Spontaneous preterm birth is the basic problem of perinatal mortality in Bulgaria. Pregnancy is characterized by physiological leukocytosis - determined by twice increased neutrophils levels, unchanged monocyte levels, and a reduction in the levels of lymphocytes, eosinophils, and basophils. The leukocytosis is particularly pronounced in the last trimester, due to physiological stress during pregnancy, and it is a result of the increased inflammatory response, a consequence of selective immune tolerance, immunosuppression, and fetus-immunomodulation. The objective of this study was to investigate and to appreciate maternal leukocytosis as potential diagnostic markers in spontaneously declared preterm birth and term birth declared cases. Methods: The case–control study was performed in UMBAL-City Hospital, Stara Zagora during 2017–2020. The study involved 200 patients, split into two groups: G1- the healthy pregnant women, n1=100 with а registered singleton pregnancy, at gestational age 37-39+6, gave birth on the term (TB); G2- the PTB patients, n2=100, confirmed clinically by cardiotocography for gestation >32 weeks. The highly sensitive Leuko-TIC-ELISA-WBC (upper limit of the normal number of Leuc during pregnancy - 15x109 per L) test was used for investigation. Blood samples were taken by puncture of the v.cubitalis in an anticoagulant at the time of hospitalization and examined within 1 hrs. Patients with systemic chronic illness and infections were excluded from the study. The results showed statistically significant Leuc reduction in SPTB patients (14.31±2.66×109/L, p>0.03), compared to the increased (14.67±3.21×109/L) (in normal-15x109L) Leuc values in the TB group.In conclusion, the results of the Leuc value confirm the presence of a systemic leukocytosis/oxidative inflammatory reaction at the time of declaring SPTB and TB.
It is estimated that every year fifteen million premature babies are born worldwide mainly due to spontaneous preterm birth (sPTB). Furthermore, in clinical settings, there still are no reliable and accurate tools to predict preterm labor. Hence, the aim of this pioneering research was to estimate the relationship between the maternal inflammatory indicator and sPTB in a case-control study between 220 South Bulgarian women. The study was conducted at UMBAL, Stara Zagora, Bulgaria (2017-2020) and enrolled a total of 220 women, determined into two groups: 1) TB (n = 110), who were to give birth at term ≥ 37 to ≤ 39 + 6 gestation weeks with active labor at the time of hospitalization; and 2) sPTB (n = 110), women with preterm birth ≤ 32–34 + 6 gestation weeks and declared active labor, who were to give birth within 5-24 hrs. The inflammatory indicators/CRP concentration was quantified in plasma by immunoturbidimetric methods within 2 hrs. in mg/l. The median maternal CRP (8.77 ± 3.91), with cutoff = 4.9 mg/l was identified as optimal inflammation with highest risk of sPTB (sensitivity = 86.6%; specificity = 53.7%, р < 0.0001). Moreover, a cutoff CRP = 4.9 mg/l was found to be most effective in determining maternal age ≤ 19 years, the sensitivity of 68.6%, and positively correlated OR = 8.122 vs. OR = 2.354, with increased total sPTB risk at ≤ 32-34 + 6 weeks, respectively (p < 0.001). In conclusion, increased CRP concentrations and a decreased maternal age were associated with increased risks of sPTB, before ≤ 32-34 + 6 weeks. Minimal inflammation and other factors in combination may also act as sPTB prognosis.
Ochratoxin A (OTA) mycotoxin affects protein synthesis, metabolic oxidative pathways and ROS mediation. In the present study, we investigate the antioxidative-therapeutic potential of Azadirachta Indica Oil (A. indica) against chronic (28 days) OTA-induced nephrotoxicity. Balb/c male mice were exposed to i.e., (1) controls; (2) A. indica treated (120 mg/kg b. wt. i.p., given every two days); (3) c) OTA treated (Isolate D2306, 1.25 mg/kg b.wt., i.p., given every two days); d) A. indica (120 mg/kg b.wt., i.p.) administered 2h prior to OTA-administration. Till the end of the 28 experimental days of chronic OTA-nephrotoxicity, the mortality rate (±0) was not observed in mice. Kidney tissue was subjected for the determination of biochemical indexes (the MDA ratio, and antioxidant capability of SOD, and GSH) and EPR- ROS productions. OTA-exposure resulted in twofold significant increases in SOD (p<0.004), GSH (p<0.05) and malondialdehyde (MDA) levels (p<0.00), compared to controls. Further, a nephro-cells experiment was performed to investigate the oxidative stress-protective action of A. indica. Our results showed that A. indica oil and A. indica +OTA combination inhibited OTA-induced nephrotoxicity via a significant reduction of lipid peroxidation (p<0.003), ROS production (p<0.005), and endogenous antioxidant activation. Thus, it can be concluded that A. indica treatment neutralized chronic OTA-induced nephrotoxicity, not only by reducing lipid peroxidation but also by improving antioxidant status. Through the present experiments, it was demonstrated that A. indica has protective potential in nephron-inflammations.
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