Rapid detection of human blood in triatomines (kissing bugs) utilizing a lateral flow immunochromatographic assay - A pilot study

BACKGROUND DNA- and proteomics-based techniques are currently used to identify a triatomine human blood meal. These methods are time consuming, require access to laboratories with sophisticated equipment, and trained personnel. OBJECTIVES We tested a rapid and specific immunochromatographic assay (that detects human blood in forensic samples) to determine if human blood was present in triatomines and their fecal excreta. METHODS We fed Triatoma rubida human blood (positive control) or mouse blood (negative control) and performed the assay on the abdominal contents and fecal excreta. Triatomine field specimens collected in and around human habitations and excreta were also tested. FINDINGS The assay was positive in triatomines fed human blood (N = 5/5) and fecal excreta from bugs known to have ingested human blood (N = 5/5). Bugs feeding on mice (N = 15/15) and their fecal excreta (N = 8/8) were negative for human blood. Human blood was detected in 47% (N = 23/49) triatomines, representing six different species, collected in the field. MAIN CONCLUSIONS The pilot study shows that this rapid and specific test may have applications in triatomine research. Further study is needed to determine the sensitivity of this assay compared to other well-established techniques, such as DNA- and proteomics-based methodologies and the assay's application in the field.

Rapid detection of oleandrin and adynerin in human blood by HPLC-MS/MS / 中国法医学杂志

Objective To establish the analytical method of oleandrin and adynerin in human blood by HPLC-MS/MS. Methods After protein sediment by acetonitril, the concentrations of oleandrin and adynerin in human blood were quantitatively determined by HPLC-MS/MS. The qualitative analysis was conducted based on retention time and MRM ions. Besides, the standard curve method was used for quantification. Results The detection limits of both oleandrin and adynerin were 0.5ng/mL, the linear range was from 1ng/mL to 1mg/mL, with a recovery rate of 75.2%~95.7%. Conclusion The detecting protocol has the advantages of high sensitivity, fast and high accuracy with a relatively wide linear range, which is especially suitable for rapid detection of oleander toxins, alexandrine and adynerin in particular, in human blood in poisoning cases.

Assessment of the DNA Damage in Human Sperm and Lymphocytes Exposed to the Carcinogen Food Contaminant Furan with Comet Assay

ABSTRACTThe aim of this work was to assess the damage of DNA in human blood cell and spermin vitro under the influence of furan. These cells were administered 0-600 μM of furan at 37 and 32oC for 30 and 60 min, respectively. A significant increase in tail DNA%, tail length and moment indicating DNA damage was observed at increasing doses when compared to the controls. The treatment with 300 and 600 μM of furan showed a maximum increase of 86.74 ± 2.43 and 93.29 ± 8.68 compared to the control tail DNA% in lymphocytes. However, only 600 μM of furan showed a maximum increase of 94.71 ± 6.24 compared to the control tail DNA% in sperm. The results suggested that furan caused DNA damage in lymphocytes at increasing doses, but appeared to have not the same effect on human sperm at the low doses. Genotoxic activity had an impact on the risk assessment of furan formed on the food for human cells. Therefore, it would be important to further investigate these properties of furan as the food mutagen.

Potential Use of Polysaccharides from the Brown Alga Undaria pinnatifida as Anticoagulants

Undaria pinnatifida (U. pinnatifida) is a highly invasive species and has caused concern all over the world because it has invaded coastal environments, has the potential to displace native species, significantly alters habitat for associated fauna, and disturbs navigation. Any attempt to eradicate it would be futile, owing to the elusive, microscopic gametophyte, and because the alga thrives in sites rich in anthropic activities. Venice Lagoon is the largest Mediterranean transitional environment and the spot of the highest introduction of non-indigenous species, including U. pinnatifida, which is removed as a waste. We demonstrated that polysaccharide extracts from U. pinnatifida have an anticoagulant effect on human blood in vitro and are not cytotoxic. The results obtained by PT (normal values 70-120%) and APTT (normal values 28-40s) assays were significantly prolonged by the polysaccharide extracts of U. pinnatifida, therefore algal extracts are ideal candidates as antithrombotic agents.

The investigation of cytogenetic and oxidative effects of diffractaic acid on human lymphocyte cultures

Diffractaic acid (DA) is a naturally occurring depside derivative found in several lichen species. It has a wide range of important biological effects such as analgesic and antiviral properties, although its cytotoxic, cytogenetic and oxidative effects have not been investigated in human blood tissue yet. Therefore, increasing concentrations (1, 5, 10, 25, 50, 100 and 200 mgL-1) of DA was added into human whole blood cultures. 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyl tetrazolium bromide (MTT) assay was used to assess the cell viability and/or cytotoxicity and genotoxic damage potential of DA using chromosome aberration (CA) and micronucleus (MN) tests were performed. In addition, oxidative alterations were determined by the total antioxidant capacity (TAC) and total oxidant status (TOS) assays. The results revealed that DA reduced cell viability at higher concentrations than 50 mgL-1. The all tested concentrations of DA were non-genotoxic. In vitro treatments with DA led to increases of TAC levels in the cultured blood cells without changing the TOS levels as compared to the control group. Consequently, DA exhibited a significant non-mutagenic and antioxidant potential in vitro.

Noninvasive measurement of human blood glucose under flow control of time gate / 国际生物医学工程杂志

Objective In order to further improve the noninvasive measurement precision of human blood glucose and achieve clinical requirements,experiments were conducted to measure human blood glucose by a novel measurement method our group proposed.Methods The blood flow of the tested parts was stopped by pressure,and dynamic distribution of transmission spectra was measured by time-correlated single-photon counting (TCSPC) technology under the dual wavelength light incident.Oral glucose tolerance test (OGTT) was conducted on three human subjects,and the early arriving photons were selected by Laplace transform.Meanwhile,the human blood glucose concentration was measured using the Roche glucose meter.Results The best curve fitting was got when the Laplace parameter was 1Gs-1 (determining parameter R2=0.0922).Conclusions The experimental results showed that better measurement accuracy can be obtained by selecting appropriate Laplace parameter,and noninvasive measurement of human blood glucose under flow control of time gate was feasible.

The relation of falciparum infection with human blood groups.

Background and Objectives: The ABO blood group is arguably the best known, yet the most functionally mysterious, genetic polymorphism in human. A number of studies have shown the susceptibility to several infectious diseases is related to patient’s blood group. Malaria is one of the diseases and Malaria remains the most complex and overwhelming health problem facing humanity especially in the vast majority of tropical and subtropical regions of the world. The resurgence of malaria is a serious public health problem in many parts of the world including India. It is therefore important to identify the factors which contribute to susceptibility of hosts. AIM: In order to study the distribution and correlate the incidence of ABO blood group in healthy individuals and Plasmodium falciparum malaria patients admitted in Civil Hospital, Ahmedabad with Plasmodium falciparum infection. To evaluate the morbidity and mortality associated with Plasmodium falciparum malaria infection. To figure out the correlation between ABO blood group and complications caused by Plasmodium Falciparum infection. To find out the blood group which confer some resistance against the plasmodium falciparum malaria infection. Methods: The study conducted from January 2009 to August-2010 (1year, 8 months) on 62 diagnosed cases of Plasmodium falciparum malaria patients admitted in Civil Hospital, Ahmadabad. Patients who were tested positive for plasmodium falciparum and treated for the same in the indoor wards of Civil Hospital, Ahmedabad were taken for study. Observation and Results: out of 62 cases 21 (33.80) were in blood group A, 19 (30.64) were in blood group B, 7(11.29) were in blood group AB and 15 (24.20) were in blood group O. Out of 660 controls selected 161 (24.39%) were of blood group A, 224 (33.93%) were of blood group B, 68 (10.30%) of blood group AB and 207 (31.36%) belonged to blood group O. Conclusion: So we see through Statistical data that although blood group B is commonest in controls but in patients plasmodium falciparum malaria blood group A (33.80%) followed by blood group B (30.64%), blood group O was at 3rd place and AB was least common in cases. By Statistical data there is high relative risk in blood group A (1.38) followed by blood group AB (1.09). Distribution of blood group in healthy individual shows blood group B is more common in India followed by group O then group A and lastly group AB, whereas falciparum malaria incidence is highest in group A (high relative risk is present) followed by group B, then group O and least in group AB. Incidence of malaria is seen higher among young age group (below 30 years) and in male sex probably due to occupational exposure.Highest morbidity and complications are observed in group A followed by group B. Group O appears to be immune to severe complications of falciparum malaria. Mortality is seen equally in group A and B. Death is not noted in group O. As blood group O reduces plasmodium rosseting.

Non-association between anti-Toxoplasma gondii antibodies and ABO blood group system

Toxoplasma gondii infects humans through the gastrointestinal tract (GIT), which elicits humoral immune response with specific antibodies. The expression of the ABO blood group glycoconjugates also occurs in this same system and may influence the human susceptibility of infection by T. gondii. The aim of the present study was to investigate the association between ABO blood group phenotypes and the presence of anti-T. gondii antibodies. Data - including age, results of serology tests for T. gondii infection and ABO blood group phenotypes - were assembled from the medical records of 1,006 pregnant women attended in the Base Hospital of the Medical School of São José do Rio Preto, Brazil, between 2001 and 2004. The chi-square test was used to compare the results with the level of significance set at 5 percent. Of the studied cases, 64.1 percent (645/1006) and 35.9 percent (391/1006) presented respectively positive and negative serology tests for anti-T. gondii antibodies. The mean age of those who tested positive was higher than those with negative serology tests (p = 0.0004). The frequencies of ABO blood group phenotypes were similar in those with and without anti-T. gondii antibodies (p = 0.35). In conclusion, the ABO blood group system is not associated with the presence or absence of anti-T. gondii antibodies.

The clinical effect and safety of human blood albumin for treating acute cerebral hemorrhage / 中国综合临床

Objective To assess the clinical effect and safety of human blood albumin in treatment of acute cerebral hemorrhage.Methods 88 patients with acute cerebral hemorrhage were randomly divided into 2 groups. Both group were given routine therapy and 45 cases in the treating group were given additional 10% human blood albumin 100 ml ivgtt once a day for 7 days.14 days served as a treatment course.Neurofunction was compared before and after treatment.Results Both the effective rate (P<0.05) and the neurofunction in the treating group were better than that of the control group (P<0.05).No advert effect happened.Conclusion Human blood albumin is an efiective and safe medicine for treatment of acute cerebral infarction.

On some toxinological aspects of the starfish Stellaster equestris (Retzius, 1805)

Whole-body extracts in methanol were obtained from the starfish Stellaster equestris. The crude toxin was fractionated stepwise using diethylaminoethyl (DEAE) cellulose column chromatography. The crude toxin was lethal to male albino mice at a dose of 1.00 mL (containing 531.0 µg/mL protein) when injected intraperitoneally (IP) but the toxicity was abolished in all cases except one upon fractionation. The crude toxin and all the adsorbed fractions exhibited potent hemolytic activity on chicken, goat and human blood. However, group B human erythrocytes were resistant to lysis by all fractions and group O by most of the fractions. Paw edema in mice was caused by the crude toxin and all fractions. Pheniramine maleate and piroxicam blocked the toxicity when administered earlier than, or along with, the crude or fractionated toxins but not when administered after the envenomation. Pretreatment with either of these drugs also blocked edema formation.(AU)

Study Progress of Raman Spectroscopy of Human Blood / 实用儿科临床杂志

The appearance and development of laser technology had made Raman spectroscopy one of the most attractive research fields in analysis using laser.Raman spectroscopy had been widely used in the analysis of material components and structure.In this review,recent advances of Raman spectroscopy on human blood were summarized,which including the basic research on red cells,white cells,platelets and the serum concentrations of various substances by Raman spectroscopy.The rewiew also intruduces the Raman spectroscopy on early diagnosis for cancer,diagnosis for blood diseases,and its future development.

Study of high throughput screening for inhibitors of inflammatory cytokine / 中国药理学通报

AIM The aim of our study was to establish drug screening models which can evaluate samples' effects on cytokines related with inflammtion, capable of fast and efficient screening of anti-inflammatory lead compounds on the release of inflammtory cytokines. METHODS Heparinized human blood leukocytes was evaluated as a model to study the effects of various classes of anti-inflammatory lead compounds on cytokine release/biosynthesis from leukocytes. Human blood leukocytes was stimulated with LPS (final concentration 0.5～50 mg?L -1), with or without test drugs (diclofenac, a cytooxygenase inhibitor, nordihydroguaiaretic acid NDGA, a 5-lipoxygenase inhibitor) for 1～4 h to induce cytokine release. RESULTS Human blood leukocytes stimulated with LPS could product IL-1, IL-8 and TNF-? in a dose-dependent manner. Human blood leukocytes was stimulated with LPS(5 mg?L -1) for 4h to induce cytokine release. TNF-?, IL-1 and IL-8 time-course profiles were determined in culture media, using bioassays and ELISA. LPS-mediated release of IL-1 and TNF-? was significantly suppressed by NDGA and Diclofenac. In LPS stimulated blood, NDGA and Diclofenac inhibited the release of TNF-?(IC 50 of 149 ?mol?L -1 and 23.88 ?mol?L -1) or IL-1 (IC 50 of 222.57 ?mol?L -1 and 126 ?mol?L -1). CONCLUSION This human blood leukocytes screening system in vitro has the potential to screen new cytokine release inhibitors and sites of action of new anti-inflammatory lead compounds, and increases the screening efficiency.

Exposure Assessment of PCDD/Fs and Monitoring of Health Effects on Workers and Residents near the Waste Incinerators in Korea / 예방의학회지

OBJECTIVES: In this study, the exposure status of the hazardous substances from incinerators, such as polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs), were studied, and the relationship between the exposure of these hazardous substances and their heath effects on the workers and residents near municipal solid waste (MSW) incinerators and an industrial incinerator investigated. METHODS: Between July 2001 and June 2002, 13 workers at two MSW incinerators, 16 residents from the area around the two MSW incinerators, 6 residents from the control area, and further 10 residents near an industrial incinerator, estimated to emit higher levels of hazardous substances, were interviewed. Information, including sociodemographic information, personal habits, and work history, detailed gynecologic and other medical history were collected through interviews. Blood samples were also collected from 45 subjects, and analyzed for PCDD/DFs, by high resolution gas chromatography - high resolution mass spectrometry, using the US EPA 1613 method. In addition to the questionnaire survey, urinary concentrations of 8-hydroxydeoxyguanosine (8-OH-dG) and malondialdehyde (MDA) were measured as oxidative injury biomarkers. The urinary concentrations of 8-OH-dG were determined by in vitro ELISA, and the MDA by HPLC, using an adduct with thiobarbituric acid. RESULTS: The PCDD/DFs concentrations in the residents near the industrial incinerator were higher than those in the controls, workers and residents near the MSW incinerators. The average TEQ (Toxic Equivalencies) concentrations of the PCDD/DFs in residents near the industrial incinerator were 53.4pg I-TEQs/g lipid. The estimated daily intakes were within the tolerable daily intake range (1-4 pg I-TEQ/Kg bw/day) suggested by WHO (1997) in only 30% to the people near the industrial incinerator. Animal studies have already shown that even a low body burden of PCDD/DFs, such as 10ng TEQ/kg bw, can cause oxidative damage in laboratory animals. Our study also showed that the same body burden of PCDD/DFs can cause oxidative damage to humans. CONCLUSIONS: The exposures to PCDD/DFs and the oxidative stress of residents near the industrial incinerator, were higher than those in the controls, workers and residents near the MSW incinerators. Proper protection strategies against these hazardous chemicals are needed. Because a lower body burden of PCDD/Fs, such as 10ng TEQ/kg bw, can cause oxidative damage, the tolerable daily intake range should be restrictedly limited to 1pg I-TEQ/kg bw/day.

Role of Mitogen-Activated Protein Kinase Pathways in the Production of TNF-alpha, IL-10, and MCP-1 by Mycobacterium tuberculosis H37Rv-Infected Human Blood Monocytes

Clinical manifestations of tuberculosis are closely associated with the initial responses of macrophages to mycobacteria. In this study, we investigated the signal transduction pathways for the secretion of cytokines and chemokines [tumor necrosis factor (TNF)-alpha, interleukin (IL)-10, IL-8, and monocyte chemotactic protein-1 (MCP-1)] in human blood monocytes infected with Mycobacterium tuberculosis H37Rv. M. tuberculosis H37Rv infection induced the secretion of significant amounts of TNF-alpha, IL-10, IL-8, and MCP-1 from human blood monocytes. Analysis of mitogen-activated protein kinase (MAPK) activation [extracellular signal-regulated kinase 1/2 (ERK) and p38 kinase] showed rapid phosphorylation of both subfamilies in response to M. tuberculosis H37Rv. Using highly specific inhibitors of p38 (SB203580) and of MEK-1 (U0126 and PD98059), we found that both p38 and ERK were essential for M. tuberculosis H37Rv-induced TNF-alpha production, whereas activation of the p38 pathway, but not that of ERK, was essential for M. tuberculosis H37Rv-induced IL-10 production. Interestingly, the ERK pathway, but not that of p38, was critical for MCP-1 secretion from human blood monocytes infected with M. tuberculosis H37Rv. However, IL-8 secretion was regulated neither by ERK1/2 nor p38 MAPK. Collectively, these results suggest that induction of the MAPK pathway is required for the expression of TNF-alpha. IL-10, and MCP-1 by human blood monocytes during M. tuberculosis H37Rv infection.

Estimatin of the Storage Time of Human Blood Superoxide Dismutase in Two Preparation Forms at the Room Temperature / 中国输血杂志

Estimation of the storage time of human blood superoxide dismutase (hSOD) in two preparation forms (freeze-dried and soluble states) at room temperature by the heating and accelerating experiment was reported. The freeze-dried hSOD had kept its 90% activity for as long as 3 years, while the soluble hSOD could have preserved its 90％ activity for only 20 hours. The methodhad proved to be relatively reliable when used for many series of specimens.

Níveis de pesticidas organoclorados em soro sanguíneo de pessoas expostas à doença de chagas no Brasil / Levels of hexachlorocyclorhexane (HCH) in exposed people from a Chagas' disease area in Brazil

Níveis de pesticidas organoclorados foram determinados em 51 amostras de sangue de moradores numa área onde a doença de Chagas é controlada pela pulverização das casas com hexaclorocicloexano (HCH) para eliminar insetos vetores. Foram coletadas 28 amostras de sangue de pessoas cujas casas foram tratadas com HCH (grupo 1) e 23 de moradores em casas não tratadas (grupo 2). Os resultados encontrados mostraram diferença significativa entre os dois grupos. No grupo 1, os níveis de HCH variaram de 1 a 35 mg/dl(ppb), com mediana de 10m/dl(ppb) e, no grupo 2, variaram de < 1mg/dl a 5m/dl, com mediana de 1m/dl. Foram detectados pp'DDE em 100% das amostras e Dieldrin em 43,1% das mesmas (AU).

THE APPLICATION OF DNA AMPLIFICATION TECHNIQUE IN VITRO IN FORENSIC MEDICNE PRACTICE / 中国法医学杂志

This paper reports the determination of sex of human blood stains in threecriminal cases by the DNA amplification technique.The results fit the cases de-tails and gave the scientific evidence for the justice.The blood stains were dige-sted with proteinase K.The protein was extracted with the phenol-chloroform.The DNA was precipitated with NaCl and ethyl alcohol.Amplification of DNAwas carried out using two pairs of primer Y1.1 Y1.2 and Alu9.1 Alug.2 and heatstable FD polymerase.The amplified products were subjected to agarose gel(con-taining ethidium bromide)electrophoresis.Sex of blood stains was determinedaccording to the amplified products.PCR technique is rapid to perform,sensitiveand simple.No special equipments and isotope labelled probe are required.

Corynebacterium diphtheriae isolada de sangue / Corynebacterium diphtheriae isolated of blood

Foi isolado C. diphtheriae de 3 amostras de sangue de um paciente internado no Hospital Emílio Ribas, que não apresentava suspeita clínica de difteria. Analisou-se o aspecto morfológico, comportamento bioquímico e toxigênico destas cepas e verificou-se a presença dos biotipos intermediu8, mitis e mitis variedade belfanti. O comportamento bioquímico foi idêntico para as cepas isoladas, com exceção da presença de nitrato redutase do tipo A nos biotipos intermedius e mitis. Esses dois biotipor mostraram não ser toxigênicos quando testados pelo método de Elek, enquanto o biotipo mitis varo belfanti mostrou-se toxigênico quando empregado o mesmo método. Foi observado não haver variações na sensibilidade dos diferentes antibióticos frente aos agentes antimicrobianos testados (AU).

Clinical Significance of Serum Glutathione S-transferase Activity in Viral Hepatitis / 第三军医大学学报

The clnical significance of serum glutathione S-transferase (GST) activity determination was evaluated by parallel measurements of GST and GPT activity inthe seraof 192 patients with liver diseases and 721 patients without liver diseas es. TheGST activity was significantly increased in the former, but it hadn't anychanges in patients without liver diseases. The highest GST activity was found in patients with severe hepatitis. In contrast to survival, the GST activity was persistently increased in those patients died later,and there appeared a "GST/GPT dissociation phenomenon". Although GPT activity had been normal in the convalescent stage of patients with hepatitis, the GST level in most of them was still high and different degree of pathological changes were also present in liver biopsy. Our data suggested that GST activity measurements might be a more sensitive parameter of liver cell damage than usual GPT measurements, and it might be valuable in evaluation of prognosis of severe hepatitis, and differentiation of hepa-tocellular jaundice from obstructive and hemolytic jaundice.