Qualitative analysis of quercetin present in ethanolic leaf extract of Blumea balsamifera L. DC (Sambong) using TLC-bioautography and HPLC-PDA

The objective of the study is to measure the amount of quercetin present in DCM fraction of ethanolic extract of Blumea balsamifera L. leaves using TLC-bioautography and HPLC-PDA. Furthermore, the study compared TLC-bioautography versus HPLC in quantification of quercetin in the plant sample. Powdered B. balsamifera leaves were macerated with 95% ethanol and the filtrate was subjected to liquid-liquid partitioning. The extract was partitioned with an equal amount of n-hexane. Recovered ethanol partition was subjected to rotary evaporation and remaining aqueous fraction was successively partitioned with dichloromethane, chloroform and ethyl acetate. The residue obtained from n-hexane, dichloromethane, chloroform, ethyl acetate and water were screened using Bate-Smith and Metcalf method and Wilstater “cyanidin” test. All fractions tested were negative for Bate-Smith and Metcalf method. Dichloromethane, chloroform and ethyl acetate fractions showed positive results for Wilstate test indicating possible presence of gamma-benzopyrone. Dichloromethane fraction was used for column chromatography because in comparison with ethyl acetate and chloroform fraction, the intensity of color was best. The residue obtained from dichloromethane was purified by column chromatography. Each fraction was spotted in the TLC plate and sprayed with DPPH. All fractions that tested positive for DPPH test were pooled. Thin Layer Chromatography method was developed to carry out the bioautography process utilizing the anti-oxidant property of quercetin. Using the developed method, the amount of quercetin in the DCM fraction was determined. High performance liquid chromatography method was developed to measure the amount of quercetin present in the pooled fractions. The final concentration of quercetin in the pooled fractions for HPLC is 2.022 mg/mL and TLC-bioautography method is 2.25 mg/ mL. Comparison of two methods: Mean: 0.356 mg/mL, variance: 0.000722 mg2/mL2, standard deviation: 0.02687 mg/mL, % relative standard deviation: 7.55%, and the standard error: 0.019 mg/mL. The quantitation of quercetin using TLC-bioautography and HPLC is significantly similar

Sexual dishonesty in the Philippines: Filipino college males and females lie about their sexual experiences

There have been numerous studies that aimed to delineate the factors on sexual behavior as some evolutionary theorists claim (Buss, 1995 Darwin, 1871 Greiling & Buss, 1995) that it has always been the most reproductively adaptive individual who is likely to have more benefits on the sexual competition arena. Individuals then respond to this nuance by conforming to the sexual requisites necessary to be more dominant in order to attain the most plausible potential mate. Sexual strategies are then employed and executed to achieve their desired mates (Buss & Schmitt, 1993). Hence, this is when individuals result into processing and altering themselves into manifesting these desirable characteristics that their sex counterpart prefers. This study aims to determine and gather significant empirical basis and evidences necessary concerning the reasons why men and women alter their self-reported information about their respective sexual experiences based on their sexual histories, and to whom they are most likely to report the accurate information regarding their sexual experiences

Protective effect of pre-existing natural immunity in a nonhuman primate reinfection model of congenital cytomegalovirus infection.

Congenital cytomegalovirus (cCMV) is the leading infectious cause of neurologic defects in newborns with particularly severe sequelae in the setting of primary CMV infection in the first trimester of pregnancy. The majority of cCMV cases worldwide occur after non-primary infection in CMV-seropositive women; yet the extent to which pre-existing natural CMV-specific immunity protects against CMV reinfection or reactivation during pregnancy remains ill-defined. We previously reported on a novel nonhuman primate model of cCMV in rhesus macaques where 100% placental transmission and 83% fetal loss were seen in CD4+ T lymphocyte-depleted rhesus CMV (RhCMV)-seronegative dams after primary RhCMV infection. To investigate the protective effect of preconception maternal immunity, we performed reinfection studies in CD4+ T lymphocyte-depleted RhCMV-seropositive dams inoculated in late first / early second trimester gestation with RhCMV strains 180.92 (n = 2), or RhCMV UCD52 and FL-RhCMVΔRh13.1/SIVgag, a wild-type-like RhCMV clone with SIVgag inserted as an immunological marker, administered separately (n = 3). An early transient increase in circulating monocytes followed by boosting of the pre-existing RhCMV-specific CD8+ T lymphocyte and antibody response was observed in the reinfected dams but not in control CD4+ T lymphocyte-depleted dams. Emergence of SIV Gag-specific CD8+ T lymphocyte responses in macaques inoculated with the FL-RhCMVΔRh13.1/SIVgag virus confirmed reinfection. Placental transmission was detected in only one of five reinfected dams and there were no adverse fetal sequelae. Viral whole genome, short-read, deep sequencing analysis confirmed transmission of both reinfection RhCMV strains across the placenta with ~30% corresponding to FL-RhCMVΔRh13.1/SIVgag and ~70% to RhCMV UCD52, consistent with the mixed human CMV infections reported in infants with cCMV. Our data showing reduced placental transmission and absence of fetal loss after non-primary as opposed to primary infection in CD4+ T lymphocyte-depleted dams indicates that preconception maternal CMV-specific CD8+ T lymphocyte and/or humoral immunity can protect against cCMV infection

The Mutationathon highlights the importance of reaching standardization in estimates of pedigree-based germline mutation rates

In the past decade, several studies have estimated the human per-generation germline mutation rate using large pedigrees. More recently, estimates for various nonhuman species have been published. However, methodological differences among studies in detecting germline mutations and estimating mutation rates make direct comparisons difficult. Here, we describe the many different steps involved in estimating pedigree-based mutation rates, including sampling, sequencing, mapping, variant calling, filtering, and appropriately accounting for false-positive and false-negative rates. For each step, we review the different methods and parameter choices that have been used in the recent literature. Additionally, we present the results from a ‘Mutationathon,’ a competition organized among five research labs to compare germline mutation rate estimates for a single pedigree of rhesus macaques. We report almost a twofold variation in the final estimated rate among groups using different post-alignment processing, calling, and filtering criteria, and provide details into the sources of variation across studies. Though the difference among estimates is not statistically significant, this discrepancy emphasizes the need for standardized methods in mutation rate estimations and the difficulty in comparing rates from different studies. Finally, this work aims to provide guidelines for computational and statistical benchmarks for future studies interested in identifying germline mutations from pedigrees

Gating strategy for PBMC immunophenotyping analysis in acute RhCMV reinfection.

(A) Gating strategy used for the innate immune cell compartment. (B) The gating strategy used to identify T cell subsets. (C) Representative plots of side scatter high (SSChi) population following RhCMV reinfection in CMV-seropositive rhesus macaque dams. (TIF)</p

Improved study outcome in dams with pre-existing immunity.

The main readouts of the study are described as a frequency of total number of animals.</p

CMV-specific CD8+ T lymphocyte memory responses to RhCMV immediate early (IE) proteins and exogenous SIV Gag protein in CD4+ T lymphocyte depleted RhCMV reinfected macaques.

(A) Paired IE-specific responses by CD107a expression and secretion of IFN-γ, IL-2, and TNF-α in four CMV-seropositive macaques reinfected with RhCMV UCD52 and FL-RhCMVΔRh13.1/SIVgag (n = 3) or RhCMV 180.92 (n = 1). Pre-reinfection responses were compared with responses at week 8–10 post RhCMV reinfection using paired t-test. (B) Polyfunctional SPICE analysis of IE-specific responses pre vs post RhCMV reinfection showing the proportion of four-functional, three-funtional, two-functional and single function responses. CD107a (blue arc), IFN-γ (red arc), IL-2 (orange arc), and TNF-α (green arc). Four-functional responses are displayed in white, three-functional responses in dark grey, two-functional response in light grey, and mono-functional responses in black. (C) Bar graph of the polyfunctional responses pre (grey) and post (black) RhCMV reinfection (n = 4) showing the frequency of memory CD8+ T lymphocytes responding to RhCMV IE peptides. The RhCMV IE-specific response was measured by intracellular cytokine staining after stimulation with RhCMV IE1 and / or IE2 peptide pools depending on the baseline immunodominant response in individual animals. Comparison of pre- and post reinfection Boolean responses were compared with the Wilcoxon rank sum test using SPICE v6 software.</p

Evidence of reinfection in CD4+ T lymphocyte depleted FL-RhCMVΔRh13.1/SIV<i>gag</i> inoculated dams.

(A) RhCMV-specific (black line) and SIVgag-specific (grey line) real time PCR results in the saliva of one CMV-seropositive reinfected animal (KB91). All other animals were found negative for SIVgag DNA in saliva and urine. (B) Detection of SIV Gag-specific T lymphocyte responses measured longitudinally against a SIVmac239 Gag peptide pool in CMV-seropositive reinfected macaques (KB91, KK24, and JP01) inoculated with RhCMV UCD52 and FL-RhCMVΔRh13.1/SIVgag. Horizontal stippled line shows negative cut-off based on pre-reinfection values.</p