Simultaneous Determination of Two Tyrosine Kinase Inhibitors in Tablets by HPLC-MS analysis.

The class of tyrosine kinase inhibitors (TKIs) is represented by a group of compounds which are currently used in the treatment of different types of cancer. These oral medicines present a narrow therapeutic index and a large inter-and intra-individual variability. Within this work, a simple, accurate and rapid reversed phase ultra-high-performance liquid chromatographic (RP-UHPLC) method with mass spectrometric (MS) detection for simultaneous analysis of two TKIs, ibrutinib and ruxolitinib, using pentoxifylline as internal standard (IS) in tablet dosage forms is presented. The separation was carried out on a Waters (Milford, Massachusetts, USA) Arc System coupled with a Waters QDa mass detector. The column used was a Waters CORTECS C18 (4.6×50mm, 2.7µm); a gradient elution was carried out using a mixture of ammonium formate 10 mM aqueous solution and acetonitrile. The flow rate of the mobile phase was set to 0.5mL/min. The column temperature was equilibrated to 40°C. The injected volume was 5µL. All samples were kept at 20°C during the entire analysis. Mass spectra were recorded in positive ionization mode in the range of m/z 100-400 for ruxolitinib and m/z 100-500 for ibrutinib. Quantification was established in single ion recording (SIR) mode for each compound, using pentoxifylline as internal standard. The method was validated according to International Guidelines in terms of stability, limit of detection, limit of quantitation, linearity, precision and accuracy. The validated method can be successfully applied for simultaneous determination of TKIs in tablet dosage forms.

The Ability of Doppler Uterine Artery Ultrasound to Predict Premature Birth.

OBJECTIVE: Demonstration of the predictive capacity of Doppler Uterine Artery (UtA) on preterm birth (PB) by serial measurement at various ages of pregnancy. METHODS: The prospective study included a group of 116 pregnant women, of whom 85 gave birth prematurely and 31 pregnant women gave birth at term, constituting the control group. UtA Doppler measurement was performed by the abdominal approach. Quantitative wave evaluations were performed by the pulsatility index (PI), the systole/diastole ratio (S/D), as well as the qualitative analysis of the flow rate waveform (notch). UtA Doppler evaluation was performed in 3 pregnancy periods: 18.0-22.6 weeks, 28.0-31.6 weeks, and 32.0-35.6 weeks. RESULTS: Only at the third examination, at 32.0-35.5 weeks of gestation, was there a statistically significant difference between the S/D-UtA ratio and PI-UtA correlated with the risk of premature birth (p<0.05). Although there was an association between UtA Doppler and late preterm birth, the predictive ability was low. Also, UtA Doppler was not statistically significant for preterm birth before 32 weeks of gestation. CONCLUSIONS: Although we did not find a statistical association between second-trimester UtA Doppler and preterm birth, we do suggest a closer look at women with abnormal UtA Doppler in the second trimester. We believe that, according to the results obtained, UtA Doppler can predict especially iatrogenic premature birth depending on the prediction of the most severe complications, severe preeclampsia, and SGA.

Fetal-maternal incompatibility in the Rh system. Rh isoimmunization associated with hereditary spherocytosis: case presentation and review of the literature.

Next to A and B antigens, agglutinogen D exhibits the highest immunogenicity. Following the transfusion of D-positive red blood cells (RBCs), almost 80% of D-negative recipients develop anti-D antibodies (Abs). Subsequently, anti-D immunization further promotes the synthesis of Abs towards other blood group antigens in or outside the Rh system. The D antigen is also involved in 95% of cases of hemolytic disease of the newborn. Transfusions, hemotherapy, grafts, and obstetric history (abortions, ectopic pregnancy, births) are all risk factors for Rh isoimmunization. In the case of ABO compatibility between mother and fetus, Rh-positive fetal RBCs that have reached the maternal bloodstream are not destroyed by group agglutinins, and Rh antigenic sites are not hidden by the maternal immune system. But a Rh-negative mother with a homozygous Rh-positive husband will certainly have a Rh-positive fetus. As it has an irreversible evolution, the Rh isoimmunization once installed cannot be influenced in the sense of decreasing the Ab titer, therefore, injectable globulin has no effect. A particular case was that of a newborn with Rh system incompatibility associated with hereditary spherocytosis The clinical balance at birth reflects the severe jaundice of the female newborn of 3140 g, gestational age 38∕39 weeks, extracted by lower-segment transverse Caesarean section, with a double loop nuchal cord, Apgar score 8. Because the jaundice was severe and atypical (face and upper chest), we considered the possibility of coexistence of hemolytic disease of the newborn by Rh blood group incompatibility associated with hereditary spherocytosis, as it turned out to be true and mentioned. Changes in genes encoding proteins in the structure of the RBC membrane have amplified hemolysis induced by maternal-fetal isoimmunization in the Rh system. Massive hemolysis accentuated by congenital spherocytosis, confirmed later, imposed blood transfusion and dynamic monitoring.

Prediction of Preterm Birth with Serial Measurements of Ultrasound Markers.

To compare the ability of cervical length (CL), anterior cervical angle (ACA), and cervical consistency index (CCI) to predict premature birth. METHODS: This prospective study involved 85 pregnant women who gave birth prematurely and a control group of 31 pregnant women who gave birth at term. The study was performed in the Obstetrics and Gynecology Clinic of the Municipal Clinical Hospital Filanthropia Craiova between January 1, 2019, and January 1, 2022. Cases were examined using transvaginal ultrasonography (TVU) in the second and third trimesters of pregnancy, and cervical length (CL), Anterior Cervical Angle (ACA), and Cervical Consistency Index (CCI) were measured. RESULTS: The mean value from the three measurements at all three parameters was statistically significant with preterm birth (p<0.05). Cervical length <25mm, was highly significant in the prediction of preterm labor with a sensitivity of 99%, specificity of 61%, positive predictive value (PPV) of 78%, negative predictive value (NPV) of 97 %, and a positive likelihood ratio (LR+) of 2.54 and negative likelihood ratio (LR-) of 0.02. CCI also remains, despite low specificity and PPV values, a potential predictive parameter in the prediction of preterm birth, with a sensitivity of 73%, NPV of 92% and a LR+of 1.32 and LR- of 0.6 also correlated with CL, CCI being more difficult to interpret as an independent predictive parameter. CONCLUSIONS: CL remains the standard parameter for predicting the preterm birth, but in combination with other parameters, the prediction rate can increase significantly.

The Analysis of Blood Inflammation Markers as Prognostic Factors in Parkinson's Disease.

Parkinson's disease is a chronic, progressive, and neurodegenerative disease, and yet with an imprecise etiopathogenesis. Although neuroinflammation was initially thought to be a secondary condition, it is now believed that microglia-induced inflammation could also contribute to the degeneration of the nigrostriatal pathway. Here, we aimed to establish the feasibility of basic inflammatory biomarkers as prognostic factors in PD. The study was based on retrospective analyses of blood samples taken from patients diagnosed with PD, as well as from healthy subjects. Complete medical records, total leukocyte count with subpopulations, and erythrocyte sedimentation rate (ESR) were analyzed. We calculated the serum neutrophils-to-lymphocytes ratio (NLR) and platelet-to lymphocytes ratio (PLR), and also compared the laboratory data between the PD group and the control group. Only PLR and NLR showed statistically significant differences (p < 0.001 and 0.04, respectively). In our study, ESR did not show statistically significant correlations with motor score or with disability. In our research, ESR was correlated with the disease duration (p = 0.04), and PLR showed a significant correlation with disease stage (p = 0.027) and disease duration (p = 0.001), but not with motor state. These biomarkers could prove to be effective tools for a primary evaluation of inflammation in PD, but further tests are required to properly investigate the neuroinflammatory status of these patients.

Impact of COVID-19 on fertility and assisted reproductive technology (ART): a systematic review.

The appearance of coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been a major obstacle for the performing of current medical activities throughout the world. COVID-19 has affected humanity in many ways, thus causing a great medical, social, economic, and political instability. The aim of this study was to make an analysis of the scientific data obtained by so far to highlight the impact that COVID-19 has had on fertility and assisted reproductive technology (ART). Infection with SARS-CoV-2 alters the normal immune response by local and systemic damage to tissues and organs. After the virus enters the body, the first lesions are produced in the respiratory tract. Extrapulmonary lesions specific to COVID-19 include acute renal lesions/acute kidney damage, hepatocellular lesions, neurological diseases, myocardial dysfunction and arrhythmia, gastrointestinal diseases but also genital impairment. The possible impairment of the male reproductive system is because angiotensin-converting enzyme 2 (ACE2) receptors are in an increased number in the testes, seminiferous duct cells, spermatogonia, Leydig cells and Sertoli cells. Many published studies to date have pointed out that COVID-19 could also affect female fertility and disrupt the functions of the female reproductive system. The theory that this virus can also be transmitted sexually and can cause infertility or testicular damage is supported by the fact that the virus can be isolated in the semen of COVID-19 patients but only during the disease. Choosing the best method of treating infertility during the COVID-19 pandemic is multifactorial, but the risk of infection and compliance with specific ART hygiene protocols must always be considered. Currently, there is no scientific basis regarding the fact that the COVID-19 vaccination would influence fertility.

Clinical and morphopathological assay in vulvovaginal candidiasis.

Candida vulvovaginitis is characterized by the appearance of inflammatory changes in the vaginal and vulvar epithelium secondary to infection with Candida species. The purpose of this study was to analyze and compare the clinical, microbiological, and histopathological aspects of pregnant and non-pregnant patients, symptomatic or asymptomatic in the case of candida vaginitis and to correlate the microscopic aspects with the symptoms before applying the local treatment with Nystatin. The study presents a retrospective analysis of the management of vaginitis in 166 pregnant or non-pregnant patients during 2021-2022. We observed the structure of the Malpighian squamous epithelium without keratinization present on the vaginal mucosa and the structure of the subepithelial connective tissue, which shows increased numerical values of inflammatory and vascular cellularity in the case of candida vaginitis symptomatic compared to asymptomatic ones. We noticed also in the microscopic study that in cases of asymptomatic patients before treatment, the number of inflammatory cells and blood vessels situated immediately under the epithelium was significantly lower compared to their number in symptomatic patients before treatment. Analyzing the results obtained after the administration of the treatment proposed by us, we can say that local Nystatin treatment is beneficial and safe for pregnant and non-pregnant patients and is a good alternative for patients with recurrent vulvovaginal candidiasis.

Analysis of the relationship between placental histopathological aspects of preterm and term birth.

OBJECTIVES: This study aims to establish a correlation between placental histopathological and immunohistochemical (IHC) changes and preterm birth with fetal growth restriction (FGR, formerly called intrauterine growth restriction - IUGR). PATIENTS, MATERIALS, AND METHODS: This prospective study was performed on a group of 30 parturients, with singleton gestation, of which 15 patients gave birth at term, and the other 15 patients gave birth prematurely. After the statistical correlation of the clinical and demographic data with premature birth (PB) and term birth (TB), we performed histological and IHC research on the respective placentae. To observe normal and pathological microscopic placental structures, we used the Hematoxylin-Eosin (HE) and Periodic Acid Schiff-Hematoxylin (PAS-H) classical stainings, but also special immunostaining with anti-cluster of differentiation 34 (CD34) and anti-vascular endothelial growth factor (VEGF) antibodies. RESULTS: We found a statistically significant difference between the TB∕PB categories and the age of the patients, their antepartum weight, the weight of the newborns, and the placenta according to the sex of the newborn. Histological analysis revealed in the case of TB, small areas of perivillous amyloid deposition, with the significant extension of these areas both intravillous and perivillous in the case of PB. Massive intravillous calcifications, syncytial knots, and intravillous vascular thrombosis were also frequently present in PB. With PAS-H staining were highlighted the intra∕extravillous vascular basement membranes, but especially the massive fibrin deposits rich in glycosaminoglycans. By the IHC technique with the anti-CD34 antibody, we noticed the numerical vascular density, higher in the case of TB, but in the case of PB, there were large areas of placental infarction, with a lack of immunostaining in these areas. Through the anti-VEGF antibody, we observed the presence of signal proteins that determined and stimulated the formation of neoformation vessels in the areas affected by the lack of post-infarction placental vascularization. We observed a highly significant difference between placental vascular density between TB∕PB and newborn weight, sex, or placental weight. CONCLUSIONS: Any direct proportional link between the clinical maternal-fetal and histological elements yet studied must be considered. Thus, establishing an antepartum risk group can prevent a poor pregnancy outcome.

The procoagulant status. Hypercoagulability as a risk factor of primary and secondary infertility.

Thrombophilia is a disorder that makes patients susceptible to intravascular thrombosis that may increase the risk of developing a pregnancy on a known pathology. The female patient diagnosed with hypoplastic uterus and hereditary thrombophilia had a favorable evolution under properly administered anticoagulant treatment. The homozygous status for the C677T mutation may lead to an increase in plasma homocysteine levels, especially in pregnant women, being an associated risk factor for thrombosis. The risk of developing intravascular thrombosis requires primary prevention measures by adding D-dimers in the early diagnostic algorithm, being the most accurate marker of hypercoagulability and endogenous fibrinolysis. The corroboration of the hypercoagulability status with the results of genotyping, the frequencies of the minor/major alleles studied, single mononucleotide polymorphisms (SNPs) and the establishment of preventive therapy, aims to prevent intravascular thrombosis and thromboembolic phenomena.

The perioperative effect of anesthetic drugs on the immune response in total intravenous anesthesia in patients undergoing minimally invasive gynecological surgery.

BACKGROUND: The specific mechanism of action of each anesthetic drug on the immune system is still incompletely known. It is important to know how the various anesthetics used in minimally invasive surgery (MIS) act on the inflammatory response because the choice of the anesthetic agent can influence the patient's immune system. AIM: Evaluation of the effect of anesthetic drugs used for total intravenous anesthesia (Propofol and Midazolam) on the inflammatory response after minimally invasive gynecological surgery. PATIENTS, MATERIALS AND METHODS: The inflammatory response in 20 female patients who underwent minimally invasive gynecological surgery under which intravenous anesthesia was performed. Depending on the combination of anesthetics used, we subdivided the study group into two groups, Group 1 consisting of the patients (n=10) who were given for total intravenous anesthesia, the combination with Midazolam+Fentanyl, and Group 2 (n=10) the patients who received the combination of Propofol+Fentanyl, respectively. Surgical interventional procedures included day surgery: diagnostic and operative hysteroscopy, endometrial ablation, surgical treatment of vulvar disorders. Serological profiling of patients was performed by dosing the serum concentration of nucleotide-binding domain (NOD) and leucine-rich repeat protein 3 (NLRP3) inflammasomes, interleukin (IL)-6, tumor necrosis factor-alpha (TNF-α), IL-10 before and two hours after the surgical procedure. RESULTS: In our study, we found that in both groups of patients (Midazolam+Fentanyl - Group 1, Propofol+Fentanyl - Group 2), NLRP3 and cytokines concentrations in the serum were higher after MIS than those before MIS. CONCLUSIONS: It appears that both Midazolam and Fentanyl and Propofol and Fentanyl have an immunomodulatory action due to the anti-inflammatory effect of both anesthetics. Therefore, anesthesiologists must choose an anesthetic method that uses individualized anesthetic agents, depending on the patient's immune status and disease.

Histopathological findings in the advanced natural evolution of the SARS-CoV-2 infection.

We are reporting a case of natural evolution and pathological data from a young person that was diagnosed with coronavirus disease 2019 (COVID-19). All data has been collected from the autopsy of a 30-year-old female, which was performed by the Department of Forensic Medicine from Emergency County Hospital, Drobeta Turnu Severin, Mehedinti County, Romania. The infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was confirmed by reverse transcription polymerase chain reaction (RT-PCR) on the lung tissue which was obtained during autopsy. This case provides the opportunity to study the natural evolution of COVID-19 pneumonia in a young person with clinical signs of pneumonia but without associated comorbidities. The patient had not received any treatment. The histopathological examination of the lung revealed a process of productive proliferation, proteinaceous and fibrin-macrophagic interalveolar spaces exudate, and lesions consistent with vasculitis. In the heart, we identified a cardiac thrombus. These changes are likely to suggest an advanced natural evolution of SARS-CoV-2 virus infection.

Evaluation of iNOS -2087A>G polymorphism in recurrent pregnancy loss.

BACKGROUND: Due to its role in angiogenesis, the inducible nitric oxide synthase (iNOS) gene promoter polymorphism may have a presumed role in recurrent spontaneous abortions (RSA). It is an intensely studied protein, a biological mediator, a modulator and an effector molecule by implication in numerous physiological processes: vasodilatation, angiogenesis, immunity, tissue remodeling, smooth muscle activity. AIM: Our study aims to investigate a possible association between iNOS -2087A>G (rs2297518) polymorphism and the occurrence of idiopathic recurrent pregnancy loss (RPL). PATIENTS, MATERIALS AND METHODS: In this study, as in the previously published one, 169 women, diagnosed with RPL, in the Clinics of Obstetrics and Gynecology, "Filantropia" Municipal Hospital, Craiova, Romania, were subjected to the analysis, from October 2009 to October 2016. As a control group, we used 145 women. Subjects from both groups were genotyped using specific probes for TaqMan polymerase chain reaction (PCR), allelic discrimination technique. RESULTS: We evaluated in this study a possible association between iNOS -2087A>G (rs2297518) polymorphism and the occurrence of idiopathic RPL. The chi-square test showed no significant association between the presence of this polymorphism and the increased risk to develop RPL. When we performed a comparative analysis of the frequency of genotypes and our statistical data, it was observed that this polymorphism, iNOS -2087A>G (rs2297518), has not been associated with an increased risk of developing RPL. Also, when one genotype was compared with another, we did not obtain any association that would have statistical significance, between the presence of this polymorphism and the increased risk for patients to develop RPL [in dominant - A allele carriers, iNOS 2087 AG+AA vs. GG: odds ratio (OR) 1.31, 95% confidence interval (CI) 0.83-2.07, p=0.24]. Analyzing the overall risk of developing RPL by iNOS 2087 single-nucleotide polymorphism (SNP) genotype frequencies, between controls and RPL patients (which were stratified by number of consecutive PLs), taking into account the number of consecutive pregnancies, the chi-square test showed no association between the presence of this polymorphism and the increased risk for developing RPL in all three subgroups we analyzed (in a dominant model - A allele carriers, iNOS 2087 AG+AA vs. GG: the first subgroup, OR 1.31, 95% CI 0.83-2.07, p=0.24; the second subgroup, OR 1.26, 95% CI 0.76-2.11, p=0.37; the three subgroup, OR 1.4, 95% CI 0.77-2.53, p=0.272). CONCLUSIONS: The iNOS -2087A>G (rs2297518) gene polymorphism does not influence RPL in the study area of Dolj County, Romania.

Apert syndrome - clinical case.

Apert syndrome - acrocephalosyndactyly - is a rare autosomal dominant disorder representing 1:65 000 cases of living newborns. Characteristic malformations of the Apert syndrome are early craniostenosis, microviscerocranium and II-V finger syndactyly of hand and toes with proximal phalanx of the bilateral thumb "in delta". It is difficult to determine prenatal diagnosis in the second quarter, when examining the morphology of fetal signs; the dysmorphism signs appeared in the third pregnancy quarter. We present here the case of a newborn with Apert syndrome that was born prematurely in our Clinic after a monitored pregnancy, where there was issued a suspicion of cranio-facial dysmorphism, malposition and malformation of the feet and hands in the third quarter of prenatal pregnancy. The diagnosis of Apert syndrome was placed on clinical signs, laboratory and genetic tests. The clinical outcome of the baby in the maternity was favorable, the therapeutic management being established by a multidisciplinary team. Immediate complications were due to the case of prematurity: respiratory distress syndrome and the characteristics of the syndrome: micrognathia and naso-facial dysmorphism, syndactyly, bilateral foot metatarsus adductus.

The prognostic role of Bcl-2, Ki67, c-MYC and p53 in diffuse large B-cell lymphoma.

Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous group of lymphoid malignancies, which counts for more than a third of non-Hodgkin's lymphoma cases. The aim of the current study is to evaluate the prognostic role of several immunohistochemical (IHC) markers involved in the pathological process of DLBCL. This is a retrospective analysis of the 97 de novo DLBCL patients admitted between January 2007 and December 2016 in the Department of Hematology, "Filantropia" Municipal Hospital, Craiova, Romania. The expression of Bcl-2, Ki67, c-MYC and p53 was assessed by immunohistochemistry. A significant level of association was observed between high prognostic index values and Bcl-2, Ki67, c-MYC and p53 positive cases. Moreover, overall survival and disease-free survival were higher in patients with negative expression for these markers. Bcl-2, Ki67, c-MYC and p53 could make important diagnostic and therapeutic targets; therefore, their routine assessment should be mandatory.

Correlation of clinical and biological evidence - a dominant therapeutic element of succeeding in ectopic pregnancy.

Tubal pathology, smoking, pelvic inflammatory disease, miscarriage, medical or surgical abortion, usage of intrauterine devices (IUDs) for women with salpingitis latent injuries, older than 40 years, are risk factors for ectopic pregnancy. The objective of this study concerns the correlation of the clinical and biological evidence for the early diagnosis of the ectopic pregnancy and, as soon as possible, for the estimation for eventual risk of complications that may appear. The transvaginal ultrasound test, minimal increases in serum beta-human chorionic gonadotropin (ß-hCG) dynamics and blood counts are investigations of choice in achieving our objective. Overcoming ß-hCG critical level (>1198 IU÷mL), the decrease of platelets and changes in platelet constants announce the imminent risk of ectopic pregnancy rupture and the need to take a quick decision on the course of treatment.

Velamentous insertion of the umbilical cord vessels with vasa praevia--a case report.

Among the abnormal cord insertion pathology, velamentous cord insertion associated with rupture of vasa praevia is the most severe condition related to the outcome of the newborn. In velamentous cord insertion, the fetal vessels run freely through the fetal membranes without protection from Wharton's jelly, umbilical vessels diverging as they traverse the membranes. When the membranes are ruptured, complete tearing of fetal vessels through the torn membranes or partial rupture near the site of membrane rupture may occur. Velamentous insertion occurs in approximately 1% of singleton gestations, but is observed in as many as 15% of monochorionic twin gestations. The risk of perinatal death was doubled in pregnancies with velamentous cord insertion relative to normal cord insertion. This condition can be diagnosed by ultrasonography with a sensitivity of 67% and specificity of 100% in the second trimester. We report a case of a newborn who came from a velamentous cord insertion condition associated with rupture of vasa praevia after the spontaneously membranes rupture. After a difficult resuscitation and stabilization, the newborn survived with a good outcome after the follow-up.

Prognostic factors in anterior skull base meningiomas.

AIM: Histopathological and immunohistochemical study of prognostic factors in anterior skull base meningiomas in order to determine the post-operative management. MATERIALS AND METHODS: The studied material consisted in resection specimens from 65 patients with anterior skull base meningiomas hospitalized in Clinic of Neurosurgery, National Institute of Neurology and Neurovascular Diseases, Bucharest, Romania, and diagnosed in the Department of Pathology of the same Institute, between 2007 and 2013. The biological material was processed by standard histological technique with Hematoxylin and Eosin staining which allowed the classification of tumors according to WHO 2007 system and the assessment of the morphological parameters of known prognostic value. Subsequently, the tumor fragments were submitted to immunohistochemistry to evaluate the proliferative activity (Ki-67 labeling index) and progesterone hormone receptor (PR) status. RESULTS: 83.07% of the 65 anterior skull base meningiomas were WHO grade I tumors; the grade II tumors accounted 15.38%, while the grade III tumors were rare (1.53%). Mitotic activity was variable, reaching up to 14 mitoses/10 HPF (high-power field) in atypical and anaplastic tumors; mitoses were absent in 64.81% of grade I tumors; the average mitotic index in grade II tumors was 5.15 mitoses/10 HPF. Both mitotic activity and infiltrative and invasive tumor growth (the latter found in 36.92% of cases) were correlated with tumor grade. Ki-67 labeling index ranged between 1.1% and 7.7%, with the highest value found in anaplastic tumor; progesterone receptors (PR) were expressed with variable index in 84.61% of cases. The immunonegative PR tumors were represented by 16.66% of grade I tumors and by the only grade III tumor. In contrast to PR, Ki-67 expression was statistically correlated with tumor grade. The comparison between the expression of Ki-67 and PR revealed an inverse relationship between the level of PR expression and the proliferative activity intensity. CONCLUSIONS: We found that PR expression decreases as the biological behavior of tumor becomes more aggressive; it may be related with an increased risk of recurrence, making the postoperative surveillance more rigorous in these patients.