scholarly journals Plasma parasitemia as assessed by quantitative PCR in relation to clinical disease severity in African adults with falciparum malaria with and without HIV co-infection

Infection ◽  
2020 ◽  
Vol 48 (3) ◽  
pp. 367-373
Author(s):  
Aase Berg ◽  
Sam Patel ◽  
Marit G. Tellevik ◽  
Christel G. Haanshuus ◽  
Ingvild Dalen ◽  
...  
Keyword(s):  
Blood Cell ◽  
Severe Malaria ◽  
Falciparum Malaria ◽  
Disease Severity ◽  
Quantitative Pcr ◽  
Univariate Analysis ◽  
Cell Fraction ◽  
Malaria Parasitemia ◽  
Tertiary Teaching ◽  
The Impact

Abstract Purpose When considering malaria disease severity, estimation of parasitemia in erythrocytes is important, but sometimes misleading, since the infected erythrocytes may be sequestered in peripheral capillaries. In African children and Asian adults with falciparum malaria, parasitemia as assessed by quantitative PCR (qPCR) in plasma seems to be a valuable indicator of disease severity, but data on African adults as well as the impact of co-infection with HIV is lacking. Methods In 131 patients with falciparum malaria in a public tertiary teaching hospital in Mozambique, plasma malaria parasitemia as assessed by qPCR, compared to qualitative malaria PCR in blood cell fraction, was related to malaria disease severity and HIV co-infection. Results Of the 131 patients with falciparum malaria, based on positive qualitative PCR in the blood cell fraction, 93 patients (72%) had positive malaria qPCR in plasma. Patients with severe malaria as defined by the WHO criteria had higher malaria quantitative plasma parasitemia (median 143 genomes/µL) compared to those with uncomplicated malaria (median 55 genomes/µL, p = 0.037) in univariate analysis, but this difference was attenuated after adjusting for age, sex and HIV co-infection (p = 0.055). A quarter of the patients with severe malaria had negative qPCR in plasma. Conclusions This study of adult African in-patients with falciparum malaria with and without HIV co-infection, neither confirms nor rejects previous studies of malaria qPCR in plasma as an indicator of disease severity in patients with falciparum malaria. There is a need for further and larger studies to clarify if parasitemia as assessed malaria qPCR in plasma could be a surrogate marker of disease severity in falciparum malaria.

Ovarian Cancer: Sites of Recurrence

2013 ◽  
Vol 23 (9) ◽  
pp. 1590-1596 ◽  
Author(s):  
Pascale Amate ◽  
Cyrille Huchon ◽  
Anne Lucie Dessapt ◽  
Chérazade Bensaid ◽  
Jacques Medioni ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Univariate Analysis ◽  
Progression Free Survival ◽  
International Federation ◽  
Peritoneal Recurrence ◽  
Disease Spread ◽  
Free Survival ◽  
Platinum Based Chemotherapy ◽  
Tertiary Teaching ◽  
The Impact

IntroductionImproved knowledge of recurrence sites after contemporary surgical management of ovarian cancer is needed.Material and MethodsWe retrospectively reviewed consecutive patients managed for epithelial ovarian or tubal cancer with surgery and platinum-based chemotherapy between January 1, 2005, and December 31, 2009, in a tertiary teaching hospital. The site of first recurrence was recorded. Univariate analysis was performed to identify factors associated with site-specific recurrence. Overall survival and progression-free survival were computed using the Kaplan-Meier method, and log-rank tests were performed to assess the impact on survival of the variables of interest.ResultsRecurrences were noted in 3 (20%) of 15 International Federation of Gynecologists and Obstetricians stage I to IIa patients and 36 (62.1%) of 58 International Federation of Gynecologists and Obstetricians IIb to IV patients, and the median progression-free survival was 21.6 (2.5–71) and 19.3 (1.8–67.6) months, respectively. In the advanced-disease group, 75% of recurrences involved the peritoneum and 40% were confined to the peritoneum; peritoneal recurrences developed at both treated and untreated sites. Peritoneal recurrence was associated with greater initial peritoneal involvement (Sugarbaker score, 12.1 ± 8.2 vs 7.1 ± 7.4; P = 0.01) and residual postoperative tumor. Nodal recurrences were noted in 38% of all recurrences, usually in combination with peritoneal recurrence and in the abdominal territories. Isolated distant metastasis was a rare mode of recurrence (8%).ConclusionsThe peritoneum is the main recurrence site in both early and advanced ovarian cancer. Initial disease spread and extent of surgery are associated with the recurrence risk. This article supports the view that more attention should be directed toward extensive treatment of the peritoneum.

scholarly journals Detection of Transgenes in Gene Delivery Model Mice by Adenoviral Vector Using ddPCR

2019 ◽  
Author(s):  
Takehito Sugasawa ◽  
Kai Aoki ◽  
Koichi Watanabe ◽  
Koki Yanazawa ◽  
Tohru Natsume ◽  
...  
Keyword(s):  
Blood Cell ◽  
Quantitative Pcr ◽  
Whole Blood ◽  
Digital Pcr ◽  
Cell Fraction ◽  
Detection Methods ◽  
Gene Doping ◽  
World Anti Doping Agency ◽  
Injection Methods ◽  
Tail Tip

With the rapid progress of genetic engineering and gene therapy, World Anti-Doping Agency has alerted to gene doping and prohibited its use in sports. However, there is no standard method available yet for detection of transgenes delivered by recombinant adenoviral (rAdV) vectors. Here we aimed to develop a detection method for transgenes delivered by rAdV vectors in a mouse model that mimics gene doping. rAdV vectors containing mCherry gene was delivered in mice through intravenous injection or local muscular injection. After five days, stool and whole blood samples were collected, and total DNA was extracted. As additional experiments, whole blood was also collected from mouse tail tip until 15 days from injection of the rAdv vector. Transgene fragments from different DNA samples were analyzed using semi-quantitative PCR (sqPCR), quantitative PCR (qPCR), and droplet digital PCR (ddPCR). In the results, transgene fragments could directly be detected from blood cell fraction-DNA, plasma-cell free DNA and stool-DNA by qPCR and ddPCR, depending on specimen type and injection methods. We observed that a combination of blood cell fraction-DNA and ddPCR was more sensitive than other combinations used in this model. These results could accelerate the development of detection methods for gene doping.

scholarly journals Detection of Transgenes in Gene Delivery Model Mice by Adenoviral Vector Using ddPCR

Genes ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 436 ◽  
Author(s):  
Takehito Sugasawa ◽  
Kai Aoki ◽  
Koichi Watanabe ◽  
Koki Yanazawa ◽  
Tohru Natsume ◽  
...  
Keyword(s):  
Blood Cell ◽  
Quantitative Pcr ◽  
Whole Blood ◽  
Digital Pcr ◽  
Cell Fraction ◽  
Detection Methods ◽  
Gene Doping ◽  
World Anti Doping Agency ◽  
Injection Methods ◽  
Tail Tip

With the rapid progress of genetic engineering and gene therapy, the World Anti-Doping Agency has been alerted to gene doping and prohibited its use in sports. However, there is no standard method available yet for the detection of transgenes delivered by recombinant adenoviral (rAdV) vectors. Here, we aim to develop a detection method for transgenes delivered by rAdV vectors in a mouse model that mimics gene doping. These rAdV vectors containing the mCherry gene was delivered in mice through intravenous injection or local muscular injection. After five days, stool and whole blood samples were collected, and total DNA was extracted. As additional experiments, whole blood was also collected from the mouse tail tip until 15 days from injection of the rAdv vector. Transgene fragments from different DNA samples were analyzed using semi-quantitative PCR (sqPCR), quantitative PCR (qPCR), and droplet digital PCR (ddPCR). In the results, transgene fragments could be directly detected from blood cell fraction DNA, plasma cell-free DNA, and stool DNA by qPCR and ddPCR, depending on specimen type and injection methods. We observed that a combination of blood cell fraction DNA and ddPCR was more sensitive than other combinations used in this model. These results could accelerate the development of detection methods for gene doping.

scholarly journals The impact of delayed treatment of uncomplicated P. falciparum malaria on progression to severe malaria: A systematic review and a pooled multicentre individual-patient meta-analysis

PLoS Medicine ◽  
2020 ◽  
Vol 17 (10) ◽  
pp. e1003359
Author(s):  
Andria Mousa ◽  
Abdullah Al-Taiar ◽  
Nicholas M. Anstey ◽  
Cyril Badaut ◽  
Bridget E. Barber ◽  
...  
Keyword(s):  
Systematic Review ◽  
Severe Malaria ◽  
Falciparum Malaria ◽  
Meta Analysis ◽  
Delayed Treatment ◽  
The Impact

Falciparum malaria parasitemia index for predicting severe malaria

2012 ◽  
Vol 34 (3) ◽  
pp. 320-327 ◽  
Author(s):  
N. TANGPUKDEE ◽  
S. KRUDSOOD ◽  
S. KANO ◽  
P. WILAIRATANA
Keyword(s):  
Severe Malaria ◽  
Falciparum Malaria ◽  
Malaria Parasitemia

scholarly journals Standardisation is necessary in urogenital and extragenital Chlamydia trachomatis bacterial load determination by quantitative PCR: a review of literature and retrospective study

2019 ◽  
Vol 95 (8) ◽  
pp. 562-568 ◽  
Author(s):  
Jeanne A M C Dirks ◽  
Christian J P A Hoebe ◽  
Geneviève A F S van Liere ◽  
Nicole H T M Dukers-Muijrers ◽  
Petra F G Wolffs
Keyword(s):  
Chlamydia Trachomatis ◽  
Disease Severity ◽  
Quantitative Pcr ◽  
Human Cell ◽  
Inflammatory Cells ◽  
Molecular Techniques ◽  
International Standards ◽  
The Impact ◽  
Pcr Targeting ◽  
Anal Site

ObjectivesPathogen load has been linked to disease severity in patients infected with HIV, resulting in international standards to adequately and reproducibly quantify load. Chlamydia trachomatis (CT) load has been inconsistently linked to disease severity since extensive differences exist in quantification methods (14 methods in 28 articles). Differences include normalisation for human cell load due to CT’s intracellular nature, despite the inability to distinguish inflammatory cells from epithelial cells with molecular techniques. We compared the human cell load in CT-positive men and women at the genital and anal site to a CT-negative control group to estimate the impact of inflammatory cells in these samples.Methods188 women (tested at genital and anal site) and 519 men (207 tested at the anal site and 312 tested at the urogenital site) were included from our STI-clinic in the Netherlands. Specimens were self-collected vaginal swabs, anal swabs and urine samples. Quantitative-PCR targeting the HLA-gene quantified human cell load. Mann-Whitney-U-test was used for statistical analyses.ResultsThe genital cell load had a similar range and median (6.5 log10) between CT-negative and CT-positive women . The urogenital cell load was significantly higher than the anal cell load (median 3.6 log10). The anal cell load was significantly higher in men with- than without anal CT infection (median 4.5 versus 3.9 respectively). The anal cell load is significantly higher in CT-positive men than in women. Both Neisseria gonorrhoeae-co-infections and reported anal intercourse significantly increased the human cell load in anal samples.ConclusionStandardisation in CT load studies is necessary as current studies show 14 different quantification methods in 28 studies . In this study we demonstrate the inappropriateness of normalising the CT load for the human cell load using molecular techniques, as the presence of inflammatory cells cannot be excluded.

scholarly journals Circulating Neutrophil Extracellular Traps and Neutrophil Activation Are Increased in Proportion to Disease Severity in Human Malaria

2018 ◽  
Vol 219 (12) ◽  
pp. 1994-2004 ◽  
Author(s):  
Steven Kho ◽  
Gabriela Minigo ◽  
Benediktus Andries ◽  
Leo Leonardo ◽  
Pak Prayoga ◽  
...  
Keyword(s):  
Severe Malaria ◽  
Falciparum Malaria ◽  
Disease Severity ◽  
Severe Disease ◽  
Plasmodium Species ◽  
Neutrophil Extracellular Traps ◽  
Falciparum Infection ◽  
Neutrophil Activation ◽  
Extracellular Traps

AbstractBackgroundNeutrophil activation results in Plasmodium parasite killing in vitro, but neutrophil products including neutrophil extracellular traps (NETs) mediate host organ damage and may contribute to severe malaria. The role of NETs in the pathogenesis of severe malaria has not been examined.MethodsIn Papua, Indonesia, we enrolled adults with symptomatic Plasmodium falciparum (n = 47 uncomplicated, n = 8 severe), Plasmodium vivax (n = 37), or Plasmodium malariae (n = 14) malaria; asymptomatic P falciparum (n = 19) or P vivax (n = 21) parasitemia; and healthy adults (n = 23) without parasitemia. Neutrophil activation and NETs were quantified by immunoassays and microscopy and correlated with parasite biomass and disease severity.ResultsIn patients with symptomatic malaria, neutrophil activation and NET counts were increased in all 3 Plasmodium species. In falciparum malaria, neutrophil activation and NET counts positively correlated with parasite biomass (Spearman rho = 0.41, P = .005 and r2 = 0.26, P = .002, respectively) and were significantly increased in severe disease. In contrast, NETs were inversely associated with parasitemia in adults with asymptomatic P falciparum infection (r2 = 0.24, P = .031) but not asymptomatic P vivax infection.ConclusionsAlthough NETs may inhibit parasite growth in asymptomatic P falciparum infection, neutrophil activation and NET release may contribute to pathogenesis in severe falciparum malaria. Agents with potential to attenuate these processes should be evaluated.

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