Many of the currently used PN code acquisition algorithms detect the phase of the incoming PN signal on the basis of ML estimation principle and utilize statistics grounded in taking inner products. As an extension of PN code acquisition algorithm using one auxiliary code introduced by Salih in 1996, we propose a more and optimal (hardware / time / space complexity wise) algorithm by using a vector space approach. We outline some important differences between our algorithm and that introduced by Salih and in the process point out some advantages of our algorithm.
Purpose: This study aimed to evaluate the value of serum carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) levels to detect gastric cancer recurrence. Materials and Methods: We retrospectively reviewed 154 patients who developed recurrence within 2 years after curative gastric cancer surgery and analyzed the relationship between postoperative CEA and CA19-9 levels and recurrence. We readjusted the cut-off values to improve the detection of recurrence. Subgroup analysis according to clinicopathologic variables was performed to further investigate the relationship between recurrence and CEA and CA19-9 levels. Results: The sensitivity and specificity for elevated CEA levels to detect recurrence were 40.6% and 89.5%, respectively, and those for CA19-9 were 34.2% and 93.6%, respectively. The sensitivity and specificity for elevation of either tumor marker were 54.3% and 84.0%, respectively; those for elevation of both tumor markers were 19.2% and 98.4%, respectively. By readjusting the cut-off values from 5.0 ng/ml to 5.2 ng/ml for CEA and from 37.00 U/ml to 30.0 U/ml for CA19-9, the sensitivity was increased from 34.2% to 40.2% for CA19-9, while there was no increase in sensitivity for CEA. In subgroup analysis, the sensitivity of CEA was higher in patients with elevated preoperative CEA levels than in patients with normal preoperative CEA levels (86.7% versus 33.7%; P<0.001). Furthermore, the sensitivity of CA19-9 was higher in patients with elevated preoperative CA19-9 levels than in patients with normal preoperative CA19-9 levels (82.61% versus 26.83%; P<0.001). Conclusions: CEA and/or CA19-9 measurement with the readjusted cut-off values allows for more effective detection of gastric cancer recurrence.
This paper demonstrates a microfluidic electrochemical sensor for detecting endocrine disruptor such as estradiol at a very low concentration by using preconcentration technique. In addition, self-assembled monolayer(SAM) was also employed on the working electrode of the electrochemical sensor in order to increase the estradiol capture efficiency of the sensor. SAM treatment on the working electrode enhanced the specific binding between the surface of the working electrode and the estradiol antibody. The estradiol antibody was applied on the working electrode at different concentrations(10, 20, 50, 100, 200 pg/ml) for observing the concentration dependency. The measured electrochemical redox current changed with the amount of the bound estradiol on the Au working electrode surface and the sensor can detect all the target material when the immobilized antibody amount is more than the estradiol amount in the water. The elecrochemical estradiol sensor without SAM treatment showed a low current of 7.79 nA, while the sensor treated with SAM resulted in 339 nA at 200 pg/ml, which is more than 40 fold higher output current. When combining the preconcentration technique and the SAM-treated electrode, the measured current became more than 100 fold higher than that of the sensor without neither SAM treatment nor preconcentration technique. The combination of these two techniques can would enable the proposed microfluidic electrochemical sensor to detect a very low concentration endocrine disruptor.
Streptococcus mutans is the primary causative agent of caries, which is one of the most common human diseases. Thus, rapid and early detection of cariogenic bacteria is critical for its prevention. This study investigated the combination of loop-mediated isothermal amplification (LAMP) and microfluid technology to quantitatively detect S. mutans. A low-cost, rapid microfluidic chip using LAMP technology was developed to amplify and detect bacteria at 2.2-2.2 × 106 colony-forming units (CFU)/ml and its detection limits were compared to those of standard polymerase chain reaction. A visualization system was established to quantitatively determine the experimental results, and a functional relationship between the bacterial concentration and quantitative results was established. The detection limit of S. mutans using this microfluidic chip was 2.2 CFU/ml, which was lower than that of the standard approach. After quantification, the experimental results showed a good linear relationship with the concentration of S. mutans, thereby confirming the effectiveness and accuracy of the custom-made integrated LAMP microfluidic system for the detection of S. mutans. The microfluidic system described herein may represent a promising simple detection method for the specific and rapid testing of individuals at risk of caries.
A qualitative and quantitative analytical method was developed for detection of methamphetamine (MA) and its main metabolite amphetamine (AM) in oral fluid. Oral fluids of eleven drug abusers were provided by Police, specimens were collected by stimulation with a cotton swab treated with 20 mg of citric acid ($Salivette^{(R)}$; Sarstedt, USA). As the preliminary test, oral fluid samples were screened for amphetamines by Fluorescence Polarization Immunoassay (TDxFLx, Abbott Co.). Extraction for MA was performed using solid-phase extraction (SPE) by $RapidTrace^{TM}$ (Zymark, USA) with mixed mode cation exchange cartridge, CLEAN $SCREEN^{(R)}$ (130 mg/3 ml, UCT) after dilution with phosphate buffer. Samples were evaporated and derivatized by pentafluoropropionic acid anhydride (PFPA). Quantitation of MA and AM was performed by gas chromatography-mass spectrometry (GC-MS) using selective ion monitoring (SIM), the quantitation ions were m/z 204 (MA), 208 (MA-$D_5$), 190 (AM) and 194 (AM-$D_5$). The selectivity, linearity of calibration, limit of detection (LOD) and quantification (LOQ) within- and between day precision, accuracy and recoveries were examined as parts of the method validation. All oral fluid samples gave positive results to immunoassay for MA (cut-off level, 50 ng/ml as d-amphetamine). Concentrations of MA and AM by GC-MS in eleven samples were ranged 104.2${\sim}$4603.3 ng/ml and 32.4${\sim}$268.6 ng/ml, respectively. Extracted calibration curves of MA and AM were linear over the two concentration range of 1${\sim}$100 and 50${\sim}$1000 ng/ml with correlation coefficient of above 0.999. LOQ of MA and AM was 1 and 3 ng/ml, respectively. The intraand inter-day run precisions (CV) for MA and AM were less than 10%, and the accuracies (bias) for MA and AM were also less than 10% at the two different concentrations 5 and 100 ng/ml at low calibration range, 50 and 1000 ng/ml at high calibration range. The absolute recoveries of MA and AM at low and high calibration ranges were more than 82% and 75%, respectively. In this study the qualitative and quantitative analytical method of MA in oral fluid was established. Oral fluid testing may detect drug use in past hours because of its shorter detection window than urine, and be useful in post-accident situations. So oral fluids will be most useful for testing drug abuse in the driving under the influence of drug (DUID) as the alternative specimens of urine.
To date, there has been no employment of a magnetoelastic (ME) biosensor method to detect Salmonella enterica serovar Typhimurium in soil. The ME biosensor method needs to be investigated and modified for its successful performance. The filtration method, cation-exchange resin method, and combinations of both methods were employed for the extraction of S. Typhimurium from soil. The number of S. Typhimurium and the resonant frequency shift of the ME sensor were then compared using a brilliant green sulfa agar plate and an HP 8751A network analyzer. A blocking study was performed using bovine serum albumin (BSA), polyethylene glycol (PEG), and casein powder suspension. Finally, the modified ME biosensor method was performed to detect S. Typhimurium in soil. The number of S. Typhimurium was significantly decreased from 7.10 log CFU/soil to 4.45-4.72 log CFU/soil after introduction of the cation-exchange resin method. The greatest resonant frequency shift of the measurement sensor was found when employing centrifugation and filtration procedures. The resonant frequency shift of the PEG-blocked measurement sensor was $3,219{\pm}755Hz$, which was significantly greater than those of the BSA- and casein-blocked ME sensor. The optimum concentration of PEG was determined to be 1.0 mg/ml after considering the resonant shift and economic issue. Finally, the modified ME biosensor method was able to detect S. Typhimurium in soil in a dose-response manner. Although these modifications of the ME biosensor method sacrificed some advantages, such as cost, time effectiveness, and operator friendliness, this study demonstrated a novel approach of the ME biosensor method to detect S. Typhimurium in soil.
This study was conducted to detect the toxoplasma specific-DNA in circulating blood and organs collected from slaughtered pigs at slaughtering house and experimentally infected pigs with Toxoplasma gondii tachyzoites by polymerase chain reaction(PCR), and also PCR was applied to diagnose for acute phase of swine toxoplasmosis as a newly developed diagnostic test. The sensitivity of oligonucleotide primer, T-1 & T-2, designed from toxoplasma B1 gene amplification method was compared with Tp parasite detection by mouse inoculation(MI). On the other hand, latex agglutination test(LAT) was conducted to detect the serum antibodies comparing with the detection of toxoplasma by PCR and MI. The results obtained were summarized as follows. PCR was able to determine at the lowest level of $10^0/ml$ T. gondii in blood samples which were blended with a serial diluted T gondii in vitro. On the other hand, $10^2/5g$ of T gondii could detect from a variety of tissues including lung, diaphragm, liver, heart, spleen and brain in vitro. The primer was proved to specifically determine T gondii in blood and tissues in vitro but it did not detect Neospora caninum used as a negative control. DNA of T. gondii was effectively extracted by freezing, thawing and grinding twice both tissues mixed with T gondii in vitro and in experimentally infected pig's tissues. PCR detected specific DNA in the blood of experimentally infected pigs at 108 hrs and 120 hrs post-infection, it was the same time that the pigs showed fever and parasitaemia. In case of tissue, specific DNA was, however, detected only lung from experimentally infected pigs. Even though the duration of acute phase was from 3 to 7 days post-infection, but the latex agglutination test (LAT) results appeared from 8 days post-infection. A comparison of sensitivity in determining T gondii in blood samples between PCR and MI, PCR positive rate ranged from 25 to 33.3%, but that of MI covered from 75 to 100%.
Mastitis is a major problem for the global dairy industry and causes substantial economic losses from decreasing milk production and considerable compositional changes in milk, reducing milk quality. The potential of near infrared (NIR) spectroscopy in the region from 1100 to 2500nm and chemometric method for classification to detect milk from mastitic cows was investigated. A total of 189 milk samples from 7 Holstein cows were collected for 27 days, consecutively, and analyzed for somatic cells (SCC). Three of the cows were healthy, and the rest had mastitis periods during the experiment. NIR transflectance milk spectra were obtained by the InfraAlyzer 500 spectrophotometer in the spectral range from 1100 to 2500nm. All samples were divided into calibration set and test set. Class variable was assigned for each sample as follow: healthy (class 1) and mastitic (class 2), based on milk SCC content. The classification of the samples was performed using soft independent modeling of class analogy (SIMCA) and different spectral data pretreatment. Two concentration of SCC - 200 000 cells/ml and 300 000 cells/ml, respectively, were used as thresholds fer separation of healthy and mastitis cows. The best detection accuracy was found for models, obtained using 200 000 cells/ml as threshold and smoothed absorbance data - 98.41% from samples in the calibration set and 87.30% from the samples in the independent test set were correctly classified. SIMCA results for classes, based on 300 000 cells/ml threshold, showed a little lower accuracy of classification. The analysis of changes in the loading of first PC factor for group of healthy milk and group of mastitic milk showed, that separation between classes was indirect and based on influence of mastitis on the milk components. The accuracy of mastitis detection by SIMCA method, based on NIR spectra of milk would allow health screening of cows and differentiation between healthy and mastitic milk samples. Having SIMCA models, mastitis detection would be possible by using only DIR spectra of milk, without any other analyses.
Competitive direct ELISA was developed to detect gentamicin residues. Mice immunized with gentamicin-keyhole limpet hemocyanin (KLH) conjugate developed good antiserum titers, which gradually increased with booster injections, indicating immunization was successfully processed. Monoclonal antibody against gentamicin was prepared using hybridoma cells cloned by limit dilution of fused cells. IgG was purified from ascites fluid of hybridoma cell-injected mice through ammonium sulfate precipitation and Sephadex G-25 gel filtration. After the gel filtration, fractions of high antibody titer were further purified through affinity chromatography on protein A/G column. Monoclonal antibody against gentamicin was confirmed as IgG1, which has kappa light chain. Cross-reactivities ($CR_{50}$) of gentamicin monoclonal antibody to other aminoglycosides (kanamycin, neomycin, and streptomycin) were less than 0.005%, indicating the monoclonal antibody was highly specific for gentamicin. Standard curve constructed through competitive direct ELISA showed measurement range (from 80 to 20% of B/$B_0$ ratio) of gentamicin was between 1 and 40 ng/ml, and 50% of B/$B_0$ ratio was about 4 ng/ml. The gentamicin concentration rapidly increased to 1,300 ng/ml after the intramuscular administration up to 2 h, then sharply decreased to less than 300 ng/ml after 4 h of withdrawal, during which the elimination half-life ($t_{1/2}$) of gentamicin in the rabbit plasma was estimated to be 1.8 h. Competitive direct ELISA method developed in this study using the prepared monoclonal antibody is highly sensitive for gentamicin, and could be useful for detecting gentamicin residues in plasma of live animals.
Plectranthus amboinicus Benth (Lamiaceae) is a medicinal plant native to India, and its leaves are widely used in several traditional medicinal preparations. The purpose of this study was to detect and quantify phenolics present in ethyl acetate and acetone extracts of P. amboinicus leaves, and evaluate their antioxidant, antibacterial, antimutagenic and anticancer activities. The HPLC chromatograms of crude leaf extracts indicated the presence of phenolics like caffeic acid, coumaric acid, rutin, quercetin and gallic acid, which were present in the range of 0.01 - 1.41 mg/g in ethyl acetate and 0.03 - 1.93 mg/g in the acetone extract. The acetone extract showed statistically (p < 0.05) higher antioxidant activity ($IC_{50}$, 99.59 ${\mu}g/ml$) than ethyl acetate extract ($IC_{50}$, 149.96 ${\mu}g/ml$). Statistically (p < 0.05) higher antimutagenicity was shown by acetone extract (46.16%) as compare to ethyl acetate extract (12.16%) at 500 ${\mu}g/plate$ concentration. The acetone extract showed higher antibacterial activity than ethyl acetate extract, and both the extracts showed highest activity against B. cereus (375 and 625 ${\mu}g/ml$, respectively) and lowest activity against Y. enterocolitica (1000 and 1125 ${\mu}g/ml$, respectively). Both the extracts also showed inhibitory effect on cancer cell lines HCT-15 and MCF-7. These results suggest that the leaves of P. amboinicus possess various biological activities, and validate the traditional use of the leaves of P. amboinicus against cold, infection and ulceration.
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