Enumeration Results Research Articles

Analysis of the Occurrence of Antibiotic Resistant Bacteria in the Hospital's Effluent and its Receiving Environment.

The use of antibiotics on a regular and excessive basis is a major factor in the spread of antibiotic-resistant bacteria. Patients discharge un-metabolized or relatively low doses of non-metabolized antibiotics through urine and stool, which might enter into the environment through sewage disposal and promote the emergence of antibiotic resistant bacteria. This study is designed to investigate how excessive use of antibiotics in the hospital sector and their release into hospital wastes contribute to the spread of antibiotic-resistant bacteria in different environmental settings. In this study, liquid hospital waste was collected from the sewage of Chittagong Medical College Hospital (CMCH), Bangladesh as well as from its distribution position in Chittagong city, Bangladesh. A total of 5 samples were collected from different positions in Chittagong city, including CMCH liquid waste. After collection, total bacteria and total cefixime resistant bacteria were counted by the total viable count (TVC) method. The result of bacteriological enumeration showed that a high magnitude of cefixime-resistant bacteria were available in all the hospital’s associated waste samples. The highest proportion of cefixime resistant bacteria (23.35%) was found in sample 2, whereas 17.4%, 7.6%, 5%, and 1.32% were found in samples 1, 3, 4, and 5, respectively. The total number of cefixime-resistant bacteria decreased with the increase in distance between the sample collection site and the hospital drain. This means that resistant bacteria developed in the hospital effluent are transferred to the environmental distribution sites.

Localization of Mixed Far-field and Near-field Incoherently Distributed Sources Using Two-stage RARE Estimator

In this paper, a mixed source localization method utilizing a two-stage rank-reduction (RARE) estimator is investigated. Different from the existing methods, the proposed one is built on the incoherently distributed (ID) source model, which is more appropriate for multipath and fast time-varying channels. Firstly, a general array manifold (GAM) model is established, where nominal DOAs and nominal ranges are extracted from the initial array manifold. By exploiting the shift invariance property of the far-field (FF) GAM and combining virtual source enumeration result, nominal FF DOA estimation is achieved by a one-dimensional (1-D) RARE spectral search. Secondly, the oblique projection operation is adopted to separate near-field (NF) sources, and the nominal DOA and range parameters of NF sources are subsequently obtained by jointly utilizing the manifold separation technique (MST) and another two 1-D spectral searches. With the estimated nominal DOA and range parameters, the angular spread and range spread are then successfully estimated. Moreover, the Cramér-Rao bound (CRB) for the considered case is also derived. Simulation results are presented to validate the effectiveness of the proposed method.

On the structure of random graphs with constant $r$-balls

We continue the study of the properties of graphs in which the ball of radius r around each vertex induces a graph isomorphic to the ball of radius r in some fixed vertex-transitive graph F , for various choices of F and r . This is a natural extension of the study of regular graphs. More precisely, if F is a vertex-transitive graph and r \in \mathbb{N} , we say a graph G is r -locally F if the ball of radius r around each vertex of G induces a graph isomorphic to the graph induced by the ball of radius r around any vertex of F . We consider the following random graph model: for each n \in \mathbb{N} , we let G_n = G_n(F,r) be a graph chosen uniformly at random from the set of all unlabelled, n -vertex graphs that are r -locally F . We investigate the properties possessed by the random graph G_n with high probability, i.e. with probability tending to 1 as n \to \infty , for various natural choices of F and r . We prove that if F is a Cayley graph of a torsion-free group of polynomial growth, and r is sufficiently large depending on F , then the random graph G_n = G_n(F,r) has largest component of order at most n^{5/6} with high probability, and has at least \exp(n^{\delta}) automorphisms with high probability, where \delta>0 depends upon F alone. Both properties are in stark contrast to random d -regular graphs, which correspond to the case where F is the infinite d -regular tree. We also show that, under the same hypotheses, the number of unlabelled, n -vertex graphs that are r -locally F grows like a stretched exponential in n , again in contrast with d -regular graphs. In the case where F is the standard Cayley graph of \mathbb{Z}^d , we obtain a much more precise enumeration result, and more precise results on the properties of the random graph G_n(F,r) . Our proofs use a mixture of results and techniques from geometry, group theory and combinatorics. We make several conjectures regarding what happens for Cayley graphs of other finitely generated groups.

Twenty Vertex Model and Domino Tilings of the Aztec Triangle

We show that the number of configurations of the 20 Vertex model on certain domains with domain wall type boundary conditions is equal to the number of domino tilings of Aztec-like triangles, proving a conjecture of the author and Guitter. The result is based on the integrability of the 20 Vertex model and uses a connection to the U-turn boundary 6 Vertex model to re-express the number of 20 Vertex configurations as a simple determinant, which is then related to a Lindström-Gessel-Viennot determinant for the domino tiling problem. The common number of configurations is conjectured to be $2^{n(n-1)/2}\prod_{j=0}^{n-1}\frac{(4j+2)!}{(n+2j+1)!}=1, 4, 60, 3328, 678912...$ The enumeration result is extended to include refinements of both numbers.

The role of salinity and Total Suspended Solids (TSS) to abundance and structure of phytoplankton communities in estuary Saddang Pinrang

Phytoplankton is organisms that have a sensitivity to changes in the characteristics of the waters. Some determining factors of its existence are salinity, total suspended solids (TSS), and other compounds. Research has been conducted on the role of salinity and TSS in the abundance and structure of phytoplankton communities in the estuary of Saddang River Pinrang Regency. Research implementation in September to December 2020. The result of phytoplankton enumeration is found 25 Genera in 4 classes namely Bacillariophyceae, Chlorophyceae, Cyanophyceae, and Dinophyceae. Salinity has a real effect and shows a negative relationship to phytoplankton abundance. In contrast, TSS showed no real link to the abundance of phytoplankton in the estuary waters of Saddang Pinrang.

CHARACTERISTICS OF TUNA, SMALL TUNA AND SKIPJACK (TTS) FISHERY IN KUTARAJA OCEAN FISHING PORT, ACEH

Kutaraja Ocean Fishery Port is the oldest port in Banda Aceh City, Lampulo. Fishery resources are a new potential in the regional economic development of Aceh province. Fish resources that have great potential are from the large pelagic fish groups, including Tuna, Small Tuna and Skipjack (TTS) . Fishery characteristics are used to carry out sustainable fishing. This research was conducted at PPS Kuta Raja, Aceh for one year. Data collection was carried out directly through site surveys and interviews with related fisheries actors. Biological data and capturing aspects were carried out using a stratifield random sampling system by placing enumerators. The results showed that the production of TCT at PPS Kutaraja was dominated by 2 fishing gears, purse seine and hand lines. Based on the results of enumeration in 2020, the estimated total production of tuna skipjack and small tuna at PPS Kuta Raja reaches 11,520 tons. The highest total estimated fish catch is skipjack tuna (Katsuwonus pelamis) (SKJ) 5,802 tons, Bullet tuna (Auxis rochei)(BLT) 2814 tons, Yellow fin (Thunnus albacares) (YFT) 2,452 tons, Krai Tongkol (Auxis thazard) (FRI) 448 tons, and Eastern little tuna (Euthynnus affinis) (KAW) 1,745 tonnes. All of the fish caught were dominated by immature fish. So it is very necessary to evaluate the use of fishing gear or the licensing of FADs. The monthly CPUE distribution pattern is the same, where in August it reaches the highest CPUE and in October it reaches the lowest CPUE value.

Exfoliated tumor cells in bile as a promising indicator of disease status in cholangiocarcinoma

Cholangiocarcinoma (CCA) is highly metastatic, extremely difficult to diagnose and is consequently characterized by a low 5-year survival rate. Herein we investigated the potential of using exfoliated tumor cells (ETCs) in bile to serve as an early-warning CCA metastasis indicator. ETCs were isolated from bile using a CCA-specific aptamer conjugated to magnetic beads and identified via immunostaining (CK17+CK7+Hoechst+). A conventional protein biomarker, epithelial cell adhesion molecule (EpCAM), was used for comparison. Circulating tumor cell (CTC) isolation in blood was achieved by a microfluidic system from samples from five CCA patients undergoing chemotherapy, and the enumeration results of ETCs and CTCs were compared with diagnoses derived from computed tomography scans. The CCA-specific aptamer-conjugated magnetic beads effectively bound ETCs from bile samples of 40/40 CCA patients (>1 ETC in 3 ml of bile), and the ETC and CTC count data from patients corresponded well with cancer progression and tumor size. ETCs in bile could serve as promising indicators of disease status in early-advanced stage CCA while CTCs in blood could be representative of late-advanced stage CCA. The developed technique could consequently be a powerful tool in CCA diagnostics and prognostics.

Probiotic Lactobacilli in Fermented Dairy Products: Selective Detection, Enumeration and Identification Scheme.

A selection of 36 commercial probiotic fermented dairy products from UK and Europe markets were evaluated for the numbers, types, and viability of Lactobacillus strains against the stated information on their packages. A comparative study was carried out on selectivity of MRS-Clindamycin, MRS-Sorbitol, and MRS-IM Maltose, to select the right medium for enumeration of probiotic Lactobacillus. Based on selectivity of medium for recovery of the targeted lactobacilli, and also simplicity of preparation, MRS-Clindamycin was chosen as the best medium for enumeration of probiotic Lactobacillus in fermented milks. The results of enumeration of lactobacilli showed that 22 out of a total 36 tested products contained more than 106 colony-forming units/g at the end of their shelf life, which comply with the recommended minimum therapeutic level for probiotics. Rep-PCR using primer GTG-5 was applied for initial discrimination of isolated strains, and isolates, which presented different band profile, were placed in different groups. The isolated Lactobacillus spp. were identified mainly as Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus paracasei by analysis of partial sequences of the 16S ribosomal RNA and rpoA genes.

Gut Health Function of Instant Dehydrated Rice Sticks Substituted with Resistant Starch Types 2 and 4.

The purpose of this study was to analyze the effects of instant dehydrated rice sticks (IDRS) which were substituted with resistant starch (RS) types 2 and 4 whose gut health function targets gut microbiota. IDRS are a type of rice noodles that were developed by two formulations. The first formulation had substitution of rice flour with 20% RS type 2 and 0.15% carboxymethyl cellulose (CMC) (RSc-2), and the second formulation had 25% RS type 4 and 0.15% CMC (RSc-4). RSc-2 and RSc-4 were investigated for gut health function by human fecal fermentation in a pH-controlled batch culture. The results of gut microbiota enumeration by fluorescent in situ hybridization confirmed that significantly (P < 0.05) higher numbers of bifidobacteria were obtained with RSc-2 (10.06 ± 0.09 log cells/mL) and RSc-4 (10.00 ± 0.06 log cells/mL) compared to the control (100% rice flour formula) at 24h fermentation. Additionally, the prebiotic indexes of RSc-2 and RSc-4 were 3.8 and 2.8 -fold higher than that of the control at 24h fermentation. The short-chained fatty acids, acetic, propionic and butyric acid were analyzed by gas chromatography-flame ionization detector. The butyric acids were significantly (P < 0.05) higher with RSc-2 (43.56 ± 0.01mM) and RSc-4 (43.63 ± 0.07mM) compared to the control at 24h. Thus, RSc-2 and RSc-4 showed butyrogenic, bifidogenic and prebiotic potential to support gut health and could aid in prevention of colon cancer.

Enumerating Non-Stable Vector Bundles

Abstract In this article, we establish a motivic analog of an enumeration result of James–Thomas [ 28] on non-stable vector bundles in topological setting. Using this, we obtain results on enumeration of projective modules of rank $d$ over a smooth affine $k$-algebra $A$ of dimension $d$, recovering in particular results of Suslin and Bhatwadekar on cancellation of such vector bundles. Admitting a conjecture of Asok and Fasel, we prove cancellation of such vector bundles of rank $d-1$ if the base field $k$ is algebraically closed. We also explore the cancellation properties of symplectic vector bundles.

RESOURCES OF WOODY PLANTS OF NORTH CAUCASUS BEECH FORESTS

The aim of this study is to investigate material and monetary valuation of forest resources in the beech forests of the North Caucasus. The plots were selected in the Central part of the North Caucasus, on the territory of the Republic of North Ossetia-Alania (Kartсinsky ridge), between 1200-1350 m above sea level. Two objects are located on the northern macroslopes and two on the southern macroslope. The characteristics of phytocenoses on the experimental plots were established based on the results of a continuous enumeration of trees on a test area of 50x100 m. The number of undergrowth and undergrbrush, their composition and distribution by height groups, are established on circular areas of 10 m2. The average wood volume is about 300 m3/ha, and the average stand density is 248 trees per 1 hctare. The average number of undergrowth is 8988 per hectare, and underbrush 1998 per hectare. The average height of the young generation is 0.65 m, and the undergrowth is 0.86 m. The weight of one plant of average height in the air-dry state is 0.24 kg for undergrowth and 0.55 kg for undergrbrush. Phytomass of undergrowth and underbrush by species was set by weighing in a fresh and airdry state. Prices for all types of wood resources used average market prices as of 2019. The main income can be obtained from the sale of wood – about 650 thousand RUB/ha. Undergrowth and underbrush can give in a total about 43 thousand RUB/ha. At the same time, the period of maturation for the harvest of beech and related species is 100-140 years. During this period, it can be carried out about 15-20 harvests of undergrowth and underbrush, which together can generate income comparable to the amount from the sale of wood.

Six-cycle systems

AbstractIn this article, we attempt to survey all that has been written about 6-cycle systems. In addition, several new results, including many enumeration results, are included here for the first time. Our survey concludes with a list of open problems.

Exploiting Reuse for GPU Subgraph Enumeration

Subgraph enumeration is important for many applications such as network motif discovery, community detection, and frequent subgraph mining. To accelerate the execution, recent works utilize graphics processing units (GPUs) to parallelize subgraph enumeration. The performances of these parallel schemes are dominated by the set intersection operations which account for up to 95 percent of the total processing time. (Un)surprisingly, a significant portion (as high as 99 percent) of these operations is actually redundant, i.e., the same set of vertices is repeatedly encountered and evaluated. Therefore, in this article, we seek to salvage and recycle the results of such operations to avoid repeated computation. Our solution consists of two phases. In the first phase, we generate a reusable plan that determines the opportunity for reuse. The plan is based on a novel reuse discovery mechanism that can identify available results to prevent redundant computation. In the second phase, the plan is executed to produce the subgraph enumeration results. This processing is based on a newly designed reusable parallel search strategy that can efficiently maintain and retrieve the results of set intersection operations. Our implementation on GPUs shows that our approach can achieve up to 5 times speedups compared with the state-of-the-art GPU solutions.

Development and application of a DNA metabarcoding method for comprehensive analysis of soil nematode communities

Nematodes mediate soil ecosystem functions together with other soil biota and are employed as effective ecological indicators. Among nematodes, plant-parasitic nematodes are economically important due to the damage they cause to many crops. While morphological identification of nematodes after elutriation from soil has been widely used to assess their diversity and for diagnostic analyses, high-throughput sequencing techniques are becoming more widely available. In this study, highly sensitive primer sets using the 18S rDNA region as metabarcoding markers were developed specifically for soil nematode fauna using next generation sequencing (NGS). In silico analysis using Primer BLAST identified a primer set (F548_A/R1912) specific to nematodes and is effective in application to DNA directly extracted from soil as well as from elutriated nematode populations. In silico analysis also indicated that three primer sets (F548_A/R915, F548_A/SSU26R and F548_A/R1912) developed in this study were expected to properly amplify the target sequences of 97–98% of 88 selected free-living and 83–92% of 60 selected plant-parasitic nematode species. All the results of NGS using three primer sets applied to DNA from a nematode suspension and the nematode specific primer set applied to soil DNA showed significant (p < 0.01, linear mixed-effects model) correlations with the results of nematode enumeration using a morphological approach. Furthermore, all the NGS assays indicated population structures were 3–9% more diverse than the microscopic approach. The current NGS protocol detects a proper level of sequence reads to represent nematode fauna for diversity and diagnostic analyses in 5 soil sampling sites in Japan, based on the rarefaction curves. As a result, this study revealed that the primer set F548_A/R1912 is highly specific to nematode species and applicable to soil DNA, and further F548_A/R915 and F548_A/SSU26R may extend the coverage to detect more nematode species than that using only F548_A/R1912 with soil DNA. The three primer sets and the metabarcoding protocol examined appear suitable for comprehensive studies of nematode community structure.

Parking function varieties for combinatorial tree models

We study the enumeration problem for different kind of tree parking functions introduced recently, called tree parking functions, tree parking distributions, prime tree parking functions, and prime tree parking distributions, for rooted labelled trees of important combinatorial tree families including labelled ordered, unordered and binary trees. Using combinatorial decompositions of the underlying structures yields, after solving the resulting equations, implicit characterizations of suitable generating functions of the total number of such tree parking functions for trees of size n and n successful drivers, from which we obtain exact and asymptotic enumeration results. The approach can be extended to the general situation of tree parking functions for trees of size n and m<n drivers for which we are also able to characterize the generating functions solutions, which allow, by applying analytic combinatorics techniques, a study of the asymptotic behaviour of the total number of tree parking functions and distributions for n→∞ depending on the load factor 0<α=mn<1.

Abstract GS4-08: Clinical utility of repeated circulating tumor cell (CTC) enumeration as early treatment monitoring tool in metastatic breast cancer (MBC) - a global pooled analysis with individual patient data

Abstract Background: Several studies suggest clinical utility of serial circulating tumor cell (CTC) enumeration as a means of assessing response status in metastatic breast cancer (MBC). The aim of this study is to conduct a comprehensive pooled analysis comprising globally available data to further define and explore the role of CTC enumeration as a tool for early treatment monitoring in patients with MBC with a focus on the predictive power in different breast cancer subtypes and clinical settings. Methods: In a global effort, peer-reviewed published studies with data on repeated CTC assessments (CellSearch® technology; Menarini Silicon Biosystems; Bologna, Italy) in MBC patients were screened and investigators were asked to provide individual patient data for this pooled analysis. 2761 cases from 32 data sets with data on both baseline and one follow up CTC assessments were included in the analysis (median time interval between the two CTC assessments 35 days). Data were analyzed using log rank tests and Cox regressions to evaluate the association between serial CTC enumeration results and overall survival (OS) in the full patient cohort and defined subgroups. Results: 588 (21.3%) patients had no CTCs at both time points (neg/neg), 236 (8.5%) patients were CTC negative at baseline and CTC positive at follow up (neg/pos), 712 (25.8%) patients converted from CTC positive at baseline to CTC negative (pos/neg), and 1225 (44.4%) patients had at least one CTC at both time points (pos/pos). Log rank tests showed significant differences in OS between these four CTC change groups (p &amp;lt; 0.0001 for all pairwise comparisons except for the comparison between neg/pos and pos/neg, p = 0.015). Median OS for the neg/neg, neg/pos, pos/neg and pos/pos group was 45.6, 26.1, 34.6, and 17.6 months, respectively. Hazard ratios (HR) (reference group neg/neg) were 1.38 (95% CI 1.16 - 1.64) for the pos/neg group, 1.78 (95% CI 1.43 - 2.22) for the neg/pos group, and 3.06 (95% CI 2.63 - 3.56) for the pos/pos group. Results were similar if a cutoff of 5 CTCs was used for CTC positivity (pos/neg group: HR 1.43, 95% CI 1.25 - 1.63; neg/pos group: HR 2.39, 95% CI 1.91 - 2.99; pos/pos group: HR 3.54, 95% CI 3.12 - 4.02). In total, 2586 patients could be assigned to different tumor subtypes based on known hormone receptor (ER) and HER2 status of the primary tumor: 1513 (58.5%) patients had a luminal-like tumor (ER positive, HER2 negative), 682 (26.4%) patients had a HER2-positive tumor, and 391 (15.1%) patients had a triple-negative tumor. In patients with luminal-like tumors, the hazard ratios were 1.67 (95% CI 1.29 - 2.17), 2.01 (95% CI 1.45 - 2.77), and 3.87 (95% CI 3.09 - 4.83) for the pos/neg, neg/pos, and pos/pos group, respectively. In patients with HER2-positive tumors, the neg/pos group (HR 1.68, 95% CI 1.12 - 2.53) and the pos/pos group (HR 2.11, 95% CI 1.58 - 2.83) showed significantly worse OS compared to the neg/neg group, while in triple-negative patients, the pos/pos group had a significantly shorter OS compared to the neg/neg group (HR 2.99, 95% CI 2.11 - 4.24). The results will be up-dated by inclusion of additional large data sets (CALGB 40502, CALGB 40503, COMET, SWOG S0500, TBCRC 001) for the analysis to be presented at SABCS 2020. Conclusion: This large pooled analysis confirms that at a median of 35 days after treatment initiation, follow-up CTC assessments strongly predict overall survival. These results suggest potential clinical utility of CTC monitoring as early response marker in MBC, especially in luminal-like tumors. Citation Format: Wolfgang J Janni, Tracy C. Yab, Daniel F. Hayes, Massimo Cristofanilli, Francois-Clement Bidard, Michail Ignatiadis, Meredith M. Regan, Catherine Alix-Panabières, William E. Barlow, Carlos Caldas, Lisa A. Carey, Luc Dirix, Tanja Fehm, Jose A. Garcia-Saenz, Paola Gazzaniga, Daniele Generali, Lorenzo Gerratana, Rafael Gisbert-Criado, William Jacot, Zefei Jiang, Evi Lianidou, Mark J.M. Magbanua, Luis Manso, Dimitrios Mavroudis, Volkmar Müller, Elisabetta Munzone, Klaus Pantel, Jean-Yves Pierga, Brigitte Rack, Sabine Riethdorf, Hope S. Rugo, Kostandinos Sideras, Stefan Sleijfer, Jeffrey Smerage, Justin Stebbing, Leon W.M.M. Terstappen, José Vidal-Martínez, Rita Zamarchi, Karthik Giridhar, Thomas W.P. Friedl, Minetta C. Liu. Clinical utility of repeated circulating tumor cell (CTC) enumeration as early treatment monitoring tool in metastatic breast cancer (MBC) - a global pooled analysis with individual patient data [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr GS4-08.

Image capture device based on Internet of Thing (IoT ) technology

Tracing stolen cattle in the Lombok Tengah regency of NTB province is carried out by tracing footprints and waiting at certain location points which are considered as the route to be traversed or the final destination of cattle theft which is called “Ngendeng”. Ngendeng is carried out day and night from the time the livestock is declared missing until the livestock is found or the potential to be found is considered lost. The problem that occurs in Ngendeng’s activities is that it endangers the safety of life if there is a clash with thieves and a decrease in health for the people of Ngendeng. To replace the role of the community in doing Ngendeng, an Internet of Things (IoT) devices for image capturing was developed. The device can take a picture if someone passes through the front of the device then it is sent to the Telegram account of the District Police and the Community telegram account. The development of devices uses the Research and Development (R&D) method to produce products, and test the product’s effectiveness. The placement of the devices will be distributed in three (3) sub-districts in Lombok Tengah Regency based on the results of data analysis obtained from the results of data enumeration. The method used in tracing the point of placement of devices in each sub-district was using the Participatory Rural Appraisal (PRA) method. So that it is found the most frequently traversed point (very potential) to place IoT-based image capture devices as an effort to reduce the crime of cattle theft in Lombok Tengah Regency.

Lactic acid bacteria diversity and dynamics during ripening of traditional Turkish goatskin Tulum cheese produced in Mut region assessed by culturing and PCR-DGGE

The study aims to identify the lactic acid bacteria diversity of Tulum cheese which is made from raw milk and ripened in the goatskin for 180 days. The samples were produced in Mut located in the Central Taurus Mountains of Turkey. For the determination of the lactic acid bacteria population of Tulum cheese samples, the culture-dependent and culture-independent (PCR-DGGE) methods were used, and cheese samples were sampled on days 7, 15, 30, 60, 90, and 180. According to the enumeration results, Lactococci and Streptococci counts of Tulum cheese samples decreased during ripening. Furthermore, the DGGE profile of the population consisted of Streptococcus gallolyticus, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus lutetiensis, Streptococcus spp., and Enterococcus hirae at the end of the ripening period. On the other hand, Enterococcus spp. and Streptococcus spp. were found as prevalent species in the culture-based method at the end of the maturation period of Tulum cheese samples. Results revealed that the culture-dependent and culture-independent methods presented a piece of complementary information for the lactic acid bacteria population of Tulum cheese.

A Direct Comparison between the Lateral Magnetophoretic Microseparator and AdnaTest for Isolating Prostate Circulating Tumor Cells.

Circulating tumor cells (CTCs) are important biomarkers for the diagnosis, prognosis, and treatment of cancer. However, because of their extreme rarity, a more precise technique for isolating CTCs is required to gain deeper insight into the characteristics of cancer. This study compares the performance of a lateral magnetophoretic microseparator (“CTC-μChip”), as a representative microfluidic device, and AdnaTest ProstateCancer (Qiagen), as a commercially available specialized method, for isolating CTCs from the blood of patients with prostate cancer. The enumeration and genetic analysis results of CTCs isolated via the two methods were compared under identical conditions. In the CTC enumeration experiment, the number of CTCs isolated by the CTC-μChip averaged 17.67 CTCs/mL, compared to 1.56 CTCs/mL by the AdnaTest. The number of contaminating white blood cells (WBCs) and the CTC purity with the CTC-μChip averaged 772.22 WBCs/mL and 3.91%, respectively, whereas those with the AdnaTest averaged 67.34 WBCs/mL and 1.98%, respectively. Through genetic analysis, using a cancer-specific gene panel (AR (androgen receptor), AR-V7 (A\\androgen receptor variant-7), PSMA (prostate specific membrane antigen), KRT19 (cytokeratin-19), CD45 (PTPRC, Protein tyrosine phosphatase, receptor type, C)) with reverse transcription droplet digital PCR, three genes (AR, AR-V7, and PSMA) were more highly expressed in cells isolated by the CTC-μChip, while KRT19 and CD45 were similarly detected using both methods. Consequently, this study showed that the CTC-μChip can be used to isolate CTCs more reliably than AdnaTest ProstateCancer, as a specialized method for gene analysis of prostate CTCs, as well as more sensitively obtain cancer-associated gene expressions.

Optimization of Viability Treatment Essential for Accurate Droplet Digital PCR Enumeration of Probiotics.

Improvements offered by viability droplet digital PCR (v-ddPCR) include increased precision, specificity and decreased time to results making for an attractive alternative method to traditional plate count enumeration of probiotic products. A major hurdle faced in v-ddPCR, however, is distinguishing between live and dead cells. The objective of this study was to evaluate a combination of PMA and EMA (PE51) for viability treatment of freeze-dried probiotic powders. Lactobacillus acidophilus La-14 and Bifidobacterium animalis subsp. lactis Bi-07 were analyzed over a 2-log PE51 concentration gradient to investigate the efficiency across genus and assay targets. Results suggest a need to optimize viability dye concentration based on the genera of the organism, but also the assay target, even when analyzing the same organism. When optimized for PE51 concentration, strain specific v-ddPCR assays for both La-14 and Bi-07 were demonstrated to agree with plate count enumeration results. In conclusion, while these v-ddPCR assays require highly specific optimization, they are better suited for the future of the probiotic industry and are suggested to be implemented in probiotic product testing.