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Unlocking the growth potential: harnessing the power of synbiotics to enhance cultivation of Pleurotus spp

Patil R, Ramli ANM, Xuan AS, Xin NZ, Azelee NIW, Bhuyar P

J Zhejiang Univ Sci B, 2024 Apr 15;25(4):293-306.
PMID: 38584092 DOI: 10.1631/jzus.B2300383

The oyster mushroom (Pleurotus spp.) is one of the most widely cultivated mushroom species globally. The present study investigated the effect of synbiotics on the growth and quality of Pleurotus ostreatus and Pleurotus pulmonarius. Different synbiotics formulations were applied by spraying mushroom samples daily and measuring their growth parameters, yield, biological efficiency, proximate composition, mineral content, total phenolic content (TPC), and diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging activity. Results demonstrated that the most significant yield of oyster mushrooms was harvested from synbiotics sprayed with inulin and Lactobacillus casei (56.92 g). Likewise, the highest biological efficiency obtained with a similar synbiotic was 12.65%. Combining inulin and L. casei was the most effective method of improving the mushrooms' growth performance and nutrient content in both samples. Furthermore, synbiotics that combined inulin and L. casei resulted in the highest TPC (20.550 mg gallic acid equivalent (GAE)/g dry extract (DE)) in white oyster mushrooms (P. ostreatus). In comparison, in grey mushroom (P. pulmonarius) the highest TPC was yielded by L. casei (1.098 mg GAE/g DE) followed by inulin and L. casei (1.079 mg GAE/g DE). The DPPH results indicated that the oyster mushroom could be an efficient antioxidant. The results revealed that applying synbiotics improved the mushrooms' quality by increasing their antioxidant capacity with higher amounts of phenolic compounds and offering better health benefits with the increased levels of mineral elements. Together, these studies demonstrated the potential of using synbiotics as a biofertilizer, which is helpful for mushroom cultivation; therefore, it might solve the challenge of inconsistent quality mushroom growers face.
Ethical and legal implications of gene editing in plant breeding: a systematic literature review

Idris SH, Mat Jalaluddin NS, Chang LW, 曾立纬

J Zhejiang Univ Sci B, 2023 Jul 06.
PMID: 37423764 DOI: 10.1631/jzus.B2200601

Biotechnology policies and regulations must be revised and updated to reflect the most recent advances in plant-breeding technology. New Plant Breeding Techniques (NPBT) such as gene editing have been applied to address the myriad of challenges in plant breeding, while the use of NPBT as emerging biotechnological tools raises legal and ethical concerns. This study aims to highlight how gene editing is operationalized in the existing literature and examine the critical issues of ethical and legal issues of gene editing for plant breeding. We carried out a systematic literature review (SLR) to provide the current states of ethical and legal discourses surrounding this topic. We also identified critical research priority areas and policy gaps that must be addressed when designing the future governance of gene editing in plant breeding.
Fulltext Electromagnetic field exposure as a plausible approach to enhance the proliferation and differentiation of mesenchymal stem cells in clinically relevant scenarios

Hamid HA, Sarmadi VH, Prasad V, Ramasamy R, Miskon A

J Zhejiang Univ Sci B, 2022 Jan 15;23(1):42-57.
PMID: 35029087 DOI: 10.1631/jzus.B2100443

Mesenchymal stem/stromal cell (MSC)‍-based therapy has been regarded as one of the most revolutionary breakthroughs in the history of modern medicine owing to its myriad of immunoregulatory and regenerative properties. With the rapid progress in the fields of osteo- and musculoskeletal therapies, the demand for MSC-based treatment modalities is becoming increasingly prominent. In this endeavor, researchers around the world have devised new and innovative techniques to support the proliferation of MSCs while minimizing the loss of hallmark features of stem cells. One such example is electromagnetic field (EMF) exposure, which is an alternative approach with promising potential. In this review, we present a critical discourse on the efficiency, practicability, and limitations of some of the relevant methods, with insurmountable evidence backing the implementation of EMF as a feasible strategy for the clinically relevant expansion of MSCs.
Fulltext Detection of human papillomavirus in oropharyngeal squamous cell carcinoma

Fauzi FH, Hamzan NI, Rahman NA, Suraiya S, Mohamad S

J Zhejiang Univ Sci B, 2021 4 13;21(12):961-976.
PMID: 33843162 DOI: 10.1631/jzus.B2000161

Worldwide there has been a significant increase in the incidence of oropharyngeal squamous cell carcinoma (OPSCC) etiologically attributed to oncogenic human papillomavirus (HPV). Reliable and accurate identification and detection tools are important as the incidence of HPV-related cancer is on the rise. Several HPV detection methods for OPSCC have been developed and each has its own advantages and disadvantages in regard to sensitivity, specificity, and technical difficulty. This review summarizes our current knowledge of molecular methods for detecting HPV in OPSCC, including HPV DNA/RNA polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), p16 immunohistochemistry (IHC), and DNA/RNA in situ hybridization (ISH) assays. This summary may facilitate the selection of a suitable method for detecting HPV infection, and therefore may help in the early diagnosis of HPV-related carcinoma to reduce its mortality, incidence, and morbidity.
Fulltext Hydroxychavicol, a polyphenol from Piper betle leaf extract, induces cell cycle arrest and apoptosis in TP53-resistant HT-29 colon cancer cells

Rajedadram A, Pin KY, Ling SK, Yan SW, Looi ML

J Zhejiang Univ Sci B, 2021 Feb 15;22(2):112-122.
PMID: 33615752 DOI: 10.1631/jzus.B2000446

This study aims to elucidate the antiproliferative mechanism of hydroxychavicol (HC). Its effects on cell cycle, apoptosis, and the expression of c-Jun N-terminal kinase (JNK) and P38 mitogen-activated protein kinase (MAPK) in HT-29 colon cancer cells were investigated. HC was isolated from Piper betle leaf (PBL) and verified by high-performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR), and gas chromatography-mass spectrometry (GC-MS). The cytotoxic effects of the standard drug 5-fluorouracil (5-FU), PBL water extract, and HC on HT-29 cells were measured after 24, 48, and 72 h of treatment. Cell cycle and apoptosis modulation by 5-FU and HC treatments were investigated up to 30 h. Changes in phosphorylated JNK (pJNK) and P38 (pP38) MAPK expression were observed up to 18 h. The half maximal inhibitory concentration (IC50) values of HC (30 μg/mL) and PBL water extract (380 μg/mL) were achieved at 24 h, whereas the IC50 of 5-FU (50 μmol/L) was obtained at 72 h. Cell cycle arrest at the G0/G1 phase in HC-treated cells was observed from 12 h onwards. Higher apoptotic cell death in HC-treated cells compared to 5-FU-treated cells (P<0.05) was observed. High expression of pJNK and pP38 MAPK was observed at 12 h in HC-treated cells, but not in 5-FU-treated HT-29 cells (P<0.05). It is concluded that HC induces cell cycle arrest and apoptosis of HT-29 cells, with these actions possibly mediated by JNK and P38 MAPK.
Fulltext Amino acid compounds released by the giant freshwater prawn Macrobrachium rosenbergii during ecdysis: a factor attracting cannibalistic behaviour?

Juneta-Nor AS, Noordin NM, Azra MN, Ma HY, Husin NM, Ikhwanuddin M

J Zhejiang Univ Sci B, 2020 10 13;21(10):823-834.
PMID: 33043647 DOI: 10.1631/jzus.B2000126

Ecdysis is a common phenomenon that happens throughout the life phase of the giant freshwater prawn Macrobrachium rosenbergii. It is vital to better understand the correlation between cannibalism and biochemical compound that exists during the moulting process. The objective of the present study was to determine the amino acid profile released by M. rosenbergii during the ecdysis process that promotes cannibalism. To accomplish this, changes in amino acid levels (total amino acid (TAA) and free amino acid (FAA)) of tissue muscle, exoskeleton, and sample water of culture medium from the moulting (E-stage) and non-moulting (C-stage) prawns were analysed using high-performance liquid chromatography (HPLC). Comparison study revealed that among the TAA compounds, proline and sarcosine of tissues from moulting prawn were found at the highest levels. The level of FAA from water that contains moulting prawns (E-stage) was dominated by tryptophan and proline. Significant values obtained in the present study suggested that these amino acid compounds act as a chemical cue to promote cannibalism in M. rosenbergii during ecdysis. The knowledge of compositions and compounds that were released during the moulting process should be helpful for better understanding of the mechanism and chemical cues that play roles on triggering cannibalism, and also for future dietary manipulation to improve feeding efficiencies and feeding management, which indirectly impacts productivity and profitability.
Fulltext Understanding and reducing the fear of COVID-19

Ng KH, Kemp R

J Zhejiang Univ Sci B, 2020 9 8;21(9):752-754.
PMID: 32893533 DOI: 10.1631/jzus.B2000228

The world is now plagued by a pandemic of unprecedented nature caused by a novel, emerging, and still poorly understood infectious disease, coronavirus disease 2019 (COVID-19) (Wu and McGoogan, 2020). In addition to the rapidly growing body of scientific and medical literature that is being published, extensive public reports and stories in both the traditional media and social media have served to generate fear, panic, stigmatization, and instances of xenophobia (Zarocostas, 2020).
Fulltext Anti-migratory effects of Piper betle leaf aqueous extract on cancer cells and its microtubule targeting properties

Looi ML, Wong AKH, Gnapragasan SA, Japri AZ, Rajedadram A, Pin KY

J Zhejiang Univ Sci B, 2020 9 8;21(9):745-748.
PMID: 32893531 DOI: 10.1631/jzus.B2000278

Piper betle (PB), also known as "betel" in Malay language, is a tropical Asian vine. PB leaves are commonly chewed by Asians along with betel quid. It contains phenols such as eugenol and hydroxychavicol along with chlorophyll, β-carotene, and vitamin C (Salehi et al., 2019). Extracts from PB leaves have various medicinal properties including anticancer, antioxidant, anti-inflammatory, and antibacterial effects (Salehi et al., 2019). Previous research has shown that PB induces cell cycle arrest at late S or G2/M phase and causes apoptosis at higher doses (Wu et al., 2014; Guha Majumdar and Subramanian, 2019). A combination of PB leaf extract has also been shown to enhance the cytotoxicity of the anticancer drug, 5-fluorouracil (5-FU), in cancer cells (Ng et al., 2014).
Fulltext Purification and identification of novel cytotoxic oligopeptides from soft coral Sarcophyton glaucum

Quah Y, Mohd Ismail NI, Ooi JLS, Affendi YA, Abd Manan F, Teh LK, et al.

J Zhejiang Univ Sci B, 2019 1 8;20(1):59-70.
PMID: 30614230 DOI: 10.1631/jzus.B1700586

Globally, peptide-based anticancer therapies have drawn much attention. Marine organisms are a reservoir of anticancer peptides that await discovery. In this study, we aimed to identify cytotoxic oligopeptides from Sarcophyton glaucum. Peptides were purified from among the S. glaucum hydrolysates produced by alcalase, chymotrypsin, papain, and trypsin, guided by a methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay on the human cervical cancer (HeLa) cell line for cytotoxicity evaluation. Purification techniques adopted were membrane ultrafiltration, gel filtration chromatography, solid phase extraction (SPE), and reversed-phase high-performance liquid chromatography (RP-HPLC). Purified peptides were identified by de novo peptide sequencing. From papain hydrolysate, three peptide sequences were identified: AGAPGG, AERQ, and RDTQ (428.45, 502.53, and 518.53 Da, respectively). Peptides synthesized from these sequences exhibited cytotoxicity on HeLa cells with median effect concentration (EC50) values of 8.6, 4.9, and 5.6 mmol/L, respectively, up to 5.8-fold stronger than the anticancer drug 5-fluorouracil. When tested at their respective EC50, AGAPGG, AERQ, and RDTQ showed only 16%, 25%, and 11% cytotoxicity to non-cancerous Hek293 cells, respectively. In conclusion, AERQ, AGAPGG, and RDTQ are promising candidates for future development as peptide-based anticancer drugs.
Fulltext Towards precision medicine: from quantitative imaging to radiomics

Acharya UR, Hagiwara Y, Sudarshan VK, Chan WY, Ng KH

J Zhejiang Univ Sci B, 2018 1 9;19(1):6-24.
PMID: 29308604 DOI: 10.1631/jzus.B1700260

Radiology (imaging) and imaging-guided interventions, which provide multi-parametric morphologic and functional information, are playing an increasingly significant role in precision medicine. Radiologists are trained to understand the imaging phenotypes, transcribe those observations (phenotypes) to correlate with underlying diseases and to characterize the images. However, in order to understand and characterize the molecular phenotype (to obtain genomic information) of solid heterogeneous tumours, the advanced sequencing of those tissues using biopsy is required. Thus, radiologists image the tissues from various views and angles in order to have the complete image phenotypes, thereby acquiring a huge amount of data. Deriving meaningful details from all these radiological data becomes challenging and raises the big data issues. Therefore, interest in the application of radiomics has been growing in recent years as it has the potential to provide significant interpretive and predictive information for decision support. Radiomics is a combination of conventional computer-aided diagnosis, deep learning methods, and human skills, and thus can be used for quantitative characterization of tumour phenotypes. This paper discusses the overview of radiomics workflow, the results of various radiomics-based studies conducted using various radiological images such as computed tomography (CT), magnetic resonance imaging (MRI), and positron-emission tomography (PET), the challenges we are facing, and the potential contribution of radiomics towards precision medicine.
Fulltext A comparative study of conventional and supercritical fluid extraction methods for the recovery of secondary metabolites from Syzygium campanulatum Korth

Memon AH, Hamil MS, Laghari M, Rithwan F, Zhari S, Saeed MA, et al.

J Zhejiang Univ Sci B, 2016 Sep;17(9):683-982.
PMID: 27604860 DOI: 10.1631/jzus.B1600019

Syzygium campanulatum Korth is a plant, which is a rich source of secondary metabolites (especially flavanones, chalcone, and triterpenoids). In our present study, three conventional solvent extraction (CSE) techniques and supercritical fluid extraction (SFE) techniques were performed to achieve a maximum recovery of two flavanones, chalcone, and two triterpenoids from S. campanulatum leaves. Furthermore, a Box-Behnken design was constructed for the SFE technique using pressure, temperature, and particle size as independent variables, and yields of crude extract, individual and total secondary metabolites as the dependent variables. In the CSE procedure, twenty extracts were produced using ten different solvents and three techniques (maceration, soxhletion, and reflux). An enriched extract of five secondary metabolites was collected using n-hexane:methanol (1:1) soxhletion. Using food-grade ethanol as a modifier, the SFE methods produced a higher recovery (25.5%‒84.9%) of selected secondary metabolites as compared to the CSE techniques (0.92%‒66.00%).
Fulltext Effects of temperature and diet on length-weight relationship and condition factor of the juvenile Malabar blood snapper (Lutjanus malabaricus Bloch & Schneider, 1801)

Mazumder SK, Das SK, Bakar Y, Ghaffar MA

J Zhejiang Univ Sci B, 2016 Aug;17(8):580-90.
PMID: 27487803 DOI: 10.1631/jzus.B1500251

In this study we aimed to analyze the effects of water temperature and diet on the length-weight relationship and condition of juvenile Malabar blood snapper Lutjanus malabaricus over a 30-d experimental period. The experiment was conducted in the laboratory using a flow-through-sea-water system. The fish were subjected to four different temperatures (22, 26, 30, and 34 °C) and two diets (commercial pellet and natural shrimp). Fish were fed twice daily. L. malabaricus exhibited negative allometric growth (b<3) at the beginning of the experiment (Day 0) at all temperatures and both diets except for 22 °C fed with shrimp, which showed isometric growth (b=3). Conversely, at the end of the experiment (Day 30) fish showed isometric growth (b=3) at 30 °C fed with the pellet diet, indicating that the shape of the fish did not change with increasing weight and length, and a positive allometric growth (b>3) at 30 °C fed with shrimp diet, which indicated that fish weight increases faster than their length. The rest of the temperatures represented negative allometric growth (b<3) on both diet, meaning that fish became lighter with increasing size. The condition factors in the initial and final measurements were greater than 1, indicating the state of health of the fish, except for those fed on a pellet diet at 34 °C. However, the best condition was obtained at 30 °C on both diets. Nevertheless, diets did not have a significant effect on growth and condition of juvenile L. malabaricus. The data obtained from this study suggested culturing L. malabaricus at 30 °C and feeding on the pellet or shrimp diet, which will optimize the overall production and condition of this commercially important fish species.
Fulltext Therapeutic radionuclides in nuclear medicine: current and future prospects

Yeong CH, Cheng MH, Ng KH

J Zhejiang Univ Sci B, 2014 Oct;15(10):845-63.
PMID: 25294374 DOI: 10.1631/jzus.B1400131

The potential use of radionuclides in therapy has been recognized for many decades. A number of radionuclides, such as iodine-131 ((131)I), phosphorous-32 ((32)P), strontium-90 ((90)Sr), and yttrium-90 ((90)Y), have been used successfully for the treatment of many benign and malignant disorders. Recently, the rapid growth of this branch of nuclear medicine has been stimulated by the introduction of a number of new radionuclides and radiopharmaceuticals for the treatment of metastatic bone pain and neuroendocrine and other malignant or non-malignant tumours. Today, the field of radionuclide therapy is enjoying an exciting phase and is poised for greater growth and development in the coming years. For example, in Asia, the high prevalence of thyroid and liver diseases has prompted many novel developments and clinical trials using targeted radionuclide therapy. This paper reviews the characteristics and clinical applications of the commonly available therapeutic radionuclides, as well as the problems and issues involved in translating novel radionuclides into clinical therapies.
Fulltext Piper betle leaf extract enhances the cytotoxicity effect of 5-fluorouracil in inhibiting the growth of HT29 and HCT116 colon cancer cells

Ng PL, Rajab NF, Then SM, Mohd Yusof YA, Wan Ngah WZ, Pin KY, et al.

J Zhejiang Univ Sci B, 2014 Aug;15(8):692-700.
PMID: 25091987 DOI: 10.1631/jzus.B1300303

OBJECTIVE: The combination effect of Piper betle (PB) and 5-fluorouracil (5-FU) in enhancing the cytotoxic potential of 5-FU in inhibiting the growth of colon cancer cells was investigated.
METHODS: HT29 and HCT116 cells were subjected to 5-FU or PB treatment. 5-FU and PB were then combined and their effects on both cell lines were observed after 24 h of treatment. PB-5-FU interaction was elucidated by isobologram analysis. Apoptosis features of the treated cells were revealed by annexin V/PI stain. High-performance liquid chromatography (HPLC) was performed to exclude any possible chemical interaction between the compounds.
RESULTS: In the presence of PB extract, the cytotoxicity of 5-FU was observed at a lower dose (IC50 12.5 µmol/L) and a shorter time (24 h) in both cell lines. Both cell lines treated with 5-FU or PB alone induced a greater apoptosis effect compared with the combination treatment. Isobologram analysis indicated that PB and 5-FU interacted synergistically and antagonistically in inhibiting the growth of HT29 and HCT116 cells, respectively.
CONCLUSIONS: In the presence of PB, a lower dosage of 5-FU is required to achieve the maximum drug effect in inhibiting the growth of HT29 cells. However, PB did not significantly reduce 5-FU dosage in HCT116 cells. Our result showed that this interaction may not solely contribute to the apoptosis pathway.
KEYWORDS: 5-Fluorouracil; Herb-drug interaction; Isobologram analysis; Piper betle L.; Piperaceae
Fulltext Anti-adipogenic effects of extracts of Ficus deltoidea var. deltoidea and var. angustifolia on 3T3-L1 adipocytes

Woon SM, Seng YW, Ling AP, Chye SM, Koh RY

J Zhejiang Univ Sci B, 2014 Mar;15(3):295-302.
PMID: 24599694 DOI: 10.1631/jzus.B1300123

This study examined the anti-adipogenic effects of extracts of Ficus deltoidea var. deltoidia and var. angustifolia, a natural slimming aid, on 3T3-L1 adipocytes.
Fulltext Immunocytochemical localization of carbonic anhydrase in the pseudobranch tissue of the rainbow trout Oncorhynchus mykiss

Rahim SM, Mazlan AG, Simon KD, Delaunoy JP, Laurent P

J Zhejiang Univ Sci B, 2014 Feb;15(2):194-200.
PMID: 24510712 DOI: 10.1631/jzus.B1200297

Pseudobranch function has long interested scientists, but its role has yet to be elucidated. Several studies have suggested that pseudobranchs serve respiratory, osmoregulatory, and sensory functions. This work investigated the immunolocalization of pseudobranch carbonic anhydrase (CA) in the teleost fish species rainbow trout (Oncorhynchus mykiss) to clarify its physiological function. CA was purified from rainbow trout gills O. mykiss and specific antibodies were raised. Immunoblotting between tissue homogenates of pseudobranch and gill CA antibodies showed specific immunostaining with only one band corresponding to CA in the pseudobranch homogenate. Results of immunohistochemical technique revealed that CA was distributed within pseudobranch cells and more precisely in the apical parts (anti-vascular) of cells. The basal (vascular) parts of cells, tubular system, blood capillaries, and pillar cells were not immunostained. Immunocytochemistry confirmed these results and showed that some CA enzyme was cytoplasmic and the remainder was linked to membranous structures. The results also showed that the lacunar tissue layers did not display immunoperoxidase activity. Our results indicated that pseudobranch CA may have a function related to the extracellular medium wherein CA intervenes with the mechanism of stimulation of afferent nerve fibers.
Fulltext Morphological and biochemical responses of Oryza sativa L. (cultivar MR219) to ion beam irradiation

Ling AP, Ung YC, Hussein S, Harun AR, Tanaka A, Yoshihiro H

J Zhejiang Univ Sci B, 2013 Dec;14(12):1132-43.
PMID: 24302713 DOI: 10.1631/jzus.B1200126

Heavy ion beam, which has emerged as a new mutagen in the mutation breeding of crops and ornamental plants, is expected to result in the induction of novel mutations. This study investigates the morphological and biochemical responses of Oryza sativa toward different doses of carbon ion beam irradiation.
Fulltext Comparative effects of plant growth regulators on leaf and stem explants of Labisia pumila var. alata

Ling AP, Tan KP, Hussein S

J Zhejiang Univ Sci B, 2013 Jul;14(7):621-31.
PMID: 23825148 DOI: 10.1631/jzus.B1200135

OBJECTIVE: Labisia pumila var. alata, commonly known as 'Kacip Fatimah' or 'Selusuh Fatimah' in Southeast Asia, is traditionally used by members of the Malay community because of its post-partum medicinal properties. Its various pharmaceutical applications cause an excessive harvesting and lead to serious shortage in natural habitat. Thus, this in vitro propagation study investigated the effects of different plant growth regulators (PGRs) on in vitro leaf and stem explants of L. pumila.
METHODS: The capabilities of callus, shoot, and root formation were evaluated by culturing both explants on Murashige and Skoog (MS) medium supplemented with various PGRs at the concentrations of 0, 1, 3, 5, and 7 mg/L.
RESULTS: Medium supplemented with 3 mg/L indole-3-butyric acid (IBA) showed the optimal callogenesis from both leaf and stem explants with (72.34 ± 19.55)% and (70.40 ± 14.14)% efficacy, respectively. IBA was also found to be the most efficient PGR for root induction. A total of (50.00 ± 7.07)% and (77.78 ± 16.47)% of root formation were obtained from the in vitro stem and leaf explants after being cultured for (26.5 ± 5.0) and (30.0 ± 8.5) d in the medium supplemented with 1 and 3 mg/L of IBA, respectively. Shoot formation was only observed in stem explant, with the maximum percentage of formation ((100.00 ± 0.00)%) that was obtained in 1 mg/L zeatin after (11.0 ± 2.8) d of culture.
CONCLUSIONS: Callus, roots, and shoots can be induced from in vitro leaf and stem explants of L. pumila through the manipulation of types and concentrations of PGRs.
Acute and subacute oral toxicity of Litsea elliptica Blume essential oil in rats

Budin SB, Siti Nor Ain SM, Omar B, Taib IS, Hidayatulfathi O

J Zhejiang Univ Sci B, 2012 Oct;13(10):783-90.
PMID: 23024045

Litsea elliptica Blume has been traditionally used to treat headache, fever, and stomach ulcer, and has also been used as an insect repellent. The acute and subacute toxicities of L. elliptica essential oil were evaluated orally by gavage in female Sprague-Dawley rats. For the acute toxicity study, L. elliptica essential oil was administered in doses from 500 to 4000 mg/kg (single dose), and in the subacute toxicity test, the following doses were used: 125, 250, and 500 mg/kg, for 28 consecutive days. In the acute toxicity study, L. elliptica essential oil caused dose-dependent adverse behaviours and mortality. The median lethal dose value was 3488.86 mg/kg and the acute non-observed-adversed-effect level value was found to be 500 mg/kg. The subacute toxicity study of L. elliptica essential oil did not reveal alterations in body weight, and food and water consumptions. The haematological and biochemical analyses did not show significant differences between control and treated groups in most of the parameters examined, except for the hemoglobin, mean cell hemoglobin concentration, mean cell volume, mean cell hemoglobin, serum albumin, and serum sodium. However, these differences were still within the normal range. No abnormalities or histopathological changes were observed in the liver, pancreatic islet of Langerhans, and renal glomerulous and tubular cells of all treated groups. In conclusion, L. elliptica essential oil can be classified in the U group, which is defined as a group unlikely to present an acute hazard according to World Health Organization (WHO) classification.
Absence of postzygotic isolating mechanisms: evidence from experimental hybridization between two species of tropical sea urchins

Rahman MA, Uehara T, Arshad A, Yusoff FM, Shamsudin MN

J Zhejiang Univ Sci B, 2012 Oct;13(10):797-810.
PMID: 23024047

Two reef margin species of tropical sea urchins, Echinometra sp. C (Ec) and Echinometra oblonga (Eo), occur sympatrically on Okinawa intertidal reefs in southern Japan. Hybridization between these species was examined through a series of cross-fertilization experiments. At limited sperm concentrations, where conspecific crosses reached near 100% fertilization, both heterospecific crosses showed high fertilization rates (81%-85%). The compatibility of the gametes demonstrated that if gamete recognition molecules are involved in fertilization of these species, they are not strongly species-specific. We found that conspecific crosses reached peak fertilization levels much faster than did heterospecific crosses, indicating the presence of a prezygotic barrier to hybridization in the gametes. Larval survival, metamorphosis, and juvenile and adult survival of hybrid groups were nearly identical to those of their parent species. Hybrids from crosses in both directions developed normally through larval stages to sexually mature adults, indicating that neither gametic incompatibility nor hybrid inviability appeared to maintain reproductive isolation between these species. In adults, Ec×Ec crosses gave the highest live weight, followed by Eo (ova)×Ec (sperm), Ec (ova)×Eo (sperm), and Eo×Eo. Other growth performance measures (viz., test size, Aristotle's lantern length, and gonad index) of hybrid groups and their parental siblings showed the same trends. The phenotypic color patterns of the hybrids were closer to the maternal coloration, whereas spine length, tube-foot and gonad spicule characteristics, pedicellaria valve length, and gamete sizes showed intermediate features. Adult F(1) hybrids were completely fertile and displayed high fertilization success in F(1) backcrosses, eliminating the likelihood that hybrid sterility is a postzygotic mechanism of reproductive isolation. Conversely, intensive surveys failed to find hybrid individuals in the field, suggesting the lack or rarity of natural hybridization. This strongly suggests that reproductive isolation is achieved by prezygotic isolating mechanism(s). Of these mechanisms, habitat segregation, gamete competition, differences in spawning times, gametic incompatibility or other genetic and non-genetic factors appear to be important in maintaining the integrity of these species.

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