Phylo-geographic analysis of whitefly on the basis of mitochondrial cytochrome oxidase 1 gene / Análise filogeográfica da mosca-branca com base no gene citocromo oxidase 1 mitocondrial

Bemisia tabaci is a species complex that causes damage to its broad range of plant hosts through serious feeding. It transmits plant viruses of different groups to important agricultural crops. Some important cash crops of Pakistan are sugar cane, rice, tobacco and seed oil. It shows high genetic variability and is differentiated as races or biotypes. Biotypes are, biotype Q, biotype B, biotype B2, biotype M, biotype L, biotype A, biotype H, biotype C, biotype K, biotype N, biotype R, biotype E, biotype P, biotype J, biotype S, biotype AN. Although the current report based on the Bayesian study of mitochondrial cytohrome oxidase gene1 (CO1) DNA sequences has classified the different populations of whiteflies into twelve genetic groups which are Mediterranean, Sub-Saharan Africa silverleafing, Indian Ocean, Asia II, Asia I, Australia, New World, Italy, China, Sub-Saharan Africa non-silverleafing, Mediterranean/Asia Minor/Africa and Uganda sweet potato. Begomoviruses is largest group of viruses transmitted by B. tabaci and cause major diseases of crops such as tomato and chili leaf curl disease, cassava mosaic disease; yellow mosaic disease of legumes and cotton leaf curl disease. The main objective of current study is to inculpate knowledge regarding genetic diversity of whitefly in cotton fields across Pakistan via analysis of partial DNA sequence of mitochondrial gene Cytochrom Oxidase I (mtCO1).

Bemisia tabaci é um complexo de espécies que causa danos a uma ampla gama de hospedeiros vegetais por meio de alimentação séria. Ele transmite vírus de plantas de diferentes grupos para importantes safras agrícolas. Algumas safras comerciais importantes do Paquistão são cana-de-açúcar, arroz, tabaco e óleo de semente. Apresenta alta variabilidade genética e é diferenciado em raças ou biótipos. Os biótipos são: biótipo Q, biótipo B, biótipo B2, biótipo M, biótipo L, biótipo A, biótipo H, biótipo C, biótipo K, biótipo N, biótipo R, biótipo E, biótipo P, biótipo J, biótipo S, biótipo AN . Embora o relatório atual baseado no estudo bayesiano das sequências de DNA do gene 1 da oxidase do citocromo mitocondrial (CO1) tenha classificado as diferentes populações de moscas-brancas em doze grupos genéticos, que são Mediterrâneo, África Subsaariana com folha de prata, Oceano Índico, Ásia II, Ásia I, Austrália, Novo Mundo, Itália, China, África Subsaariana sem folha prateada, Batata-doce Mediterrâneo / Ásia Menor / África e Uganda. Os begomovírus são o maior grupo de vírus transmitidos por B. tabaci e causam as principais doenças de culturas, como a doença do cacho do tomate e da pimenta-malagueta, doença do mosaico da mandioca, doença do mosaico amarelo de leguminosas e doença do enrolamento da folha do algodão. O principal objetivo do presente estudo é inculpar conhecimento sobre a diversidade genética da mosca-branca em campos de algodão em todo o Paquistão por meio da análise da sequência parcial de DNA do gene mitocondrial Citocromo Oxidase I (mtCO1).

Phylo-geographic analysis of whitefly on the basis of mitochondrial cytochrome oxidase 1 gene

Abstract Bemisia tabaci is a species complex that causes damage to its broad range of plant hosts through serious feeding. It transmits plant viruses of different groups to important agricultural crops. Some important cash crops of Pakistan are sugar cane, rice, tobacco and seed oil. It shows high genetic variability and is differentiated as races or biotypes. Biotypes are, biotype Q, biotype B, biotype B2, biotype M, biotype L, biotype A, biotype H, biotype C, biotype K, biotype N, biotype R, biotype E, biotype P, biotype J, biotype S, biotype AN. Although the current report based on the Bayesian study of mitochondrial cytohrome oxidase gene1 (CO1) DNA sequences has classified the different populations of whiteflies into twelve genetic groups which are Mediterranean, Sub-Saharan Africa silverleafing, Indian Ocean, Asia II, Asia I, Australia, New World, Italy, China, Sub-Saharan Africa non-silverleafing, Mediterranean/Asia Minor/Africa and Uganda sweet potato. Begomoviruses is largest group of viruses transmitted by B. tabaci and cause major diseases of crops such as tomato and chili leaf curl disease, cassava mosaic disease; yellow mosaic disease of legumes and cotton leaf curl disease. The main objective of current study is to inculpate knowledge regarding genetic diversity of whitefly in cotton fields across Pakistan via analysis of partial DNA sequence of mitochondrial gene Cytochrom Oxidase I (mtCO1).

Resumo Bemisia tabaci é um complexo de espécies que causa danos a uma ampla gama de hospedeiros vegetais por meio de alimentação séria. Ele transmite vírus de plantas de diferentes grupos para importantes safras agrícolas. Algumas safras comerciais importantes do Paquistão são cana-de-açúcar, arroz, tabaco e óleo de semente. Apresenta alta variabilidade genética e é diferenciado em raças ou biótipos. Os biótipos são: biótipo Q, biótipo B, biótipo B2, biótipo M, biótipo L, biótipo A, biótipo H, biótipo C, biótipo K, biótipo N, biótipo R, biótipo E, biótipo P, biótipo J, biótipo S, biótipo AN . Embora o relatório atual baseado no estudo bayesiano das sequências de DNA do gene 1 da oxidase do citocromo mitocondrial (CO1) tenha classificado as diferentes populações de moscas-brancas em doze grupos genéticos, que são Mediterrâneo, África Subsaariana com folha de prata, Oceano Índico, Ásia II, Ásia I, Austrália, Novo Mundo, Itália, China, África Subsaariana sem folha prateada, Batata-doce Mediterrâneo / Ásia Menor / África e Uganda. Os begomovírus são o maior grupo de vírus transmitidos por B. tabaci e causam as principais doenças de culturas, como a doença do cacho do tomate e da pimenta-malagueta, doença do mosaico da mandioca, doença do mosaico amarelo de leguminosas e doença do enrolamento da folha do algodão. O principal objetivo do presente estudo é inculpar conhecimento sobre a diversidade genética da mosca-branca em campos de algodão em todo o Paquistão por meio da análise da sequência parcial de DNA do gene mitocondrial Citocromo Oxidase I (mtCO1).

Expression features of cytochrome C oxidase subunit Ⅰ, BCL2 interacting protein 3 and interleukin-1β in MRL/lpr lupus mice liver tissue / 中华风湿病学杂志

Objective:To explore the expression features of cytochrome C oxidase subunit Ⅰ (MT-CO1), BCL2 interacting protein 3 (BNIP3) and interleukin (IL)-1β in the liver of MRL/lpr lupus mice.Methods:The mRNA and protein levels of MT-CO1, BNIP3, IL-1β, p16 and p21 in lupus mice and control mice were detected by polymerase chain reaction (PCR) and Western blot, the IL-1β expression site were detected by hematoxylin and eosin (HE) staining and immunohistochemical method, and themalondialdehyde (MDA) was detected by colorimetry. Hepatocytes and macrophages were stimulated with lipopolysaccharide (LPS), while hepatocytes were also cultured with supernatants obtained after macrophages stimulated with LPS, and the mRNA and protein levels of MT-CO1, BNIP3 and LC3B, as well as p16 and p21 expression, were determined by qPCR and Western blot. The expression of mitochondrial reactive oxygen species (mtROS) was detected by immunofluorescence. One way Analysis of Variance (ANOVA) was used to compare the mean of each group, and LSD method was used to compare the means of multiple samples, and Tamhane's T2 method was used to compare the means of multiple samples when the variance was uniform. Results:The results of PCR showed that the mRNA levels of MT-CO1 and BNIP3 in the liver tissue of the lupus group (0.14±0.04; 0.16±0.05) were significantly lower than those of the control group (0.11±0.04; 0.16±0.06), and the differences were statistically significant ( t=7.16, P<0.001; t=4.54, P<0.001). The expression levels of IL-1β, p16 and p21 in the lupus group (2.06±0.69; 0.37±0.14; 0.16±0.06) were significantly higher than those of the control group (0.23±0.06; 0.25±0.08; 0.11±0.04) ( t=9.58, P<0.001; t=24.35, P<0.001; t=22.36, P<0.001). The results of Western blot were consistent with those of PCR. HE staining showed lymphocyte infiltration in the liver tissue of lupus mice, and immunohistochemistry showed IL-1β in the liver tissue of lupus mice. The positive cells were mainly concentrated in the sinusoids, and the expression of hepatic parenchymal cells was not rearkable. The content of MDA in liver tissue of the lupus group (0.19±0.10) was higher than that of the control group (0.17±0.09), and the difference was statistically significant ( t=4.33, P=0.005). LPS directly stimulated AML12 hepatocytes (0.069±0.028; 0.17±0.07). The PCR results showed that compared with the control group (0.176±0.072; 0.08±0.03), the expression of MT-CO1, and BNIP3 were not significantly different ( t=1.01, P=0.337; t=0.88, P=0.399). The expression of IL-1β was significantly higher when incubated with the supernatants of LPS stimulated macrophages (0.28±0.09) compared than that of the control group (0.15±0.05) ( t=28.26, P<0.001). The results of PCR showed that the mRNA levels of MT-CO1 and BNIP3 in the LPS stimulated group (0.046±0.026; 0.17±0.05) were significantly lower than those in the control group (0.143±0.083; 0.18±0.06), and the differences were statistically significant ( t=7.52, P<0.001; t=4.24, P<0.001), The expression of p16 and p21 in LPS stimulated group (0.29±0.09; 0.27±0.09) were significantly higher than those in the control group (0.18±0.06; 0.22±0.07) ( t=13.54, P<0.001; t=8.69, P<0.001). The results of Western blot were consistent with those of PCR. Immunofluorescence showed that the fluorescence intensity of mtROS in LPS stimulated group (0.25±0.10) was higher than that in the control group (0.08±0.03), and the difference was statistically significant ( t= 4.86, P<0.001). Conclusion:Immune-mediated inflammation in the liver tissue of lupus mice can stimulate liver parenchymal cells to cause intracellular mitochondrial dysfunction. However, the mechanism of liver organ damage in lupus mice is not limited to the immune-mediated inflammation of immune active cells, but also include parenchymal cell mitochondrial dysfunction.

Effect of curcumin on mitochondrial gene and high mobility group box 1 protein in sepsis myocardial injury of rats / 中华急诊医学杂志

Objective@#To observe the effect of curcumin on myocardial mitochondrial related gene (mtATP6), high mobility group box 1(HMGB1) mRNA expression and plasma HMGB1 level in rats with sepsis, and to explore a new strategy for protection of myocardial injury in sepsis.@*Methods@#A rat sepsis model was established by cecal ligation and puncture (CLP). Thirty male Sprague-Dawley rats were randomly (random number) divided into the sham operation group (sham group), sepsis group (CLP group), and curcumin intervention group (Cur group), with 10 rats in each group. Rats in the Cur group were treated with curcumin by intraperitoneal injection at 100 mg/kg, and normal saline was injected into rats in the sham and CLP groups. Five rats were randomly (random number) sacrificed at 6 h and 24 h after modeling. The pathological changes of myocardial tissue were observed under light microscope. The levels of CK and CK-MB in serum were detected. The HMGB1 level in plasma was detected by ELISA. Real-time PCR was used to detect the expression of HMGB1 mRNA and mtATP6 in myocardial tissue. One-way ANOVA were performed in multigroup mean comparison, LSD-t test was used to compare the mean values of two samples, and P<0.05 was considered statistically significant.@*Results@#The pathological score of myocardial tissue in the Cur group was lower than those in the CLP group at 6 h and 24 h (P<0.05). The CK and CK-MB in the Cur group were lower than those in the CLP group at 6 h and 24 h (P<0.05). The levels of HMGB1 in the Cur group were lower than those in the CLP group at 6 h and 24 h (P<0.05). The expression levels of HMGB1 mRNA and mtATP6 in myocardial tissue of the CLP group were lower than those in the sham group at 6 h (P<0.05), while the expression levels of HMGB1 mRNA and mtATP6 in myocardial tissue of the Cur group were higher than those in the CLP group at 6 h, but the difference was not statistically significant (P>0.05).@*Conclusion@#Curcumin protects acute myocardial injury by affecting HMGB1 release and translocation down-regulation of extracellular levels and regulation of mtATP6 expression in CLP model rats.

Study progress of mitochondrial DNA depletion syndromes / 中华实用儿科临床杂志

Mitochondrial DNA(mtDNA)depletion syndromes(MDS)is a type of autosomal recessive genetic disease characterized by a severe reduction in mtDNA content caused by mutations in the nuclear gene,which results in impaired energy production in affected tissues and organs. According to phenotype,MDS are usually classified as 4 forms:myopathic,encephalomyopathic,hepa - tocerebarl and neurogastrointestinal. The following 9 types of related genes have been reported:a myopathic form associated with mutations in TK2;an encephalomyopathic form associated with mutations in SUCLA2,SUCLGl,or RRM2B;a hepa-tocerebral form associated with mutations in DGUOK,MPVl7, POLG,or Cl0orf2;and a neurogastrointestinal form associated with mutations in TYMP. Some MDS can lead to early death in newborns and infants,so early identification is very important. Combination of biochemical testing,histopatholo-gy,respiratory chain complex testing and mtDNA quantification is needed for the diagnosis. The final diagnosis requires genetic testing.

Effect of curcumin on mitochondrial gene and high mobility group box 1 protein in sepsis myocardial injury of rats / 中华急诊医学杂志

Objective To observe the effect of curcumin on myocardial mitochondrial related gene (mtATP6), high mobility group box 1(HMGB1) mRNA expression and plasma HMGB1 level in rats with sepsis, and to explore a new strategy for protection of myocardial injury in sepsis.Methods A rat sepsis model was established by cecal ligation and puncture (CLP). Thirty male Sprague-Dawley rats were randomly (random number) divided into the sham operation group (sham group), sepsis group (CLP group), and curcumin intervention group (Cur group), with 10 rats in each group. Rats in the Cur group were treated with curcumin by intraperitoneal injection at 100 mg/kg, and normal saline was injected into rats in the sham and CLP groups. Five rats were randomly (random number) sacrificed at 6 h and 24 h after modeling. The pathological changes of myocardial tissue were observed under light microscope. The levels of CK and CK-MB in serum were detected. The HMGB1 level in plasma was detected by ELISA. Real-time PCR was used to detect the expression of HMGB1 mRNA and mtATP6 in myocardial tissue. One-way ANOVA were performed in multigroup mean comparison, LSD-t test was used to compare the mean values of two samples, andP<0.05 was considered statistically significant.Results The pathological score of myocardial tissue in the Cur group was lower than those in the CLP group at 6 h and 24 h (P<0.05). The CK and CK-MB in the Cur group were lower than those in the CLP group at 6 h and 24 h (P<0.05). The levels of HMGB1 in the Cur group were lower than those in the CLP group at 6 h and 24 h (P<0.05). The expression levels of HMGB1 mRNA and mtATP6 in myocardial tissue of the CLP group were lower than those in the sham group at 6 h (P<0.05), while the expression levels of HMGB1 mRNA and mtATP6 in myocardial tissue of the Cur group were higher than those in the CLP group at 6 h, but the difference was not statistically significant (P>0.05).Conclusion Curcumin protects acute myocardial injury by affecting HMGB1 release and translocation down-regulation of extracellular levels and regulation of mtATP6 expression in CLP model rats.

Mitochondrial DNA and childhood obesity / 国际儿科学杂志

Childhood obesity is one of the major public health problems in the 21st century.The specific pathogenesis of obesity in children is still not very clear.It is currently believed that it may be caused by multiple factors including heredity,diet,and physical activity.The mitochondrial dysfunction plays an important role in the development of obesity.Mitochondria are important sites for energy metabolism in human body,and the normal function depends on the regulation of both nuclear and mitochondrial genes.In addition,due to the maternal inheritance,mitochondrial DNA can be passed to the next generation,which explains why some studies show larger correlation for BMI between mothers and their children than between fathers and their children.Therefore,mitochondrial genes are likely to be an important part of the development of childhood obesity.This article reviews the association and possible mechanisms of mitochondrial DNA and childhood obesity.

Prevalence of Hymenolepis nana and H. diminuta from Brown Rats (Rattus norvegicus) in Heilongjiang Province, China

Hymenolepis nana and Hymenolepis diminuta are globally widespread zoonotic cestodes. Rodents are the main reservoir host of these cestodes. Brown rats (Rattus norvegicus) are the best known and most common rats, and usually live wherever humans live, especially in less than desirable hygiene conditions. Due to the little information of the 2 hymenolepidid species in brown rats in China, the aim of this study was to understand the prevalence and genetic characterization of H. nana and H. diminuta in brown rats in Heilongjiang Province, China. Total 114 fecal samples were collected from brown rats in Heilongjiang Province. All the samples were subjected to morphological examinations by microscopy and genetic analysis by PCR amplification of the mitochondrial cytochrome c oxidase subunit 1 (COX1) gene and the internal transcribed spacer 2 (ITS2) region of the nuclear ribosomal RNA gene. In total, 6.1% (7/114) and 14.9% (17/114) of samples were positive for H. nana and H. diminuta, respectively. Among them, 7 and 3 H. nana isolates were successfully amplified and sequenced at the COX1 and ITS2 loci, respectively. No nucleotide variations were found among H. nana isolates at either of the 2 loci. Seventeen H. diminuta isolates produced 2 different COX1 sequences while 7 ITS2 sequences obtained were identical to each other. The present results of H. nana and H. diminuta infections in brown rats implied the risk of zoonotic transmission of hymenolepiasis in China. These molecular data will be helpful to deeply study intra-specific variations within Hymenolepis cestodes in the future.

Molecular phylogeny of Toxoplasmatinae: comparison between inferences based on mitochondrial and apicoplast genetic sequences / Filogenia molecular de Toxoplasmatinae: comparação entre inferências baseadas em sequências genéticas mitocondriais e de apicoplasto

Abstract Phylogenies within Toxoplasmatinae have been widely investigated with different molecular markers. Here, we studied molecular phylogenies of the Toxoplasmatinae subfamily based on apicoplast and mitochondrial genes. Partial sequences of apicoplast genes coding for caseinolytic protease (clpC) and beta subunit of RNA polymerase (rpoB), and mitochondrial gene coding for cytochrome B (cytB) were analyzed. Laboratory-adapted strains of the closely related parasites Sarcocystis falcatula and Sarcocystis neurona were investigated, along with Neospora caninum, Neospora hughesi, Toxoplasma gondii (strains RH, CTG and PTG), Besnoitia akodoni, Hammondia hammondiand two genetically divergent lineages of Hammondia heydorni. The molecular analysis based on organellar genes did not clearly differentiate between N. caninum and N. hughesi, but the two lineages of H. heydorni were confirmed. Slight differences between the strains of S. falcatula and S. neurona were encountered in all markers. In conclusion, congruent phylogenies were inferred from the three different genes and they might be used for screening undescribed sarcocystid parasites in order to ascertain their phylogenetic relationships with organisms of the family Sarcocystidae. The evolutionary studies based on organelar genes confirm that the genusHammondia is paraphyletic. The primers used for amplification of clpC and rpoB were able to amplify genetic sequences of organisms of the genus Sarcocystisand organisms of the subfamily Toxoplasmatinae as well.

Resumo A filogenia da subfamília Toxoplasmatinae tem sido amplamente investigada com diversos marcadores moleculares. Neste estudo, a filogenia molecular da subfamília Toxoplasmatinae foi analisada através de genes de apicoplasto e mitocondriais. Foram analisadas sequências parciais de genes de apicoplasto codificadores da protease caseinolítica (clpC), e da subunidade beta da RNA polimerase (rpoB) e de gene mitocondrial codificador de citocromo B (cytB). Foram investigadas cepas adaptadas em laboratório de Sarcocystis neurona eSarcocystis falcatula, parasitos estreitamente relacionados, além de Neospora caninum, Neospora hughesi, Toxoplasma gondii (cepas RH, CTG e PTG),Besnoitia akodoni, Hammondia hammondi e duas linhagens geneticamente divergentes de Hammondia heydorni. A análise molecular, baseada em genes de organelas, não diferenciou claramenteN. caninum de N. hughesi, porém foi possível confirmar as duas linhagens de H. heydorni. Foram encontradas pequenas diferenças entre as cepas adaptadas em laboratório deS. falcatula e S. neurona em todos os marcadores moleculares avaliados. Concluindo, filogenias congruentes foram reconstruídas com os três diferentes genes que podem ser úteis em triagem de parasitos sarcocistídeos não identificados, para identificar sua relação com organismos da família Sarcocystidae. Os estudos evolutivos com genes organelares confirmam que o gênero Hammondia é parafilético. Osprimers utilizados para amplificação declpC e rpoB foram capazes de amplificar sequências genéticas de organismos do gênero Sarcocystis e da subfamília Toxoplasmatinae.

The analysis of common mutation in deafness-associated gene in 111 neonates who failed to pass newborn hearing screening / 临床儿科杂志

Objective To explore the genetic mutation in neonates who failed to pass hearing screening.Methods A total of 111 cases of neonates who failed to pass hearing screening and were conifrmed sensorineural deafness by auditory brainstem evoked potential (ABR) were randomly selected. The heel blood was collected and DNA was extracted.GJB2, SLC26A4, and 11 mutation hotspots in mitochondria gene12SrRNA were tested. The relationship between degree of hearing loss and gene mutation was analyzed.Results In 111 neonates, mutation in deafness gene were found in 24 cases (21.6%) . Among them 14 cases (12.6%) hadGJB2 gene mutation including 5 cases of 235delC single heterozygous mutation, 5 cases of 235delC, and 1 case each of 299_300delAT compound heterozygous mutation, 235delC homozygous mutation, 299_300delAT single heterozygous mutation, 176_191del16 and 235delC compound heterozygous mutation, and 299_300delAT and 508_511dupAACG compound heterozygous mutation respectively. Ten cases (9.0%) hadSLC26A4 gene mutation including 2 cases of IVS7-2A>G single heterozygous mutation, 3 cases of 1226G>A single heterozygous mutation, 2 cases of 2168A>G single heterozygous mutation, and 3 cases of IVS7-2A>G and 2168A>G compound heterozygous mutation. Mitochondrial gene mutations were not detected. Conclusions Deafness gene mutation is detected in more than 1/5 neonates who failed to pass newborn hearing screening. GJB2 gene mutation is the most commons. The implementation of hotspots deafness gene detection can improve the diagnostic rate of deafness.

Mitochondrial diseases with the main manifestations of cardiomyopathy and respiratory muscle damage / 中华实用儿科临床杂志

Objective The 3243A ＞ G mutation in mitochondrial DNA is a common cause of the classical mitochondrial diseases characterized by neuro-muscular disorders.This study reports a rare case with the main manifestations of mitochondrial disease in children of mitochondrial cardiomyopathy and respiratory muscle damage.Methods The clinical characteristics,diagnosis and treatment,biochemical,pathological and genetic features of a 10-year-old girl were studied.Results The girl was admitted because of heart failure and respiratory failure at the age of 10.Ragged red fibers in skeletal muscles had been noticed.On her mitochondrial gene,3243A ＞ G mutation,Leu tRNA (UUR),was detected.The mutation rate in the peripheral blood cells was 94％.After the treatment with a high dose of creatine phosphate sodium,coenzyme Q10 and L-carnitine with assisted ventilation,the patient improved rapidly.The child was followed up for 2 years without recurrence.Meanwhile the growth,development and daily life were normal.Conclusions Cardiac and respiratory muscle impairments that appeared at the same time as the first manifestations of the children's mitochondrial disease is not common,and it is rare to have cardiomyopathy based mitochondrial gene 3243A ＞ G mutation is seldom seen clinically.Skeletal muscle biopsy and genetic test is the key for accurate diagnosis.Improving mitochondrial metabolism and assisted ventilation appear to be helpful treatments.

Human Infections with Spirometra decipiens Plerocercoids Identified by Morphologic and Genetic Analyses in Korea

Tapeworms of the genus Spirometra are pseudophyllidean cestodes endemic in Korea. At present, it is unclear which Spirometra species are responsible for causing human infections, and little information is available on the epidemiological profiles of Spirometra species infecting humans in Korea. Between 1979 and 2009, a total of 50 spargana from human patients and 2 adult specimens obtained from experimentally infected carnivorous animals were analyzed according to genetic and taxonomic criteria and classified as Spirometra erinaceieuropaei or Spirometra decipiens depending on the morphology. Morphologically, S. erinaceieuropaei and S. decipiens are different in that the spirally coiled uterus in S. erinaceieuropaei has 5-7 complete coils, while in S. decipiens it has only 4.5 coils. In addition, there is a 9.3% (146/1,566) sequence different between S. erinaceieuropaei and S. decipiens in the cox1 gene. Partial cox1 sequences (390 bp) from 35 Korean isolates showed 99.4% (388/390) similarity with the reference sequence of S. erinaceieuropaei from Korea (G1724; GenBank KJ599680) and an additional 15 Korean isolates revealed 99.2% (387/390) similarity with the reference sequences of S. decipiens from Korea (G1657; GenBank KJ599679). Based on morphologic and molecular databases, the estimated population ratio of S. erinaceieuropaei to S. decipiens was 35: 15. Our results indicate that both S. erinaceieuropaei and S. decipiens found in Korea infect humans, with S. erinaceieuropaei being 2 times more prevalent than S. decipiens. This study is the first to report human sparganosis caused by S. decipiens in humans in Korea.

Karyotype of the invasive species Pterois volitans (Scorpaeniformes: Scorpaenidae) from Margarita Island, Venezuela / Cariotipo de la especie invasora Pterois volitans (Scorpaeniformes: Scorpaenidae) de Isla Margarita Island, Venezuela

The genus Pterois includes nine valid species, native to the Red Sea and Indian Ocean throughout the Western Pacific. P. volitans and P. miles are native to the Indo-Pacific, and were introduced into Florida waters as a result of aquarium releases, and have been recently recognized as invaders of the Western Atlantic and Caribbean Sea (Costa Rica to Venezuela). Thus far, cytogenetic studies of the genus Pterois only cover basic aspects of three species, including P. volitans from Indo-Pacific Ocean. Considering the lack of more detailed information about cytogenetic characteristics of this invasive species, the objective of the present study was to investigate the basic and molecular cytogenetic characteristics of P. volitans in Venezuela, and compare the results with those from the original distribution area. For this, the karyotypic characteristics of four lionfish caught in Margarita Island, Venezuela, were investigated by examining metaphase chromosomes by Giemsa staining, C-banding, Ag-NOR, and two-colour-Fluorescent in situ hybridization (FISH) for mapping of 18S and 5S ribosomal genes. Comparing the sequences of the 16S gene of the specimens analyzed, with sequences already included in the Genbank, we corroborated that our specimens identified as P. volitans are in fact this species, and hence exclude the possibility of a misidentification of P. miles. The diploid number was 2n=48 (2m+10sm+36a) with FN=60. Chromosomes uniformly decreased in size, making it difficult to clearly identify the homologues except for the only metacentric pair, and the pairs number two, the largest of the submetacentric series. C-banding revealed only three pairs of chromosomes negative for C-band, whereas all remaining chromosomes presented telomeric and some interstitial C-positive blocks. Only two chromosomes were C-banding positive at the pericentromeric regions. Sequential staining revealed Ag-NOR on the tips of the short arms of chromosome pair number two and the FISH assay revealed that 18S rDNA and 5S rDNA genes are co-located on this chromosome pair. The co-localization of 5S rDNA and 45S rDNA is discussed. Both constitutive heterochromatin and NOR location detected in samples examined in this study, differ from those reported for P. volitans in previous analysis of specimens collected in Indian Ocean (Java), suggesting the occurrence of chromosome microrearrangements involving heterochromatin during the spread of P. volitans.Rev. Biol. Trop. 62 (4): 1365-1373. Epub 2014 December 01.

El género Pterois contiene nueve especies válidas, nativas del Mar Rojo y el Océano Índico en el Pacífico occidental. P. volitans y P. miles son nativas del Indo-Pacífico, y fueron introducidas en las aguas de Florida como resultado de la liberación de peces confinados en acuario y han sido reconocidas recientemente como invasoras en el Atlántico Occidental y Mar Caribe (Costa Rica hasta Venezuela). Los estudios citogenéticos realizados hasta ahora en el género Pterois cubren solamente aspectos básicos de tres especies que incluyen a P. volitans del océano Indo-Pacífico. Debido a la ausencia de información detallada sobre las características cromosómicas de esta especie invasora, el objetivo del presente estudio fue investigar las características citogenéticas en ejemplares de Venezuela mediante técnicas convencionales y moleculares y comparar los resultados con los reportados para el área de distribución original. Para ello, se investigaron las características cariotípicas mediante tinción con Giemsa, bandeo-C, impregnación con Nitrato de Plata (Ag-NOR) e hibridación fluorescente in situ (FISH) dual para localizar los genes ribosomales 18S rDNA y 5S rDNA en cuatro ejemplares de pez león capturados en la Isla Margarita, Venezuela. La comparación de secuencias del gen 16S de los especímenes analizados con secuencias ya incluidas en el Genbank permitieron corroborar la identificación de P. volitans excluyendo así la posibilidad de una identificación errónea de P. miles. El número diploide fue 2n=48 (2m+10sm+36a) con un FN=60. Los cromosomas presentaron tamaños que disminuyen de manera uniforme dificultando la identificación de homólogos, excepto el único par metacéntrico y el par cromosómico número 2. El bandeo-C reveló tres pares de cromosomas bandas-C negativos, mientras que los restantes presentaron bloques bandas-C positivos en posición telomérica y, en algunos casos, intersticial. Sólo dos cromosomas mostraron bandas-C pericentroméricas. La tinción secuencial reveló las Ag-NOR localizadas en los extremos de los brazos cortos del par número dos y el ensayo FISH demostró que los genes 18S rDNA y 5S rDNA se localizan en ese mismo par. Se discute la co-localización de los genes 5S rDNA y 18S rDNA. La distribución de la heterocromatina constitutiva y localización de las NORs en los peces examinados difirió de la reportada para ejemplares de P. volitans del Océano Índico (Java), sugiriendo que durante la propagación de P. volitans han ocurrido reorganizaciones cromosómicas que involucran la heterocromatina.

Prevalence and clinical characteristics of mitochondrial mutations in Chinese patients with type 2 diabetes / 中国糖尿病杂志

Objective To assess the prevalence of the A to G mutation at position 3243 of the mitochondrial tRNALeu(UUR) gene in type 2 diabetes in a Chinese population. Methods We screened 716 randomly selected, unrelated patients with type 2 diabetes for the mutation with a PCR-RFLP technique. Results Three individuals with this mutation were identified, representing approximately 0. 4％ of the type 2 patients screened. Further screening of first degree relatives of these 3 patients identified another 4 affected carriers. In comparison with type 2 diabetic patients without the mutation, these 7 carriers of the mt3243 mutation had:①an earlier onset of diabetes (38. 0±10.1 yr vs 53. 4±10.0 yr, P ＜0. 001) ;②lower BMI (19.5±2.0 vs 24. 9±10. 9, P ＜0. 0001) ;and ③ lower post-challenge insulin levels (Area under the curve of insulin levels during the OGTT, 2946± 1647.2 vs 7469±6647.7, P ＜ 0. 01). In addition, we screened the same 716 patients with type 2 diabetes, as well as 181 controls with normal glucose tolerance,for a newly described mt 3316 G→A mutation. This mutation was found in 16 patients with type 2 diabetes (2.20％) and 5 controls (2.7％). Therefore, the frequency of the mutation was not different between patients and controls.Moreover, clinical characteristics such as age of onset of diabetes, BMI, and insulin levels were not different between diabetic patients with the mt3316 mutation and those without it. Concision The mt3316 G→ A mutation is a polymorphism unrelated to diabetes.

Familial Hearing Loss Associated with mtDNA A1555G Mutation in Korea: 1 Pedigree / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Familial aminoglycoside-induced deafness has been described in a number of Chinese and Japanese pedigrees. Recently, the familial aminoglycoside-induced ototoxicity is proved to be associated with a mutation in mitochondrial (mt) 12S ribosomal RNA (rRNA) gene at nucleotide position 1555 in some families. In this study, we analyzed mt 12S rRNA gene to find out this particular mutation in Korean pedigrees who had a family history of hearing loss. MATERIALS AND METHODS: Peripherial blood was obtained from 91 individuals of 30 families, and total genomic DNA (gDNA) was extracted. A fragment of DNA including a part of mt 12S rRNA gene was amplified by polymerase chain reaction (PCR). The PCR products were analyzed by restriction digestion with Bsm A1 and DNA sequencing. RESULTS: We found one family of mtDNA A1555G. Six family members had mutant genotype and three of them showed severe sensorineural hearing loss or deafness. The mutation was homoplasmic in all affected family members, and the genotype revealed maternal transmission. CONCLUSION: We found the first case of familial hearing loss genetically proved to be associated with the mt 12S rRNA gene mutation, in Korea. Because it is possible that an individual with this mutation shows a progressive sensorineural hearing loss, a screening of mtDNA A1555G mutation for the familial members who have a maternal inheritant hearing loss might be necessary.