Direct Count Method Research Articles

Seasonal variation in the population of Black kite (Milvus migrans), inhabiting the Arid Zone of Rajasthan

The population of Black kite (Milvus migrans) inhabiting rural and urban area of Churu city, Rajasthan was seasonally determined by the direct count method. The counting of the Black kite was made at roosting sites of Churu city (28o 15’ N and 74o 55’ E, 286 msl) and it’s out skirts area of 400 km2. Total 46 roosting sites were identified in the study area of which 34 were located in the urban area and 12 in the rural areas. It was found that number was relatively higher from June to September 2012, 2013 and 2014. However, the maximum numbers of Black kite were during August 2012 and July 2014 and minimum were during November 2012 and February 2012. The maximum number of adult was noticed in the rainy months of every year. A sharp decline in the juvenile population was observed from September 2014 to December 2014. Maximum 28 and 22 juvenile birds were observed in the month of August 2012 and July 2013 while the minimum 4 juvenile were observed in February 2012, March 2013 and November 2014. An average 25.41 ±SD12.4 and 42.33 ±SD 18.82in 2102, 27.33 ±SD 9.19 and 54.25 ±SD 14.9 in 2013, 20.90 ±SD 5.05and 43.08 ±SD 12.75in 2014 Black kites were observed in rural and urban areas, respectively. The maximum number of Black kites i.e. 134 individuals, occurred in the month of August 2012. But in general, the population remained more or less constant with seasonal variations involving dispersion during the monsoons and aggregation during the winters. The winter season adversely affects the population of Black kite in the study area because of the non-availability of preferential food.

Banco de semillas de retamo espinoso Ulex europaeus L. en bordes del matorral invasor en un ecosistema zonal de bosque altoandino, Colombia

Se estableció la densidad de semillas de retamo espinoso (Ulex europaeus) en bordes de matorral invasor alsur de los Cerros Orientales de Bogotá. A través del método de conteo directo se estableció la abundancia desemillas presente en seis puntos de muestreo en sitios ubicados entre los 2.700 y 3.200 m de altitud, y cuatrointervalos de profundidad entre los 0 y 20 cm. Se establecieron densidades de semillas variables para los seispuntos de muestreo entre 109 y 3.384 semillas m-2. A nivel de profundidad, la mayor densidad de semillasse encuentra en el intervalo de 0 a 5 cm para todos los sitios muestreados. En los puntos de muestreo seencontraron diferencias significativas en la cantidad de semillas a diferentes altitudes (p=0,0416) y intervalosde profundidad evaluados (p=0,0392). Se determina que existe un alto potencial en los bordes de matorral parala extensión de las áreas invadidas, por activación del banco de semillas de la especie invasora, lo que sugierela necesidad de una estrategia prioritaria para el control de estas áreas de borde en el marco de procesos derestauración ecológica en ecosistemas altoandinos.

Suitability of Two Sampling Methods for Determining the Population Trends of Certain Sap-Sucking Species Inhabiting Tomato and Cucumber Plantations

Faunistic composition and seasonal abundance of certain sap-sucking species inhabiting tomato and cucumber plantations were determined by using sweep net and direct count methods, during summer seasons of 2015 and 2016 in Assiut Governorate, Northern Upper Egypt. Twelve species belonging to 8 families and 3 orders in addition to the acarid mite were recorded. Heteropteran species were found to be constituted 69.24% of the gathered species. However, Homopteran species were found to be constituted 15.38%. Thysanoptera and Tetranichidae were presented by 7.69% for each. Concerning to the sampling method, 76.92% of the collected species were captured by the sweep net method, while 23.08% were gathered by the direct count method. The recovered species were classified as predominantly predaceous, predominantly phytophagous, predaceous in part and phytophagous in part species. Seasonal abundance of the recorded species was determined. The distinct predators, Coranus aegyptius (Fabricius, 1775) and Orius spp. showed relatively low abundance percentages. The predatory-phytophagous mirid species [Campylomma unicolor Poppius, 1914; Nesidiocoris tenuis (Reuter, 1895); Creontiades pallidus (Rambur, 1839). and Deraeocoris serenus (Douglas & Scott, 1868)], showed different abundance percentages. Amongst these taxa, N. tenuis revealed the highest abundance percentage on tomato plantations with an average of 63.00 and 75.00% during 2015 and 2016 seasons, respectively. The distinct phytophagous species, Thrips tabaci Lindeman, 1889; Bemisia tabaci (Gennadius, 1889) and Tetranychus urticae Koch, 1836 presented the greatest abundance percentages on cucumber when estimated by the direct count method.So, this work can be reflect the importance of sweep net as a suitable sampling method for determine the faunistic composition of the flight predatory insects inhabiting tomato and cucumber. However, direct count can be consider the most suitable method for determine the population trends of mobile or non-mobile arthropods inhabiting the same crops. Also, use of more than one sampling method could be successful to clarify the relation between useful and harmful arthropod species.

Hematologic reference intervals and age effect in European Strigiformes.

The clinical importance of hematologic testing in avian veterinary medicine is reflected in the increasing number of studies for the establishment of hematologic RIs of Strigiformes and other species. Age is an important physiologic factor in birds and the effect on hematology variable should be understood. The objective of this study was to determine baseline data of hematologic variables in 5 species of Iberian Strigiformes in different age classes. Nocturnal birds of prey were sampled at Wildlife Health Centers. Packed cell volume was determined by the microhematocrit centrifugation method, and RBC and WBC counts were determined using the direct hemocytometer count method with Natt and Herrick solution. Hemoglobin concentration was measured spectrophotometrically. The MCV, MCHC, and MHC were calculated using the standard formulas. The differential WBC count was performed by the routine microscopic evaluation of 200 cells on a blood smear manually stained with Wright stain. Thrombocyte blood count estimate was obtained from the blood film. No differences were observed between juveniles and adults for any variable evaluated in Tawny owl, Little owl, Scops owl, Long-eared owl, and Barn owl. In addition, PCV, RBC, and HGB of chicks were statistically significantly lower than in juveniles and adults, and total WBC was significantly higher in Tawny owl, Little owl, Scops owl, and Long-eared owl. Our findings provide evidence that laboratory data from chicks of Strigiformes are different compared to juveniles and adults; therefore, separate RIs were defined.

The perimeter of words

We define [k]={1,2,…,k} to be a (totally ordered) alphabet on k letters. A wordw of length n on the alphabet [k] is an element of [k]n. A word can be represented by a bargraph (i.e., by a column-convex polyomino whose lower edges lie on the x-axis) in which the height of the ith column equals the size of the ith part of the word. Thus these bargraphs have heights which are less than or equal to k. We consider the perimeter, which is the number of edges on the boundary of the bargraph. By way of Cramer’s method and the kernel method, we obtain the generating function that counts the perimeter of words. Using these generating functions we find the average perimeter of words of length n over the alphabet [k]. We also show how the mean and variance can be obtained using a direct counting method.

Abstract 5929: A novel image cytometric analysis method for T-cell-mediated cytotoxicity of 3D tumor spheroids

Abstract Cell-mediated cytotoxicity assays have been frequently performed to characterize cancer cytotoxic potential of immune cells, antibodies, and drug compounds. Traditionally, these assays are performed using release assays such as Cr51 (radioactivity), Calcein (fluorescence), or LDH (enzymatic). However, release assays have limitations such as the handling of hazardous material, the indirect measurement of cell death leading to an under estimation of cytotoxicity, the requirement for a large volume of cell sample, and the inability to visually confirm, image ad track the assay kinetically. In the recent years, Celigo image cytometry has been used to perform high-throughput cell-mediated cytotoxicity assays using a direct cell counting method where cancer cells (Target) are stained with Calcein AM. Upon the addition of effector immune cells with the fluorescently stained Target cells in the presence or absence of antibody or drug compounds, the Celigo image cytometer is used to capture bright-field and fluorescent images and analyze the change in Target cell count over time to determine the cytotoxicity percentage. In general, these assays are performed in a 2D cultures. The 2D assays for cancer drug discovery have had some difficulty in identifying more qualified drug candidates for clinical testing, thus there has been an increase in interest of performing cytotoxicity assays in 3D tumor models. Traditional 3D spheroid analysis methods require the use of standard microscopy, which is time-consuming and subjective. Celigo imaging cytometer has also been used to rapidly analyze drug effects on 3D tumor spheroids. In this work, we demonstrate a novel method of analyzing T cell-mediated cytotoxicity on 3D tumor spheroids in the presence of absence of ImmTAC molecules, which can promote higher T cell killing. In this experiment, MDA-MB-453 GFP expressing breast cancer cells are used to form tumor spheroids in an ultra-low attachment plate. The spheroids are then treated with primary T cells at 1:10 and 1:50 E:T ratios, as well as 10, 1, 0.1, 0.01, and 0.001 nM ImmTAC. The results showed a dose response effect of T cell killing with the addition of ImmTAC molecules by measuring spheroid size in GFP fluorescence and viability using propidium iodide. The image cytometry method was able to monitor changes in spheroid size and fluorescence over time and rapidly quantify T cell killing, which was as high as 75%. The captured bright-field and fluorescent images also clearly showed the cytotoxicity effect from the combination of T cells and ImmTAC. The ability to screen cytotoxic effects of immune cells, antibody, and drug compounds on 3D tumor spheroids can provide an alternative tumor model for identifying more qualified cancer drug candidates for drug discovery campaigns. Citation Format: Leo Li-Ying Chan, Laure Humbert, Scott Cribbes. A novel image cytometric analysis method for T-cell-mediated cytotoxicity of 3D tumor spheroids [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5929. doi:10.1158/1538-7445.AM2017-5929

Bio-corrosion of carbon steel by sulfate reducing bacteria consortium in oil and gas pipelines

This research is aimed to give an overview of the impact of bio-corrosion on carbon steel grade API 5L X-70 immersed in growth medium and exposed to SRB consortium. Simulation of anaerobic corrosion conditions was carried out in a laboratory for 28 days. Raw crude oil gathered from the Baram Delta Operation Terminal was cultured in broth number 1249 (Modified Barr’s Medium) to study the effect of bacteria growth upon metal loss. Carbon steel coupons grade X70 were cut to approximately 10mm x 20mm x 5mm and immersed in the cultured broth. During the experiment, the planktonic SRB were enumerated using a counting chamber (direct cell count method) under the electronic microscope at 200x magnification. Results indicated that the optimum pH and temperature for the respected SRB consortium genes were 8.5 and 37 °C, respectively. Metal loss of the corrosion specimen was measured and recorded after retrieval from the immersion period in the medium on a weekly basis prior to SRB inoculation for further analysis. The metal loss values supported that SRB activity can increase the metal loss of carbon steel against time of exposure. Additionally, the FESEM image showed the biofilm formations on the corrosion specimen. Thus, the results could conclude that biocorrosion caused by particular local SRB consortium can be considered as a threat to carbon steel pipelines. Besides, the effect of SRB activity and response towards metallic materials in a dynamic environment is an interesting topic to be studied upon in the future.

Sixty-six Biallelic Genetic Markers on Y chromosome by MALDI-TOF-MS

To analyse the genetic polymorphisms of 66 biallelic genetic markers on Y chromosome in Eastern Chinese Han population, and evaluate their values in forensic application. Genotyping of 66 biallelic genetic markers on Y chromosome was studied in 205 unrelated males of Eastern Chinese Han population by multiplex PCR combined matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS）. The allele frequencies on the loci to be tested were calculated by direct counting method, and the gene diversity （GD） and haplotype diversity （HD） were calculated by corresponding formulas. The haplotypes of this system were tested by software Arlequin v3.5.2.2 and the comparison of population genetics were analyzed. A total of 60 biallelic genetic markers on Y chromosome were polymorphic in males of Eastern Chinese Han population, and the ranges of GD were from 0.038 5 to 0.501 9. Eighty-five different haplotypes were observed and the HD was 0.970 3. The differences of partial SNP loci between the Han population of Eastern China and that of Xinjiang and Guangdong were statistically significance. Sixty biallelic genetic markers and the detection system can complementally provide genetic information in kinship testing and individual identification. The MALDI-TOF-MS technology is able to type biallelic genetic markers.

The site-perimeter of words

We define $[k]={1‎, ‎2‎, ‎3,ldots,k}$ to be a (totally ordered) {em alphabet} on $k$ letters‎. ‎A {em word} $w$ of length $n$ on the alphabet $[k]$ is an element of $[k]^n$‎. ‎A word can be represented by a bargraph which is a family of column-convex polyominoes whose lower edge lies on the $x$-axis and in which the height of the $i$-th column in the bargraph equals the size of the $i$-th part of the word‎. ‎Thus these bargraphs have heights which are less than or equal to $k$‎. ‎We consider the site-perimeter‎, ‎which is the number of nearest-neighbour cells outside the boundary of the polyomino‎. ‎The generating function that counts the site-perimeter of words is obtained explicitly‎. ‎From a functional equation we find the average site-perimeter of words of length $n$ over the alphabet $[k]$‎. ‎We also show how these statistics may be obtained using a direct counting method and obtain the minimum and maximum values of the site-perimeters‎.

(Invited) Recent Advances in SWCNT Analytical Spectroscopy

Since the 2002 assignment of absorption and emission transitions to specific (n,m) structures, optical spectroscopy has offered the potential for simple, detailed structure-resolved characterization of SWCNT samples. However, this goal has not been fully realized because of incomplete spectroscopic and photophysical knowledge and a lack of specialized instrumentation. We describe here recent progress in overcoming these problems to enable quantitative sample analysis through combined fluorescence and absorption spectroscopy. One important step was determining absolute E11 absorption cross sections (molar absorptivities) for a number of common (n,m) species. This was achieved by using a direct counting method to calibrate the absolute concentrations of reference samples, and then by extending the list of measured cross sections through variance spectroscopy determinations of (n,m)-resolved particle concentrations in mixed samples. To properly interpret fluorimetric data, we have measured, interpreted, and modeled emission spectra and excitation spectra for a range of semiconducting (n,m) species. Combinations of single particle and ensemble spectroscopies were used for these studies. The models were extrapolated to other (n,m) species to predict E11 line widths and shapes plus the positions, amplitudes, and widths of spectral sidebands. When combined with data on structure-dependent fluorimetric efficiencies, the results allow fluorescence spectra of nanotube mixtures to be simulated as a sum of contributions from multiple (n,m) species of determined relative concentrations. To convert those concentrations from relative into absolute values, the absorption spectrum of a mixed sample is simulated as a background function plus components determined from the fluorescence emission and excitation profiles for the (n,m) species that were found to be present. Then the separate E11 (n,m) absorbance components are divided by their structure-specific absorption cross sections to find absolute species concentrations. The overall process allows quantitation of semiconducting (n,m) concentrations from simple and quick spectral measurements.

An update of the breeding population status of the critically endangered Mariana Crow Corvus kubaryi on Rota, Northern Mariana Islands 2013–2014

SummaryThe critically endangered Mariana Crow now exists in a single population on the island of Rota, Northern Mariana Islands. Targeted management requires an accurate measure of the population status of this species. In Mariana Crows the breeding population is both the easiest cohort to accurately survey and the most important segment of the population in terms of population recovery. The total number of Mariana Crow territorial pairs was estimated on the island of Rota using a direct count method, and total population size was calculated using a Chapman estimate. From September 2013 to April 2014, 46 crow pairs were found and up to an additional eight pairs were estimated in unsearched areas. The total population was estimated to be 178 individuals. This represents a 10–23% decline in pairs in the six years since 2007 and a 46–53% decline since 1998. This number is also considerably lower than the minimum 75 pairs recommended to maintain a viable population on Rota.

Blocking autophagy enhances the pro-apoptotic effect of bufalin on human gastric cancer cells through endoplasmic reticulum stress.

ABSTRACTBufalin has been used to treat cancer for several years. However, the molecular mechanisms for its anti-tumor function are not fully understood. This work aimed to investigate the effect of bufalin on the proliferation and apoptosis of human gastric cancer (HGC) cells and the roles of endoplasmic reticulum (ER) stress and autophagy in bufalin-induced apoptosis. HGC cell lines, SGC7901 and BGC823, were treated with different concentrations of bufalin or 80 nmol/l bufalin for 1, 2, 3 and 4 days. Cell counting kit-8 (CCK-8) assay and direct cell counting method were used to detect proliferation. Cell cycle arrest and apoptosis was detected using flow cytometry. Protein levels of caspase-3, -8, Bax/Bcl-2, Beclin-1, LC3, inositol-requiring enzyme 1 (IRE1) and C/EBP homologous protein (CHOP) were determined using western blotting. Autophagy was blocked using 3-methyladenine (3MA) or Atg5 siRNA to evaluate the effect of autophagy on bufalin-induced apoptosis. The IRE1 and CHOP were knocked down using specific siRNA to determine the pathway involved in bufalin-induced autophagy. It was found that bufalin significantly suppressed proliferation of SGC7901 and BGC823 cells and induced apoptosis in a time- and dose-dependent manner. The mechanism responsible for bufalin-induced apoptosis was the formation of ER stress via the IRE1-JNK pathway. Moreover, autophagy was activated during ER stress, and blocking autophagy significantly exacerbated bufalin-induced apoptosis.

Detection of Acute Radiation Sickness: A Feasibility Study in Non-Human Primates Circulating miRNAs for Triage in Radiological Events.

Development of biomarkers capable of estimating absorbed dose is critical for effective triage of affected individuals after radiological events. Levels of cell-free circulating miRNAs in plasma were compared for dose-response analysis in non-human primates (NHP) exposed to lethal (6.5 Gy) and sub-lethal (1 and 3 Gy) doses over a 7 day period. The doses and test time points were selected to mimic triage needs in the event of a mass casualty radiological event. Changes in miRNA abundance in irradiated animals were compared to a non-irradiated cohort and a cohort experiencing acute inflammation response from exposure to lipopolysaccharide (LPS). An amplification-free, hybridization-based direct digital counting method was used for evaluation of changes in microRNAs in plasma from all animals. Consistent with previous murine studies, circulating levels of miR-150-5p exhibited a dose- and time-dependent decrease in plasma. Furthermore, plasma miR-150-5p levels were found to correlate well with lymphocyte and neutrophil depletion kinetics. Additionally, plasma levels of several other evolutionarily and functionally conserved miRNAs were found altered as a function of dose and time. Interestingly, miR-574-5p exhibited a distinct, dose-dependent increase 24 h post irradiation in NHPs with lethal versus sub-lethal exposure before returning to the baseline level by day 3. This particular miRNA response was not detected in previous murine studies but was observed in animals exposed to LPS, indicating distinct molecular and inflammatory responses. Furthermore, an increase in low-abundant miR-126, miR-144, and miR-21 as well as high-abundant miR-1-3p and miR-206 was observed in irradiated animals on day 3 and/or day 7. The data from this study could be used to develop a multi-marker panel with known tissue-specific origin that could be used for developing rapid assays for dose assessment and evaluation of radiation injury on multiple organs. Furthermore this approach may be utilized to screen for tissue toxicity in patients who receive myeloablative and therapeutic radiation.

HLA-A, HLA-B and HLA-DRB1 allele and haplotype frequencies of 10918 Koreans from bone marrow donor registry in Korea.

The human leucocyte antigen (HLA) system is the most polymorphic genetic system in humans, and HLA matching is crucial in organ transplantation, especially in hematopoietic stem cell transplantation. We investigated HLA-A, HLA-B and HLA-DRB1 allele and haplotype frequencies at allelic level in 10918 Koreans from bone marrow donor registry in Korea. Intermediate resolution HLA typing was performed using Luminex technology (Wakunaga, Japan), and additional allelic level typing was performed using PCR-single-strand conformation polymorphism method and/or sequence-based typing (Abbott Molecular, USA). Allele and haplotype frequencies were calculated by direct counting and maximum likelihood methods, respectively. A total of 39 HLA-A, 66 HLA-B and 47 HLA-DRB1 alleles were identified. High-frequency alleles found at a frequency of ≥5% were 6 HLA-A (A*02:01, *02:06, *11:01, *24:02, *31:01 and *33:03), 6 HLA-B (B*15:01, *35:01, *44:03, *51:01, 54:01 and *58:01) and 8 HLA-DRB1 (DRB1*01:01, *04:05, *04:06, *07:01, *08:03, *09:01, *13:02 and *15:01) alleles. At each locus, A*02, B*15 and DRB1*14 generic groups were mostdiverse at allelic level, consisting of 9, 12 and 11 different alleles, respectively. A total of 366, 197 and21 different HLA-A-B-DRB1 haplotypes were estimated with frequencies of ≥0.05%, ≥0.1% and ≥0.5%, respectively. The five most common haplotypes with frequencies of ≥2.0% were A*33:03-B*44:03-DRB1*13:02 (4.97%), A*33:03-B*58:01-DRB1*13:02, A*33:03-B*44:03-DRB1*07:01, A*24:02-B*07:02-DRB1*01:01 and A*24:02-B*52:01-DRB1*15:02. Among 34 serologic HLA-A-B-DR haplotypes with frequencies of ≥0.5%, 17 haplotypes revealed allele-level diversity and majority of the allelic variation was arising from A2, A26, B61, B62, DR4 and DR14 specificities. Haplotype diversity obtained in this study is the most comprehensive data thus far reported in Koreans, and the information will be useful for unrelated stem cell transplantation as well as for disease association studies.

Population organization and ranging pattern of Rhesus macaque (Macaca mulatta) at Bandar, Narayanganj

A study was conducted on population organization and ranging pattern of rhesus macaque at Bandar Upazila, Narayanganj, Dhaka from December, 2009 to September, 2010. Direct count method was used to study the group size and composition, and ranging was determined by GPS coordination plotting on the map. A total of 66 monkeys (29 to 37 individuals) were found in two groups with a mean of 33±5.6. Among the population 7.6% adult male, 33.3% adult female, 9.1% sub-adult male, 7.6% sub-adult female, 25.8% juvenile and 16.7% was infant. The ratio between adult male and adult female was 1:4.4 and between adult and nonadults was 1:1.4. Day range length varied from 1543m to1716m (mean=1638.43±56.7). Home range was completely overlapped and varied negligibly from 45.2 ha. to 45.5 ha. (mean=45.32±0.2). They minimized conflictions by using different time to move. But the growing dependence of these urban macaques on human resources is a matter of concern. Therefore, conservation strategies should be directed to minimize this dependence.Jahangirnagar University J. Biol. Sci. 3(1): 27-35, 2014 (June)

Fabrication, optical properties and laser outputs of Nd:YAG ceramics based on laser ablated and pre-calcined powders

Transparent Nd:YAG ceramic was fabricated by the solid-state reaction method with an additional round of pre-calcining using nanopowders of 1 at.% Nd:Y2O3 and Al2O3 synthesized by laser ablation. The pre-calcining step and addition of tetraethyl orthosilicate was found crucial for fabricating high optical quality Nd:YAG ceramic from such nanoparticles. The transmittance of the obtained 2-mm-thick Nd:YAG ceramic was 83.6% at the wavelength of 1064 nm, which is very close to the theoretical value. The uniformity of the optical quality of ceramic was mapped by analyzing the point-by-point transmission of a focused laser beam. The average volume of the scattering centers in the obtained ceramic was evaluated by direct count method to be 17 ppm. The Nd distribution was determined by fluorescence imaging to be homogeneous throughout the sample. Output power of 4.9 W with a slope efficiency of 52.7% was obtained in 1.5-mm-thick Nd:YAG ceramic under a quasi-continuous wave (QCW) laser diode end pumping at 805 nm.

Campylobacter in broiler slaughter samples assessed by direct count on mCCDA and Campy-Cefex agar.

Campylobacter spp. cause foodborne illnesses in humans primarily through the consumption of contaminated chicken. The aim of this study was to evaluate the United States Department of Agriculture's (USDA) recommended methodology, protocol MLG 41.02, for the isolation, identification and direct plate counting of Campylobacter jejuni and C. coli samples from the broiler slaughtering process. A plating method using both mCCDA and Campy-Cefex agars is recommended to recover Campylobacter cells. It is also possible to use this method in different matrices (cloacal swabs and water samples). Cloacal swabs, samples from pre-chiller and post-chiller carcasses and samples of pre-chiller, chiller and direct supply water were collected each week for four weeks from the same flock at a slaughterhouse located in an abattoir in southern Brazil. Samples were analyzed to directly count Campylobacter spp., and the results showed a high frequency of Campylobacter spp. on Campy-Cefex agar. For the isolated species, 72% were identified as Campylobacter jejuni and 38% as Campylobacter coli. It was possible to count Campylobacter jejuni and Campylobacter coli from different samples, including the water supply samples, using the two-agar method. These results suggest that slaughterhouses can use direct counting methods with both agars and different matrices as a monitoring tool to assess the presence of Campylobacter bacteria in their products.

HLA-A, HLA-B, HLA-DRB1 allele and haplotype frequencies of 14,529 Chinese Han bone marrow donors living in Dalian, China.

We investigated the allele and haplotype frequencies of HLA-A, HLA-B and HLA-DRB1 loci in Dalian Chinese Han population using blood samples of unrelated marrow donors who live in Dalian. The genetic relationship between Dalian and different regions worldwide was further explored based on HLA status of different populations. A total of 14,529 samples were genotyped at 2-digit level only by sequence-specific oligonucleotide and sequence-based typing methods. Allele frequencies of HLA-A, HLA-B and HLA-DRB1 were calculated by the direct counting method. Haplotype frequencies and linkage disequilibrium (LD) values were calculated by the maximum likelihood method. F(ST) values were calculated by allele frequency data of each locus. Phylogeny tree of Nei's DA genetic distances was constructed by the UPGMA method. HLA-A*02 was the most frequent allele at HLA-A locus followed by A*11 and A*24. Alleles at HLA-B locus ranked in decreasing order by frequency were B*40, B*15 and B*13. The three highest frequency alleles were DRB1*15, DRB1*09 and DRB1*12 at HLA-DRB1 locus. A*30-B*13-DRB1*07 was the most frequent three-locus haplotype. For the population relationships, Dalian had a relative close genetic relationship with Liaoning and Yantai-Weihai and a relative distant genetic relationship with Australia. The information obtained in this study may provide useful information for anthropological studies, for disease-association studies and helping bone marrow transplantation patients to search HLA-matched donors.

Effect of White Turmeric Extract (Curcuma zedoaria) Using Zam-zam Solvent Compare with Ethanol Solvent Against Breast Cancer Cell T47D

Introduction: Curcuma zedoaria is one of the herbal plants potentially protects and cures many diseases, particularly as anti-cancer and anti-tumor. Anti-cancer active compounds of it are flavonoid, phenolic, and saponin. Objectives: This study aimed to explore the cytotoxicity of Curcuma zedoria extract (CZE). Methods: Experimental Quasi with post test non equivalent control group design on T47D cell line at Biology laboratory, Faculty of Medicine, Sultan Agung Islamic University, Semarang. The research was consisted of 2 groups, namely: intervention group with CZE zam-zam solvent and intervention group with CZE ethanol solvent given 10 different dosages each for 1.000 μg/mL; 500 μg/mL; 250 μg/mL; 125 μg/mL; 62.5 μg/mL; 31.25 μg/mL; 15.62 μg/mL; 7.81 μg/mL; 3.90 μg/mL; 1.95 μg/mL. Cytotoxicity test of IC50 using direct counting method and analyzed by probit analysis. Results: IC50 value of CZE in zam-zam and ethanol solvent were 28.24 μg/ml and 13.71 µg/ml respectively at the same 24 hours incubation period. Chi-square test revealed p value was 0.000 (α = 0.05), meaning that there was significant relationship. Conclusion: CZE activity using ethanol is highly active while CZE using zam-zam water is moderate and both of them have the toxicity on breast cancer cell. As the result, apoptosis process may occur.

Effect of White Turmeric Extract (Curcuma zedoaria) Using Zam-zam Solvent Compare with Ethanol Solvent Against Breast Cancer Cell T47D

Introduction: Curcuma zedoaria is one of the herbal plants potentially protects and cures many diseases, particularly as anti-cancer and anti-tumor. Anti-cancer active compounds of it are flavonoid, phenolic, and saponin. Objectives: This study aimed to explore the cytotoxicity of Curcuma zedoria extract (CZE). Methods: Experimental Quasi with post test non equivalent control group design on T47D cell line at Biology laboratory, Faculty of Medicine, Sultan Agung Islamic University, Semarang. The research was consisted of 2 groups, namely: intervention group with CZE zam-zam solvent and intervention group with CZE ethanol solvent given 10 different dosages each for 1.000 μg/mL; 500 μg/mL; 250 μg/mL; 125 μg/mL; 62.5 μg/mL; 31.25 μg/mL; 15.62 μg/mL; 7.81 μg/mL; 3.90 μg/mL; 1.95 μg/mL. Cytotoxicity test of IC50 using direct counting method and analyzed by probit analysis. Results: IC50 value of CZE in zam-zam and ethanol solvent were 28.24 μg/ml and 13.71 µg/ml respectively at the same 24 hours incubation period. Chi-square test revealed p value was 0.000 (α = 0.05), meaning that there was significant relationship. Conclusion: CZE activity using ethanol is highly active while CZE using zam-zam water is moderate and both of them have the toxicity on breast cancer cell. As the result, apoptosis process may occur.