Extraction, PCR amplification, and sequencing of mitochondrial DNA from scent mark and feces in the giant panda

To expand the feasibility of applying simple, efficient, non-invasive DNA preparation methods using samples that can be obtained from giant pandas living in the wild, we investigated the use of scent markings and fecal samples. Giant panda-specific oligonucleotide primers were used to amplify a portion of the mitochondrial DNA control region as well as a portion of the mitochondrial DNA cytochrome b gene and tRNA(Thr) gene region. A 196 base pair (bp) fragment in the control region and a 449 bp fragment in the cytochrome b gene and tRNA(Thr) gene were successfully amplified. Sequencing of polymerase chain reaction (PCR) products demonstrated that the two fragments are giant panda sequences. Furthermore, under simulated field conditions we found that DNA can be extracted from fecal samples aged as long as 3 months. Our results suggest that the scent mark and fecal samples are simple, efficient, and easily prepared DNA sources. (C) 1998 Wiley-Liss, Inc.

Stabilizing effect of surrounding gas flow on a plane liquid sheet

The stability of a plane liquid sheet is studied experimentally and theoretically, with an emphasis on the effect of the surrounding gas. Co-blowing with a gas velocity of the same order of magnitude as the liquid velocity is studied, in order to quantify its effect on the stability of the sheet. Experimental results are obtained for a water sheet in air at Reynolds number Rel = 3000 and Weber number W e = 300, based on the half-thickness of the sheet at the inlet, water mean velocity at the inlet, the surface tension between water and air and water density and viscosity. The sheet is excited with different frequencies at the inlet and the growth of the waves in the streamwise direction is measured. The growth rate curves of the disturbances for all air flow velocities under study are found to be within 20 % of the values obtained from a local spatial stability analysis, where water and air viscosities are taken into account, while previous results from literature assuming inviscid air overpredict the most unstable wavelength with a factor 3 and the growth rate with a factor 2. The effect of the air flow on the stability of the sheet is scrutinized numerically and it is concluded that the predicted disturbance growth scales with (i) the absolute velocity difference between water and air (inviscid effect) and (ii) the square root of the shear from air on the water surface (viscous effect).

Global linear and nonlinear stability of viscous confined plane wakes with co-flow

The global stability of confined uniform density wakes is studied numerically, using two-dimensional linear global modes and nonlinear direct numerical simulations. The wake inflow velocity is varied between different amounts of co-flow (base bleed). In accordance with previous studies, we find that the frequencies of both the most unstable linear and the saturated nonlinear global mode increase with confinement. For wake Reynolds number Re = 100 we find the confinement to be stabilising, decreasing the growth rate of the linear and the saturation amplitude of the nonlinear modes. The dampening effect is connected to the streamwise development of the base flow, and decreases for more parallel flows at higher Re. The linear analysis reveals that the critical wake velocities are almost identical for unconfined and confined wakes at Re ≈ 400. Further, the results are compared with literature data for an inviscid parallel wake. The confined wake is found to be more stable than its inviscid counterpart, whereas the unconfined wake is more unstable than the inviscid wake. The main reason for both is the base flow development. A detailed comparison of the linear and nonlinear results reveals that the most unstable linear global mode gives in all cases an excellent prediction of the initial nonlinear behaviour and therefore the stability boundary. However, the nonlinear saturated state is different, mainly for higher Re. For Re = 100, the saturated frequency differs less than 5% from the linear frequency, and trends regarding confinement observed in the linear analysis are confirmed.

Stabilizing effect of surrounding gas flow on a plane liquid sheet

The stability of a plane liquid sheet is studied experimentally and theoretically, with an emphasis on the effect of the surrounding gas. Co-blowing with a gas velocity of the same order of magnitude as the liquid velocity is studied, in order to quantify its effect on the stability of the sheet. Experimental results are obtained for a water sheet in air at Reynolds number Rel = 3000 and Weber number We = 300, based on the half-thickness of the sheet at the inlet, water mean velocity at the inlet, the surface tension between water and air and water density and viscosity. The sheet is excited with different frequencies at the inlet and the growth of the waves in the streamwise direction is measured. The growth rate curves of the disturbances for all air flow velocities under study are found to be within 20% of the values obtained from a local spatial stability analysis, where water and air viscosities are taken into account, while previous results from literature assuming inviscid air overpredict the most unstable wavelength with a factor 3 and the growth rate with a factor 2. The effect of the air flow on the stability of the sheet is scrutinized numerically and it is concluded that the predicted disturbance growth scales with (i) the absolute velocity difference between water and air (inviscid effect) and (ii) the square root of the shear from air on the water surface (viscous effect).

The extraction of mimosine from ipil-ipil (Leucaena leucocephala) by soaking in water

The study aimed to find a cheap and practical method of extracting mimosine from Leucaena leucocephala, otherwise known as ipil-ipil in the Philippines. L. leucocephala leaves are used in cattle, poultry and swine feed and have been tried as a food ingredient in some fish diets. While it contains relatively high amount of protein, its use as feed has been limited because of the presence of toxic substance, mimosine. Findings revealed that soaking the leaves in water was highly efficient for the extraction of mimosine, the longer the duration of soaking the more mimosine was extracted. On the other hand, 87 % of the juveniles Penaeus monodon fed with diets containing L. leucocephala leaves soaked for 24 hours survived, much higher compared to those that were fed with unsoaked leaves for eight weeks.

Extraction, purification and analysis of conotoxin of Conus textile captured from Persian Gulf and the investigation of analgesic effects of conotoxins in an animal model

Identification of venomous species of Persian Gulf cone snails and characterization of venom composition and their features is so important from the point of medical importance. Marine cone snails from the genus Conus are estimated to consist of up to 700 species. The venom of cone snails has yielded a rich source of novel neuroactive peptides or conotoxins. The present study was aimed to study the analgesic effect of Persian Gulf Conus textile and its comparison with morphine in mouse model. The specimens of Conus textile were collected of Larak Island from depth of 7 m. The collected samples were transferred to laboratory alive and were stored at -700 c. he veno s ducts were separated and ho ogenized with deionized water he ixture centrifuged at rp for inutes upernatant was considered as extracted veno and stored at - C after lyophylization. The protein profile of venom determined by using SDS-PAGE and HPLC used to investigate the extracted venom and to evaluate the analgesic activity, formalin test was carried out. SDS-PAGE indicated several bands ranged between 6 and 250 kDa. Chromatogram of the venom demonstrated more than 44 large and small fractions. The amount of 10 ng of Conus crude venom and analgesic peptide showed the best anti-pain activity in formalin test. No death observed up to 100 mg/kg, which is 250,000 times higher than the effective dose.Venom characterization of Persian Gulf Conus textile may be of medical importance and potential for new pharmaceutical drugs as well.

Venomics and biological analysis of Scatophagus argus argus (Spotted scat) venom isolated in Persian Gulf, extraction and purification of hemolytic protein

Green scat namely as Scatophagus argus argus is a venomous aquarium fish belonging to Scatophagidae family. It can induce painful wounds in injured hand with partial paralysis to whom that touch the spines. Dorsal and ventral rough spines contain cells that produce venom with toxic activities. According to unpublished data collected from local hospitals in southern coastal region of Iran, S. argus is reported as a venomous fish. Envenomation induces clinical symptoms such as local pain, partial paralysis, erythema and itching. In the present study green scat (spotted scat) was collected from Persian Gulf coastal waters. SDS-PAGE indicated 12 distinct bands in the venom ranged between 10-250 KDa. The crude venom had hemolytic activity on human erythrocytes (1%) with an LC100 (Lytic Concentration) of about 1.7 μg. The crude venom can release 813 μg proteins from 0.5% casein. Phospholipase C activity was recorded at 3.125 μg of total venom. Our findings showed that the edematic activity remained over 48 h after injection. The purification of the venom was done by HPLC and 30 peaks were obtained within 80 min but only one peak in 68 min retention time showed hemolytic activity at 90% acetonitril was isolated. The area percentage of the hemolytic protein showed that this hemolytic protein consist of 32 percent of total proteins and its molecular weight was 72 KDa in SDS_PAGE. The results demonstrated that crude venom extracted from Iranian coastal border has different toxic and enzymatic activities.

Extraction and structure elucidation of natural compounds of some gorgonians of the Persian Gulf and also physiological and pharmacological effects of gorgonian extract on some selected organisms

In the present research, investigations were carried out for structure elucidation of natural compounds and also for studing biological and teratogenical effects of two Genus of soft corals named as " Echinogorgia cf. indica" and "Sinularia erecta" in Persian Gulf. First, 350 gr Echinogorgia was extracted by Acetone, then, the extract was separated by ether from aqueos phase to give 4.5 gr oil. The oil eluted with Petrol - ether Et2 o (9:1) which was recovered Linderazulene and it's derivative as purple Cristals (350 mg/ca 0.1 %). In order to determine molecular structure, the Samples were used for spectroscopic method as: H1- NMR , C13- NMR and 2D NMR. Also, for extraction and structure elucidation of natural compounds, the soft coral " sinularia erecta " were used 1187/37 gr and extracted by Aceton. The extract was concentrated and resulting aqueous suspension and extracted by using ether to give 8.41 gr oil. The oil , was Chromatographed on a column of silica gel and some different fractions were gathered. Initial fraction (1-11) which were nonpolar compounds were seprated by GC/MS. Mass spectrum were prepared and much compounds were recognized.