New evidence on oral L. plantarum P17630 product in women with history of recurrent vulvovaginal candidiasis (RVVC): a randomized double-blind placebo-controlled study.

OBJECTIVE: To evaluate activity and tolerability of an oral product containing the specific probiotic strain Lactobacillus plantarum P17630 on vaginal colonization of lactic acid bacteria. PATIENTS AND METHODS: L. plantarum P17630 (5x109 CFU/capsule) or placebo were formulated in capsules and administrated orally to a total of 93 women with a history of recurrent vulvovaginal candidiasis over a period of 3 treatment cycles (15 days/cycle) separated by 15-day wash-out intervals. Vulvovaginal examinations and vaginal mycological culture, including Nugent score and Lactobacillary grade (LBG), were assessed at baseline, at day 45 and at day 90 (end of the study). RESULTS: Probiotic intake significantly improved lactobacilli colonization on vaginal epithelial cells. The results demonstrated a significant difference in LBG score when comparing day 0 to day 45 (p=0.000016) and to day 90 (p=0.001415) in women treated with L. plantarum P17630. No evidence of improvement in LBG score was recorded in women who received placebo (p<0.005). The increased lactobacilli adhesion was associated with the improvement of clinical signs such as redness, swelling, and discharge. CONCLUSIONS: This study demonstrates that the administration of oral probiotic Lactobacillus plantarum P17630 improves vagina colonization of acid lactic bacteria (an increase of vaginal LBG score) and suggests the use of this oral product to successfully prevent episodes of vulvovaginal candidiasis. The clinical study described in this article is registered in the ISRCTN registry with trial registration ID ISRCTN12669692.

Histological changes in the vulva and vagina from ovariectomised rats undergoing oestrogen treatment.

BACKGROUND: The purpose of this study was to assess the histological changes occurring in the vagina and vulva in ovariectomised female rats, as well as the response to the administration of injectable oestrogens. MATERIAL AND METHODS: We used 30 female Wistar white rats, distributed as follows: group 1 - the control group, group 2 - the operated but untreated rats, and groups 3, 4 and 5 - operated rats, to which oestrogenic treatment was administered (Estradiol, Estradurin, Sintofolin) at a dosage of 0.2 mg/rat/day. After 14 days of treatment, all animals were sacrificed and vaginal and vulvar biopsies were taken from all groups. RESULTS: In group 2, we encountered structural changes of the vaginal mucosa, with severe atrophy and alterations in the thickness of the vagina and vulva. In groups 3, 4 and 5 we found marked hyperplasia of the vaginal and vulvar epithelium, eosinophilic and mast cell infiltration in the chorion. CONCLUSIONS: Our study proves that the histopathological changes during anoestrus after administration of oestrogens are cell hyperplasia, thickening of the superficial mucosal layer, eosinophilic and mast cells infiltrations, and chorionic congestion. Furthermore, we demonstrated that Estradiol therapy induces the most evident histological changes when compared to synthetic oestrogens such as Estradurin or Sintofolin.

VASCULAR ENDOTHELIAL GROWTH FACTOR SERUM LEVELS IN WOMEN WITH ADVANCED ENDOMETRIOSIS.

CONTEXT: Endometriosis is a common gynecological disease, characterized by ectopic deposits of endometrial tissue outside of the uterine cavity, and it is associated with pelvic pain and infertility, with an important impact on the quality of life. At this point there is a controversy regarding the etiology and pathophysiology of endometriosis and it seems that pro-angiogenic growth factors might be involved, but their role is not completely understood. OBJECTIVE: To evaluate the serum concentration of the main growth factors in patients with diagnosed endometriosis compared to healthy controls. SUBJECTS AND METHODS: A total of 157 women were divided into two study groups (Group I - endometriosis; Group 2 - healthy women). Serum levels of VEGF, G-CSF, GM-CSF, b-FGF, EGF, and HGF were measured with Human Multiplex Cytokine Panels. RESULTS: VEGF serum levels were significantly lower in women with endometriosis compared to controls (1.924±0.145 compared to 1.806±0.078 pg/mL, p<0.001). Serum levels of GM-CSF, b-FGF, EGF, and HGF respectively did not differ significantly between patients with endometriosis and healthy controls. G-CSF had a very low detection rate. CONCLUSIONS: The present study showed that VEGF serum levels are significantly lower in endometriosis patients compared to healthy controls, indicating a possible role in endometriosis pathogenesis.

Degenerative alterations of the cementum-periodontal ligament complex and early tooth loss in a young patient with periodontal disease.

Premature exfoliation of primary or permanent teeth in children or adolescents is extremely rare and it can be a manifestation of an underlying systemic disease. This study aims to present the histological aspects associated with early tooth loss in a case of periodontal disease developed without local inflammation and with minimal periodontal pockets and attachment loss. The maxillary left second premolar was extracted together with a gingival collar attached to the root surface. The histological analysis recorded the resorption of the cementum in multiple areas of the entire root surface with the connective tissue of the desmodontium invading the lacunae defects. The connective tissue rich in cells occupied the periodontal ligamentar space and the resorptive areas. No inflammation was obvious in the periodontal ligament connective tissue. This report may warn clinicians about the possibility of the association of cemental abnormalities with early tooth loss.

[Amniotic membrane in conjunctivoplasty]. / Membrana amniotica în conjunctivoplastie.

The basic principles of conjunctival reconstruction are similar in all pathologies that inflict conjunctival tissue loss. Large conjunctival defects are difficult to treat, with little conjunctival reserve that can be used to close the defect. The study had the objective to find alternatives to conjunctival autograft. Frozen amniotic membrane was investigated. From 27 cases, 22 cases were reconstructed using amniotic membrane alone, but in 5 cases conjunctival autograft was also needed. Bulbar conjunctiva was reconstructed in 23 cases, of which 19 with pterygium, 3 with conjunctival tumors, one operatory plague. Fornix reconstruction was carried out in 4 cases, one tumor and 3 symblepharons (2 after conjunctival burns and one pemfigus). The rate of pterygium recurrence was 7,14%. Amniotic membrane can heal large defects of the conjunctiva, either bulbar or forniceal.

Pancreatic exocrine adult cells and placental stem cells co-culture. Working together is always the best way to go.

BACKGROUND AND AIM: The progress made in the last few years have managed to come up withy the possibility of using different stem cell types in an endeavor to correct the alterations that appear in different degenerative diseases. The pancreas, an organ with extremely low regenerative capacity, both for the endocrine and for the exocrine component, is an organ perfect for cell therapy in the hope of restoring its function and cure diabetes mellitus or chronic pancreatitis. One main issue in the stem cell transplantation problem is represented by the influence of the cellular niche, formed by completely differentiated cells, on the phenotype and function of the transplanted cells. In this study, we challenge current knowledge in the field by evaluating the influence of exocrine pancreatic cells on placental stem-like cells using the co-culture technique. METHODS: In our experiments, we used two different protocols in which adult pancreatic cells were cultured together with mesenchymal stem cells isolated from human placenta. In the case of the first protocol, we seeded pancreatic cells on a pre-adhered single-cell layer of mesenchymal stem cells and in the second one, the seeding of two cell populations in suspension was done at the same time, after passage. During the experiment, we evaluated the alteration of the morphology of the placental cells using and inverted phase microscope and reverse transcriptase-PCR. RESULTS AND CONCLUSIONS: Based on morphology, in both cases the interaction between epithelial pancreatic cells and placental ones have determined a change in phenotype from mesenchymal to epithelial-like. Taking into consideration the gene expression, placental stem cells have maintained pluripotency gene expression throughout the study. They also expressed pancreatic amylase. These experiments bring out the plasticity of placental stem cells, the cell microenvironment with a decisive part in phenotype and the level of gene expression. The results obtained in vitro can bring a new picture on the effects of the pancreatic stem cell niche.

Aspects of placental morphogenesis and angiogenesis.

Placental morphology and vascularization are important stages in the evolution of pregnancies. Placental morphogenesis and angiogenesis processes are studied by two-dimensional, three-dimensional and Doppler ultrasound. Ultrasound methods provide important data on the physiology and pathophysiology of fetal-placental exchange. The macroscopic and microscopic study of the placenta brings valuable information on the possible structural changes and implicitly allows assessing fetal-placental circulation. The ultrasound and microscopic evaluation of the placenta are complementary means of examination for the assessment of fetal-maternal exchange. These methods of investigation can be applied in the context of a strict knowledge of placental morphogenesis and angiogenesis.

Isolation and characterization of mesenchymal stem cells from the amniotic membrane.

The aim of the study was the isolation and the genotypic and phenotypic characterization of mesenchymal stem cells from the amniotic membrane. The placentas included in the study were derived from pregnancies with a normal evolution. Along with the placentas, umbilical cord blood and maternal peripheral blood samples were taken. The isolation and the culture of cells from the amniotic membrane was followed by the determination of the markers of these cells. The cells expressed markers characteristic of stem cells. Immunofluorescence and evaluation of the gene expression evidenced the pluripotential properties of these cells. HLA expression provides information that might help explain the immunological mechanisms of tolerance between the maternal organism and fetal structures.

[Ocular changes in preeclampsia]. / Modificari oculare induse de preeclampsie.

The pregnancy is associated with modifications involving multiple systems and organs, including the eye. The retinal vascular changes are present in 30-100% of the PE cases. The most frequent ocular modification is the vasoconstriction of the retinal arterioles. The exudative retinal detachment is a rare cause (under 1%) of visual loss in the PE-E syndrome, being produced by the involvement of the choroidal vascularization. Most cases of blindness during pregnancy have been determined by cortical, occipital anomalies, the used terminology in these circumstances being cortical blindness. The pathogenesis of the Purtscher retinopathy is the embolism of the retinal arterioles, by leucocytic aggregation, as a response to the activation of the complement. The evaluation of the ophthalmic arterial flow by eco Doppler might offer new perspectives regarding the understanding of the physiopathology, the diagnosis and the quantification of the PE severity. The decrease of the IP values and the increase of the median velocity in the ophthalmic artery in pregnant women with PE, suggest a hyperperfusion process and the decrease of the vascular resistances at the level of the orbital vessels, but also in the CNS. In the severe forms of PE, the increase of the impedance of the orbital vessels has been noted. The described vascular changes cannot disappear completely 6 weeks after birth; a residual arteriolar constriction might persist, as a permanent stigma of the PE. The evaluation, monitoring, therapeutical approach of the patients with PE must be performed by a complete team: obstetrician, ophthalmologist, neurologist, radiologist, in order to decrease the materno-fetal risks and improve the prognosis of PE.

Isolation and characterization of stem cells from the placenta and the umbilical cord.

In addition to its essential role in the development, nutrition and immunological tolerance of the product of conception, human placenta is an important source of stem cells. Over the past years, scientific research has been aimed at isolating and characterizing mesenchymal cells and amniocytes, which show a high plasticity and are found in the chorionic villi and the membranes. At the level of the umbilical cord, two types of stem cells can be found: hematopoietic and mesenchymal. The blood of the umbilical cord is already in the focus of attention of researchers, as an important source of hematopoietic stem cells that can be used for transplantation.

Infertility in a new 46, XX male with positive SRY confirmed by fluorescence in situ hybridization: a case report.

The 46, XX male syndrome (de la Chapelle syndrome or 46, XX testicular disorder of sex development) is a rare form of sex reversal with complex mechanisms leading to a large spectrum of clinical manifestations ranging from ambiguous genitalia in the newborn to normal male phenotype. Therefore, diagnosis is established either pre- or early postnatal, or in adult life due to male infertility. In some cases, subtle clinical signs during childhood and puberty may be overlooked. A 28-year-old married man presented with azoospermia without erectile dysfunction. Between 9-14 years he was examined for the small testes and under-masculinized external genitalia but the diagnosis was not further clarified. At presentation, hormonal laboratory evaluation revealed hypergonadotropic hypogonadism. Chromosome analysis showed a 46, XX karyotype and translocation of SRY (testis-determining factor) from chromosome Y to chromosome X was identified by fluorescence in situ hybridization (FISH). Despite early subtle clinical signs of abnormal sexual development in this new 46, XX male syndrome, medical investigations were triggered by infertility.

New method for determination of dydrogesterone in human plasma for therapeutic drug monitoring in gynecological disorders.

UNLABELLED: A new simple, sensitive and selective liquid chromatography coupled with mass spectrometry (LC/MS) method for quantification of dydrogesterone in human plasma was validated. MATERIAL AND METHOD: The analytes was eluted in 1.3 minutes on a reversed phase column (Zorbax SB-C18, 100 mm x 3.0 mm I.D., 3.5 microm) under isocratic conditions using a mobile phase of a 20 : 80 (v/v) mixture of ammonium acetate 1 mM and acetonitrile. The flow rate was 1 mL/min at the column temperature of 35 degrees C. The detection of the analyte was in MS/ MS mode using an atmospheric pressure chemical ionization source (APCI+, m/z 313 > m/z 295). The sample preparation was very simple and rapid and consisted in plasma protein precipitation from 0.2 mL plasma using 0.6 mL methanol. RESULTS: Calibration curves were generated over the range of 5-150 ng/mL with values for coefficient of determination greater than 0.997 and by using a weighted (1/y) linear regression. The values of precision and accuracy were less than 12.5% and 7.5%, respectively, both for within- and between-run analysis. The mean recovery of the analyte was 99.8%. This is the first reported method for analysis dydrogesterone in human plasma that uses protein precipitation as sample processing procedure. The validated LC/MS method could be applied for determination of dydrogesterone in human plasma for therapeutic drug monitoring in gynecological disorders.

C-reactive protein, marker for evaluation of systemic inflammatory response in preeclampsia.

AIM: Determination by a high sensitivity technique of serum C-reactive protein (CRP), a sensitive marker of inflammation in women with preeclampsia compared to normal pregnancy and investigation of the relationship between CRP and the severity of the preeclamptic syndrome. MATERIAL AND METHOD: The study included 40 women with preeclampsia and 40 control subjects with normal pregnancies in the last trimester of pregnancy. The serum CRP concentration was determined using the universal high sensitivity immunoturbidimetric assay. RESULTS: The serum CRP concentration was significantly higher (p < 0.001) in preclampsia (5.69 +/- 1.8 mg/L) compared to normal pregnancy (2.89 +/- 1.2 mg/L). In women with preeclampsia, CRP correlated positively and significantly with diastolic blood pressure, proteinuria and uric acid levels. Maternal CRP values also correlated negatively and significantly with fetal weight at birth. CONCLUSIONS: Our results demonstrate that serum CRP is increased in preeclampsia and represents a marker of the severity of the preeclamptic syndrome and of fetal weight at birth. Taking into consideration these observations and the fact that CRP testing is rapid and relatively inexpensive, we recommend the use of this acute phase reagent in clinical practice, in all women with preeclampsia in order to establish the prognosis of the disease.