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Abstract Background Cotton is an important economic crop and a host of Liriomyza sativae. Pectin methylesterase (PME)-mediated pectin metabolism plays an indispensable role in multiple biological processes in planta. However, the pleiotropic functions of PME often lead to unpredictable effects on crop resistance to pests. Additionally, whether and how PME affects susceptibility to Liriomyza sativae remain unclear. Results Here, we isolated GhPME36, which is located in the cell wall, from upland cotton (Gossypium hirsutum L.). Interestingly, the overexpression of GhPME36 in cotton caused severe susceptibility to Liriomyza sativae but increased leaf biomass in Arabidopsis. Cytological observations revealed that the cell wall was thinner with more demethylesterified pectins in GhPME36-OE cotton leaves than in WT leaves, whereas the soluble sugar content of GhPME36-OE cotton leaf cell walls was accordingly higher; both factors attracted Liriomyza sativae to feed on GhPME36-OE cotton leaves. Metabolomic analysis demonstrated that glucose was significantly differentially accumulated. Transcriptomic analysis further revealed DEGs enriched in glucose metabolic pathways when GhPME36 was overexpressed, suggesting that GhPME36 aggravates susceptibility to Liriomyza sativae by affecting both the structure and components of cell wall biosynthesis. Moreover, GhPME36 interacts with another pectin-modifying enzyme, GhC/VIF1, to maintain the dynamic stability of pectin methyl esterification. Conclusions Taken together, our results reveal the cytological and molecular mechanisms by which GhPME36 aggravates susceptibility to Liriomyza sativae. This study broadens the knowledge of PME function and provides new insights into plant resistance to pests and the safety of genetically modified plants.

Lead is a toxic metal known to be harmful once within the body. Chronic lead accumulation causes the emergence of free radicals, thereby causing damage to organ systems, one of which is the reproductive system. Chitosan can chelate lead metal ions which behave as free radicals. In nanostructures, chitosan's ability to chelate heavy metal ions is expected to be more effective. The use of nano-chitosan is expected to facilitate its distribution so that its ability to chelate lead ions is more effective. This study aims to analyze the effect of nano-chitosan on the histology of seminiferous tubules and spermatogenesis of rats induced by lead acetate (PbAc). Animals are divided into 3 groups; Naive Group (NG: aquadest), Positive Control (PC: PbAc 200 mg kg-1 BW), Treatment Group (TG: PbAc 200 mg kg-1 BW and nano-chitosan 64 mg kg-1 BW) with a treatment duration of 35 days, and 6 rats in each group. Rats were exposed to lead by oral administration. At the end of the treatment period, animals were euthanized by dislocation of the cervical spine after being lightly anesthetized using 10% chloroform. Then, testicular organs were dissected, and histological preparations of the seminiferous tubules were analyzed. Measurements include epithelial thickness, tubule diameter and spermatogenesis scoring based on the Johnson criteria. The results obtained proved that administration of nano-chitosan after PbAc induction increased epithelial thickness, seminiferous tubule diameter and spermatogenesis score. In conclusion, nano-chitosan can overcome lead toxicity in the rat reproductive system induced by PbAc with indicators of improvement in the histology structure of seminiferous tubules and increased spermatogenesis scores.

The pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae) is a serious insect pest of cotton crop. The studies to evaluate the impact of abiotic factors on cotton pests’ biology are limited. The current study was undertaken to determine the impact of abiotic factors (temperature, humidity, photoperiod) and an insecticide (lambda-cyhalothrin) on the biological aspects of P. gossypiella. The results revealed that all the treatments showed a significant impact on different life parameters of P. gossypiella. The temperature exposure at 27 °C revealed a total life span of about 33 days. Maximum mortality for larvae (51.11%), pupae (59.04%) and adults (61.92%) were recorded at 33 °C exposure. Both low and high humidity levels caused negative impacts on life parameters of P. gossypiella. The life span was completed in about 30 days at 60% relative humidity (RH). Maximum mortality for larvae (75.00%) and pupae (49.59%) were recorded at 80% RH level exposure, while adult mortality was maximum (63.09%) at 40% RH level followed by 80% RH level (55.52%). The P. gossypiella exhibited a life span of about 32 days at 14:10 light-dark period. The larval mortality was maximum (14.83%) at 12:12 light-dark period while pupal (47.36%) and adult (48.84%) mortality was maximum at 16:08 light-dark period. Lambdacyhalothrin (LC) showed dose dependent negative impacts on biological aspects of P. gossypiella. The P. gossypiella exhibited a life span of about 26 days at 0.5 ppm LC concentration. The P. gossypiella exposure to highest concentration (LC) revealed maximum mortality of larval (80.22%), pupal (64.63%) and adult (70.74%) stages. Conclusively, the best suited abiotic factor ranges were 27 °C (temperature), 60% (RH) and 14:10 (light-dark) which can be used for successful rearing and bioassay activities of P. gossypiella.

Abstract Objectives In recent years, there has been much discussion and research on electronic nose (e-nose). This topic has developed mainly in the medical and food fields. Typically, e-nose is combined with machine learning algorithms to predict or detect multiple sensory classes in each tea sample. Therefore, in e-nose systems, e-nose signal processing is an important part. In many situations, a comprehensive set of experiments is required to ensure the prediction model can be generalized well. This data set specifically focuses on two main goals such as classification of green tea quality and prediction of organoleptic score. In this experiment, Gambung dry green tea samples were used. The challenge is that dry tea does not emit as strong an aroma as tea infusions, making it more difficult for the e-nose system to detect and identify the aromas. This data set offers a valuable resource for researchers and developers to conduct investigations and experiments by classifying and detecting organoleptic scores that aim to categorize and identify organoleptic ratings. This enables a deeper understanding of the quality of dry green tea and encourages further integration of e-nose technology in the tea industry. Data description This experiment focused on analyzing green tea aroma using six gas sensors. Seventy-eight green tea samples were tested, each observed three times, using a tea chamber connected to a sensor chamber via a hose and an intake micro air pump. Air flowed from the tea chamber to the sensor chamber for 60 s, followed by 60 s of aroma data recording. This data was saved into CSV files and labeled according to the Indonesian National Standard (SNI) 3945:2016, which includes special and general requirements for green tea quality. An organoleptic test by a tea tester further labeled the data set into “good” or “quality defect” for classification and provided organoleptic scores based on dry appearance, brew color, taste, aroma, and dregs of brewing for continuous label.

Dengue virus (DENV), a common and prevalent mosquito-borne endemic disease, is caused by four serotypes (DENV-1–4) and has spread rapidly on a global scale over the past decade. A crucial step in the development of antiviral therapeutics requires the utilization of in vitro cell-based techniques, such as plaque assays and focus-forming assays (FFA) for virus quantification. Vero cells have been widely used for FFA and plaque assay; however, there are instances when their efficacy and efficiency in the detection of certain clinical DENV isolates are low. Here, we showed that BHK-21 cells are more sensitive than Vero cells in the detection of all DENV-1–4 plaques and foci. In addition, we developed an improved FFA protocol for the quantification of all four DENV serotypes. Using a pan-flavivirus envelope (E) antibody, we reduce the possibility of false positives by defining a focus to consist of a minimum of eight infected cells. We outlined a protocol using the Operetta® high-content imaging system to automate the digital capture of these infected cells. A pipeline was also designed using the CellProfilerTM automated image analysis software to detect these foci. We then compare the results of the improved FFA with plaque assay. Notably, the improved FFA detected clear foci of the DENV-4 strain that does not form distinct plaques. We subsequently demonstrated the potential application of the improved FFA protocol in antiviral testing, utilizing a nucleoside inhibitor of DENV, NITD008 as a control. The protocol is amenable to a diverse array of applications, including high-throughput compound screening (HTS).

Background Understanding human stem cell differentiation into osteoblasts and osteoclasts is crucial for bone regeneration and disease modeling. Numerous morphological techniques have been employed to assess this differentiation, but a comprehensive review of their application and effectiveness is lacking. Methods Guided by the PRISMA framework, we conducted a rigorous search through the PubMed, Web of Science and Scopus databases, analyzing 254 articles. Each article was scrutinized against pre-defined inclusion criteria, yielding a refined selection of 14 studies worthy of in-depth analysis. Results The trends in using morphological approaches were identified for analyzing osteoblast and osteoclast differentiation. The three most used techniques for osteoblasts were Alizarin Red S (mineralization; six articles), von Kossa (mineralization; three articles) and alkaline phosphatase (ALP; two articles) followed by one article on Giemsa staining (cell morphology) and finally immunochemistry (three articles involved Vinculin, F-actin and Col1 biomarkers). For osteoclasts, tartrate-resistant acid phosphatase (TRAP staining) has the highest number of articles (six articles), followed by two articles on DAPI staining (cell morphology), and immunochemistry (two articles with VNR, Cathepsin K and TROP2. The study involved four stem cell types: peripheral blood monocyte, mesenchymal, dental pulp, and periodontal ligament. Conclusion This review offers a valuable resource for researchers, with Alizarin Red S and TRAP staining being the most utilized morphological procedures for osteoblasts and osteoclasts, respectively. This understanding provides a foundation for future research in this rapidly changing field.

Osteoarthritis (OA) is a chronic disease affecting joints and further causing disabilities. This disease affects around 240 million people worldwide. It is a multifactorial disease, and its etiology is difficult to determine. Although numerous therapeutic strategies are available, the therapies are aimed at reducing pain and improving patients’ quality of life. Hence, there is an urgent need to develop disease-modifying drugs (DMOAD) that can reverse or halt OA progression. Apoptosis is a cell removal process that is important in maintaining homeostatic mechanisms in the development and sustaining cell population. The apoptosis of chondrocytes is believed to play an important role in OA progression due to poor chondrocytes self-repair abilities to maintain the extracellular matrix (ECM). Hence, targeting chondrocyte apoptosis can be one of the potential therapeutic strategies in OA management. There are various mediators and targets available to inhibit apoptosis such as autophagy, endoplasmic reticulum (ER) stress, oxidative stress, and inflammation. As such, this review highlights the importance and potential targets that can be aimed to reduce chondrocyte apoptosis.

Abstract Drought stress is a significant environmental factor that adversely affects the growth and development of carrot (Daucus carota L.), resulting in reduced crop yields and quality. Drought stress induces a range of physiological and biochemical changes in carrots, including reduced germination, hindered cell elongation, wilting, and disrupted photosynthetic efficiency, ultimately leading to stunted growth and decreased root development. Recent research has focused on understanding the molecular mechanisms underlying carrot's response to drought stress, identifying key genes and transcription factors involved in drought tolerance. Transcriptomic and proteomic analyses have provided insights into the regulatory networks and signaling pathways involved in drought stress adaptation. Among biochemical processes, water scarcity alters carrot antioxidant levels, osmolytes, and hormones. This review provides an overview of the effects of drought stress on carrots and highlights recent advances in drought stress-related studies on this crop. Some recent advances in understanding the effects of drought stress on carrots and developing strategies for drought stress mitigation are crucial for ensuring sustainable carrot production in the face of changing climate conditions. However, understanding the mechanisms underlying the plant's response to drought stress is essential for developing strategies to improve its tolerance to water scarcity and ensure food security in regions affected by drought.

The chair-mounted passive trunk orthosis (CMPTO) is designed to enhance wheelchair safety for individuals with dystrophinopathy during their daily activities. Given the disease’s progressive nature, it is crucial to ensure that assistive devices are carefully evaluated to prevent overexertion. This study aims to assess the CMPTO’s user experience and its impact on fatigue-related psychophysiological measurements. We conducted electromyography (EMG) evaluations of four trunk muscles and assessed perceived exertion using the Borg CR-10 scale in 40 healthy subjects while they performed seated maximal reaching tasks with the CMPTO. Additionally, fifteen dystrophinopathy patients evaluated the CMPTO for usability with the System Usability Scale. Paired t-tests were employed to compare the median frequency (MDF) of EMG signals, the Wilcoxon signed-rank test for evaluating exertion, and the Mann–Whitney U test to compare the usability reported by patients to those of healthy subjects. The 4-way ANOVA revealed that MDF patterns were significantly influenced by task orientation for each muscle. The CMPTO did not cause a significant reduction in the MDF. Tasks requiring greater trunk rotation were perceived as more exhaustive. Patients reported acceptable usability with the CMPTO, with scores higher than those of healthy subjects. The CMPTO’s usability was comprehensively evaluated in both healthy subjects and patients with dystrophinopathy. Our findings indicate that the CMPTO can be safely used by individuals with dystrophinopathy as an assistive device to improve seated comfort and functional abilities.

Traumatic damage to the spinal cord (SCI) frequently leads to irreversible neurological deficits, which may be related to apoptotic neurodegeneration in nerve tissue. The MLC901 treatment possesses neuroprotective and neuroregenerative activity. This study aimed to explore the regenerative potential of MLC901 and the molecular mechanisms promoting neurogenesis and functional recovery after SCI in rats. A calibrated forceps compression injury for 15 s was used to induce SCI in rats, followed by an examination of the impacts of MLC901 on functional recovery. The Basso, Beattie, and Bresnahan (BBB) scores were utilized to assess neuronal functional recovery; H&E and immunohistochemistry (IHC) staining were also used to observe pathological changes in the lesion area. Somatosensory Evoked Potentials (SEPs) were measured using the Nicolet® Viking Quest™ apparatus. Additionally, we employed the Western blot assay to identify PI3K/AKT/GSK-3β pathway-related proteins and to assess the levels of GAP-43 and GFAP through immunohistochemistry staining. The study findings revealed that MLC901 improved hind-limb motor function recovery, alleviating the pathological damage induced by SCI. Moreover, MLC901 significantly enhanced locomotor activity, SEPs waveform, latency, amplitude, and nerve conduction velocity. The treatment also promoted GAP-43 expression and reduced reactive astrocytes (GFAP). MLC901 treatment activated p-AKT reduced p-GSK-3β expression levels and showed a normalized ratio (fold changes) relative to β-tubulin. Specifically, p-AKT exhibited a 4-fold increase, while p-GSK-3β showed a 2-fold decrease in T rats compared to UT rats. In conclusion, these results suggest that the treatment mitigates pathological tissue damage and effectively improves neural functional recovery following SCI, primarily by alleviating apoptosis and promoting neurogenesis. The underlying molecular mechanism of this treatment mainly involves the activation of the PI3K/AKT/GSK-3β pathway.

This study is an ex post facto research that aims to discover the influence of knowledge on environmental changes, receiving information, on environmental awareness and the influence of environmental changes, receiving information on environmental awareness by involving naturalistic intelligence. The study population was all students of grade XI at SMAN in East Banggae subdistrict in Majene district, namely SMAN 1 Majene, SMAN 2 Majene, and SMAN 3 Majene. The research instrument employed a test of environmental change knowledge, a questionnaire for information reception, naturalistic intelligence and environmental awareness. The results showed that, 1) environmental changes are in the very low category, information reception is in the medium category, naturalistic intelligence is in the high category.

تعرضت ترب الغابات والنبت الطبيعي في الساحل السوري إلى انتهاكات واضحة من خلال القطع الجائر والحرائق المفتعلة وغير المفتعلة، انعكس ذلك بشكل سلبي على خواص تلك الترب التي تعرضت للاستنزاف وعلى الأخص الحرائق التي حدثت في العام 2020 التي حوّلت الجبال الساحلية إلى رماد. اعتماداً على تحميل صورتين فضائيتين Landsat8 المركبتين بعد دمج نطاقاتهما وتعديلهما لتحديد مكان الحريق دون ضباب أو دخان، الأولى قبل الحريق في شهر تموز عام 2020 والثانية بعد الحريق في شهر تشرين أول 2020، صُممت خرائط شدة الحرائق لمنطقة الباير والبسيط لتقييم المناطق التي تأثرت بسبب حرائق الغابات. أُنتجت خارطة مؤشر شدة الحريق وتصنيفها اعتماداً على تصنيف مستويات شدة الحريق وبالتالي المساحات المحترقة التي شكّلت مستوى خطورة عالية بلغت 1499.72 هكتار من منطقة الباير والبسيط في حين بلغت المساحات المحترقة التي شكّلت مستوى خطورة متوسطة 10820.73 هكتار. أُخذت ثلاث عينات تربة من الغابات المحروقة في زغرين وبللوران ورأس البسيط وقُورنت مع ثلاث عينات تربة من نفس المواقع غير المحروقة. تربة المواقع غنية بالمادة العضوية قبل الحريق بينما انخفضت تقريباً الى النصف بعد الحريق، أما قيمة الـPH نجد أنها بالمواقع المحروقة كانت أعلى من المواقع غير المحروقة بسبب الرماد الناتج عن حرق الغطاء النباتي فوق أرض الغابة، وإن نسبة الآزوت في المواقع المحروقة لا تتجاوز 0.3 %، كما أن كمية الفوسفور بالمواقع المحروقة كانت أكبر منها بالمواقع غير المحروقة، وتتزايد كمية البوتاسيوم والكالسيوم والمنغنيز والحديد والزنك والنحاس المتاحة بعد الحريق، وعلى الرغم من كل هذه الزيادات في الكاتيونات فإن الحرائق قد تؤدي لفقدانها أيضاً وذلك عن طريق انجراف التربة والسيلان السطحي وانغسال العناصر أو عن طريق تطاير الرماد.

Highlights • Patients with breast cancer who have distant metastases are frequently impacted by pleural effusion. • HER2 status was found to be substantially correlated with glucose levels in pleural effusion fluid. Abstract Background: Patients diagnosed with breast cancer who have also been affected by distant metastases often suffer from pleural effusion. The prognosis of malignant pleural effusions worsens when glucose levels in the pleural fluid are low. Elevated levels of anaerobic glycolysis due to HER2 overexpression cause breast cancer cells to take in more glucose. Objective: This study aimed to identify any correlation between glucose levels in pleural fluid and HER2 status in breast cancer patients. Material and Method: In this research, 35 female patients with thoracocentesis pleural metastatic breast cancer participated in an analytical observational study using a cross-sectional design. Patients who had undergone thoracocentesis had their HER2 status determined based on the information included in their medical records in the Integrated Oncology Clinic (Poli Onkologi Satu Atap (POSA)) Dr. Soetomo General Academic Hospital, Surabaya, Indonesia. After obtaining data on glucose levels in pleural fluid and HER2 status, statistical analysis was carried out to determine the relationship between the two parameters. Results: The statistical test results with a 95% significance level obtained p=0.004 (p

Piper sarmentosum or ‘kaduk’ is a well-known herb plant in Malaysia. Its extracts were found to exhibit a bone-protective effect against osteoporotic rats. Our study aim is to morphologically observe the effect of P. sarmentosum ethyl acetate extract on the differentiation of human peripheral blood stem cells (PBSCs) into osteoblasts. P. sarmentosum extracts (1-900 µg/mL) prepared using 1% dimethyl sulfoxide (DMSO) were used in the cytotoxicity assay. Then, the differentiation assay was performed using concentrations of 1-50 µg/mL. The untreated cells acted as the negative control, while the cells cultured in 50 µg/mL ascorbic acid and 10 mM β-glycerophosphate was a positive control. The cytotoxicity effect and proliferation capacity of the cells were analyzed using Trypan Blue exclusion method, while the differentiation of PBSCs was observed using von Kossa staining and ALP gene expression analysis. The result showed a decrease in cells’ viability in a dose-dependent manner during cytotoxicity assay. After 14 days of the differentiation, a constant rate of proliferation could be observed in the treated cells and positive control, while the untreated cells showed an increase in proliferation. The mineralization of extract-treated cells showed significant differences (p

Pain is a multifaceted, multisystem disorder that adversely affects neuro-psychological processes. This study compares the effectiveness of central stimulation (transcranial direct current stimulation—tDCS over F3/F4) and peripheral stimulation (transcutaneous electrical nerve stimulation—TENS over the median nerve) in pain inhibition during a cognitive task in healthy volunteers and to observe potential neuro-cognitive improvements. Eighty healthy participants underwent a comprehensive experimental protocol, including cognitive assessments, the Cold Pressor Test (CPT) for pain induction, and tDCS/TENS administration. EEG recordings were conducted pre- and post-intervention across all conditions. The protocol for this study was categorized into four groups: G1 (control), G2 (TENS), G3 (anodal-tDCS), and G4 (cathodal-tDCS). Paired t-tests (p < 0.05) were conducted to compare Pre-Stage, Post-Stage, and neuromodulation conditions, with t-values providing insights into effect magnitudes. The result showed a reduction in pain intensity with TENS (p = 0.002, t-value = −5.34) and cathodal-tDCS (p = 0.023, t-value = −5.08) and increased pain tolerance with TENS (p = 0.009, t-value = 4.98) and cathodal-tDCS (p = 0.001, t-value = 5.78). Anodal-tDCS (p = 0.041, t-value = 4.86) improved cognitive performance. The EEG analysis revealed distinct neural oscillatory patterns across the groups. Specifically, G2 and G4 showed delta-power reductions, while G3 observed an increase. Moreover, G2 exhibited increased theta-power in the occipital region during CPT and Post-Stages. In the alpha-band, G2, G3, and G4 had reductions Post-Stage, while G1 and G3 increased. Additionally, beta-power increased in the frontal region for G2 and G3, contrasting with a reduction in G4. Furthermore, gamma-power globally increased during CPT1, with G1, G2, and G3 showing reductions Post-Stage, while G4 displayed a global decrease. The findings confirm the efficacy of TENS and tDCS as possible non-drug therapeutic alternatives for cognition with alleviation from pain.

Abstract Background Excitotoxicity-induced in vivo injury models are vital to reflect the pathophysiological features of acute spinal cord injury (SCI) in humans. The duration and concentration of chemical treatment controls the extent of neuronal cell damage. The extent of injury is explained in relation to locomotor and behavioural activity. Several SCI in vivo methods have been reported and studied extensively, particularly contusion, compression, and transection models. These models depict similar pathophysiology to that in humans but are extremely expensive (contusion) and require expertise (compression). Chemical excitotoxicity-induced SCI models are simple and easy while producing similar clinical manifestations. The kainic acid (KA) excitotoxicity model is a convenient, low-cost, and highly reproducible animal model of SCI in the laboratory. The basic impactor approximately cost between 10,000 and 20,000 USD, while the kainic acid only cost between 300 and 500 USD, which is quite cheap as compared to traditional SCI method. Methods In this study, 0.05 mM KA was administered at dose of 10 µL/100 g body weight, at a rate of 10 µL/min, to induce spinal injury by intra-spinal injection between the T12 and T13 thoracic vertebrae. In this protocol, detailed description of a dorsal laminectomy was explained to expose the spinal cord, following intra-spinal kainic acid administration at desired location. The dose, rate and technique to administer kainic acid were explained extensively to reflect a successful paraplegia and spinal cord injury in rats. The postoperative care and complication post injury of paraplegic laboratory animals were also explained, and necessary requirements to overcome these complications were also described to help researcher. Results This injury model produced impaired hind limb locomotor function with mild seizure. Hence this protocol will help researchers to induce spinal cord injury in laboratories at extremely low cost and also will help to determine the necessary supplies, methods for producing SCI in rats and treatments designed to mitigate post-injury impairment. Conclusions Kainic acid intra-spinal injection at the concentration of 0.05 mM, and rate 10 µL/min, is an effective method create spinal injury in rats, however more potent concentrations of kainic acid need to be studied in order to create severe spinal injuries.

Mount Lawu is an inactive volcano located on the border of Central Java and East Java. Geographical conditions that are between these two areas make Mount Lawu unique in its biodiversity. Orchid or familia Orchidaceae is a group of flowering plants with the second-highest species diversity with the number of species reaching of 25.000 worldwide. Several previous studies have shown that orchids in the Mount Lawu area are unique and have high diversity. Documentation of the diversity of orchids in the Mount Lawu area needs to be done as a first step in collecting data on potential flora and can be used as a basis for formulating orchid conservation policies. This research was conducted on the Mount Lawu Cemara Kandang Hiking Track, Karanganyar Regency, Central Java to take an inventory of the diversity of orchids along with their abundance and distribution along the Cemara Kandang Hiking Track. Research was conducted using the exploration method shows that along the Cemara Kandang Hiking Track there are 14 species of orchids. The fourteen species consist of Bulbophyllum schefferi, Bulbophyllum sect. Aphanobulbon, Bulbophyllum sp.1, Bulbophyllum sp.2, Coelogyne miniata, Crepidium koordersii, Liparis montana, Microtis unifolia, Pholidota carnea, Pholidota globosa, Pinalia multiflora, Schoenorchis juncifolia, Taeniophyllum glandulosum, dan Thelymitra javanica. The total abundance of orchids found in this study was 642 species and had a clustered distribution divided into two main zones along the hiking track.

Chayote plants (Sechium edule) with monoecious characters require a pollination process. The pollination process requires pollinating agents to increase its productivity, one of which is insects. This research aimed to determine the diversity and abundance of insects pollinator on chayote plants. Observation of diversity and abundance used a scan sampling method. Pollinator insects observations were carried out in 3 time periods, morning, afternoon, and evening on male and female flowers. We measured environmental parameters such as temperature, humidity, wind speed, and light intensity. Eight species of wild insects pollinated chayote flowers, Apis cerana, Apis dorsata, Lasioglossum leucozonium, Polistes sagittarius, Phimenes flavopictus, Campsomeriella annulata, Lucilia sericata, and Musca domestica. The insect pollinators community had moderate diversity (1.23), a relatively dynamic community (0.59), and moderate dominance (0.62), with A. cerana, which had been the dominant insect pollinator with a relative abundance of 61.63%. Musca domestica and L. sericata were (0,58%) the least dominant insect pollinator with a relative abundance of 0.58%. This research concludes that the insects pollinator of chayote has a moderate level of diversity, relatively dynamic community, and average dominance.

Abstract Background Functional bioengineered tooth regeneration using autologous or allogeneic alternative differentiated cells sources are thought to have a great potential in replacing conventional dentures. This study investigated the potential of dental pulp stem cells (DPSCs) conditioned medium for odontoblastic differentiation of Wharton’s jelly mesenchymal stem cells (WJMSCs). The DPSCs derived from healthy adult permanent first molars were cultured at high confluence prior to conditioned medium collection. The WJMSCs were cultured in six different treatments, with varying ratios of culture media to DPSCs-conditioned medium. MTT assay was used to measure the rate of proliferation of WJMSCs, while immunocytochemistry staining was utilised to detect the expression of dental matrix protein 1 (DMP-1). The deposited calcium was detected and analysed via Alizarin-Red Staining (ARS). Results It was found that the proliferation of WJMSCs cultured under the mixture of complete medium and DPSCs conditioned medium showed significantly lower than the control; presumably the cells started to exit proliferative state prior differentiation. In 14 days of induction, the cells in all treatments showed osteoblastic-like morphology, calcium compound deposits were observed at day 7, 10 and 14 of differentiation suggested that DPSCs conditioned medium could lead to osteoblastic/odontoblastic differentiation. However, the DMP-1 protein can be seen only expressed minimally at day 14 of conditioned medium induction. Conclusions In conclusion, DPSCs conditioned medium appeared as a potential odontoblastic induction approach for WJMSCs. To further investigate the stimulatory effects by DPSCs conditioned medium, specific signalling pathway need to be elucidated to enhance the differentiation efficiency.

Background and Objective: Chitinase-3-like protein 1 (CHI3L1) is an upcoming biomarker for the diagnosis of liver fibrosis. The reference intervals (RIs) for CHI3L1 have not been established in the Pakistani population. Thus, this study aimed to determine the RIs in our population and the cut-off value for diagnosis of hepatic fibrosis. Methods: This is a cross-sectional study. A total of 408 participants (202 healthy and 206 diagnosed liver fibrosis cases) were recruited. Serum CHI3L1 level was measured on CHI3L1 kits (Proprium Biotech Co. Ltd) by manual enzyme-linked immunosorbent assay. The RIs were estimated by percentile and working normal method. Results: The distribution of CHI3L1 values showed no remarkable variation with gender and age. The 95% RI of CHI3L1 was 12.80-81.80 ng/ ml in healthy Pakistani subjects and the cut-off for the diagnosis was 102.12 ng/ml in hepatic fibrosis cases. Conclusion: The RIs in healthy adults and the cut-off for the diagnosis of hepatic fibrosis of serum CHI3L1 were determined in a selective adult Pakistani population. This will be a useful reference for further local and international studies.

Abstract Background In the area of oral and maxillofacial surgery, regenerative endodontics aims to present alternative options to conventional treatment strategies. With continuous advances in regenerative medicine, the source of cells used for pulp tissue regeneration is not only limited to mesenchymal stem cells as the non-mesenchymal stem cells have shown capabilities too. In this review, we are systematically assessing the recent findings on odontoblastic differentiation induction with scaffold and non-scaffold approaches. Methods A comprehensive search was conducted in Pubmed, and Scopus, and relevant studies published between 2015 and 2020 were selected following the PRISMA guideline. The main inclusion criteria were that articles must be revolving on method for osteoblast differentiation in vitro study. Therefore, in vivo and human or animal clinical studies were excluded. The search outcomes identified all articles containing the word “odontoblast”, “differentiation”, and “mesenchymal stem cell”. Results The literature search identified 99 related studies, but only 11 articles met the inclusion criteria. These include 5 odontoblastic differentiation induction with scaffold, 6 inductions without scaffolds. The data collected were characterised into two main categories: type of cells undergo odontoblastic differentiation, and odontoblastic differentiation techniques using scaffolds or non-scaffold. Conclusion Based on the data analysis, the scaffold-based odontoblastic induction method seems to be a better option compared to the non-scaffold method. In addition of that, the combination of growth factors in scaffold-based methods could possibly enhance the differentiation. Thus, further detailed studies are still required to understand the mechanism and the way to enhance odontoblastic differentiation.

Squamocin is one of the annonaceous acetogenins produced by the Annonaceae family and displays potent anti-cancer activity against cancer cell lines. This study aimed to investigate the growth inhibition activity of squamocin coupled with nanodiamond on rats (Rattus norvegicus)-induced breast cancer. Twenty-five female R. norvegicus were divided into five groups (n = 5), including normal control (without any treatment), negative control, group treated with nanodiamond only (ND), group treated with squamocin only (SQ), and the group treated with squamocin coupled with nanodiamond (NDSQ). All of the animal models were induced for breast cancer, except for the normal control group. Breast cancer induction was performed using two doses of N-nitroso-N-methylurea (NMU) injection (50 and 30 mg/kg body weight) intraperitoneally and waited for 22 weeks until the tumor was detected to formed. Nanodiamond coupled with squamocin were administered by intraperitoneal injection (1.5 mg/kg body weight) for 5 weeks, one injection per 3 days. This study showed that the treatment with squamocin coupled with nanodiamond (NDSQ) significantly reduced the proliferation (Ki-67) and induced apoptosis (Caspase-3) of breast cancer cells, corresponding to the reduction of the thickness of the mammary ductal epithelium (p

A large number of species of marine invertebrates that have similar morphology cause misidentification very possible. Molecular species identification is important in taxonomy study accurately. This study aims to determine the identity of Aquatic Papua marine invertebrates with DNA barcoding technique using markers COI gene (Cytochrome c oxidase I). The study identified 29 individuals of marine invertebrates, which came from sea cucumbers (6 samples), lobsters (6 samples), octopus (6 samples), chitons (5 samples), and sea urchins (6 samples). Molecular identification methods consist of DNA isolation, PCR COI gene fragments, sequencing, and DNA sequence analysis using bioinformatics software. The results showed that the sea cucumber came from the Bohadschia marmorata species, the lobster came from the Panulirus versicolor species, the octopus came from the Octopus cyanea species, the chiton came from the Ischnochitonaustralis species, and the sea urchins came from the Tripneustesgratilla species. The similarity of samples and references are in the range of 84.58 to 100.00%. The disparity index, genetic distance, and phylogenetic tree supported this result.

Dental caries is a biofilm-mediated, sugar-driven, multifactorial, dynamic disease that results in the phasic demineralization and remineralization of dental hard tissues. Despite scientific advances in cariology, dental caries remains a severe global concern. The aim of this study was to determine the optimization of microbial and molecular techniques for the detection of cariogenic pathogens in dental caries patients, the prevalence of cariogenic bacteria on the basis of socioeconomic, climatological, and hygienic factors, and in vitro evaluation of the antimicrobial activity of selected synthetic antibiotics and herbal extracts. In this study, oral samples were collected from 900 patients for bacterial strain screening on a biochemical and molecular basis. Plant extracts, such as ginger, garlic, neem, tulsi, amla, and aloe vera, were used to check the antimicrobial activity against the isolated strains. Synthetic antimicrobial agents, such as penicillin, amoxicillin, erythromycin, clindamycin, metronidazole, doxycycline, ceftazidime, levofloxacin, and ciprofloxacin, were also used to access the antimicrobial activity. Among 900 patients, 63% were males and 37% were females, patients aged between 36 and 58 (45.7%) years were prone to disease, and the most common symptom was toothache (61%). For oral diseases, 21% used herbs, 36% used antibiotics, and 48% were self-medicated, owing to sweets consumption (60.66%) and fizzy drinks and fast food (51.56%). Staphylococcus mutans (29.11%) and Streptococcus sobrinus (28.11%) were found as the most abundant strains. Seven bacterial strains were successfully screened and predicted to be closely related to genera S. sobrinus, S. mutans, Actinomyces naeslundii, Lactobacillus acidophilus, Eubacterium nodatum, Propionibacterium acidifaciens, and Treponema Pallidum. Among plant extracts, the maximum zone of inhibition was recorded by ginger (22.36 mm) and amla (20.01 mm), while among synthetic antibiotics, ciprofloxacin and levofloxacin were most effective against all microbes. This study concluded that phyto extracts of ginger and amla were considered suitable alternatives to synthetic antibiotics to treat dental diseases.

Background and Objective: COVID-19 took the world by a storm and impacted not only the general population but the doctors as well. This study was done in order to determine the financial and economic burden faced by the doctors working in COVID-19 pandemic in Pakistan. Methods: This longitudinal study was carried out from May 9th to May 23rd, 2020 in Pakistan. A self-designed validated Questionnaire was distributed to doctors working in different hospitals of Pakistan through online Google form. Questions regarding emotional and financial stress in Pakistani doctors dealing with COVID-19 patients were included. Results: One hundred and twenty doctors responded to the current study. Majority of them 97 (80.6%) reported stress during duty hours. Stress level was high among 78 (65.3%) doctors to the extent they thought of leaving the job because of risk of getting infected. Less salaries and family pressure were the other demotivating factors. Most of the doctors 71 (59.2%)] were younger than 30 years of age. Eighty-five (71%) doctors reported being pressurized by the families to leave the job. Shortage of protective gears was also the challenge. A total of 65 (54.2%) doctors got their gears from their institutes, 27 (22.2%) bought their own gears, 23 (19.4%) worked without gears, and 5 (4.8%) got gears from non-governmental organizations (NGOs). Conclusion: Majority of the doctors reported stress during duty hours regardless of level or experience in that job; the contributing factors included risk of getting infected, lesser salaries and family pressure. During COVID-19 pandemic, doctors feel that they are not only emotionally exhausted but also tried to cope financial challenges while endangering themselves to save the lives of patients.

Background and Objective: COVID-19 took the world by a storm and impacted not only the general population but the doctors as well. This study was done in order to determine the financial and economic burden faced by the doctors working in COVID-19 pandemic in Pakistan. Methods: This longitudinal study was carried out from May 9th to May 23rd, 2020 in Pakistan. A self-designed validated Questionnaire was distributed to doctors working in different hospitals of Pakistan through online Google form. Questions regarding emotional and financial stress in Pakistani doctors dealing with COVID-19 patients were included. Results: One hundred and twenty doctors responded to the current study. Majority of them 97 (80.6%) reported stress during duty hours. Stress level was high among 78 (65.3%) doctors to the extent they thought of leaving the job because of risk of getting infected. Less salaries and family pressure were the other demotivating factors. Most of the doctors 71 (59.2%)] were younger than 30 years of age. Eighty-five (71%) doctors reported being pressurized by the families to leave the job. Shortage of protective gears was also the challenge. A total of 65 (54.2%) doctors got their gears from their institutes, 27 (22.2%) bought their own gears, 23 (19.4%) worked without gears, and 5 (4.8%) got gears from non-governmental organizations (NGOs). Conclusion: Majority of the doctors reported stress during duty hours regardless of level or experience in that job; the contributing factors included risk of getting infected, lesser salaries and family pressure. During COVID-19 pandemic, doctors feel that they are not only emotionally exhausted but also tried to cope financial challenges while endangering themselves to save the lives of patients.

Orchid is an important ornamental plant in Indonesia due to their natural beauty of flowers. In the tropical forest, orchids are being acquired for trading and commercial market. Thus, the effort is required to proliferate orchid in large quantities for conservation and improve the floral variation for plant breeding. The purpose of this study is to develop a firmed methodology of molecular breeding of orchids using CRISPR/Cas9 KO system. The plant material used was Phalaenopsis amabilis protocorms growth on NP medium+pepton (2 g/L). Protocorm were submerged in the culture of Agrobacterium tumefaciens that Ti‐plasmid had been filled with a T‐DNA construct of a pRGEB32 vector harboring sgRNA with PDS3 sequence. Detection for transformants was confirmed by PCR using HPT primers (545 bp), Cas9 primers (402 bp), PDS primers (280 bp) and trnL‐F (1200 bp) as an internal control. The results showed that 0.96% PDS transformants were obtained from PDS3T2 lines. Several transformant showed pale leaf color compared to non‐transformant plants. This study suggests that the target gene has successfully edited by CRISPR/Cas9 system and could be applied for that functional gene editing in orchids.

Abstract Stem cell-based regenerative medicine is a promising approach for tissue reconstruction. However, a large number of cells are needed in a typical clinical study, where conventional monolayer cultures might pose a limitation for scale-up. The purpose of this review was to systematically assess the application of microcarriers in Mesenchymal Stem Cell cultures. A comprehensive search was conducted in Medline via Ebscohost, Pubmed, and Scopus, and relevant studies published between 2015 and 2019 were selected. The literature search identified 53 related studies, but only 14 articles met the inclusion criteria. These include 7 utilised commercially available microcarriers, while the rest were formulated based on different surface characteristics, all of which are discussed in this review. Current applications of microcarriers were focused on MSC expansion and induction of MSCs into different lineages. These studies demonstrated that MSCs could proliferate in a microcarrier culture system in-fold compared to monolayer cultures, and the culture system could simulate a three-dimensional environment which induces cell differentiation. However, detailed studies are still required before this system were to be adapted into the scale of GMP manufacturing.

Cryptic organisms that live within the interstices of reef habitats contribute significantly to coral reef biodiversity. One example of this cryptic biodiversity is the high abundance of decapods in dead coral heads that associated with various biota. However, this diversity poorly studied especially species identification and species richness. This study aims to estimate the decapods diversity in Raja Ampat, Papua, using Pocillopora dead coral head method as semi-quantitative sampling approach. Raja Ampat in Papua is chosen because it situated in the center of Coral Triangle marine hotspot. Data were collected from 10 dead coral heads of genus Pocillopora from 10-meter depth near the islands of Kri and Misool. This study observed a total of 205 individuals Decapoda from Kri Island and 672 Individual from Misool Islands. Species richness observation (Chao1 and ACE) of the total samples reports only 11 families of decapoda detected in this study. Rarefaction curve reach an asymptote after all after all ten dead coral were analyzed; indicating that additional sample collection would not change estimates of diversity found in this study. The Shannon-Wiener index diversity on the Kri Island showed lower diversity value (2.09) compared to Misool Island (2.18). In the future, this research can be used as a basis for understanding the diversity of coral reefs as well as for management and conservation of coral reef ecosystems.

Pseudomonas aeruginosa is an opportunistic Gram-negative bacterium implicated in acute and chronic nosocomial infections and a leading cause of patient mortality. Pseudomonas aeruginosa infections are frequently associated with the development of biofilms, which give the bacteria additional drug resistance and increase their virulence. The goal of this study was to find strong compounds that block the Anthranilate-CoA ligase enzyme made by the pqsA gene. This would stop the P. aeruginosa quorum signaling system. This enzyme plays a crucial role in the pathogenicity of P. aeruginosa by producing autoinducers for cell-to-cell communication that lead to the production of biofilms. Pharmacophore-based virtual screening was carried out utilizing a library of commercially accessible enzyme inhibitors. The most promising hits obtained during virtual screening were put through molecular docking with the help of MOE. The virtual screening yielded 7/160 and 10/249 hits (ZINC and Chembridge). Finally, 2/7 ZINC hits and 2/10 ChemBridge hits were selected as potent lead compounds employing diverse scaffolds due to their high pqsA enzyme binding affinity. The results of the pharmacophore-based virtual screening were subsequently verified using a molecular dynamic simulation-based study (MDS). Using MDS and post-MDS, the stability of the complexes was evaluated. The most promising lead compounds exhibited a high binding affinity towards protein-binding pocket and interacted with the catalytic dyad. At least one of the scaffolds selected will possibly prove useful for future research. However, further scientific confirmation in the form of preclinical and clinical research is required before implementation.

Background: Soymilk is a plant-based protein-rich beverage that is produced from soybeans. It is a steady emulsion of water, oil, and protein. Along with similar vegetable-based milk, soymilk can be utilized as an alternative for dairy milk by individuals who are lactose intolerant or vegan. It can also be a decent swap for cow's milk where cow milk is not accessible in adequate amounts. Objectives: To evaluate the sensory characteristics of soymilk under different storage conditions i.e., refrigerator, chiller, and room temperature. Methodology: Examined soybean seeds were rinsed and soaked in the water multiple times the volume of the beans, for around 10hrs. The soaked beans were parboiled (partial cooking) in boiling water for a few moments with steady fomentation. The boiled beans were cooled, de-husked, washed, and after that, the seeds were homogenized with clean water into a stable paste. After that obtained paste was drawn out, (sieved) by utilizing a perfect muslin cloth to isolate the filtrate from the paste. Then the obtained milk was boiled to eliminate the beany flavor and was packed in sterilized glass bottles, stored, and coded as T1 (refrigerator temperature, 7°C ±1°C), T2 (chiller temperature, 4°C ±1°C), and T3 (room temperature, 25°C ±2°C). Results: The results of the study showed that the maximum score of color, flavor, taste, aroma, and overall acceptability of soymilk recorded as 8.06, 8.22, 8.45, 8.54, and 8.57, respectively on chiller temperature (4°C ±1°C). While the lowest score of color, flavor, taste, aroma, and overall acceptability of soymilk was recorded 7.36, 7.55, 7.78, 7.65, and 7.65, respectively at room temperature (25°C ±2°C). Conclusion: It was observed from the present study that in between the stored samples, sensory attributes such as color, flavor, taste, aroma, and overall acceptability perceived better scoring for T2, T1, and T3, respectively. Also, during storage, a little decrease in sensory attributes was observed among all treatments mostly from treatments stored at room temperature.

In Southeast Asian nations, cultured hybrid groupers are facing serious infestation by the marine parasitic leech Zeylanicobdella arugamensis (Annelida, Hirudinea). They attach to the hybrid groupers by sucking and biting on the surface of the skin, paving the way for secondary infection upon the fish. The objective of the study is to utilize the locally available seaweed to control the infestation of parasitic leeches. The methanol extracts of the brown alga Sargassum polycystum C. Agardh, 1824 (Phaeophyceae) from Sabah were prepared and investigated for antiparasitic efficacy against Z. arugamensis through in vitro bioassay. A total of 126 adult leeches from the host hybrid groupers were obtained from the fish hatchery. The parasitic leeches were treated with the methanol extracts of S. polycystum for 180 min by preparing five different dosages at concentrations of 100, 50, 25, 12.5, and 6.25 mg/mL. The brown alga was found to have high antiparasitic efficacy, resulting in 100% leech mortality over a short period of time. It showed the highest antiparasitic efficacy (total mortality of leeches) in a short time limit of 0.96 ± 0.44 min, for 100 mg/mL of the extract. Observations on leech behavior in the positive control and the seaweed extract treatments showed vigorous swimming before mortality. LC-MS/MS analysis was used to reveal the phytochemical composition of the extract to understand the nature of the main components responsible for its antiparasitic activities. A total of 29 metabolites were identified via Q Exactive HF Orbitrap mass spectrometry, including two flavonoids (ephedrannin A and hinokiflavone), two organoarsenics (1-dimethylarsinoyl-heptadecane and cacodylic acid), four heterocyclic compounds, and two chlorophyll breakdown products. The presence of bioactive compounds could increase the mortality rate of parasitic leeches. Thus, this study concludes that the brown alga showed high efficacy in its antiparasitic activities and can be effectively applied for treatment in grouper aquaculture farms for sustainable aquaculture.

Advances in data acquisition via high resolution genomic, transcriptomic, proteomic and metabolomic platforms have driven the discovery of the underlying factors associated with metabolic disorders (MD) and led to interventions that target the underlying genetic causes as well as lifestyle changes and dietary regulation. The review focuses on fourteen of the most widely studied inherited MD, which are familial hypercholesterolemia, Gaucher disease, Hunter syndrome, Krabbe disease, Maple syrup urine disease, Metachromatic leukodystrophy, Mitochondrial encephalopathy lactic acidosis stroke-like episodes (MELAS), Niemann-Pick disease, Phenylketonuria (PKU), Porphyria, Tay-Sachs disease, Wilson’s disease, Familial hypertriglyceridemia (F-HTG) and Galactosemia based on genome wide association studies, epigenetic factors, transcript regulation, post-translational genetic modifications and biomarker discovery through metabolomic studies. We will delve into the current approaches being undertaken to analyze metadata using bioinformatic approaches and the emerging interventions using genome editing platforms as applied to animal models.

Dendrobium capra is an Indonesian endemic orchid species that live in Java. It grows on low altitude with warm climate. D. capra has beautiful small yellow greenish flower that grow in raceme inflorescence. This orchid faces a threat in its natural habitat due to having a long life cycle and a forestry main commodity as a main host thus categorized as Appendix II on CITES list. To address that problem, ex situ conservation approach using in vitro culture method is necessary. Germination enhancement effort using complex organic substances found that 200 ml/l tomato extract gave best germination result. Analysis on D. capra plantlet growth also showed that MS medium produced better plantlet size than NP, VW and KC medium. Supplementing medium with a combination of NAA and TDZ has also successfully induced early flowering within 11 month of culture period. This information is important to achieve successful in vitro culture of D. capra for various purposes.

Malaysia has a long coastline surrounded by various islands, including North Borneo, that provide a suitable environment for the growth of diverse species of seaweeds. Some of the important North Bornean seaweed species are Kappaphycus alvarezii, Eucheuma denticulatum, Halymenia durvillaei (Rhodophyta), Caulerpa lentillifera, Caulerpa racemosa (Chlorophyta), Dictyota dichotoma and Sargassum polycystum (Ochrophyta). This review aims to highlight the therapeutic potential of North Bornean seaweeds and their nutraceutical profiling. North Bornean seaweeds have demonstrated anti-inflammatory, antioxidant, antimicrobial, anticancer, cardiovascular protective, neuroprotective, renal protective and hepatic protective potentials. The protective roles of the seaweeds might be due to the presence of a wide variety of nutraceuticals, including phthalic anhydride, 3,4-ethylenedioxythiophene, 2-pentylthiophene, furoic acid (K. alvarezii), eicosapentaenoic acid, palmitoleic acid, fucoxanthin, β-carotene (E. denticulatum), eucalyptol, oleic acid, dodecanal, pentadecane (H. durvillaei), canthaxanthin, oleic acid, pentadecanoic acid, eicosane (C. lentillifera), pseudoephedrine, palmitic acid, monocaprin (C. racemosa), dictyohydroperoxide, squalene, fucosterol, saringosterol (D. dichotoma), and lutein, neophytadiene, cholest-4-en-3-one and cis-vaccenic acid (S. polycystum). Extensive studies on the seaweed isolates are highly recommended to understand their bioactivity and mechanisms of action, while highlighting their commercialization potential.

Abstract Breast cancer remains a worldwide public health dilemma and is currently the most common tumour in the globe. Awareness of breast cancer, public attentiveness, and advancement in breast imaging has made a positive impact on recognition and screening of breast cancer. Breast cancer is life-threatening disease in females and the leading cause of mortality among women population. For the previous two decades, studies related to the breast cancer has guided to astonishing advancement in our understanding of the breast cancer, resulting in further proficient treatments. Amongst all the malignant diseases, breast cancer is considered as one of the leading cause of death in post menopausal women accounting for 23% of all cancer deaths. It is a global issue now, but still it is diagnosed in their advanced stages due to the negligence of women regarding the self inspection and clinical examination of the breast. This review addresses anatomy of the breast, risk factors, epidemiology of breast cancer, pathogenesis of breast cancer, stages of breast cancer, diagnostic investigations and treatment including chemotherapy, surgery, targeted therapies, hormone replacement therapy, radiation therapy, complementary therapies, gene therapy and stem-cell therapy etc for breast cancer.

Background: Ziarat juniper (Juniperus excelsa M.Bieb) is an evergreen and dominant species of Balochistan juniper forests. This forest is providing many benefits to regional ecosystems and surrounding populations. No functional genomics study is reported for this important juniper plant. This research is aimed to characterize the Ziarat juniper functional genome based on the analyses of 1500 transcripts. Methods: Total RNA from shoot of Juniperus excelsa was extracted and subjected for transcriptome sequencing using Illumina HiSeq 2000 with the service from Macrogen, Inc., South Korea. The Illumina sequenced data was subjected to bioinformatics analysis. Quality assessment and data filtration was performed for the removal of low-quality reads, ambiguous reads and adaptor sequences. The high-quality clean reads data was deposited in the Sequence Read Archive (SRA) at NCBI, and used for downstream processes. Fifteen hundred transcripts were randomly chosen and used for functional characterization. Results: As a result of homology search 80.3% transcripts showed significant similarities and were placed in significant similarities category, 19.3% transcripts showed low similarities and assigned to the ‘‘unclassified’’ category while 0.4% transcripts are defined as no hits. The functional characterization results showed that most (18%) of the transcripts are involved in metabolism, followed by 11.7% in transcription and 11.5% as structural protein. 8.8% transcripts are engaged in stress response, whereas the transcripts involved in growth and development constituted 6.7%. Transcripts involved in signal transduction represented 5.6%, while 3.5% facilitating transport and 34.1% are involved in hypothetical functions. Conclusion: The functional annotation data produced in this study will be very useful for future functional genome analysis of Juniperus excelsa.

Here we describe a novel method of using green fluorescence protein (GFP) as a reporter gene for early detection of an integrated T­DNA containing the orchid flowering gene, PaFT1 (Phalaenopsis aphrodite Flowering locus T1) in the tobacco genome. Functional assays that report the presence of exogenous DNA early in development are especially useful in plants where the desired phenotype is only apparent after long periods of vegetative growth. The objective of this study is to establish a method for detecting an inserted Phalaenopsis orchid flowering gene and examining its function in tobacco. The p35S::PaFT1­ 35S::GFP construct was introduced into Agrobacterium tumefaciens strain EHA101. Transformed tobacco leaves were cultured on MS medium with addition of 1 mgL-1 NAA+3 mgL-1 BAP+50 mgL-1 Kanamycin+300 mgL-1 timentin for selection. Results showed bright green GFP fluorescent signals in 11 out of 15 (73%) tobacco leaf cells at a 2­month time point after transformation. GFP and PaFT1 fragments were amplified in genomic PCR using GFP and PaFT1 specific primers. The accumulated PaFT1 transcripts were observed in 3 month­old transgenic tobacco plants containing p35S::PaFT1­35S::GFP. Green florescence was observed only in the transgenic plants at the 5 month­old stage but not in the wild type controls.

Vehicular ad hoc network (VANET) is a special form of mobile ad hoc network (MANET), which plays a key role in the intelligent transportation system (ITS). Though many outstanding geographic routing protocols are designed for VANETs, a majority of them use parameters that only affect routing performance. In this article, we propose an intersection routing based on fuzzy multi-factor decision (IRFMFD), which utilizes several features. The scheme is divided into two parts, namely vehicular decision management and intersection decision management. In the vehicular component, candidate vehicles between two static nodes (SNs) located at two intersections derive potential routing paths considering distance, neighbor quantity, and relative velocity. In the intersection component, the candidate SN was chosen from the current intersection’s 2-hop neighbors which were connected with the current intersection by a route that was decided on in part one. To get the best scheme, we also introduced other factors to estimate the number of hops in each link and link lifetime. The simulation shows that the IRFMFD outperforms on delivery ratio and end-to-end delay compared with AODV (Ad hoc on-demand distance vector), GPSR (Greedy perimeter stateless routing) and GeOpps (Geographical opportunistic routing).

MicroRNAs (miRNAs) are tiny, non-coding and regulatory RNAs approximately 21 nucleotides in length.They are reported in various plants but still needs discovery in important plant species. The 12 plant species ofFabaceae were subjected this time to identify their miRNAs. The comparative genomics approaches withcombination of various bioinformatics’ tools were applied to find the novel miRNAs. This research leads to thefinding of 29 miRNAs belonging to 13 miRNA families. From the 29 miRNAs, nine belongs to Arachis duranensis,four to Lotus japonicas, three to each Pisum sativum and Arachis hypogaea, two to each Arachis ipaensis andPhaseolus vulgaris and one to each Cicer arietinum, Phaseolus acutifolius, Lupinus luteus, Glycyrrhiza uralensis,Robinia pseudoacacia and Lathyrus odoratus. These findings will be useful in the future to design and developdesirable traits in the 12 plant species of Fabaceae.

Human body temperature is an important sign of physical condition in terms of comfort, heat or cold stresses, and of performance. This paper presents the preliminary investigation into the design, manufacturing and testing of the textile based temperature sensor. This sensing fabric may be employed to measure the temperature of the human body on a continuous basis over extensive periods of time, outside the clinical environment. The sensing fabric was manufactured on an industrial scale flat-bed knitting machine by laying-in the sensing element (in the form of fine metal wire) into the double layer knitted structure. The operational principle of the sensing fabric is based on the inherent tendency of metal wire to change in its electrical resistance because of the change in its temperature. An experimental resistance-temperature relationship showed promising validation in comparison with their modeled counterparts.

its chemical structure than curcumin. As a curcumin analogue, PGV-1 was considered to have anticanceractivities. This research was conducted to study the effect of PGV-1 on the cycle progression of T47D cells. Cytotoxiceffects of PGV-1 on T47D cells were determined using MTT assay, and the the effect on cell cycle progressionwas carried out using flowcytometry. Western blot analysis was used to analyze protein expression correspondingto cell cycle progression. The result showed that at the concentration of 2.5 μM PGV-1 inhibited cell cycleprogression through G2/M arrest and induced of cells hyperploidy formation. The hyperploidy formation inducedby PGV-1 was related to the increase of cdc-2 expression. PGV-1 2.5 μM elevated the level of p21 CIP/KIPthrough p53- independent manner. Apoptosis was also induced by PGV-1 at early phase of treatment indicated byPARP cleavage due to activation of caspase-3/7 after 12 h treatment. The results above suggest that PGV-1 inhibitsthe growth of T47D cells targeted on microtubules.Keywords: PGV-1, G2/M arrest, apoptosis, p21

Carcinogenesis may resulted from the malfunctioning of programmed cell death. Most of the anticancer drugs incurrent use induce apoptosis in susceptible cells. The fact that disparate agent interacting with different targets seemto induce cell death through some common mechanisms suggest that anticancer activity is determined by the abilityof inhibiting cell growth. Pentagamavunon-1 (PGV-1) is one of the curcumin analogues which showed to havepotency in inhibiting proliferation of T47D human breast carcinoma cells. The effects on T47D cells growth isassociated with cell cycle arrest in G2/M phase at the concentration of 2.5 ?M, followed by hyperploidy. The data onpolymerization assay, indicated that PGV-1 interact with tubulin in different manner from taxol. PGV-1 inhibittubulin polymerization on cell culture while taxol stabilized tubulin polymerization. Immunostainning data onPGV-1 treated cells showed slightly tubulin condensation, while taxol treated cells showed tubulin condensationdistinctly at 12 minutes after releasing from depolymerizing agent.In conclusion, PGV-1 represent a new microtubule inhibitor and has the potential to be developed for antimitoticdrugKey words: Pentagamavunon-1, T47D, tubulin

This research was a quasi experimental aimed to investigate the influenced of inquiry learning model to generic science skills of students class XI IPA SMAN 1 Pangkajene Sidrap on buffer solution subject matter. Research design used “Nonequivalent Control Group Design” that consist of two variable there are inquiry learning model as dependent variable and generic science skills as independent variable. Population in this research is students class XI IPA SMAN 1 Pangkajene Sidrap academic year 2012/2013 which amount 240 students. The sample choosen by class random sampling and have choosen XI IA 1 as experiment class consist of 34 students and XI IA 2 as control class consist of 35 students. Instrument that used is generic science skills test (KGS) were integrated into buffer solution subject matter that includes indirect observation, sense of scale, logical frame, causality, mathematical modeling and concept formation. The data was collected by given generic science skills test in pretest and posttest by objective test. Data were analyzed by descriptive statistics and inferential statistics. The descriptive results showed that generic science skills student of experiment class get N-gain average 0.71 in high categories and control class get N-gain average 0.31 in medium categories. The inferential statistics analysis showed that tcalculated 7,17 > ttable 1,68 in significant level α = 0.05, so H0 failed or H1 accepted. The result of analysis describe that inquiry learning model have positive influenced to generic skills science of students on buffer solution subject matter. Kata Kunci: Inquiry, Keterampilan Generik Sains

This paper reports the uncertainty analysis of the temperature–resistance (TR) data of the newly developed temperature sensing fabric (TSF), which is a double-layer knitted structure fabricated on an electronic flat-bed knitting machine, made of polyester as a basal yarn, and embedded with fine metallic wire as sensing element. The measurement principle of the TSF is identical to temperature resistance detector (RTD); that is, change in resistance due to change in temperature. The regression uncertainty (uncertainty within repeats) and repeatability uncertainty (uncertainty among repeats) were estimated by analysing more than 300 TR experimental repeats of 50 TSF samples. The experiments were performed under dynamic heating and cooling environments on a purpose-built test rig within the temperature range of 20–50 °C. The continuous experimental data was recorded through LabVIEW-based graphical user interface. The result showed that temperature and resistance values were not only repeatable but reproducible, with only minor variations. The regression uncertainty was found to be less than ±0.3 °C; the TSF sample made of Ni and W wires showed regression uncertainty of ±0.18 °C). The cooling TR data showed considerably reduced values (±0.07 °C) of uncertainty in comparison with the heating TR data (±0.24 °C). The repeatability uncertainty was found to be less than ±0.5 °C. By increasing the number of samples and repeats, the uncertainties may be reduced further. The TSF could be used for continuous measurement of the temperature profile on the surface of the human body.

The teaching of the educational levels including at the university level is still emphasized on the aspect of cognitive ability. The emphasis on the aspect of empowering the metacognitive is not touched yet. As a matter of fact, there are some techniques of teaching that can be applied to empower their metacognitive ability such Problem-based Learning (PBL) and cooperative Group Investigation (GI) strategy applies. This study aims to explain the effect of PBL and cooperative GI strategy to the metacognition and this study use “Pretest-Posttest Nonequivalent Control Group Design”. This study indicates that students that learned with PBL and GI strategy have scores of metacognition from pretest to posttest higher than students that learned with conventional strategy. Based on the ANACOVA test showed that there were significance differences of students’ metacognition ability between the students who were thought through PBL strategy and the students who were thought through conventional and GI strategy. Kata kunci: PBL, kooperatif GI, metakognisi

Carcinogenesis may resulted from the malfunctioning of programmed cell death. Most of the anticancer drugs in current use induce apoptosis in susceptible cells. The fact that disparate agent interacting with different targets seem to induce cell death through some common mechanisms suggest that anticancer activity is determined by the ability of inhibiting cell growth. Pentagamavunon-1 (PGV-1) is one of the curcumin analogues which showed to have potency in inhibiting proliferation of T47D human breast carcinoma cells. The effects on T47D cells growth is associated with cell cycle arrest in G2/M phase at the concentration of 2.5 ?M, followed by hyperploidy. The data on polymerization assay, indicated that PGV-1 interact with tubulin in different manner from taxol. PGV-1 inhibit tubulin polymerization on cell culture while taxol stabilized tubulin polymerization. Immunostainning data on PGV-1 treated cells showed slightly tubulin condensation, while taxol treated cells showed tubulin condensation distinctly at 12 minutes after releasing from depolymerizing agent. In conclusion, PGV-1 represent a new microtubule inhibitor and has the potential to be developed for antimitotic drug Key words: Pentagamavunon-1, T47D, tubulin

its chemical structure than curcumin. As a curcumin analogue, PGV-1 was considered to have anticancer activities. This research was conducted to study the effect of PGV-1 on the cycle progression of T47D cells. Cytotoxic effects of PGV-1 on T47D cells were determined using MTT assay, and the the effect on cell cycle progression was carried out using flowcytometry. Western blot analysis was used to analyze protein expression corresponding to cell cycle progression. The result showed that at the concentration of 2.5 μM PGV-1 inhibited cell cycle progression through G2/M arrest and induced of cells hyperploidy formation. The hyperploidy formation induced by PGV-1 was related to the increase of cdc-2 expression. PGV-1 2.5 μM elevated the level of p21 CIP/KIP through p53- independent manner. Apoptosis was also induced by PGV-1 at early phase of treatment indicated by PARP cleavage due to activation of caspase-3/7 after 12 h treatment. The results above suggest that PGV-1 inhibits the growth of T47D cells targeted on microtubules. Keywords: PGV-1, G2/M arrest, apoptosis, p21