Mitochondrial Cyt Research Articles

Hydrogen sulfide negatively regulates cd-induced cell death in cucumber (Cucumis sativus L) root tip cells

BackgroundHydrogen sulfide (H2S) is a gas signal molecule involved in regulating plants tolerance to heavy metals stress. In this study, we investigated the role of H2S in cadmium-(Cd-) induced cell death of root tips of cucumber seedlings.ResultsThe results showed that the application of 200 μM Cd caused cell death, increased the content of reactive oxygen species (ROS), chromatin condensation, the release of Cytochrome c (Cyt c) from mitochondria and activated caspase-3-like protease. Pretreatment of seedlings with 100 μM sodium hydrogen sulfide (NaHS, a H2S donor) effectively alleviated the growth inhibition and reduced cell death of root tips caused by Cd stress. Additionally, NaHS + Cd treatment could decrease the ROS level and enhanced antioxidant enzyme activity. Pretreatment with NaHS also inhibited the release of Cyt c from the mitochondria, the opening of the mitochondrial permeability transition pore (MPTP), and the activity of caspase-3-like protease in the root tips of cucumber seedling under Cd stress.ConclusionH2S inhibited Cd-induced cell death in cucumber root tips by reducing ROS accumulation, activating the antioxidant system, inhibiting mitochondrial Cyt c release and reducing the opening of the MPTP. The results suggest that H2S is a negative regulator of Cd-induced cell death in the root tips of cucumber seedling.

Completing the genetic puzzle of the reed warbler complex: insights from Italy

ABSTRACT Capsule Analysis of mitochondrial DNA from reed warblers sampled in Italy reveals that they are most similar to the nominate race of European Reed Warbler Acrocephalus scirpaceus scirpaceus. Aim To fill gaps in our knowledge of the distribution of European/African Reed Warblers Acrocephalus scirpaceus/baeticatus in the Mediterranean region through genetic analysis of populations from the key biogeographic areas of Sicily, Sardinia and mainland Italy. Methods We assessed the genetic placement of Italian reed warbler populations within a comprehensive dataset of this species complex, by sampling these birds from principal peninsular and insular breeding populations (for a total of 17 samples) and by comparing their mitochondrial cyt b sequences with those available in GenBank-NCBI. Results The final dataset included 171 cyt b sequences (763 base pair long). The analysis showed samples from Italy had a high similarity with the European Reed Warbler A. s. scirpaceus subspecies. This fills an important information gap in the phylogeography of the group. Conclusion The genetic cohesion shown between Italian and other European populations may denote a structuring in at least three distinct refugia in the Western Palearctic: Iberia for A. baeticatus ambiguus, the Caucasus Mountains for A. s. fuscus, and both Italy and the Balkans for A. s. scirpaceus.

Old and Cosmopolite: Molecular Phylogeny of Tropical–Subtropical Kites (Aves: Elaninae) with Taxonomic Implications

Kites of the Elaninae group are small and medium-sized, mostly tropical raptors traditionally considered as an early diverged subfamily of the Accipitridae. We used nucleotide sequences of three genetic markers (mitochondrial Cyt b and COI, nuclear RAG-1) to reconstruct the phylogenetic relationships of the Elaninae, other kites, and representatives of different families of diurnal raptors. Our results confirm the basal position of Elaninae, separated the latest in Early Miocene, including Chelictinia riocourii, which was not sequenced before and belongs to this group. Not only DNA data but also cytological, morphological, and ecological data show the singularity of Elaninae. We suggest elevating this group to family level as Elanidae within the order Accipitriformes. It includes Gampsonyx swainsonii as a monotypic subfamily because of distinctive traits and DNA sequence data. Taxonomic implications for other macrogroups of Accipitriformes are discussed.

Linker Histone H1.2 Directly Activates BAK through the K/RVVKP Motif on the C-Terminal Domain

H1.2 is a key mediator of apoptosis following DNA double-strand breaks. The link between H1.2 and canonical apoptotic pathways is unclear. One study found that H1.2 stimulates cytochrome c (Cyt c) release; in contrast, apoptosis-inducing factor was found to be released in another study. The C-terminal domain (CTD) of H1.2 has been implicated in the latter pathway, but activation of the proapoptotic protein BCL-2 homologous antagonist/killer (BAK) is a common denominator in both pathways. This study aimed to determine whether the CTD of H1.2 is also responsible for mitochondrial Cyt c release and whether a previously identified K/RVVKP motif in the CTD mediates the response. This study investigated if H1.2 mediates apoptosis induction through direct interaction with BAK. We established that the CTD of H1.2 stimulates mitochondrial Cyt c release in vitro in a mitochondrial permeability transition-independent manner and that the substitution of a single valine with threonine in the K/RVVKP motif abolishes Cyt c release. Additionally, we showed that H1.2 directly interacts with BAK with weak affinity and that the CTD of H1.2 mediates this binding. Using two 20-amino acid peptides derived from the CTD of H1.2 and H1.1 (K/RVVKP motif inclusive), we determined the main residues involved in the direct interaction with BAK. We propose that H1.2 operates through the K/RVVKP motif by directly activating BAK through inter- and intramolecular interactions. These findings expand the view of H1.2 as a signal-transducing molecule that can activate apoptosis in a BAK-dependent manner.

Melatonin improves cryopreservation of ram sperm by inhibiting mitochondrial permeability transition pore opening.

Cryopreservation damages permeability of sperm mitochondrial membranes, with formation of a mitochondrial permeability transition pore (mPTP). Mitochondria are both a primary synthesis site and principle target for melatonin, which can directly inhibit mPTP formation. The objective was to determine effects of melatonin on mPTP opening of frozen-thawed ram sperm and elucidate underlying pathways by antagonist and agonists of melatonin receptors (MTs), and antagonists of PI3K and GSK 3β treatments; furthermore, plasma membrane integrity, mitochondrial membrane potential (ΔΨm), mitochondrial cytochrome c (Cytc) release and fertilization were analysed to assess the effect of mPTP status mediated by melatonin on quality of frozen-thawed sperm. Fresh ram semen was diluted in glucose-egg yolk buffer with 0 or 10-7 M melatonin (frozen and frozen+melatonin groups, respectively) and slow-frozen. In frozen-thawed sperm, melatonin added at initiation of 4°C equilibration was most effective for inhibiting mPTP opening, decreasing peptidyl-prolyl-cis/trans isomerase activity of cyclophilin D and increasing plasma membrane integrity, ΔΨm, mitochondrial Cyt c concentration and fertilizing ability (p<.05). In a mechanistic study, the melatonin receptor (MT)1 antagonist eliminated inhibition of melatonin on mPTP opening, whereas MT1 agonist had opposite effects (p<.05). Neither MT2 antagonist nor agonist had significant effect, but PI3K and/or GSK 3β antagonist decreased inhibition of MT1 agonist on mPTP opening (p<.05). In conclusion, melatonin improved sperm cryopreservation, perhaps by acting on MT1 via the PI3K-Akt-GSK 3β pathway to inhibit mPTP opening.

Ancient DNA of northern China Hystricidae sub-fossils reveals the evolutionary history of old world porcupines in the Late Pleistocene

BackgroundOld World porcupines (Family: Hystricidae) are the third-largest rodents and inhabit southern Europe, Asia, and most regions of Africa. They are a typical indicator of warm climate and their distribution is restricted to tropical and subtropical zones. In China, porcupines are widely distributed in southern areas of the Yangtze River. However, fossil remains have been identified in a few sites in northern China, among which Tianyuan Cave—near Zhoukoudian site—represents the latest known porcupine fossil record. So far, studies have focused mainly on porcupines’ husbandry and domestication but little is known about their intrafamilial phylogenetic relationships and evolutionary history.ResultsIn this study, we sequence partial mitochondrial 12S rRNA and cyt b genes for seven Late Pleistocene porcupine individuals from Northern, Southern and Central China. Phylogenetic analyses show that the Tianyuan Cave porcupines, which had been morphologically identified as Hystrix subcristata, have a closer relationship to Hystrix brachyura.ConclusionTogether with morphological adaptation characteristics, associated fauna, and climate change evidence, the molecular results reveal that a Late Quaternary extirpation has occurred during the evolutionary history of porcupines.

Molecular systematics and phylogeography of the genus Alburnus Rafinesque, 1820 (Teleostei, Leuciscidae) in Turkey

In this study, the phylogeny of Alburnus genus distributed in Turkish freshwaters was performed by analyzing mitochondrial cyt b gene (1141 bp) and COI gene (1551 bp) sequences from 1172 samples representing 112 populations of 24 species through their geographical distribution. According to our findings, 20 valid species are distributed in Turkey of which 18 have already been known. While six Alburnus species (A. battalgilae, A. istanbulensis, A. carinatus, A. schischkovi, A. nasreddini ve A. adanensis) have been synonomized, two new species (Alburnus sp.1 and Alburnus sp.2) from Dicle River and Çapraz Stream/Susurluk River have been identified. Extinct species such as A. akili and A. nicaeensis have not been observed in situ. Phylogenetic tree topologies and haplotype network of the 119 cyt b and 80 COI haplotypes detected in Alburnus species have indicated a consensus tree topology containing twenty lineages, each of corresponding to one species, and three Alburnus haplogroups corresponding to the geographical origins: Eastern Anatolia (I), Mediterranean (2) and Western & Northern Anatolia (3). The results indicate that the divergence between those haplogroups may have occurred during the Middle Miocene-Middle Pleistocene periods (from 14.9 to 5.29 million years).

Phylogeography of the genus Chondrostoma Agassiz, 1835 (Teleostei: Leuciscidae) in Anatolia, as inferred from mitochondrial DNA analysis

The molecular phylogeny of the Anatolian species of the genus Chondrostoma (Cypriniformes: Leuciscidae) was studied by sequencing of the mitochondrial cyt b gene. A total of 144 tissue samples from 14 described species were collected from 35 different localities in Turkey. In total, 74 cyt b haplotypes were identified. Haplotype network construction and phylogenetic analysis allocated all 14 species to two main haplogroups that are congruent with those currently accepted on the basis of morphological characteristics. The first haplogroup, the Nasus lineage, is distributed across Caucasia and the Black Sea and Marmara regions and is also represented in the Aegean basin. The second haplogroup, the Regium lineage, is distributed from eastern Anatolia to the Mediterranean Sea basin. Based on estimates generated in this study, the radical divergence between the main Chondrostoma haplogroups in Turkey occurred during the late Miocene (Tortonian stage) and late Pliocene Period, approximately 5.82 Mya (8.48–3.45 Mya). The splitting of the Nasus lineage occurred approximately 5.11 Mya, in the early Pliocene, and the Regium Lineage diverged from its sister group, the Nasus lineage approximately 4.77 Mya. The primary diversification events for the species belonging to these two haplogroups occurred in the Late Pliocene to Early Pleistocene which coincides with the uplifting of the Anatolian plateau about 2.5–3.0 Mya.

Scoulerine promotes cell viability reduction and apoptosis by activating ROS-dependent endoplasmic reticulum stress in colorectal cancer cells

Scoulerine, an isoquinoline alkaloid isolated from Corydalis plants, has been reported to possess potent anti-proliferative and pro-apoptotic function in cancer cells. However, the effects and underlying mechanisms of scoulerine on colorectal cancer (CRC) progression remain elusive. CCK-8 and LDH assays were used to evaluate cell viability. Apoptosis was assessed by flow cytometry analysis, caspase-3/7 activity assay, and Western blot analysis of Bax, Bcl-2 and cytochrome c (Cyt C) expression. Oxidative stress level was examined by measuring reactive oxygen species (ROS) and glutathione (GSH) contents and superoxide dismutase (SOD) activity. Endoplasmic reticulum (ER) stress activation was detected by Western blot analysis of glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) expression. Results showed that scoulerine dose-dependently suppressed CRC cell viability. Scoulerine induced apoptosis and increased caspase-3/7 activity in CRC cells. Bax and cytosolic Cyt C expression was enhanced while Bcl-2 and mitochondrial Cyt C expression was reduced in scoulerine-treated CRC cells. Additionally, scoulerine induced oxidative damage in CRC cells by increasing ROS generation and reducing GSH content and SOD activity. Scoulerine activated ER stress, as evidenced by the increased GRP78 and CHOP expression in CRC cells. Interestingly, blocking ROS production by ROS scavenger N-acetyl-cysteine (NAC) attenuated scoulerine-induced ER stress. Inhibition of ER stress by 4-phenyl butyric acid (4-PBA) abolished scoulerine-induced ROS generation in CRC cells. Blockage of ROS and ER stress attenuated scoulerine-induced cell viability reduction and apoptosis in CRC cells. In conclusion, scoulerine promoted cell viability reduction and apoptosis by activating ROS-dependent ER stress in CRC cells.

Identification of hybrid green peafowl using mitochondrial and nuclear markers

Green peafowl (Pavo muticus), a representative of the Southeast Asia Phasianidae, is assessed as globally endangered with populations that are declining. It is highly sensitive to human disturbance and habitat change, two threats that are unlikely to be removed soon in present range countries, and that increase the need for long-term continuous effort for in situ conservation. Captive breeding for reintroduction is potentially an important measure for localized population restoration. But green peafowl can hybridize with blue peafowl (P. cristatus) and produce fertile offspring that can backcross with ancestral species. Hybrids backcrossed to green peafowl for a few generations are indistinguishable from pure green peafowl in appearance. Such hybrids must be excluded from captive breeding programs to avoid anthropogenic genetic contamination. In this study, we developed technology to identify hybrids using mitochondrial Cyt b and COI gene sequences, and 16 microsatellites. We studied maternal species assignment for excluding individuals of the blue peafowl maternal line and genetic clustering analysis to exclude individuals containing blue peafowl nuclear genes. Maternal species assignment was accomplished using K2P pairwise genetic distance computed using transition nucleotide substitution. Nuclear genetic clustering analysis was performed using STRUCTURE (assuming K = 2) and NewHybrids. The method was validated using reference pure green (n = 18) and blue (n = 21) peafowl and known hybrids (n = 12). Two criteria were set up to discriminate hybrids based on the probability that an individual is either pure green or pure blue peafowl at 95% confidence. This method was applied to identification of 59 peafowl of ambiguous genetic composition. Our study provides an example for establishing a purity test workflow for other species that can hybridize and has potential for use in reintroduction of genetically pure green peafowl.

Taxonomic assessment of the Malayan water shrew Chimarrogale hantu Harrison, 1958 and reclassification to the genus Crossogale

This study reassesses the taxonomic status of the Malayan water shrew, Chimarrogale hantu Harrison 1958 as a distinct species and reclassifies it as the genus Crossogale through genetic and morphological analyses. We construct phylogenetic trees using both mitochondrial (cyt b) and nuclear markers (ApoB and RAG2). Chimarrogale hantu formed a distinct clade that diverged from other Chimarrogale species and other genera in the tribe Nectogalini (Nectogale, Neomys, Pseudosoriculus, Episoriculus, Chodsigoa, and Soriculus clades). The C. hantu clade is closely related to the Bornean water shrew, Crossogale phaeura with a K2P genetic distance of 7%. High K2P genetic distance (> 15% K2P) was also observed when compared to other Chimarrogale species (C. platycephalus, C. himalayica, C. styani, C. varennei and C. leander) as well as other genera from the tribe Nectogalini (> 20% K2P). We proved that C. hantu genetically and morphologically merits species rank. We suggest classifying C. hantu in the genus Crossogale along with Cr. phaeura, while the rest of the Asiatic shrews remain in the genus Chimarrogale. Future comprehensive reviews of the Sundaic water shrew group should include Chimarrogale sumatrana from Sumatra, Indonesia, for a comprehensive understanding on the relationship of water shrews between Sundaland and the rest of the Asiatic region.

Role of mitochondrial damage in cadmium-induced cell apoptosis and DNA damage of hepatocytes

To investigate the role of mitochondrial damage mediated by reactive oxidative species(ROS) in cadmium-induced cell apoptosis and DNA damage of L02 hepatocytes, so as to provide experimental basis for the subsequent study and protection of people exposed to Cd. The L02 hepatocytes were cultured in vitro treated with 0-90 μmol/L Cd for 24 h, and the methylthiazolyldiphenyltetrazoliumbromide assay was used to detect the cell viability. The colony formation assay, flow cytometry, comet assay, 2', 7'-dichlorofluorescein diacetate, MitoTracker Red CMXRos and 10-N-nonyl-acridine-orange, mitochondrial membrane potential detection kit(JC-1) and adenosine triphosphate(ATP) assay kits and Western Blot were used to investigate cell growth and proliferation, cell apoptosis, DNA damage, ROS levels, mitochondrial morphology, mitochondrial membrane potential, mitochondrial mass, ATP content and related proteins after the cells exposed to 0, 20, 40 μmol/L Cd for 24 h. The cells were pretreated with vitamin C before adding Cd exposure, and ROS levels, mitochondrial function, cell apoptosis, DNA damage and proteins were measured. The cell viability was significantly inhibited with the increase of Cd concentration and treatment time. The cells were treated with Cd for 24 h for further study according to the result of MTT assay. Compared with control group, the colony formation rate were 8. 23% and 6. 17% respectively in 20 and 40 μmol/L Cd treatment and the apoptosis rate were 15. 85% and 26. 26%, respectively. We also found that the B cell lymphoma/leukemia(Bcl-2) gene protein was significantly reduced, while the levels of Bcl-2 associated X protein(Bax) and cleaved cysteine aspastic acid-specific protease 3(cleaved-caspase-3) were increased in a dose-dependent manner. Cd treatment also induced DNA damage and accumulation of intracellular ROS, accompanied by a mitochondrial morphological change, significant decrease in Δψm, mitochondrial mass, ATP content, mitochondrial cytochrome C(cyt c) and an increase in cytoplasmic cyt c expression(P&lt;0. 05). In addition, pretreatment with antioxidant vitamin C not only significantly increased cyt c, mitochondrial mass, ATP content and mitochondrial cyt c, but also reduced the expression of cytoplasmic cyt c(P&lt;0. 05), cell apoptosis and DNA damage induced by Cd. Cd exposure could induce ROS accumulation in L02 hepatocytes, which can lead to mitochondrial damage, and ultimately lead to cell apoptosis and DNA damage.

On-Site Detection of Tissues of Buffalo Origin by Loop-Mediated Isothermal Amplification (LAMP) Assay Targeting Mitochondrial Gene Sequences

Meat fraud and adulteration is a growing concern in the marketplace today. Species identification in food product is among primary duties of food office control. In this study, a one-step, species-specific, sensitive, and rapid loop-mediated isothermal amplification (LAMP) assay was developed for identification of tissues of buffalo origin. A set of six LAMP primers were designed by targeting mitochondrial cyt b gene. The reaction temperature of LAMP and component of reaction mixture were optimized as 64 °C amplification temperature, 6 U/μL concentration of Bst DNA polymerase, 3 mM concentration of MgSO4, 0.4 mM concentration of dNTP, 0.6 M concentration of betaine, and 1:6 ratio of outer and inner primer pair. Successful amplifications were confirmed using SYBR Green I dye and agarose gel electrophoresis. The specificity of assay was tested with DNA of cattle, goat, sheep, and pig where amplification was observed exclusively in buffalo. The analytical sensitivity of the LAMP assay for buffalo DNA detection was 0.1 pg. The applicability check of developed assay carried out on known/coded samples and binary meat admixture substantiated the accuracy of assay. Supernatant of tissue exposed to Phire Animal Tissue Direct PCR master mix treatment was observed rich enough with DNA of target species and successfully amplified under optimized LAMP reaction. The comparison of results on the samples undergone pre-standardized species-specific PCR taken as gold standard shown complete match with developed species-specific LAMP assay. Hence, the developed species-specific LAMP assays are effective in detection of tissue of buffalo origin.

Genetic diversity of wild wintering red-crowned crane (Grus japonensis) by microsatellite markers and mitochondrial Cyt B gene sequence in the Yancheng reserve

The red-crowned crane (Grus japonensis) is one of the most endangered cranes in the world, and its wild population is still declining. To characterize the genetic resources of East Asian migratory populations, we studied the genetic variation in wild red-crowned cranes at the Yancheng reserve. Based on a partial Cyt b gene sequence, 32 wild red-crowned crane samples were screened from 100 feathers with unknown sample information. Twelve haplotypes were detected using 32 wild red-crowned crane samples. Six pairs of published microsatellite primers were selected for genotyping. A total of 47 alleles were obtained, with an average of 7.8 alleles per locus. All microsatellite loci were highly polymorphic; the average polymorphic information content and expected heterozygosity were 0.721 ± 0.080 and 0.768 ± 0.071, respectively. These results show that the East Asian migratory population of wild red-crowned cranes exhibits high polymorphism. These data are useful for informing reintroduction efforts. The study results provide a basis for understanding the population genetic properties of an endangered crane.

Psilorhynchus nahlongthai, a new fish species (Teleostei: Psilorhynchidae) from the Brahmaputra drainage, northeast India.

Psilorhynchus nahlongthai, a new psilorhynchid fish, is described from the Diyung River, a tributary to the Kopili River (itself a southern tributary of the Brahmaputra drainage) in Assam, northeast India. It is placed in the Psilorhynchus balitora species group and can be easily distinguished from all other members of this group by a combination of the following characters: dense and prominent tuberculation on the head region; thick and long pre- and post-epiphyseal fontanelles on the neurocranium; 9 + 8 caudal-fin rays; and 34 (24 + 10) vertebrae. Genetic divergence between P. nahlongthai and members of the P. balitora species group from the Brahmaputra and neighbouring drainages, with K2P distances ranging 3.7%-14.7% and 7.4%-20.7% in the mitochondrial COI and cyt b gene datasets respectively, support its report as a new species.

Phylogeography of the banded butterflyfish, Chaetodon striatus, indicates high connectivity between biogeographic provinces and ecosystems in the western Atlantic

ABSTRACT Among the four butterflyfishes of the genus Chaetodon present in the western Atlantic, the banded butterflyfish Chaetodon striatus has the largest distribution range, spanning 44 degrees of latitude (from Massachusetts, USA to Santa Catarina, Brazil). Although the ecology of the banded butterflyfish has been well studied over its entire range, nothing is known about its phylogeography and how biogeographic barriers structure its populations. To assess the level of genetic connectivity among populations from distinct biogeographic provinces and environmental conditions, we collected samples from seven localities: Puerto Rico, in the Caribbean, and Tamandaré, Salvador, Abrolhos, Trindade Island, Arraial do Cabo and Florianópolis, in Brazil. One nuclear (rag 2) and two mitochondrial (control region and cyt b) molecular markers were sequenced. Our findings are consistent with a recent population expansion, around 30-120 thousand years ago, which was found for all populations. Haplotype network analyses point to the Caribbean as a refugium before the population expansion. Results show no geographic pattern of genetic diversity. Indeed, a lack of population structure was found and no isolation was observed across oceanographic barriers, as well as between coral and rocky reef ecosystems. Furthermore, no directionality in the migration pattern was found among populations. Since ecological and environmental characteristics are very diverse across such a vast geographic range, the lack of genetic differentiation suggests that C. striatus evolved ecological plasticity rather than local adaptation in the western Atlantic.

Population genetics and sympatric divergence of the freshwater gudgeon, Gobiobotia filifer, in the Yangtze River inferred from mitochondrial DNA.

The ecosystem and Pleistocene glaciations play important roles in population demography. The freshwater gudgeon, Gobiobotia filifer, is an endemic benthic fish in the Yangtze River and is a good model for ecological and evolutionary studies. This study aimed to decode the population structure of G. filifer in the Yangtze River and reveal whether divergence occurred before or after population radiation. A total of 292 specimens from eight locations in the upper and middle reaches of the Yangtze River were collected from 2014 to 2016 and analyzed via mitochondrial DNA Cyt b gene sequencing. A moderately high level of genetic diversity was found without structures among the population. However, phylogenetic and network topology showed two distinct haplotype groups, and each group contained a similar proportion of individuals from all sampled sites. This suggested the existence of two genetically divergent source populations in G. filifer. We deduced that a secondary contact of distinct glacial refugia was the main factor creating sympatric populations of G. filifer, and climate improvement promoted population expansion and colonization.

SS31 attenuates oxidative stress and neuronal apoptosis in early brain injury following subarachnoid hemorrhage possibly by the mitochondrial pathway

BackgroundSS31 has been shown to have neuroprotective effects in a number of neurological degenerative diseases. However, the mechanisms and its role of neuroprotection after subarachnoid hemorrhage (SAH) remain unexplored. The aim of the present study is to evaluate the neuroprotective effects of SS31 on early brain injury (EBI) induced by SAH in rats and the potential mechanisms of the protective effects of SS31. MethodsSprague-Dawley rats were randomly divided into four groups: Sham, SAH, SAH + vehicle, and SAH + SS31 groups. The SAH-induced prechiasmatic cistern rat model was established in this study. Neurological scores were evaluated at 24 h and 72 h after SAH. The brain edema, blood-brain barrier (BBB) permeability, neuronal apoptosis, malondialdehyde (MDA), glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities, as well as the expression of mitochondrial and cytosolic cytochrome C (Cyt C), and Bax were analyzed at 24 h after SAH. ResultsWhen compared with the vehicle-treated group, treatment with SS31 significantly reduced MDA levels and restored the activities of GPx and SOD in the temporal cortex following SAH when compared with the vehicle-treated group. In addition, the levels of mitochondrial Cyt C and Bax respectively increased and decreased by SS31 treatment. Moreover, SS31 treatment ameliorated brain edema and Evans blue dye extravasation, improved neurological deficits, and decreased neuronal apoptosis at 24 h after SAH. ConclusionOur data provides initial evidence that SS31 could alleviate EBI after SAH through its antioxidant property and ability in inhibiting neuronal apoptosis, likely by modulating the mitochondrial apoptotic pathway.

CYT B L15601 and H15748 primers for genetic identification of cetacean species

Dolphins and other cetacean are protected species whose capture is illegal in many countries. Despite the high mercury content accumulated by these animals, occasional episodes of illegal fishing occur to supply certain black markets with dolphin meat. When fishing authority inspections detect meat portions presumably of cetacean origin, it is particularly necessary to identify the species before taking the case to court. The mitochondrial cytochrome b gene has been used extensively to identify species. In forensic cases, due to the degradation of the samples, short amplicons are preferable. The universal cyt b L15601 and H15748 primers, previously tested in 63 species, have been used in this work on individuals of seven species of cetaceans: Balaenoptera physalus, Delphinus delphis, Globicephala melas, Phocoena phocoena, Stenella coeruleoalba, Tursiops truncatus and Ziphius cavirostris. These universal primers, with a short amplicon of 148 bp, have the advantage of identifying postmortem samples even when showing obvious signs of degradation. Our results showed that all tested species were correctly identified, except S. coeruleoalba and S. clymene. This is because S. clymene arose by hybrid speciation between S. coeruleoalba and S. longirostris. Therefore, their identification requires the analysis of both mitochondrial DNA and nuclear DNA. In summary, the identification of illegally obtained cetacean meat can be easily performed in forensic laboratories by analyzing a short fragment of the mitochondrial cyt b in all the studied species except in S. coeruleoalba and S. clymene and even when the remains were in an advanced stage of degradation.

Genetic Analysis of Turkish lynx (Lynx lynx) Based on Mitochondrial DNA Sequences

Genetic variability and phylogenetic relationships of the Eurasian lynx (Lynx lynx) have been reported by various researchers from species’ range. As genetic data of Turkish lynx comes from a single genetic study that only contains limited number of lynx samples from the Anatolian part of Turkey, there is still a lack of genetic information for the Eurasian lynx population distributed in Turkey. In this study, mitochondrial Cyt b (1140 bp) and COI (630 bp) sequences were obtained from eight Anatolian samples of the Eurasian lynx. The Anatolian lynx sequences were compared to those of conspecific populations published in GenBank. We found two COI (630 bp) and four Cyt b (1140 bp) haplotypes among the eight Anatolian lynx sequences. Despite the limited number of the Anatolian lynx sequences, Bayesian, Maximum Likelihood, Neighbor-Joining and Network analyses revealed that there was a significant genetic differentiation in the Eurasian lynx, in that at least there were two main mitochondrial lineages within the sampling area. Also, the present study suggested that the Anatolian lynx might have a relatively high genetic variability despite the scarce samples.