Refolding of riboflavin carrier protein as probed by biochemical and immunological parameters

The unfolding of the chicken egg white riboflavin carrier protein by disulfide reduction with dithiothreitol led to aggregation with concomitant loss of ligand binding characteristics and the capacity to interact with six monoclonal antibodies directed against surface-exposed discontinuous epitopes. The reduced protein could, however, bind to a monoclonal antibody recognizing sequential epitope. Under optimal conditions of protein refolding, the vitamin carrier protein regained its folded structure with high efficiency with simultaneous complete restoration of hydrophobic flavin binding site as well as the epitopic conformations exposed at the surface in a manner comparable to its native form.

Haptic Feedback for Injecting Biological Cells using Miniature Compliant Mechanisms

We present a real-time haptics-aided injection technique for biological cells using miniature compliant mechanisms. Our system consists of a haptic robot operated by a human hand, an XYZ stage for micro-positioning, a camera for image capture, and a polydimethylsiloxane (PDMS) miniature compliant device that serves the dual purpose of an injecting tool and a force-sensor. In contrast to existing haptics-based micromanipulation techniques where an external force sensor is used, we use visually captured displacements of the compliant mechanism to compute the applied and reaction forces. The human hand can feel the magnified manipulation force through the haptic device in real-time while the motion of the human hand is replicated on the mechanism side. The images are captured using a camera at the rate of 30 frames per second for extracting the displacement data. This is used to compute the forces at the rate of 30 Hz. The force computed in this manner is sent at the rate of 1000 Hz to ensure stable haptic interaction. The haptic cell-manipulation system was tested by injecting into a zebrafish egg cell after validating the technique at a size larger than that of the cell.

Time-Dependent Predominance of Nonhomologous DNA End-Joining Pathways during Embryonic Development in Mice

Repair of DNA double-strand breaks (DSBs) is crucial for maintaining genomic integrity during the successful development of a fertilized egg into a whole organism. To date, the mechanism of DSB repair in postimplantation embryos has been largely unknown. In the present study, using a cell-free repair system derived from the different embryonic stages of mice, we find that canonical nonhomologous end joining (NHEJ), one of the major DSB repair pathways in mammals, is predominant at 14.5 day of embryonic development. Interestingly, all four types of DSBs tested were repaired by ligase IV/XRCC4 and Ku-dependent classical NHEJ. Characterization of end-joined junctions and expression studies further showed evidences for canonical NHEJ. Strikingly, in contrast to the above, we observed noncanonical end joining accompanied by DSB resection, dependent on microhomology and ligase III in 18.5-day embryos. Interestingly, we observed an elevated expression of CtIP, MRE11, and NBS1 at this stage, suggesting that it could act as a switch between classical end joining and microhomology-mediated end joining at later stages of embryonic development. Thus, our results establish for the first time the existence of both canonical and alternative NHEJ pathways during the postimplantation stages of mammalian embryonic development. (C) 2012 Elsevier Ltd. All rights reserved.

Reproductive queue without overt conflict in the primitively eusocial wasp Ropalidia marginata

Colonies of the primitively eusocial wasp Ropalidia marginata consist of a single egg layer (queen) and a number of non-egg-laying workers. Although the queen is a docile individual, not at the top of the behavioral dominance hierarchy of the colony, she maintains complete reproductive monopoly. If the queen is lost or removed, one and only one of the workers potential queen (PQ)] becomes hyperaggressive and will become the next queen of the colony. The PQ is almost never challenged because she first becomes hyperaggressive and then gradually loses her aggression, develops her ovaries, and starts laying eggs. Although we are unable to identify the PQ when the queen is present, she appears to be a ``cryptic heir designate.'' Here, we show that there is not just one heir designate but a long reproductive queue and that PQs take over the role of egg-laying, successively, without overt conflict, as the queen or previous PQs are removed. The dominance rank of an individual is not a significant predictor of its position in the succession hierarchy. The age of an individual is a significant predictor, but it is not a perfect predictor because PQs often bypass older individuals to become successors. We suggest that such a predesignated reproductive queue that is implemented without overt conflict is adaptive in the tropics, where conspecific usurpers from outside the colony, which can take advantage of the anarchy prevailing in a queenless colony and invade it, are likely to be present throughout the year.

Virgin wasps develop ovaries on par with mated females, but lay fewer eggs

In the primitively eusocial wasp Ropalidia marginata, mating is not necessary for a female wasp to develop her ovaries, lay eggs, and even to become the sole egg layer of her colony despite the presence of other mated nestmates. Here, we show that virgin wasps do not differ from their mated counterparts in the extent and rapidity of their ovarian development, in the proportion of individuals that build a nest and laid eggs, and in the time taken to do so. However, a significantly larger proportion of virgin females showed resorbing oocytes, and laid fewer eggs as compared to mated individuals. Thus, virgin females have the ability to develop ovaries and lay eggs but also to refrain from necessarily laying all mature eggs produced, before mating opportunities arise. This dual ability would be adaptive in haplodiploid, tropical species with perennial nesting cycles and frequent opportunities for workers to become replacement queens or solitary nest foundresses throughout the year.

Epoch extraction based on integrated linear prediction residual using plosion index

Epoch is defined as the instant of significant excitation within a pitch period of voiced speech. Epoch extraction continues to attract the interest of researchers because of its significance in speech analysis. Existing high performance epoch extraction algorithms require either dynamic programming techniques or a priori information of the average pitch period. An algorithm without such requirements is proposed based on integrated linear prediction residual (ILPR) which resembles the voice source signal. Half wave rectified and negated ILPR (or Hilbert transform of ILPR) is used as the pre-processed signal. A new non-linear temporal measure named the plosion index (PI) has been proposed for detecting `transients' in speech signal. An extension of PI, called the dynamic plosion index (DPI) is applied on pre-processed signal to estimate the epochs. The proposed DPI algorithm is validated using six large databases which provide simultaneous EGG recordings. Creaky and singing voice samples are also analyzed. The algorithm has been tested for its robustness in the presence of additive white and babble noise and on simulated telephone quality speech. The performance of the DPI algorithm is found to be comparable or better than five state-of-the-art techniques for the experiments considered.

Divvying up an incubator: How parasitic and mutualistic fig wasps use space within their nursery microcosm

Differential occupancy of space can lead to species coexistence. The fig-fig wasp pollination system hosts species-specific pollinating and parasitic wasps that develop within galls in a nursery comprising a closed inflorescence, the syconium. This microcosm affords excellent opportunities for investigating spatial partitioning since it harbours a closed community in which all wasp species are dependent on securing safe sites inside the syconium for their developing offspring while differing in life history, egg deposition strategies and oviposition times relative to nursery development. We determined ontogenetic changes in oviposition sites available to the seven-member fig wasp community of Ficus racemosa comprising pollinators, gallers and parasitoids. We used species distribution models (SDMs) for the first time at a microcosm scale to predict patterns of spatial occurrence of nursery occupants. SDMs gave high true-positive and low false-positive site occupancy rates for most occupants indicating species specificity in oviposition sites. The nursery microcosm itself changed with syconium development and sequential egg-laying by different wasp species. The number of sites occupied by offspring of the different wasp species was negatively related to the risk of syconium abortion by the plant host following oviposition. Since unpollinated syconia are usually aborted, parasitic wasps ovipositing into nurseries at the same time as the pollinator targeted many sites, suggesting response to lower risk of syconium abortion owing to reduced risk of pollination failure compared to those species ovipositing before pollination. Wasp life history and oviposition time relative to nursery development contributed to the co-existence of nursery occupants.

Mud-packing frog: A novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae)

Reproductive modes are diverse and unique in anurans. Selective pressures of evolution, ecology and environment are attributed to such diverse reproductive modes. Globally forty different reproductive modes in anurans have been described to date. The genus Nyctibatrachus has been recently revised and belongs to an ancient lineage of frog families in the Western Ghats of India. Species of this genus are known to exhibit mountain associated clade endemism and novel breeding behaviours. The purpose of this study is to present unique reproductive behaviour, oviposition and parental care in a new species Nyctibatrachus kumbara sp. nov. which is described in the paper. Nyctibatrachus kumbara sp. nov. is a medium sized stream dwelling frog. It is distinct from the congeners based on a suite of morphological characters and substantially divergent in DNA sequences of the mitochondrial 16S rRNA gene. Males exhibit parental care by mud packing the egg clutch. Such parental care has so far not been described from any other frog species worldwide. Besides this, we emphasize that three co-occurring congeneric species of Nyctibatrachus, namely N. jog, N. kempholeyensis and Nyctibatrachus kumbara sp. nov. from the study site differ in breeding behaviour, which could represent a case of reproductive character displacement. These three species are distinct in their size, call pattern, reproductive behaviour, maximum number of eggs in a clutch, oviposition and parental care, which was evident from the statistical analysis. The study throws light on the reproductive behaviour of Nyctibatrachus kumbara sp. nov. and associated species to understand the evolution and adaptation of reproductive modes of anurans in general, and Nyctibatrachus in particular from the Western Ghats.

Sensitivity of polarization fluctuations to the nature of protein-water interactions: Study of biological water in four different protein-water systems

Since the time of Kirkwood, observed deviations in magnitude of the dielectric constant of aqueous protein solution from that of neat water (similar to 80) and slower decay of polarization have been subjects of enormous interest, controversy, and debate. Most of the common proteins have large permanent dipole moments (often more than 100 D) that can influence structure and dynamics of even distant water molecules, thereby affecting collective polarization fluctuation of the solution, which in turn can significantly alter solution's dielectric constant. Therefore, distance dependence of polarization fluctuation can provide important insight into the nature of biological water. We explore these aspects by studying aqueous solutions of four different proteins of different characteristics and varying sizes, chicken villin headpiece subdomain (HP-36), immunoglobulin binding domain protein G (GB1), hen-egg white lysozyme (LYS), and Myoglobin (MYO). We simulate fairly large systems consisting of single protein molecule and 20000-30000 water molecules (varied according to the protein size), providing a concentration in the range of similar to 2-3 mM. We find that the calculated dielectric constant of the system shows a noticeable increment in all the cases compared to that of neat water. Total dipole moment auto time correlation function of water < dM(W) (0)delta M-W (t) > is found to be sensitive to the nature of the protein. Surprisingly, dipole moment of the protein and total dipole moment of the water molecules are found to be only weakly coupled. Shellwise decomposition of water molecules around protein reveals higher density of first layer compared to the succeeding ones. We also calculate heuristic effective dielectric constant of successive layers and find that the layer adjacent to protein has much lower value (similar to 50). However, progressive layers exhibit successive increment of dielectric constant, finally reaching a value close to that of bulk 4-5 layers away. We also calculate shellwise orientational correlation function and tetrahedral order parameter to understand the local dynamics and structural re-arrangement of water. Theoretical analysis providing simple method for calculation of shellwise local dielectric constant and implication of these findings are elaborately discussed in the present work. (C) 2014 AIP Publishing LLC.

异硫氰酸荧光素-溶菌酶的制备及晶体生长预研究

迄今尚未有适用于光源为488 nm激光扫描共聚焦显微镜研究用的溶菌酶.为此,用异硫氰酸荧光素作为探针标记了溶菌酶,测定了溶菌酶标记物的紫外-可见吸收光谱和荧光光谱,摸索了其晶体的生长条件.实验结果表明,在标记过程中异硫氰酸荧光素没有影响溶菌酶的生化性质,标记后的溶菌酶可用于激光扫描共聚焦显微镜进行后续研究.

气相扩散法生长溶菌酶晶体的动态光散射研究

首次采用动态光散射研究了气相扩散法生长溶菌酶晶体 .实验中采用了两种溶解溶菌酶的方法，所得实验结果是有区别的 .这种区别表明了 NaCl对溶菌酶分子间相互作用产生十分重要的影响 .实验结果表明，晶体生长过程中，溶液中溶菌酶始终保持单分子与两分子聚集体的状态，这种状态是生长晶体的基础 .

Proceedings of the Second PICES Workshop on the Okhotsk Sea and Adjacent Areas [Nemuro, Japan, November 9-12, 1998]

Table of Contents [pdf, 0.01 Mb] Preface [pdf, 0.01 Mb] Masaaki Aota Long-term tendencies of sea ice concentration and air temperature in the Okhotsk Sea coast of Hokkaido [pdf, 0.05 Mb] Hajime Ito & Miki Yoshioka Geography of the seasonally ice covered seas [pdf, 0.5 Mb] George V. Shevchenko & Victor F. Putov On wind and tide induced sea-ice drift on the northeastern shelf of Sakhalin Island (analysis of radar data) [pdf, 0.96 Mb] Boris S. Dyakov, A.A. Nikitin, L. S. Muktepavel & T.A. Shatilina Variability of the Japan and Okhotsk Seas ice cover depending on geopotential field H500 over the Far-Eastern region [pdf, 0.10 Mb] Aleksandr G. Petrov & Nikolay A. Rykov Intermediate cold layer and ice cover in the Sea of Okhotsk [pdf, 0.37 Mb] Vladimir Ponomarev, Olga Trusenkova, Elena Ustinova & Dmitry Kaplunenko Interannual variations of oceanographic and meteorological characteristics in the Sea of Okhotsk [pdf, 0.16 Mb] George V. Shevchenko & Akie Kato Seasonal and interannual changes of atmospheric pressure, air and water temperature in the area of the Kuril Ridge [pdf, 0.13 Mb] George V. Shevchenko & Vladimir Yu. Saveliev Spatial variability of the wind field in the area of the Kuril Islands [pdf, 0.15 Mb] Alexander L. Figurkin & Igor A. Zhigalov Seasonal variability and specifity of the oceanological conditions in the northern Okhotsk Sea in 1997 [pdf, 1.04 Mb] Igor A. Zhabin Ventilation of the upper portion of the intermediate water in the Okhotsk Sea [pdf, 0.80 Mb] Vladimir A. Luchin & Alexander L. Figurkin Oceanographic conditions over the Kashevarov Bank [pdf, 0.61 Mb] Toshiyuki Awaji, Tomohiro Nakamura, Takaki Hatayama, Kazunori Akimoto & Takatoshi Takizawa Tidal exchange through the Kuril Straits [pdf, 2.01 Mb] Tomohiro Nakamura, Toshiyuki Awaji, Takaki Hatayama, Kazunori Akimoto, Takatoshi Takizawa & Masao Fukasawa Vertical mixing induced by tidally generated internal waves in the Kuril Straits [pdf, 0.83 Mb] Katsuro Katsumata & Ichiro Yasuda Water exchange between the Okhotsk Sea and the North Pacific Ocean estimated by simple models [pdf, 0.97 Mb] Konstantin A. Rogachev Oyashio west path culmination as the consequence of a rapid thermohaline transition in the Pacific Subarctic [pdf, 0.22 Mb] Yasuhiro Kawasaki On the year-to-year change in subarctic water characteristics around the Kuril Islands [pdf, 0.39 Mb] Alexander L. Figurkin & Evgeniy E. Ovsyannikov Influence of oceanological conditions of the West Kamchatka shelf waters on spawning grounds and on pollock egg distribution [pdf, 0.97 Mb] Igor E. Kochergin & Alexander A. Bogdanovsky Transport and turbulence characteristics for the northeastern Sakhalin shelf conditions [pdf, 0.08 Mb] Igor E. Kochergin, Alexander A. Bogdanovsky, Valentina D. Budaeva, Vyacheslav G. Makarov, Vasily F. Mishukov, S.N. Ovsienko, Victor F. Putov, L.A. Reitsema, J.W. Sciallabba, O.O. Sergucheva & P.V. Yarosh Modeling of oil spills for the shelf conditions of northeastern Sakhalin [pdf, 0.32 Mb] Valentina D. Budaeva & Vyacheslav G. Makarov A peculiar water regime of currents in the area of eastern Sakhalin shelf [pdf, 0.66 Mb] Nikolay A. Rykov The oceanographic databases on the Sakhalin shelf [pdf, 0.27 Mb] Akifumi Nakata, Iori Tanaka, Hiroki Yagi, Tomomi Watanabe, Gennady A. Kantakov & Andrew D. Samatov Formation of high-density water (over 26.8 sigma-t) near the La Perouse Strait (the Soya Strait) [pdf, 0.09 Mb] Minoru Odamaki & Kouji Iwamoto Currents and tidal observations by Hydrographic Department of Maritime Safety Agency, off the Okhotsk coast of Hokkaido [pdf, 0.16 Mb] Yasushi Fukamachi, Genta Mizuta, Kay I. Ohshima, Motoyo Itoh, Masaaki Wakatsuchi & Masaaki Aota Mooring measurements off Shiretoko Peninsula, Hokkaido in 1997-1998 [pdf, 0.19 Mb] Mikhail A. Danchenkov, David Aubrey & Stephen C. Riser Oceanographic features of the La Perouse Strait [pdf, 0.91 Mb] Iori Tanaka & Akifumi Nakata Results of direct current measurements in the La Perouse Strait (the Soya Strait), 1995-1998 [pdf, 0.06 Mb] Gennady A. Kantakov & George V. Shevchenko In situ observations of Tsushima and West-Sakhalin currents near La Perouse (Soya) Strait [pdf, 0.79 Mb] Irina Y. Bragina Geographical and biological characteristics of the net zooplankton in the southwestern part of the Sea of Okhotsk during 1987-1996 [pdf, 0.27 Mb] List of corresponding authors [pdf, 0.01 Mb] (Document pdf contains 193 pages)

Population Characteristics of Largemouth Bass Associated with Changes in Abundance of Submersed Aquatic Vegetation in Lake Seminole, Georgia

Population characteristics of largemouth bass ( Micropterus salmoides L.) including growth, body condition (relative weight), survival, and egg production were examined in relation to abundance of submersed aquatic vegetation (SAV) coverage (primarily hydrilla [ Hydrilla verticillata L.f. Royle]) in three embayments of Lake Seminole, GA, and compared to a previous study conducted in 1998. (PDF has 8 pages.)

Effects of shear on eggs and larvae of striped bass, morone saxatilis, and white perch, M. americana

Shear stress, generated by water movement, can kill fish eggs and larvae by causing rotation or deformation. Through the use of an experimental apparatus, a series of shear (as dynes/cm2)-mortality equations for fixed time exposures were generated for striped bass and white perch eggs and larvae. Exposure of striped bass eggs to a shear level of 350 dynes/cm2 kills 36% of the eggs in 1 min; 69% in 2 min, and 88% in 4 min; exposure of larvae to 350 dynes/cm2 kills 9.3% in 1 min, 30.0% in 2 min, and 68.1% in 4 min. A shear level of 350 dynes/cm2 kills 38% of the white perch eggs in 1 min, 41% in 2 min, 89% in 5 min, 96% in 10 min, and 98% in 20 min. A shear level of 350 dynes/cm2 applied to white perch larvae destroys 38% of the larvae in 1 min, 52% in 2 min, and 75% in 4 min. Results are experimentally used in conjunction with the determination of shear levels in the Chesapeake and Delaware Canal and ship movement for the estimation of fish egg and larval mortalities in the field.

Assessment of Atlantic Red Drum for 1999: Northern and southern regions

An assessment of the status of the Atlantic stock of red drum is conducted using recreational and commercial data from 1986 through 1998. This assessment updates data and analyses from the 1989, 1991, 1992 and 1995 stock assessments on Atlantic coast red drum (Vaughan and Helser, 1990; Vaughan 1992; 1993; 1996). Since 1981, coastwide recreational catches ranged between 762,300 pounds in 1980 and 2,623,900 pounds in 1984, while commercial landings ranged between 60,900 pounds in 1997 and 422,500 pounds in 1984. In weight of fish caught, Atlantic red drum constitute predominantly a recreational fishery (ranging between 85 and 95% during the 1990s). Commercially, red drum continue to be harvested as part of mixed species fisheries. Using available length-frequency distributions and age-length keys, recreational and commercial catches are converted to catch in numbers at age. Separable and tuned virtual population analyses are conducted on the catch in numbers at age to obtain estimates of fishing mortality rates and population size (including recruitment to age 1). In tum, these estimates of fishing mortality rates combined with estimates of growth (length and weight), sex ratios, sexual maturity and fecundity are used to estimate yield per recruit, escapement to age 4, and static (or equilibrium) spawning potential ratio (static SPR, based on both female biomass and egg production). Three virtual analysis approaches (separable, spreadsheet, and FADAPT) were applied to catch matrices for two time periods (early: 1986-1991, and late: 1992-1998) and two regions (Northern: North Carolina and north, and Southern: South Carolina through east coast of Florida). Additional catch matrices were developed based on different treatments for the catch-and-release recreationally-caught red drum (B2-type). These approaches included assuming 0% mortality (BASEO) versus 10% mortality for B2 fish. For the 10% mortality on B2 fish, sizes were assumed the same as caught fish (BASEl), or positive difference in size distribution between the early period and the later period (DELTA), or intermediate (PROP). Hence, a total of 8 catch matrices were developed (2 regions, and 4 B2 assumptions for 1986-1998) to which the three VPA approaches were applied. The question of when offshore emigration or reduced availability begins (during or after age 3) continues to be a source of bias that tends to result in overestimates of fishing mortality. Additionally, the continued assumption (Vaughan and Helser, 1990; Vaughan 1992; 1993; 1996) of no fishing mortality on adults (ages 6 and older), causes a bias that results in underestimates of fishing mortality for adult ages (0 versus some positive value). Because of emigration and the effect of the slot limit for the later period, a range in relative exploitations of age 3 to age 2 red drum was considered. Tuning indices were developed from the MRFSS, and state indices for use in the spreadsheet and FADAPT VPAs. The SAFMC Red Drum Assessment Group (Appendix A) favored the FADAPT approach with catch matrix based on DELTA and a selectivity for age 3 relative to age 2 of 0.70 for the northern region and 0.87 for the southern region. In the northern region, estimates of static SPR increased from about 1.3% for the period 1987-1991 to approximately 18% (15% and 20%) for the period 1992-1998. For the southern region, estimates of static SPR increased from about 0.5% for the period 1988-1991 to approximately 15% for the period 1992-1998. Population models used in this assessment (specifically yield per recruit and static spawning potential ratio) are based on equilibrium assumptions: because no direct estimates are available as to the current status of the adult stock, model results imply potential longer term, equilibrium effects. Because current status of the adult stock is unknown, a specific rebuilding schedule cannot be determined. However, the duration of a rebuilding schedule should reflect, in part, a measure of the generation time of the fish species under consideration. For a long-lived, but relatively early spawning, species as red drum, mean generation time would be on the order of 15 to 20 years based on age-specific egg production. Maximum age is 50 to 60 years for the northern region, and about 40 years for the southern region. The ASMFC Red Drum Board's first phase recovery goal of increasing %SPR to at least 10% appears to have been met. (PDF contains 79 pages)