotic patients

Ubiquitin Fold Modifier 1 (UFM1) and Its Target UFBP1 Protect Pancreatic Beta Cells from ER Stress-Induced Apoptosis

UFM1 is a member of the ubiquitin like protein family. While the enzymatic cascade of UFM1 conjugation has been elucidated in recent years, the biological function remains largely unknown. In this report we demonstrate that the recently identified C20orf116 [1], which we name UFM1-binding protein 1 containing a PCI domain (UFBP1), andCDK5RAP3 interact with UFM1. Components of the UFM1 conjugation pathway (UFM1, UFBP1, UFL1 and CDK5RAP3) are highly expressed in pancreatic islets of Langerhans and some other secretory tissues. Co-localization of UFM1 with UFBP1 in the endoplasmic reticulum (ER)depends on UFBP1. We demonstrate that ER stress, which is common in secretory cells, induces expression of Ufm1, Ufbp1 and Ufl1 in the beta-cell line INS-1E.siRNA-mediated Ufm1 or Ufbp1knockdown enhances apoptosis upon ER stress.Silencing the E3 enzyme UFL1, results in similar outcomes, suggesting that UFM1-UFBP1 conjugation is required to prevent ER stress-induced apoptosis. Together, our data suggest that UFM1-UFBP1participate in preventing ER stress-induced apoptosis in protein secretory cells

JunD/AP-1-Mediated Gene Expression Promotes Lymphocyte Growth Dependent on Interleukin-7 Signal Transduction

Interleukin-7 (IL-7) is an essential cytokine for lymphocyte growth that has the potential for promoting immune reconstitution. This feature makes IL-7 an ideal candidate for therapeutic development. As with other cytokines, signaling through the IL-7 receptor induces the JAK/STAT pathway. However, the broad scope of IL-7 regulatory targets likely necessitates the use of other signaling components whose identities remain poorly defined. To this end, we used an IL-7 dependent T-cell line to examine how expression of the glycolytic enzyme, Hexokinase II (HXKII) was regulated by IL-7 in a STAT5-independent manner. Our studies revealed that IL-7 promoted the activity of JNK (Jun N-terminal Kinase), and that JNK, in turn, drove the expression of JunD, a component of the Activating Protein 1 (AP-1) transcription factors. Gel shifts showed that the AP-1 complex induced by IL-7 contained JunD but not c-Fos or c-Jun. Inhibition of JNK/JunD blocked glucose uptake and HXKII gene expression, indicating that this pathway was responsible for promoting HXKII expression. Because others had shown that JunD was a negative regulator of cell growth, we performed a bioinformatics analysis to uncover possible JunD-regulated gene targets. Our search revealed that JunD could control the expression of proteins involved in signal transduction, cell survival and metabolism. One of these growth promoters was the oncogene, Pim-1. Pim-1 is an IL-7-induced protein that was inhibited when the activities of JNK or JunD were blocked, showing that in IL-7 dependent T-cells JunD can promote positive signals transduced through Pim-1. This was confirmed when the IL-7-induced proliferation of CD8 T-cells was impaired upon JunD inhibition. These results show that engagement of the IL-7 receptor drives a signal that is more complex than the JAK/STAT pathway, activating JNK and JunD to induce rapid growth stimulation through the expression of metabolic and signaling factors like HXKII and Pim-1

バレイショ近縁種における種の分化 XIII. S.acaule X S.demissumより得た7倍雑種の染色体行動と両親ゲノムの類縁関係

中央アンデス産Acaulia群4倍種S. acaule (acl, 2n=48)とメキシコ産Demissa群6倍種S. demissum (dms, 2n=72)のゲノムの類縁関係を明らかにするために, 前者を母本として得た7倍雑種(2n=84)の還元分裂における染色体行動と稔性を調べた。以下その結果を要約する。両種間の交雑は極めて困難で, aclを母とした時のみ受粉花数当り0.02の低率で雑種が得られたにすぎない。得られた雑種は, 両親との形態的比較から, aclの非還元性卵とdmsの還元性花粉の受精に起因するものと推定された。この雑種の第1中期における染色体対合行動は甚だしく多様であったが, その対合型のモードは(12)_+(20)_+8_I, その平均対合頻度は(0.18)_V+(1.11)_+(11.73)_+(18.11)_+(7.26)_Iで, 著しく高頻度の3価形成を示す点が特徴的であった。このような対合行動はその後の染色体行動にも反映し, 第1後期では観察細胞のすべてに平均4.8の遅滞染色体がみられ, 第2中期では94%の細胞が分散染色体を示し, 数的平衡核板頻度は0.6%にすぎなかった。稔性は極めて低く, 調査花粉粒数の27%が一見正常であったが, 自殖及び戻交配のいずれにおいても全く種子を生じなかった。上記の観察結果, 特に高頻度で出現した3価染色体の成因を考察して次の知見を得た。すでにaclはAAA^aA^a, dmsはA^dA^dC_1C_1C_2C_2のゲノム型をもつことが知られているので, 当雑種のゲノム型はAAA^aA^aA^dC_1C_2となる。A^dゲノムは若干の構造的差異はもつもののAゲノム群に属することも知られている。したがって, 当雑種にみられる3価形成は, 主に, aclからのAAとdmsからのA^dの3ゲノム間の染色体対合に由来すると推論でき, 両種はこれらのゲノムの相同性によって相互に関係づけられているものと考えられる。 / Meiotic behavior and fertility were studied in a heptaploid F_1 hybrid (2n=84) obtained from crossing S. acaule (acl, 2n=48) with S. demissum (dms, 2n=72), with the aim of assessing a genomic relationship between the parent species. Crossability between the two species was very low, the number of hybrid plants per pollination being only 0.02. Morphological evidence indicated that the hybrid arose through the union of an unreduced egg of acl and a reduced pollen grain of dms. The hybrid had the mean pairing frequency of (0.18)_V+(1.11)_+(11.73)_+(18.11)_+(7.26)_I per cell at metaphase I, with (12)_+(20)_+8_I as the modal configuration. Its subsequent behaviors were extremely irregular, showing several laggards in all the cells and chromatid bridges in occasional cells at anaphase I and also scattered chromosomes in 94% of the cells at metaphase II. The hybrid gave only 27% stainable pollen and no seed either on selfing or on backcrossing with both parents. The pattern of chromosome pairing found in the hybrid was interpreted in terms of genomic relationship between both parent species. From this, it was suggested that one (A) of the two genomes (designated AA^a) which acl possess in its gametes seems to be closely similar to, but not identical with, one (A^d) of the three genomes (A^dC_1C_2) which dms possess in its gemetes

Reconceptualizing Policing for Cybercrime: Perspectives from Singapore

As cybercrime proliferates globally, law enforcement agencies face significant challenges in responding effectively. This essay shares perspectives from Singapore, where cybercrime accounted for about 70% of the total annual crime in 2023, with no clear data on case resolution rates. This situation reflects a broader global trend and highlights the need to reconceptualize policing objectives in cyberspace. The fundamental differences between cybercrime and physical crime necessitate a shift from emphasizing the identification and prosecution of perpetrators to adopting a harm-centric perspective. Under this perspective, structures and policies should be implemented to disrupt financial flows, ensure data security, disrupt the spread of harmful content, and prevent physical damage. Once this is done, strategies such as public–private partnerships, international cooperation, and training and building capabilities to address specific harms can be more effectively implemented to mitigate the growing threat that cybercrime poses worldwide