Phantom mutation hotspots in human mitochondrial DNA

Phantom mutations are systematic artifacts generated in the course of the sequencing process. Contra common belief these artificial mutations are nearly ubiquitous in sequencing results, albeit at frequencies that may vary dramatically. The amount of artifacts depends not only on the sort of automated sequencer and sequencing chemistry employed, but also on other lab-specific factors. An experimental study executed on four samples under various combinations of sequencing conditions revealed a number of phantom mutations occurring at the same sites of mitochondrial DNA (mtDNA) repeatedly. To confirm these and identify further hotspots for artifacts, > 5000 mtDNA electropherograms were screened for artificial patterns. Further, > 30000 published hypervariable segment 1 sequences were compared at potential hotspots for phantom mutations, especially for variation at positions 16085 and 16197. Resequencing of several samples confirmed the artificial nature of these and other polymorphisms in the original publications. Single-strand sequencing, as typically executed in medical and anthropological studies, is thus highly vulnerable to this kind of artifacts. In particular, phantom mutation hotspots could easily lead to misidentification of somatic mutations and to misinterpretations in all kinds of clinical mtDNA studies.

The fecundity of the false trevally, Lactarius lactarius (Bloch and Schneider) along the Karwar coast

A clear knowledge of the reproductive potential or fecundity of a fish is quite an essential pre-requisite for the proper management and conservation of the resources. The fecundity studies are also undertaken to determine the index of diversity dependent factor affecting the population size (Simpson, 1951). Qasim & Qayyam (1963) have detailed the various pathways by which an understanding of fecundity could be used for fishery biological work. The ability of egg production varies within the individual limits such as length, somatic weight, gonadal weight, volume of fish etc.

基因组中新遗传结构的起源与进化

生物遗传物质的多样性从根本上决定了当今地球上生命世界的丰富多 彩。然而生命在漫长的进化历程中如何从最原始的生命形态不断演变，创造 出如此巨大的多样性，是自然留给我们最吸引人的奥秘之一。因此对于分子 进化生物学或进化基因组学的研究者来说，承载生命遗传信息的基因组如何 进化一直以来都是大家关注的一个基本科学命题。本研究主要从两个方面探 讨了基因组中新遗传结构起源的分子机制和进化模式，一方面我们以实验的 方法在黑腹果蝇亚种组中大规模地对新基因进行筛选和鉴定，探讨了果蝇基 因组中新基因起源的分子机制和进化模式；另一方面我们以生物信息学手段 对啮齿类动物大鼠和小鼠进行比较基因组研究，以人和猪的转录序列作为外 群，鉴定了大量啮齿类特有的新外显子，并对这些新外显子的进化特征和产 生机制做了研究。 在果蝇基因组新基因起源与进化的研究工作中，我们综合运用了荧光原 位杂交(fluorescent in situ hybridization, FISH)，Southern 印迹 (Southern blotting)，表达转录分析，生物信息确认和进化速率分析等技术和分析手段， 通过对黑腹果蝇约7000 个基因在黑腹果蝇亚种组8 个近缘物种中同源拷贝数 分布的筛选，鉴定了17 个年轻的散在重复新基因，并对这些新基因的结构、 表达和进化进行了全面的分析。结果表明，DNA 水平的重组机制产生了大量 的新的与祖先基因结构没有冗余的散在重复基因，它们的基因结构以很高的 频率形成了嵌合结构。这些新散在重复基因形成嵌合基因的机会有可能大大 高于预期。同时，我们提供了有力证据证明，重复序列特别是DNAREP1 转 座子很可能通过了非等位同源重组方式介导了散在重复基因的形成。最后， 运用多种行之有效的分析方法，我们证明绝大部分的这些新的嵌合重复基因 是有功能的。在啮齿类新外显子的起源与进化研究中，我们首先利用有完整序列信息 的人和小鼠的基因组，通过同源比对确定了人和小鼠间12,419 个直系同源的 基因组转录单元，这些基因组转录单元中71,039 个大、小鼠共有且相位定义 清晰的外显子被用作后续的分析。通过与人的基因组序列相比较，并进一步 以猪的转录组序列作为第二外群排除掉可能是在人的基因组中丢失的外显子 后，我们共确定了2,695 个啮齿类特有的新外显子。随后对这些新外显子产 生的机制、进化速率、潜在功能以及与选择性剪接之间的关系进行了讨论。 结果显示多数新外显子来自内含子的非重复序列，存在快速的碱基非同义替 换和插入缺失，功能分布上最多的是参与细胞外结合和蛋白间相互作用，提 示这些新外显子的产生可能与啮齿类与外界环境的适应性进化相关。对这些 新外显子与选择性剪接之间关系所做的分析表明，大多数的新外显子存在于 表达量较低的选择性剪接形式中，这说明这些外显子通常参与形成执行组织 特异功能的表达形式，也从一定程度上解释了这些外显子何以能够摆脱对基 因的功能限制而产生较快的进化速度。 总之，上述对果蝇基因组中新基因和啮齿类基因组中新外显子的起源和 进化的研究结果表明，基因组中通过新基因或新外显子等基因组新材料产生 新功能的进化过程是常见的和重要的遗传机制。

Epigenetic marks in cloned rhesus monkey embryos: Comparison with counterparts produced in vitro

Until now, no primate animals have been successfully cloned to birth with somatic cell nuclear transfer (SCNT) procedures, and little is known about the molecular events that occurred in the reconstructed embryos during preimplantation development. In man

HSPC117 deficiency in cloned embryos causes placental abnormality and fetal death

Somatic cell nuclear transfer (SCNT) has been successfully used in many species to produce live cloned offspring, albeit with low efficiency. The low frequency of successful development has usually been ascribed to incomplete or inappropriate reprogramming of the transferred nuclear genome. Elucidating the genetic differences between normal fertilized and cloned embryos is key to understand the low efficiency of SCNT. Here, we show that expression of HSPC117, which encodes a hypothetical protein of unknown function, was absent or very low in cloned mouse blastocysts. To investigate the role of HSPC117 in embryo development, we knocked-down this gene in normal fertilized embryos using RNA interference. We assessed the post-implantation survival of HSPC117 knock-down embryos at 3 stages: E9 (prior to placenta formation); E12 (after the placenta was fully functional) and E19 (post-natal). Our results show that, although siRNA-treated in vivo fertilized/produced (IVP) embryos could develop to the blastocyst stage and implanted without any difference from control embryos, the knock-down embryos showed substantial fetal death, accompanied by placental blood clotting, at E12. Furthermore, comparison of HSPC117 expression in placentas of nuclear transfer (NT), intracytoplasmic sperm injection (ICSI) and IVP embryos confirmed that HSPC117 deficiency correlates well with failures in embryo development: all NT embryos with a fetus, as well as IVP and ICSI embryos, had normal placental HSPC117 expression while those NT embryos showing reduced or no expression of HSPC117 failed to form a fetus. In conclusion, we show that HSPC117 is an important gene for post-implantation development of embryos, and that HSPC117 deficiency leads to fetal abnormalities after implantation, especially following placental formation. We suggest that defects in HSPC117 expression may be an important contributing factor to loss of cloned NT embryos in vivo. (C) 2010 Elsevier Inc. All rights reserved.

The study of biological characteristics and heavy metal determination of Cd, Pb, Fe, V, Ni in the muscle, liver and gonad tissues of kingfish Scomberomorus commerson in the coastal waters of the Hormozagan Province (Persian Gulf)

Biological studies and heavy metal (Ni, v, Fe, Pb,Cd) determination in liver, gonad and muscle of Scomberomorus commerson were carried out from Oct 2006 to Sept 2007 in Hormozgan coastal waters. 599 Samples were gutted for reproduction and nutrition studies, fork length and weight were measured to nearest cm and g respectively. Meanwhile 40 samples were also investigated for heavy metal studies. All specimens were collected from two major landing sites (Bandar Lengeh & Bandar Abbas). Minimums & maximum fork length & weight were 29, 128 cm & 235 and 15350 g respectively.Isometric growth was shown according to our study and b was estimated 2.9 (overall), 2.91(male) & 2.89 (female). The average relative gut length was 0.52± 0.007 and it was determined that S. commerson is a carnivorous. More than 99 percent of gut content was different teleost fishes. Gastro somatic index had two peaks in Nov & Jan (before spawning) and with a decreased trend in July, the spawning period. Occurrence of empty stomach was estimated % 65.77. Maximum amount of condition factor was in Dec. Spawning season was started from June. The average of Absolute & relative fecundity (to weight unit) was 1217149±179315 and 178.2±15.58 respectively. Lm50% was estimated 75 cm for females. Sex ratio was 0.97: 1 (male: female). Chi- Square test showed no significant difference (p>0.05). Maximum amount of hepatosomatic index was estimated in March.Metal concentrations were determined using either Flame Atomic Absorption Spectroscopy (for Fe) or Graphite Furnace Atomic Absorption Spectroscopy (for Pb,Cd,Ni and V). The mean concentration (μg/g dry weight)of Pb,Cd,Ni,V and Fe in the liver were 0.0309, 0.0268, 0.0672, 0.0077, 2.5159 in the gonad 0.0440 ,0.0295, 0.1096, 0.0000, 1.4449 and in the muscle 0.0244, 0.0324, 0.0656, 0.0128, 1.6138 respectively. The maximum metal concentrations were below the maximum permissible limits for human consumption recommended by the USEPA, WHO and the UK. The results of Kendall's Tau-b correlation coefficient were as follows: The Liver tissue: There were significant positive linear relationships between accumulation of V, Fe, and Pb with Fork length, Pb and Fe with weight, GSI with Pb, Cd, V and 109 Fe, and a negative linear relationships between HSI with accumulation of V and Fe, Fork length, weight and GSI. The Gonad tissue: There were significant positive linear relationships between GSI with accumulation of Pb, Cd, Fe, Fork length and weight, a negative linear relationship between HSI with Fork length, weight and GSI. The Muscle tissue: There were significant positive linear relationships between accumulations of V, Fe with Fork length and weight factors and as well as GSI with Cd, V, Fe, Pb, Fork length and weight,a negative linear relationship between HSI with Fork length, weight, Cd, Fe and GSI. The results of Mann-Whitney U tests (P≤0.05) show that there were significant differences between summer and autumn from heavy metal contents in the studied tissues point of view. The only exceptions were for Ni in the liver, gonads and muscle and as well as there were significant differences between male and female from heavy metal contents in the studied tissues. The only exceptions were for Pb in muscle, Ni in liver, gonad and muscle, V in muscle, and Cd and Fe in gonads.

Study on some biological and feeding factors of spined loach Cobitis taenia from the Babolrud, Talar and Siahrud Rivers

Spined loach, Cobitis taenia, is a predominant fish in the river systems of the southern Caspian Sea basin. Although there is evidence of the geographical divergence of this taxon, but no information is available on morphological differences within the species populations. This study was designed to evaluate some biological factors including; morphometric and meristic characters, length-weight, age-growth, condition factor, diet, reproduction, variation and differentiation, in the Babolrud, Talar and Siahrud Rivers in south of the Caspian Sea basin. Age, sex ratio, fecundity, ova diameter and gonadosomatic index were estimated. Also, regression analyses was tested the relation between fecundity and fish length, weight, gonad weight, and also age. Totally 858 fish of which 721 were matures, were collected from these rivers by electrofishing. 37 morphometric characters, 9 meristic characters and 78 truss network system characters were estimated. Resulats of DFA analysis based on data of morphometric and meristic showd that these populations are highly (94.5%) varios from each other. In discriminate function analysis, the proportion of individuals correctly classified into their original groups was 61%, 65.4% and 86.5% for upstream and downstream, respectively. Clustering based on Euclidean distances among groups of centroids using an UPGMA and also principal component analysis’ results for morphometric data indicated that these populations from these three rivers were clearly distinct from each other. Regression equations between length and weight in these three populations were significantly different from Folton factor (b=3), that showed the fish has a negative Alometric growth process. Condition factor was estimated between 0.8912 to 1.2736 and 0.8131 to 1.4489 for males and females, respectively. Sex ratio (female: male) in these populations was 1.2816:1. The difference between the number of females and males was significant and females were more than males. The female and male specimens reach maturity by Tl more than 40 and 30 mm and at the age of 2+ and 1+, respectively. The mean of ova diameter was 0.5824±0.2882. The spawning took place from May to late July, at the water temperature from 18.7 to 24.0°C. The GSI values average at the beginning of the reproduction period was about 9%, with ranged from 2 to 26% in ripe mature females. The absolute and relative fecundity were 2109±792 and 579±208 respectively. The absolute fecundity was significantly related to body weight and gonads weight. Based on the pattern of gonado-somatic index, it was concluded that this fish has prolong active reproductive period, which is a type of adaptation by short-lived small fishes to environmental conditions. The macroscopic and histological results showed that the female and male have 5 and 4 stages in their maturation process, respectively. The RLG index was about 0.4732, which showed the fish is a carnivorous species. Significant difference was observed between fishes with different length and diet. The main foods of the fish were Trichoptera, Chironomidae larvae and Ephemeroptera which were their prefered food as well, however it was estimated that the food selection and diet are affected by environmental conditions.

Reproductive biology of Catla catla in the Udawalawe reservoir, Sri Lanka

The reproductive biology of Catla catla (Hamilton-Buchanan) in the Udawalawe reservoir was studied from June 2007 to December 2008. Samples of eggs from Indian major carp C. catla were collected from fish landed in the reservoir and analysed in the laboratory to assess the reproductive characteristics. C. catla. Cirrhinus mrigala, exotic Cichlids and Labeo rohita accounted for 62.2%, 21.0%, 12% and 1.0% respectively of the total landings in the Udawalawe reservoir during the study period. Gonads of C. catla were collected in the field and examined in the laboratory to determine the stage of maturity and fecundity. Data on fish length and gonad weight were collected to estimate the gonado-somatic indices (OSI). Landed catches were also examined in the field to determine the sex ratio of C. catla in the catch and was found to be 1:5.6 male to female. Results of fecundity estimates revealed that C. catla females in the Udawalwe reservoir were fully mature in June and October of the year. From the monthly variation of OSI, two recruitment pulses per year were evident. The length of the body at first maturity in female C. catla in the Udawalwe reservoir was estimated to be 74.2 cm.

Distribution and growth biology of rock oyster Saccostrea cucullata in the Oman Sea

Distribution and growth biology of rock oyster (Saccostrea cucullata) in the northern shores of Oman sea have been struied. During this one-year study, samples have been taken monthly from ten different stations. quantity of vertical distribution of this species was obseredl in the mid - intertidal zone. After determining the spread pattern, the following subjects were studied: - Growth parameters - Distionction of the "cohorts" - Determination of "spawning Season" - Condition of the "Gonado Somatic Index" - Sex ratio - Length of the species during the first year of maturation. - Identification and determination of percentage of "Biofouler Organisms." Results obtained from the above - mentioned studies show that considering a growth factor (k) of 0.52, the value of "Loo " for this species is equal to 114 (mm).Five to six different age groups were observed among the samples taken. In the areas where this study was conducted, this species grows 24 to 30 (mm) in the first year of its life this growth rate is lower in the higher - aged grpups relative to the lewer - aged groups, so that the longest size classes grow between 4 to 6 (mm) per year. • The maxinum Value of the "Condition Index" is in the pozm area and the minimum value of it belongs to Darak and Tang areas. Along with the increase in the growth of gonads the above mentioned condition index increases gradually simultaneous with the onset of spawning. Also, study of the influence of environmental factors on the maturation process suggests that the most important factors affecting maturation and spawing are temperature and salinity. The study of GSI shows that this species has a coordinated bimodal spawing trend, with its spring peak in june and its autumn peak, being still higher than the spring peak, in september. The recruitment curve confirms the above spawning peaks with its peaks occuring after a delay of one month or maximum two months in comparison to the spawning peaks. The results of calcuation of "Sex Ratio" of this species in each area show that sex ratio is 1:1. Among the first size classes that reach maturity, nearly 67% of the samples are male and the remaining 33% are female. with the increase in the shell size, the percentage of males decreases and the percentage of females increases. , The above facts prove the protandrous nature of this species the diagram showing the sizes of the first samples which reach maturity suggests that more than 50% of the samples mature after their length exceeds 36 (mm). The shortest mature sample was found to have a length of 22(mm). After studying "Biofouler Organism" nine different invertebrate groups were indentified. Barnacles and Tunicates have the highest and lowest percentages respectively. According to zonal observations, Barnacles and polychacta do the greatest damage to this species.

Fishery biology of Indian mackerel in Sri Lankan coastal waters

The objective of the project was to determine the reproductive biology of Indian Mackerel (Rastrelliger kanagurta) using Gonado-Somatic Index (GSI); length at first maturity; spawning season; morphological characteristics; and stomach contents.

Study biological characteristics related with development and reproduction in Barbus capito on the southern coast of the Caspian Sea, Gilan Province

In the present research, a total of 207 pieces of fish from 25 sampling stations in Gilan Province coasts in the years 2001-2002 were biologically studied in terms of their growth and development, reproduction and feeding. The average length and weight of the fishes are increased, as they get older. The highest index of length and weight growth is observed in the years 1 to 2. As the age increases, gradient of length and weight growth diagrams decrease. In studying the relation between length and weight, it was observed that proportionate to the total length, the weight is increased progressively. The fatness coefficient index in the initial years of life and prior to maturity is higher than the post maturity period. As the age increases, the decrease of this index is observable. The fatness coefficient index rate is directly related to index of fullness. The highest Gonadosomatic Index is seen in the months of June and July, i.e. at the times of spawning; and the lowest index rate is observed in the months of November and December. The appropriate temperature for reproduction of these species is from 18 to 22 degree centigrade. The Gonadosomatic Index is higher in spring and summer seasons as compared with autumn and winter. Besides, as the fishes become aged, the amount of the said index increases in a manner that the gradient of it in the years to maturity is less than the maturity time and thereafter. Sexual maturity stages in different months are directly related to Gonadosomatic index, and increase as the age increases. The sexual ratio of male fishes to the female fishes in terms of number is plus one prior to maturity; about one at the time of maturity and minus after maturity. In general the frequency of male fishes as compared with female fishes in all group ages is approximately two times. The fecundity mean, and the diameter and the rate of eggs will substantially increase, as the Gonadosomatic index rises. The maturity age in the male fishes is 3 to 4 years and in female fishes is 4 to 5 years. The spawning of this species in rivers occurs repeatedly and in different time intervals, and do not take place once (Asyncronous). The Gastrosomatic index is directly related to index of fullness and will decrease, as the age increases. The index of fullness is relatively the months of April and May. The underlying reason is the need of the fishes to energy for reproduction. As the spawning time commences, the index of fullness moves down and the downward direction continues. After spa g mg and reduction of the volume of energy in the body, the index of fullness rises, and it will be substantially high until the beginning of fall. In fall and winter as it gets cold, the index of fullness moves downward and the body fat deposits are used. A correlation is shown between the changes in vacuity index and fullness indices. This means that as the fullness index rises, the vacuity index decreases, and vice versa. The Hepatosomatic index prior to the reproduction is at the highest amount and after spawning is at the lowest. No correlation is observed between the fullness and Hepatosomatic indices. In other words reproduction is an inherent and instinct originated matter; and its cycle goes on, alternately and in an orderly manner, upon completion of germinal cells, even when it coincides with reduction or stoppage of somatic cell growth. The rising trend of Hepatosomatic starts in August and will continue until the next July. The volume of fat around digestive tract is severely reduced in early spring and this trend will reach its apex in summer season. In the cold seasons, i.e. the fall and winter, the accumulation of fat around digestive tract increases. Consequently, a meaningful and inverse relation is observed between index of fullness, also the progress of sexual maturity stages and the volume of fat.

A critical reassessment of the role of mitochondria in tumorigenesis

Background Mitochondrial DNA (mtDNA) is being analyzed by an increasing number of laboratories in order to investigate its potential role as an active marker of tumorigenesis in various types of cancer. Here we question the conclusions drawn in most of these investigations, especially those published in high-rank cancer research journals, under the evidence that a significant number of these medical mtDNA studies are based on obviously flawed sequencing results. Methods and Findings In our analyses, we take a phylogenetic approach and employ thorough database searches, which together have proven successful for detecting erroneous sequences in the fields of human population genetics and forensics. Apart from conceptual problems concerning the interpretation of mtDNA variation in tumorigenesis, in most cases, blocks of seemingly somatic mutations clearly point to contamination or sample mix-up and, therefore, have nothing to do with tumorigenesis. Conclusion The role of mitochondria in tumorigenesis remains unclarified. Our findings of laboratory errors in many contributions would represent only the tip of the iceberg since most published studies do not provide the raw sequence data for inspection, thus hindering a posteriori evaluation of the results. There is no precedent for such a concatenation of errors and misconceptions affecting a whole subfield of medical research.

A preliminary report on the use of methallibure in tilapia culture

The aim of this research was to retard the onset of maturation and breeding using Methallibure (I.e.I. 33,828), thereby allowing more time for somatic growth. The process of development of the gonads was studied, laboratory experiments were performed upon a breeding stock of Tilapia in an attempt to postpone spawning, and the results of these investigations were tested, on fish in ponds at the Fish Culture Research Station in Israel.

Localization and distribution of tissue type and urokinase type plasminogen activators and their inhibitors type 1 and 2 in human and rhesus monkey fetal membranes

Fetal membranes consist of 10 distinct layers including components of amnion, chorion and decidua, the latter being of maternal origin. They form mechanically integrated sheets capable of retaining amniotic fluid and play an essential role in protecting fetal growth and development in the pregnant uterus. The extracellular matrix, substrate for plasminogen activators (PAs), is an important supportive framework of the fetal membranes. :Fetal membranes from women with preterm premature rupture of membranes may differ in their protease activity compared with normal membranes. To identify the presence of PAs and their inhibitors (PAI) and their possible role in the process of fetal membrane rupture, this study in investigated the distribution and localization of both protein and mRNA for tissue (t) and urokinase (u) PA and their inhibitors type 1 (PAI-1) and type 2 (PAI-2) in amniochorion of human and rhesus monkey using conventional and. confocal immunofluorescence microscopy. In situ hybridization analysis showed that the distribution and localization of mRNAs for tPA, uPA, PAI-I and PAI-2 were similar in the fetal membranes of human and rhesus monkey; no obvious species difference was observed. Evidence of tPA mRNA was detected in amniotic epithelium, trophoblast cells and nearly all cells of the decidual layer. Strong expression of uPA mRNA was noted in the decidual cells which increased in intensity as the abscission point was approached. Weak staining in chorion laeve trophoblast was also detected. In situ hybridization experiments showed PAI-1 mRNA to be concentrated mainly in the decidual cells, some of which were interposed into the maternal-facing edge of the chorion laeve. Maximal labelling of the decidua occurred towards the zone of abscission. Weak expression of PAI-1 mRNA nas also noted in some cells of the chorion laeve. The distribution of PAI-2 mRNA in amniochorion was also concentrated in the cells of the decidual layer, maximum expression of the mRNA was in the level of abscission. No detectable amount of mRNAs for tPA, uPA, PAI-1 and PAI-2 was found in the fibroblast, reticular and spongy layers. Distribution of the proteins of tPA, uPA and PAI-1 in the fetal membranes of these two species was consistent with the distribution of their mRNA. Anti-PAI-2 immunofluorescence was found to be strongly concentrated in the amniotic epithelium, but PAI-2 mRNA was negative in this layer, suggesting that the epithelium-associated PAI-2 is not of epithelial origin. These findings suggest that a local fibrinolysis in fetal membranes generated by precisely balanced expression of PAs and their inhibitors via paracrine or autocrine mechanisms may play an essential role in fetal membrane development, maturation and in membrane rupture. Following an analysis of the distribution and synthesis of activators and inhibitors it was found that they may play a role in abscission during the third stage of labour. (C) 1998 W. B. Saunders Company Ltd.