Using Front-Face Fluorescence Spectroscopy and Biochemical Analysis of Honey to Assess a Marker for the Level of Varroa destructor Infestation of Honey Bee (Apis mellifera) Colonies

Varroa destructor is a parasitic mite responsible for the loss of honey bee (Apis mellifera) colonies. This study aimed to find a promising marker in honey for the bee colony infestation level using fluorescence spectroscopy and biochemical analyses. We examined whether the parameters of the honey samples’ fluorescence spectra and biochemical parameters, both related to proteins and phenolics, may be connected with the level of honey bee colonies’ infestation. The infestation level was highly positively correlated with the catalase activity in honey (r = 0.936). Additionally, the infestation level was positively correlated with the phenolic spectral component (r = 0.656), which was tentatively related to the phenolics in honey. No correlation was found between the diastase activity in honey and the colonies’ infestation level. The results indicate that the catalase activity in honey and the PFC1 spectral component may be reliable markers for the V. destructor infestation level of the colonies. The obtained data may be related to the honey yield obtained from the apiaries

Juvenile type granulosa cell tumor

© 2018, University of Kragujevac, Faculty of Science. All rights reserved. Granulosa cell tumor is a type of neoplasm, which represents 2-5% of all ovarian cancers. About 5% of these tumors are juvenile- type and usually occur to girls before puberty and to women younger than thirty years of age. There are signs premature puberty or premature emergence of secondary sexual characteristics with irregular vaginal bleeding that occur to these kind of patients. To the rare cases, like this, the occurrence of granulosa cell tumors can cause the appearance of hyperandrogenism with high levels of plasma testosterone, leading to virilization which happened to this female patient. We will present the female patient who was 35 years old and which was originally hospitalized to the Clinic for Haematology Clinical Center Kragujevac, because of extreme fatigue accompanied by dizziness. During diagnostics the patient underwent to the complete gynecological examination. After gynecological examinations and necessary diagnostic procedures, it was decided continuing the treatment at the Clinic of Gynecology and Obstetrics Clinical Center Kragujevac, where she underwent a total abdominal hysterectomy with bilateral salpingo- oophorectomy for suspected uterine neoplasm. Histopathological analysis of the obtained material confi rmed the presence of follicular cysts of both ovaries and juvenile type granulosa cell tumor on the right ovary; the uterus was enlarged with multiple fi broid tumors. Granulosa cell tumor should be suspected in the cases of girls and young females if there is present an ovarian cyst paired with signs of preterm puberty or hyperestrogenism. In this case, the presence of granulosa cell tumor was masked by signs of hyperandrogenism, which is not so typical, as well as the presence of uterine fi broids who have actually been the main cause for surgical treatment

Impact of In Vitro Long-Term Low-Level DEHP Exposure on Gene Expression Profile in Human Granulosa Cells

Here, we applied a model of long-term exposure of human granulosa cells to low environmentally relevant levels of di(2-ethylhexyl) phthalate (DEHP). This approach provides more relevant data regarding the impact of DEHP on the function of human granulosa cells. The immortalized human granulosa cells HGrC1 were exposed to 50 nM and 250 nM DEHP for four weeks. The cells were collected every week to analyze the basal granulosa cells’ functions. A portion of the DEHP-exposed cells was stimulated with forskolin (FOR) for 48 h. Steroidogenesis was investigated using ELISA, whereas DNBQ sequencing and RT-qPCR were used to analyze gene expression. The results show that steroidogenesis was not affected by DEHP exposure. RNAsequencing shows that DEHP caused week- and concentration-specific changes in various genes and functions in HGrC1. Sulfotransferase family 1A member 3 (SULT1A3) and 4 (SULT1A4), which are involved in catecholamine metabolism, were the most prominent genes affected by DEHP under both the basal and FOR-stimulated conditions in all four weeks of exposure. This study showed, for the first time, that SULT1A3 and SULT1A4 are expressed in human granulosa cells, are regulated by FOR, and are affected by low-level DEHP exposure. These data provide new insight into the relationship between DEHP, SULT1A3, and SULT1A4 in human granulosa cells

DEHP Decreases Steroidogenesis through the cAMP and ERK1/2 Signaling Pathways in FSH-Stimulated Human Granulosa Cells

DEHP is an endocrine disruptor that interferes with the function of the female reproductive system. Several studies suggested that DEHP affects steroidogenesis in human and rodent granulosa cells (GC). Some studies have shown that DEHP can also affect the FSH-stimulated steroidogenesis in GC; however, the mechanism by which DEHP affects hormone-challenged steroidogenesis in human GC is not understood. Here, we analyzed the mechanism by which DEHP affects steroidogenesis in the primary culture of human cumulus granulosa cells (hCGC) stimulated with FSH. Cells were exposed to DEHP and FSH for 48 h, and steroidogenesis and the activation of cAMP and ERK1/2 were analyzed. The results show that DEHP decreases FSH-stimulated STAR and CYP19A1 expression, which is accompanied by a decrease in progesterone and estradiol production. DEHP lowers cAMP production and CREB phosphorylation in FSH but not cholera toxin- and forskolin-challenged hCGC. DEHP was not able to decrease steroidogenesis in cholera toxin- and forskolin-stimulated hCGC. Furthermore, DEHP decreases FSH-induced ERK1/2 phosphorylation. The addition of EGF rescued ERK1/2 phosphorylation in FSH- and DEHP-treated hCGC and prevented a decrease in steroidogenesis in the FSH- and DEHP-treated hCGC. These results suggest that DEHP inhibits the cAMP and ERK1/2 signaling pathways, leading to the inhibition of steroidogenesis in the FSH-stimulated hCGC