Comparative advantage of anti-Mullerian hormone over other ovarian reserve metric (basal hormonal test) in prediction of fertility in women with varying menstrual cycle

Objective: Anti-Mullerian hormone has been indicated as a novel biomarker for ovarian reserve assessment. This study aimed to determine the comparative advantage of serum levels of AMH, FSH, LH, E2, and LH/FSH ratio among women with varying menstrual cycles and duration of menstruation. Material and Method: A total of 90 subjects, which consisted of sixty subjects and thirty healthy subjects as control, were recruited. Blood samples were collected on day 3 of the menstrual cycle and evaluated for ovarian markers using the ELISA technique. All data were analyzed using SPSS version 23.0. Results: AMH and LH/FSH levels were lower in women with varying menstrual cycles than in the control group. FSH, LH, and E2 levels were significantly higher in women with varying menstrual cycles and flow duration than in the control group(P<0.05). AMH was negatively correlated with age (r= -0.72), BMI( r= -0.4), FSH(r= - 0.3), LH(r = -0.2) (p<0.05) and E2(r= -0.2, p>0.05). Also age was positively correlated with FSH(r=0.5,p<0.05), E2(r=0.3, p<0.05) and BMI(r=0.4, p<0.05). The level of AMH was not significant with cycle length and days of flow(p>0.05). This implies that AMH can be measured independently of the cycle phase. This show that AMH was lower in women with varying menstrual cycle with an increase in gonadotrophin and E2. The strong negative relationship between age and AMH implies that age is determining factor of ovarian reserve. Conclusion: AMH combined with age and FSH may improve ovarian reserve evaluation, making AMH a better marker

Uji Aktivitas Ekstrak Daun Dan Akar Singawalang (Petiveria alliaceae) Terhadap Penghambatan Tirosinase

Tyrosinase is an enzyme that plays a role in the formation of skin pigments from a person because it is involved in the process of melanogenesis. Tyrosinase plays a very important role in the skin depigmentation process, there are several attempts to inhibit the skin depigmentation process, one of which is by inhibiting tyrosinase. Research on the leaves and roots of singawalang (Petiveria alliacea) was conducted to determine the potential as a tyrosinase inhibitor. Leaves and root extracts of singawalang were macerated with ethanol, then tested for identification of secondary metabolites. Singawalang leaves extract contains alkaloids, tannins and terpenoids while singawalang root extract contains alkaloids, flavonoids, tannins and terpenoids. Tyrosinase inhibitory activity used the microplate reader ELISA technique at a wavelength of 492 nm. Tests were carried out on kojic acid as a comparison and L-DOPA as a substrate. The results of the tyrosinase inhibition activity test for the extracts of singawalang leaves, singawalang roots and kojic acid, IC50 were 9.817 mg / mL, 4.987 mg / mL and 0.092 mg / mL, respectively.

Enzyme Linked Immunosorbent Assay for Fumonisin B1 Detection in Local Corn Seeds from Baghdad-Iraq

Fungi produce a series of toxic compounds on corn, especially Fumonisin B1 (FB1) toxin produced by Fusarium spp. and promoting cancer activity in humans and animals. This study aimed to the isolation and identification of fungi associated with local corn seeds and the detection for the presence of FB1 by using ELISA technique. Thirty samples of corn ears were collected from silos and markets in Baghdad city during the period from November 2018 to March 2019. The present study found that Fusarium was the dominant isolate among fungi in terms of the relative density 57.07%, followed by Aspergillus 31.17%, Rhizopus 3.36%, Alternaria 2.88%, Mucor 2.16%, Penicillium 1.92%, Trichothecium 0.96%, and Helminthosporium 0.48%. FB1 was detected in all samples of the silos and markets with a concentration range of 13.69 - 175.54 µg/kg. There were no significant differences in FB1concentration among samples collected from the silos and markets. Also, no relationship was found between the number of infected seeds by Fusarium spp. and FB1concentrations.

An Increase in TNF-α Levels in Fetus due to Prenatal Ischemic Hypoxia

BACKGROUND: Prenatal ischemic hypoxia can increase mortality and morbidity and affect the immune system. One of the immune responses is tumor necrosis factor-α (TNF-α) levels. However, the cellular mechanism of immune response abnormalities due to prenatal hypoxia remains unclear. An 11–17-day-old fetus is a sensitive period of neural development. Brain ischemia will cause cell dysfunction and can even affect TNF-α levels. Thus, how prenatal ischemic hypoxia increases TNF-α levels in the fetus remains unclear. AIM: This study aims to examine the effect of the onset and duration of prenatal ischemic hypoxia on TNF-α levels. METHODOLOGY: An experimental study with a post-test control design was conducted. Thirty Rattus norvegicus were induced with prenatal ischemic hypoxia (embryos aged 7, 12, and 17 days). The independent variable was prenatal ischemic hypoxia, while the dependent variable was TNF-α levels. TNF-α was measured using the ELISA technique and was carried out when the fetus was 19 days old (E19). The TNF-α was analyzed using ANOVA, and the limit of significance was set at p < 0.05. RESULTS: The TNF-α levels in the prenatal ischemic hypoxia group were statistically higher than in the control group (p < 0.05). The more the onset and the longer the ischemic hypoxia is, the higher the TNF-level (p < 0.05). CONCLUSION: The prenatal ischemic hypoxia increased TNF-α levels in the fetus.

Innovative application of postbiotics, parabiotics and encapsulated Lactobacillus plantarum RM1 and Lactobacillus paracasei KC39 for detoxification of aflatoxin M1 in milk powder

This study aimed to evaluate aflatoxin M1 (AFM1) level in milk powder and infant milk formulae, in addition to applying innovative methods for AFM1 & AFB1 detoxification. Fifty random samples of milk powder and infant formulae (25 of each) were collected from the Egyptian markets for assessing AFM1 level using ELISA technique. Bioactive components comprising cell free supernatants (postbiotic), acid-dead cells (parabiotic) and the encapsulated-cells of Lactobacillus plantarum RM1 and Lactobacillus paracasei KC39 were evaluated for their antifungal activity against toxigenic mold strains and their impact on AFB1 and AFM1 reduction in reconstituted milk powder. AFM1 concentration in unpacked milk powder was higher than that of packed samples and infant formulae, although these differences were not significant (P > 0.05). About 96.0, 29.4 and 25.0% of the tested infant formulae, unpacked, and packed milk powder were unacceptable in terms of the AFM1 limit defined by Egyptian and European standards, while all samples were in accordance with the USA/FDA standard. All tested mycotoxigenic strains were sensitive to the different treatments of the probiotics with the highest sensitivity regarding Fusarium strain with L. paracasei KC39 compared to other genera. The degradation ratios of AFM1 using the bioactives of the L. paracasei KC39 were higher than that of L. plantarum RM1 bioactives. Additionally, KC39 parabiotic manifested the best AFB1 reduction (60.56%). In conclusion, the positive and highly significant relationship (P < 0.05) between these effective biocompounds mirrors their major detoxification role which gives a safe solution for AFs contamination issues in milk and milk products.

Covalent Immobilization of Antibodies through Tetrazine-TCO Reaction to Improve Sensitivity of ELISA Technique

Enzyme-linked immunosorbent assay (ELISA) is routinely used to detect biomolecules related to several diseases facilitating diagnosis and monitoring of these, as well as the possibility of decreasing their mortality rate. Several methods have been carried out to improve the ELISA sensitivity through antibodies immobilization on the microtiter plates. Here, we have developed a strategy of antibodies immobilization to improve the ELISA sensitivity increasing the antibody density surface through the tetrazine (Tz)-trans-cyclooctene (TCO) reaction. For this, we prepared surfaces with tetrazine groups while the captured antibody was conjugated with TCO. The tetrazine surfaces were prepared in two different ways: (1) from aminated plates and (2) from Tz-BSA-coated plates. The surfaces were evaluated using two sandwich ELISA models, one of them using the low-affinity antibody anti-c-myc as a capture antibody to detect the c-myc-GST-IL8h recombinant protein, and the other one to detect the carcinoembryonic human protein (CEA). The sensitivity increased in both surfaces treated with tetrazine in comparison with the standard unmodified surface. The c-myc-GST-IL8h detection was around 10-fold more sensible on both tetrazine surfaces, while CEA ELISA detection increased 12-fold on surfaces coated with Tz-BSA. In conclusion, we show that it is possible to improve the ELISA sensitivity using this immobilization system, where capture antibodies bond covalently to surfaces.

INCIDENCE OF TBRV AND ArMV VIRUSES IN NEW CHERRY PLANTATIONS FROM AREA OF ROMANIA

Viral diseases can influence negatively a good evolution of cherry plantations. Two of the viral diseases that attack this species are caused by the ArMV virus, which spreads by multiplying the infected propagating material, by seed and nematodes, and by the TBRV virus, which spreads by multiplying the infected material and nematodes. In order to study the incidence of the two viruses, 10 cherry plantations located in the south of the country were evaluated, in the district of Argeș, Dolj, Ialomița, Ilfov, Călărași, Dâmbovița, Buzău. The planting material used to set up the plantations was both from Romania and from an external source: the Netherlands, Greece, Italy. Viral evaluation performed visually and serologically by DAS-ELISA technique, identified viral infections in 2 of the 10 plantations. In one plantation, TBRV was identified in 20% of the tested samples and in the second, ArMV viruses were identified in 20% of the samples and TBRV in 5% of the samples.

Intake of Vitamin D in Patients with Multiple Sclerosis in the Valencian Region and Its Possible Relationship with the Pathogenesis of the Disease

(1) Background: Multiple sclerosis (MS) is a neurodegenerative disease characterized by pronounced inflammation. Interleukin 6 (IL-6) is an accurate marker for the state of inflammation, due to the high levels of this cytokine linked to the pathogenesis of the disease. These IL-6 levels could be lowered with an adequate dietary intake of vitamin D. The objective of the study was to determine the level of vitamin D ingested in a sample of patients with MS in the Valencian region (Spain), to establish the vitamin sources, and the possible link between the intake of vitamin D and the pathogenesis of the disease through a relationship with the level of IL-6. (2) Materials and Methods: A descriptive pilot study was carried out with 39 patients with MS in the Valencian region. The dietary-nutritional anamnesis was gained through the food frequency questionnaire (FFQ) and a food diary. Diet and eating habits were analyzed through the Easy Diet (version: 2.0.1)—Consultation Management Program® software, and IL-6 levels in blood by ELISA technique. (3) Results: The results show a low intake of vitamin D, which is significantly and negatively related to the intake of proteins of vegetable origin, which are consumed in less quantity than proteins of animal origin, and significantly and negatively related with the high blood levels of IL-6, possibly as a consequence of the high intake of fats, mainly unsaturated. (4) Conclusions: MS patients in the Valencian region ingest little vitamin D related to low intake of vegetable protein, which would explain the high levels of IL-6 linked to the high intake of mainly saturated fats.

Association of TNFRII polymorphisms and IL-37 in rheumatoid arthritis Iraqi patients

Fifty RA patients and 50 healthy individuals have been participated in this study. 196 M/R polymorphism of TNFRII gene determined by PCR-RFLP, IL-37 level was measured by ELISA technique, also RF, ACCP, CRP measured by ELISA technique. Our study shows an increase in IL-37 levels in patients was suffered from rheumatoid arthritis relative to control group. (M=101.31+10.41) That shows major differences between patients and controls (p<0.01) and increasing level of IL-37 correlate significantly with increasing level of CRP (p<0.05). The frequencies of TNFRII gene polymorphism were significantly correlate with the IL-37 level (p<0.01) in RA patients compared with control group. In conclusion, IL-37 increased in RA patients associated with disease development, also significantly associated with TNFRII polymorphism.

Serum HMGB1, DKK1, and ACE2 as a Function of Lung Injuries in COVID-19 Patients

There is a need for a biomarker for lung injury in COVID-19 patients. In the present study, an attempt was carried out to examine the role of Dickkopf-related protein 1 (DKK1), High-mobility group box 1 protein (HMGB1), angiotensin-converting enzyme 2 (ACE2) as a function for the lung abnormalities in CT-scan (LACTS). To perform the goals, DKK1, HMGB1, and ACE2 were measured in patients and controls using the ELISA technique. In contrast, other parameters were measured spectrophotometrically. The results showed decreased SpO2 and albumin and an increase in the serum biochemical parameters (glucose, urea, creatinine, D-dimer, ACE2, DKK1, and HMGB1) in COVID-19 patients compared with the control group. In COVID-19 patients, the percentages of the lung abnormalities in CT-scan% are 40.67±11.84. The results showed that those patients with LACTS patients are slightly older and have lower SpO2 than the patients without the LACTS group. ACE2 shows a significant correlation with SpO2 (ρ = 0.336, p<0.01) and a negative correlation with albumin (ρ = -0.197, p<0.05). Other parameters showed no significant correlation with the measured biomarkers. In conclusion, COVID-19 patients have higher ACE, DKK1, and HMGB1 indicating the involvement of the pathways of these biomarkers in the disease progression including lung injury.