Rethinking Aspergillosis in the Era of Microbiota and Mycobiota.

Aspergillosis encompasses a wide range of clinical conditions based on the interaction between Aspergillus and the host. It ranges from colonization to invasive aspergillosis. The human lung provides an entry door for Aspergillus. Aspergillus has virulence characteristics such as conidia, rapid growth at body temperature, and the production of specific proteins, carbohydrates, and secondary metabolites that allow A. fumigatus to infiltrate the lung's alveoli and cause invasive aspergillosis. Alveolar epithelial cells play an important role in both fungus clearance and immune cell recruitment via cytokine release. Although the innate immune system quickly clears conidia in immunocompetent hosts, A. fumigatus has evolved multiple virulence factors in order to escape immune response such as ROS detoxifying enzymes, the rodlet layer, DHN-melanin and toxins. Bacterial co-infections or interactions can alter the immune response, impact Aspergillus growth and virulence, enhance biofilm formation, confound diagnosis, and reduce treatment efficacy. The gut microbiome's makeup influences pulmonary immune responses generated by A. fumigatus infection and vice versa. The real-time PCR for Aspergillus DNA detection might be a particularly useful tool to diagnose pulmonary aspergillosis. Metagenomics analyses allow quick and easy detection and identification of a great variety of fungi in different clinical samples, although optimization is still required particularly for the use of NGS techniques. This review will analyze the current state of aspergillosis in light of recent discoveries in the microbiota and mycobiota.

Expression patterns of mitochondrial OXPHOS components, mitofusin 1 and dynamin-related protein 1 are associated with human embryo fragmentation.

Developmental dysfunction in embryos, such as a lethal level of fragmentation, is assumed to be mitochondrial in origin. This study investigated the molecular basis of mitochondrial impairment in embryo fragmentation. Transcription patterns of factors that determine mitochondrial functionality: (i) components of the oxidative phosphorylation (OXPHOS) - complex I, cytochrome b, complex IV and ATP synthase; (ii) mitochondrial membrane potential (MMP); (iii) mitochondrial DNA (mtDNA) content and (iv) proteins involved in mitochondrial dynamics, mitofusin 1 (Mfn1) and dynamin related protein 1 (Drp1) were examined in six-cells Day 3 non-fragmented (control), low-fragmented (LF) and high-fragmented (HF) human embryos. Gene expression of mitochondria-encoded components of complex I and IV, cytochrome b and mtDNA were increased in HF embryos compared with control and LF embryos. In LF embryos, expression of these molecules was decreased compared with control and HF embryos. Both classes of fragmented embryos had decreased MMP compared with control. LF embryos had increased gene expression of Mfn1 accompanied by decreased expression of Drp1, while HF embryos had decreased Mfn1 expression but increased Drp1 expression. The study revealed that each improper transcriptional (in)activation of mitochondria-encoded components of the OXPHOS during early in vitro embryo development is associated with a decrease in MMP and with embryo fragmentation. The results also showed the importance of mitochondrial dynamics in fragmentation, at least in the extent of this process.

Different doses of recombinant FSH and determining parameters of oxidative stress.

Background: This study aimed to examine if there is a connection between recombinant FSH dose and OS parameters in serum and the impact on IVF outcome. Methods: This study consisted of 101 participants that went through IVF procedures. Parameter that were tested are SOD, SH groups and MDA. Serum samples were drawn before stimulation and on the last day of ovarian stimulation. Results: Two groups were formed according to the dose of gonadotropins (rFSH). In both groups there were no significant differences in live-birth rate and miscarriage. In both groups mean serum MDA and SH-groups were significantly higher after ovarian stimulation, but mean serum SOD was significantly lower when compared to values before stimulation. There were less patients without OS before stimulation. Conclusions: Our results suggest that there is a difference in serum concentration in groups SOD, SH groups and MDA at the beginning and at the end ovarian stimulation. On the other hand, dose of rFSH is not related with change of parameters for oxidative stress, quality of oocytes, embryos, fertilization, pregnancies, and miscarriage rate. Patients without oxidative stress before the IVF procedure needed lower doses of gonadotropins during stimulation.

Extreme dislocation of the cervical spine-case report.

We present the case of rare extreme dislocation of subaxial cervical spine, which was challenging regarding type and time of surgery. A 22-year-old patient was injured in a traffic accident, from very beginning with signs of spinal shock. Severe traumatic C6/C7 dislocation with resulting transection of the spinal cord was diagnosed with MDCT imaging. The main dilemmas regarding the surgical treatment of this injury referred to the timing of surgery and the choice of surgical approach. We decided to perform posterior surgery at first stage. Postoperative her condition get worsening and on the 16th postoperative day came to the fatal outcome. Despite all the available protocols, in our case, the decision had to be made on the basis of individual multidisciplinary assessment, bearing in mind the mechanism of the injury and the clinical presentation of the injured patient.

Characteristics of Patients with Sarcoidosis with Emphasis on Acute vs. Chronic Forms-A Single Center Experience.

Sarcoidosis is a granulomatous disease of unknown etiology that can affect almost any organ. Although the acute form can have spontaneous regression, a certain number of patients can have a chronic form, which leads to an increase in mortality and a decrease in the quality of life. Considering that the risk factors are still unknown, we wanted to compare the characteristics of patients with acute and chronic forms of sarcoidosis in Serbia in order to determine significant differences between them with hopes of contributing to everyday clinical practice. A total of 2380 patients treated in our clinic were enrolled in this study. They were separated into the following two groups: 1126 patients with acute form and 1254 patients with chronic form. They were further compared by gender, smoking status, radiological status, exposition, biomarkers for sarcoidosis, organ involvement, and other comorbidities; the distribution of patients according to regions of Serbia was also noted. Statistical significance was found in radiological findings (p < 0.001), biomarkers (calcium in 24 h urine p < 0.001; chitotriosidase p = 0.001), and the affliction of organs (p < 0.001). The differences noted in this paper could help improve our understanding of this disease.

Prediction of adverse maternal outcomes of early severe preeclampsia.

OBJECTIVES: Assess soluble FMS-like tyrosine kinase-1/placental growth factor ratio (sFlt-1/PlGF) diagnostic accuracy for predicting adverse maternal outcome in patients with early severe preeclampsia, and whether its predictive performance is superior to full preeclampsia integrated estimate of risk score (PIERS). STUDY DESIGN: Prospective study enrolled patients with early severe preeclampsia (defined by American College of Obstetricians and Gynaecologists 2013 guidelines) admitted to the Clinic for Obstetrics and Gynaecology, Clinical Center of Serbia intensive care unit. Patients underwent delivery to terminate preeclampsia within 48 h of admission. PIERS was generated and blood samples taken at admission. Multiple pregnancies and gestational ages outside 24-34 weeks were excluded. sFlt-1 and PlGF serum concentrations were measured using Elecsys® assays and cobas e 601 analyser. Maternal complications were recorded for seven days post-delivery. MAIN OUTCOME MEASURES: Diagnostic accuracy (sensitivity and specificity), and predictive performance (receiver operating characteristic area under curve [AUC]) vs. PIERS, of sFlt-1/PlGF for predicting adverse maternal outcome. RESULTS: Of 89 patients enrolled, 61 were evaluable. Median frequency of adverse maternal outcomes within seven days of delivery was two. Median sFlt-1/PlGF and PIERS were 521·0 and 5·0%, respectively. sFlt-1/PlGF showed greater correlation with complication number than PIERS (Spearman's rho: 0·728 [p < 0·001] and 0·134 [p = 0·304], respectively). AUC for sFlt-1/PlGF and PIERS were 0·853 and 0·628, respectively. A 377·0 sFlt-1/PlGF cut-off was optimal for predicting complications (75·0% sensitivity; 92·3% specificity). CONCLUSIONS: sFlt-1/PlGF correlated more closely with number of adverse maternal outcomes than PIERS, and was a superior predictor of maternal complications.

Pregnancy-induced hypertension is associated with down-regulation of Kir6.1 in human myometrium.

It is generally accepted that activity of K+ channels maintain resting membrane potential and uterine quiescence during pregnancy, which is, at least in part, mediated by down-regulation of ATP-sensitive K+ (KATP) channels. Pregnancy-induced hypertension (PIH) is associated with pre-term and late pre-term labour. Here, we have used real time RT-PCR to compare mRNA levels of KATP channel subunits in PIH parturient and control parturient. We have found that Kir6.1, a pore forming, myometrial KATP channel subunit is down-regulated in PIH patients. This could perfectly explain increased rate of pre-term labour in patients suffering from PIH.

Toxic Metal and Trace Element Concentrations in Blood and Outcome of In Vitro Fertilization in Women.

The aim of this study was to investigate the association of trace element and toxic metal concentrations in blood and the outcome of in vitro fertilization (IVF). The study included 104 consecutive patients that underwent assisted reproductive technology (ART) procedures. The following parameters were determined: cadmium (Cd), mercury (Hg), arsenic (As), and lead (Pb); and copper (Cu), zinc (Zn), selenium (Se), and magnesium (Mg). Serum samples were obtained before commencing stimulation. Patients with smoking habit had significantly higher Pb concentrations (P = 0.022), as well as higher concentrations of As and Hg but not significantly. All subjects were divided into groups of pregnant and nonpregnant patients. Pregnant patients had lower mean values of Mg (P = 0.009), As (P < 0.05), and Pb (P = 0.034), compared to nonpregnant, and a significant correlation between pregnancy outcome and concentrations of Mg, Cd, and Pb was found. Women who had had delivered had lower Mg (P = 0.009) and Cd (P = 0.014) concentrations. There was a significant correlation of the negative outcome of IVF procedure with higher concentrations of Pb (P = 0.046) and Cd (P = 0.012). In conclusion, our results suggest that there is a difference in Mg, Pb, and Cd concentrations between pregnant and nonpregnant women. There was no association between toxic metals and number and quality of oocytes and embryos, while there was with fertilization rate. Concerning trace elements, we did not find the correlation of trace elements with oocyte number and quality, nor with a number of fertilized oocytes, except for Cu. Patients who were pregnant had lower concentrations of Mg.

Oxidative Stress Markers in GnRH Agonist and Antagonist Protocols in IVF.

BACKGROUND: Our aim was to study the effect of GnRH agonist and antagonist protocols of ovarian stimulation on oxidative stress parameters in serum and the influence of oxidative stress parameters change on the outcome of IVF cycles. METHODS: This prospective study included 82 patients who underwent IVF procedures. We determined SOD, MDA and SH groups in serum. Serum samples were obtained between the second and fourth day of the cycle and on the day of HCG administration during ovarian stimulation. RESULTS: Patients were divided into two groups depending on the protocol of stimulation. The mean total and mature oocytes number and number of fertilized oocytes were higher in GnRH agonist group. There was no significant difference in biochemical pregnancy, miscarriage and live-birth rate in both groups. Mean serum SOD was significantly lower, while mean serum MDA and SH groups were significantly higher after ovarian stimulation. Delivery rate was higher in patients without OS while miscarriage rate was higher in patients with OS. CONCLUSIONS: Our study confirmed that there is a difference in the concentration of oxidative stress parameters before and after ovarian stimulation. IVF outcome is better in patients without OS after ovarian stimulation. However, the protocol of ovarian stimulation is neither associated with a change in oxidative stress parameters nor with the outcome of ART procedures.

KATP channels are up-regulated with increasing age in human myometrium.

It is well established that ageing is associated with decrease in myometrial efficiency and higher incidence of labour complications. In myometrium, the presence of ATP-sensitive K+ (KATP) channels has been detected and they could be a factor in regulating uterine quiescence in pregnancy and contractions during labour. Here, we have examined a possibility of ageing-mediated regulation of KATP channels in the human myometrium. Myometrial samples were taken from non-pregnant women undergoing hysterectomy (n=34) and from women undergoing caesarean section in late pregnancy (n=36). Real time RT-PCR revealed that mRNAs of all known KATP channel subunits were present in the human myometrium. In non-pregnant myometrium, ageing up-regulated SUR2B/Kir6.1, subunits forming KATP channels in this tissue, without affecting the expression of other channel subunits. In the late pregnant myometrium, the level of subunits that do not form functional KATP channels was not affected by age within 20-41 age range. However, uterine SUR2B and Kir6.1 were up-regulated in parturient over 35 years. An ageing-induced increase in those channel subunits was confirmed by Western blotting. Thus, this study suggests that KATP channels are up-regulated with increasing age in human myometrium. This may help explain, at least partially, increased rate of birth complications in women aged over 35 years.

Pregnancy in patient with Swyer syndrome.

OBJECTIVE: To report a case of successful pregnancy and delivery after IVF and ET in a patient with Swyer syndrome. DESIGN: Case report. SETTING: Unit of Assisted Reproduction, Gynecology and Obstetrics Institute, University of Belgrade. PATIENT(S): A 30-year-old patient with 46,XY gonadal dysgenesis. INTERVENTION(S): Chromosomal analysis, diagnostic laparoscopy, IVF using donor oocytes, ET, and cesarean delivery. MAIN OUTCOME MEASURE(S): Successful pregnancy and live birth. RESULT(S): Successful treatment, pregnancy, and delivery. CONCLUSION(S): A patient with 46,XY gonadal dysgenesis in a donor oocyte program, can maintain a normal pregnancy and delivery.

[Medicamentous abortion with mifepristone and misoprostol in Serbia and Montenegro].

BACKGROUND/AIM: Medicamentous abortion was first introduced in Serbia and Montenegro in September 2001. The aim of this study was to assess the efficiency, side effects, and acceptability of medicamentous abortion using mifeprostone orally (600 mg), and 48 hours later, misoprostol both orally and vaginally in different regiments in our population (400 mcg, 600 mcg, 800 mcg). METHODS: A total of 235 consecutive women with pregnancies up to 49 days of gestational age were assigned to 4 groups according to the different misoprostol regiment (group I 400 mcg, group II 600 mcg, group III 800 mcg orally, and group IV 800 mcg both orally and vaginally). The principal outcome measure was a successful abortion defined as a complete expulsion of intrauterine contents without a need for surgical intervention 14 days after the procedure. Other outcome measures were the following: drug related effects, and adverse effects related to the abortion process. RESULTS: In general, the success rate was 50%, 89.48%, 75% and 92.11% in the groups I, II, III, and IV, respectively, as judged by the complete expulsion of the intrauterine contents without surgical intervention (t1:4 = 7.005; t2:4 = 0.3872, t3:4 = 2.9784, p < 0.01). The incidence of adverse effects (vomiting, abdominal pain, bleeding, and fever) was low in general, but among our groups it occurred mostly with the higher doses of orally applied misoprostol (800 and 600 mcg). Only one case required urgent curettage for heavy vaginal bleeding, and two blood transfusions, as well. No cases of intact pregnancies were recorded in the study. CONCLUSION: Our study showed that a mifepristone dose of 600 mg orally, and misoprostol 400 mcg orally and 400 mcg vaginally were most effective. Thus, a combination of mifepristone and misoprostol for medicamentous abortion should take a higher proportion in the termination of early pregnancy in our population.

[Effects of tibolone on the risk factors for cardiovascular diseases in postmenopausal women].

BACKGROUND/AIM: Tibolone is a preparation that belongs to the group of steroidal substances. The effects of the use of the use of tibolone are the consequence of the activities of its metabolities, considering that their hormonal activity depends on the type of tissue in which they develop. The aim of this study was to evaluate the influence of the use of tibolone on risk factors for the development of cardiovascular diseases in postmenopausal women. METHODS: A prospective observational stady included 94 patients who had the concentration of 17beta estradiol < 50 pg/ml and who was in menopause more than a year. Out of the total number of patients, 63 accepted to receive tibolone 2.5 mg daily (tibolone group), while 31 of the patients refused to take tibolone (control group). We measured the concentration of lipids (cholesterol, LDL cholesterol, HDL cholesterol, triglicerides), antitrombin III, fibrinogen, and C-reactive protein, before and after the treatment within a 6-month period. Then, we compared the difference between the values of concentrations and tested the statistical significance of the difference. We also evaluated the changes of values in the concentrations of the examinated parametars inside a 6-month period in the control group. RESULTS: In 31 patients of the control group, from the control group there were no significant changes in the values of the defined parametars as compared to their initial values after six months. But there were changes of statistical significance (p < 0.001) in values of the concentrations of the exeminated parametars before and after the treatment in the tibolone group. In fact, we recorded decreases in the total cholesterol by 17.8%, HDL cholesterol by 27%, LDL cholesterol by 4% (without statistical significance p > 0.05) and triglicerids by 35%. There were no statistical differences in the concentrations of antitrombin III, fibrionogen, and C-reactive proteine in the tibolone group before and after the treatment CONCLUSION: The use of tibolone dose decrease the concentration of the total cholesterol, triglicerides, HDL cholesterol, without a significant decrease of LDL cholesterol. Also, the use of tibolone does not have any significant effect on the concentrations of antitrombin III, fibrinogen and C-reactive proteine. The number of serum parameters measured in this study was limited, thus that was the reason to discuss only about the metabolism of lipids in the patients from the tibolone group. The final condusion about the risk for cardiovascular diseases in the patients on tibolone, howerer, reqnires were extensive further clinical exeminations.