Stabilization of mRNA Expression in Whole Blood Samples
Background: Accurate quantification of mRNA in whole blood is made difficult by the simultaneous degradation of gene transcripts and unintended gene induction caused by sample handling or uncontrolled activation of coagulation. This study was designed to compare a new blood collection tube (PAXgeneTM Blood RNA System) and a companion sample preparation reagent set with a traditional sample collection and preparation method for the purpose of gene expression analysis.
Methods: We collected parallel blood samples from healthy donors into the new sample collection tubes and control EDTA tubes and performed serial RNA extractions on samples stored for 5 days at room temperature and for up to 90 days at 4 and 20 °C. Samples were analyzed by Northern blot analysis or reverse transcription-PCR (RT-PCR).
Results: Specific mRNA concentrations in blood stored in EDTA tubes at any temperature changed substantially, as determined by high-precision RT-PCR. These changes were eliminated or markedly reduced when whole blood was stored in PAXgene tubes. Loss of specific mRNAs, as measured by RT-PCR, reflected total RNA depletion as well as specific mRNA destruction demonstrated by Northern blot analysis. The salutary effects of PAXgene on mRNA stabilization extended to blood samples from eight unrelated donors.
Conclusions: Compared with whole blood collected in EDTA tubes and extracted by an organic method, the PAXgene Blood RNA System reduced RNA degradation and inhibited or eliminated gene induction in phlebotomy whole blood samples. Storage of whole blood samples in PAXgene tubes can be recommended for clinically related blood samples that will be analyzed for total or specific RNA content. 
Polybrominated diphenyl ethers in maternal and fetal blood samples.
Polybrominated diphenyl ethers (PBDEs) are widely used as flame retardants in consumer goods, such as plastics, electronics, textiles, and construction material. PBDEs have been found in human milk, fat, and blood samples. Rodent studies indicate that PBDEs may be detrimental to neurodevelopment, possibly by lowering thyroid hormone concentrations in blood. In the present study, we determined concentrations of PBDEs and thyroid hormones in human fetal and maternal serum. Patients presenting in labor to Indiana University and Wishard Memorial County hospitals in Indianapolis, who were older than 18 years, were recruited to participate. Twelve paired samples of maternal and cord blood were obtained and analyzed using gas chromatographic mass spectrometry; thyroid hormone concentrations were determined by radioimmunoassay. Six congeners of PBDE were measured in maternal and fetal serum samples. The concentrations of total PBDEs found in maternal sera ranged from 15 to 580 ng/g lipid, and the concentrations found in fetal samples ranged from 14 to 460 ng/g lipid. Individual fetal blood concentrations did not differ from the corresponding maternal concentrations, indicating that measurement of maternal PBDE blood levels is useful in predicting fetal exposure; similarly, other reports have shown a high correlation between PBDE in mother’s milk and fetal exposure. In accord with reports on other biologic samples, the tetrabrominated PBDE congener BDE-47 accounted for 53-64% of total PBDEs in the serum. The concentrations of PBDEs found in maternal and fetal serum samples were 20-106-fold higher than the levels reported previously in a similar population of Swedish mothers and infants. In this small sample, there was no apparent correlation between serum PBDEs and thyroid hormone concentrations. Our study shows that human fetuses in the United States may be exposed to relatively high levels of PBDEs. Further investigation is required to determine if these levels are specific to central Indiana and to assess the toxic potential of these exposure levels. 
Perfluorooctanesulfonate and Related Fluorochemicals in Human Blood Samples from China
Perfluorooctanesulfonylfluoride (POSF)-based compounds have been manufactured and used in a variety of industrial applications. These compounds degrade to perfluorooctanesulfonate (PFOS) which is regarded as a persistent end-stage metabolite and is found to accumulate in tissues of humans and wildlife. PFOS, perfluorohexanesulfonate (PFHxS), perfluorooctanoate (PFOA), and perfluorooctanesulfonamide (PFOSA) have been found in human sera from the United States. In this study, concentrations of PFHxS, perfluorobutanesulfonate (PFBS), PFOS, perfluorohexanoic acid (PFHxA), PFOA, perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluoroundecanoic acid (PFUnDA), perfluorododecanoic acid (PFDoDA), and PFOSA were measured in 85 samples of whole human blood collected from nine cities (eight provinces) in China, including Shenyang (Liaoning), Beijing (Hebei), Zhengzhou (Henan), Jintan (Jiangsu), Wuhan (Hubei), Zhoushan (Zhejiang), Guiyang (Guizhou), Xiamen (Fujian), and Fuzhou (Fujian). Among the 10 perfluorinated compounds (PFCs) measured, PFOS was the predominant compound. The mean concentration of PFOS was greatest in samples collected from Shenyang (79.2 ng/mL) and least in samples from Jintan (3.72 ng/mL). PFHxS was the next most abundant perfluorochemical in the samples. No age-related differences in the concentrations of PFOA, PFOS, PFOSA, and PFHxS were observed. Gender-related differences were found, with males higher for PFOS and PFHxS, and females higher in PFUnDA. Concentrations of PFHxS were positively correlated with those of PFOS, while concentrations of PFNA, PFDA, and PFUnDA were positively correlated with those of PFOA. There were differences in the concentration profiles (percentage composition) of various PFCs in the samples among the nine cities. 
Antibacterial Resistance Profile and PCR Detection of Antibiotic Resistance Genes in Salmonella serovars Isolated from Blood Samples of Hospitalized Subjects in Kano, North-West, Nigeria
Aim: The aims of the study were to determine the antibacterial resistance profile and detect the presence of antibacterial resistance genes of Salmonella isolates recovered from the blood samples of hospitalized subjects in Kano metropolis.
Study Design: The study is a descriptive cross-sectional study.
Place and Duration of Study: One milliliter of venous blood was collected from each patient with some or all clinical features of salmonellosis that sign a consent form and transfer into EDTA bottles. If daily is unavoidable blood samples were stored at 4°C. Samples were analyzed at the both Laboratories of the authors. This work was carried out between May, 2011 and March, 2013.
Methodology: The blood specimens were cultured in thioglycollate broth and sub-cultured onto deoxycholate citrate agar (DCA), Salmonella-Shigella agar (SSA) and brilliant Green agar (BGA) followed by confirmation of presumptive colonies using different biochemical tests and analytical profile index 20E. Serologic identification of Salmonella was performed by slide agglutination test using polyvalent O and H Salmonella antisera. Antibacterial drug susceptibility studies were performed by the disc diffusion method using ampicillin, chloramphenicol, ciprofloxacin, nalidixic acid and Trimethoprim-sulfamethoxazole. Out of one hundred and four salmonellae isolates obtained in this study, twenty one were subjected to DNA extraction, real-time and multiplex polymerase chain reaction (PCR) using various primer sets targeting the specific sequences of the resistance genes.
Results: Of the 104 isolates 96 (92.3%) were resistant to Ampicillin; 81 (77.9%) resisted to Nalidixic acid; 30 (30.8%) resisted to Chloramphenicol, 17(16.3%) resisted to Cotrimazole while none (1.0%) resisted to Ciprofloxacine. Among the thirteen Salmonella isolates tested by real-time PCR, Tem gene was detected with a higher frequency (38.5%) compared to the gyrB gene (30.8%). In addition, four Salmonella isolates were found to harbour both tem and gyrB genes together with 30.8%. In this study, no gyrA gene amplicon was obtained, although one Salmonella Typhi strain was resistant to ciprofloxacin phenotypically. There was significant correlation between the presence of tem and/or gyrB and resistance to ampicillin and nalidixic acid. However, in this study, out of the eight Salmonella isolates tested by multiplex PCR technique, sul2 gene was detected in two (25.0%) isolates and catP gene was detected in one (12.5%); while the amplification of these two genes failed in five Salmonella isolates (62.5%) and no isolate harboured both sul2 and catP genes. There is no significant correlation between the presence of sul2 /catP and resistance to chloramphenicol and cotrimoxazole.
Conclusion: Most Salmonella serovars isolated from patients in Kano, Nigeria resisted to Ampicillin. They also resisted to Nalidixic acid, Chloramphenicol and Cotrimazole, in decreasing order. Ciprofloxacine remained effective against all the Salmonella isolates tested. Almost all the genes tested were circulating within Kano metropolis, including tem, gyrB, sul2 and catP genes with exception of gyrA. There was only significant correlation between the presence of tem and/or gyrB and resistance to ampicillin and nalidixic acid. 
Artifactual Changes in Whole Blood and Plasma Glucose Levels of Diabetic and Non Diabetic Blood Samples Twenty Four Hours (24 h) Post Collection
Analysis of diabetic blood may be delayed for hours post collection owing to several factors. Artifactual changes may occur thereby confounding the results. This study investigated some artifactual changes in glucose levels that may occur in diabetic blood stored at room temperature for 24 h. Ten (10) male Wistar rats were assigned to two (2) groups of five (5) rats per group. Group 1 rats were made diabetic by single intraperitoneal injection of 160 mg/kg of alloxan monohydrate while group 2 rats served as normal control. Rats with blood glucose values ≥ 126 mg/dl were considered diabetic. One week (7 days), following establishment of diabetes, blood samples were collected after overnight fasting from both diabetic and non diabetic rats using heparinized capillary tubes into sample bottles. Determination of the blood glucose values were done 1 h post collection and subsequently after every 2 h for 24 h on both whole blood and plasma. Results indicated earlier significant (p<0.05) decreases in the glucose values of diabetic whole blood samples compared to the non-diabetic counterpart. Decreases in the glucose levels of whole blood sample were significantly (p<0.05) higher compared to that of the plasma. It was concluded that the blood and plasma sugar levels of diabetic rats deteriorated faster compared to the non diabetic counterpart and that significant changes in the glucose levels of both blood and plasma occurred within 2 h post collection. The plasma sugar levels of non diabetic rats were unreliable (increased rather than decrease). 
 Rainen, L., Oelmueller, U., Jurgensen, S., Wyrich, R., Ballas, C., Schram, J., Herdman, C., Bankaitis-Davis, D., Nicholls, N., Trollinger, D. and Tryon, V., 2002. Stabilization of mRNA expression in whole blood samples. Clinical chemistry, 48(11), pp.1883-1890.
 Mazdai, A., Dodder, N.G., Abernathy, M.P., Hites, R.A. and Bigsby, R.M., 2003. Polybrominated diphenyl ethers in maternal and fetal blood samples. Environmental health perspectives, 111(9), pp.1249-1252.
 Yeung, L.W., So, M.K., Jiang, G., Taniyasu, S., Yamashita, N., Song, M., Wu, Y., Li, J., Giesy, J.P., Guruge, K.S. and Lam, P.K., 2006. Perfluorooctanesulfonate and related fluorochemicals in human blood samples from China. Environmental science & technology, 40(3), pp.715-720.
 Abdullahi, M., O. Olonitola, S., J. Umoh, V. and H. Inabo, I. (2014) “Antibacterial Resistance Profile and PCR Detection of Antibiotic Resistance Genes in Salmonella serovars Isolated from Blood Samples of Hospitalized Subjects in Kano, North-West, Nigeria”, Microbiology Research Journal International, 5(3), pp. 245-256. doi: 10.9734/BMRJ/2015/9711.
 Emeka Aba, P., Nnaemeka Edeh, M. and Chukwuamachago Nweze, E. (2016) “Artifactual Changes in Whole Blood and Plasma Glucose Levels of Diabetic and Non Diabetic Blood Samples Twenty Four Hours (24 h) Post Collection”, International Blood Research & Reviews, 5(3), pp. 1-7. doi: 10.9734/IBRR/2016/26044.