IGHV Mutational Status in a Cohort of Bulgarian CLL Patients: High Unmutated CLL Prevalence in North-East Bulgaria.

Chronic lymphocytic leukemia (CLL) is the most common leukemia in adults. One of the best established CLL prognostic markers is the somatic hypermutational status of the IGHV gene which is a part of the immunoglobulin heavy chain variable region. Technology for IGHV genotyping has been optimized and has been applied in routine diagnostics for the first time in Bulgaria. A total of 105 patients with CLL from different Bulgarian regions were tested. IGHV mutational status was determined by Sanger sequencing on total genomic DNA (gDNA) or RNA extracted from mononuclear cells. All sequencing profiles were analyzed with the IMGT/V-QUEST tool. Within the course of the analysis a high percentage of IGHV unmutated status was established in the Varna district on the Black Sea (Northeast Bulgaria). In addition, the IGHV genotyping performed on gDNA revealed a rare case with multiple rearrangements. The present data from IGHV genotyping will help in choosing the proper treatment for the benefit of Bulgarian CLL patients.

Linking Cardiovascular Risk With Estradiol Level in Men.

Aim      To study the relationship between the cardiovascular risk and the level of estradiol in men of young and middle age. The main group included 71 patients with newly diagnosed hyperestrogenia (HE) (serum estradiol >41.2 pg/ ml). Using pseudorandomization, 68 men with normal estradiol level and age- and body weight index (BWI)-matched with the main group were included into the control group. Anthropometric data, bioimpedance variables, blood pressure (BP), and concentrations of estradiol, testosterone, glucose, and total cholesterol were analyzed in both groups.Results Patients of the main and control groups did not differ in age, BWI, and smoking status. Testosterone concentration was 10.18 nmol/l in the HE group and 12.18 nmol /l in the control group (p=0.006). Systolic BP was 142.0 mm Hg in the HE group and 135.2 mm Hg in the control group (p=0.011); diastolic BP was 90.3 mm Hg in the HE group and 86.2 mm Hg in the control group (p=0.008). Total cholesterol was 5.87 mmol/l in the HE group and 5.33 mmol/l in the control group (p=0.023). Blood glucose did not differ between the groups. The presence of HE in men 2.11 times (р=0.038) increased the probability of arterial hypertension. The intergroup difference by the SCORE scale did not reach statistical significance (р=0.172). BWI, waist and hip circumferences, and bioimpedance body composition parameters did not differ between the groups.Conclusion      In the studied cohort of young and middle-aged men, HE was an independent predictor for the presence of arterial hypertension. There were no significant intergroup differences in the total risk of cardiovascular diseases calculated with the SCORE scale.

[Androgenic status of men with severe COVID-19: the role of testosterone and dihydrotestosterone [within the program FOUNDER (features of a new coronavirus infection course and options therapy depending on the androgenic status)]].

PURPOSE: The aim of this study was to assess the mens androgen status influence on the severity and outcomes (transfer of patients to the ICU or death) of COVID-19 required hospital hospitalization. MATERIALS AND METHODS: The study included 151 hospitalized men with a confirmed diagnosis of COVID-19. To measure the severity of disease have been used Symptomatic Hospital and Outpatient Clinical Scale for COVID-19 (SHOCS-COVID). It includes the severity of the clinical condition (hyperthermia, shortness of breath, oxygen saturation, need for ventilation), the degree of inflammation (CRP), markers of thrombosis (D-dimer), the degree of lung damage according to CT. The patients underwent a study of full blood count, some biochemical parameters, lung CT, and a study of testosterone (T) and dihydrotestosterone (DHT) levels. RESULTS: T deficiency was observed in 46.4% of patients (70/151 men). At the same time, DHT deficiency was observed only in 14.4% of patients (18/125 men). In patients with a T level below the median, there was a significant increase in inflammatory factors (CRP, lymphocytes/CRP index), markers of thrombosis (D-dimer and fibrinogen), extensive lung damage at admission according to CT 25.75% vs. 11.95% (p<0.001), the elevated number of points for SHOCKS-COVID 7 (IQR 5-10) versus 5 (IQR 3-7) (p<0.001) and the longer duration of hospital treatment (3 days difference, p<0.001) in comparison with a group of patients with a T level above the median. At the same time, the T level had no correlation with age. The level of DHT had a weak inverse correlation with the age of patients, but not with the main markers of the severity of COVID-19, including the number of SHOCK-COVID scores. During multivariate regression analysis, it was shown that SHOCKS-COVID is the most significant predictor of admission to the ICU while no association of T and DHT levels with outcomes in COVID-19 was found. However, it was found that the concentration of T, even adjusted for age, has a significant inverse association with the severity of the course of the disease and the number of SHOCK-COVID scores (p=0.041). An analysis of the evaluation of directed acyclic graphs suggests the main role of COVID-19 severity in reducing androgenic function and T concentration, at which its anti-inflammatory effects are lost. There were no correlations between the concentration of DHT and the number of SHOCK-COVID scores and the COVID-19 prognosis. CONCLUSION: SHOCK-COVID is the most sensitive predictor of the COVID-19 outcome in hospitalized men, including adjusting to age. T and DHT do not directly affect the outcomes of the disease. The greater severity of the infection and an increase in SHOCK-COVID scores are associated with a decrease in the concentration of T, and a weakening of its anti-inflammatory and anti-cytokine effects, which indirectly worsens the prognosis of male patients with a new coronavirus infection undergoing hospital treatment. There are no such relationships for DHT.

[Features of a new corOnavirUs infection course and optioNs therapy DEpending on the andRogenic status (FOUNDER): androgenic status in men with COVID-19 and its relationship with the disease severity].

OBJECTIVE: Analysis of androgen status in men hospitalized with a moderate COVID-19 and its relationship with the severity of the disease. MATERIALS AND METHODS: The study included 152 males with a confirmed diagnosis of COVID-19 based on the results of a positive PCR for the SARS-CoV-2 virus and/or computed tomography of the lungs hospitalized at the MSU University Clinic due to the moderate and severe COVID-19. Examination of the level of biochemical blood parameters (CRP, creatinine, urea, glucose, total testosterone (T)); CT of the lungs. To objectify the severity of the clinical symptoms, the NEWS2 distress syndrome severity scales and the original scale for assessing the clinical condition of patients with COVID 19 (SHOCS-COVID) were used. RESULTS: The median T level in 152 examined patients was 2.14 [1.21; 3.40] ng/ml. In patients with a T level below the median, the CRP level was more than two times higher, and the D-dimer value was almost two times higher than in patients with T level above median. The duration of treatment in the hospital was longer in men with COVID 19 and an initial T level below the median than in patients with T about the median (13 days vs 10.5 days, p=0.003). Low T level was correlated with lung damage by lung CT. After improving the clinical condition, there was a linear increase in the level of T independent of its initial level. CONCLUSION: Among men with moderate and severe COVID-19, a decreased testosterone level is detected in 46.7% of cases. Patients with low testosterone levels on admission have more severe COVID-19. A significant increase in testosterone level was observed after successful COVID-19 treatment without any special action regarding testosterone level.

[Testosterone replacement therapy and its relationship with hyperestrogenism and obesity. Problems of laboratory diagnostics of hyperestrogenism].

Testosterone replacement therapy (TRT) is one of the most effective and common treatments for testosterone deficiency today. It is often prescribed, focusing only on the level of testosterone and the presence of patient complaints about a decrease in sexual function. Quite rarely, doctors additionally assess the level of estradiol and, as a consequence, the initial presence of a symptom of hyperestrogenism. One of the likely consequences of the appointment of TRT may be an excess of estradiol, the excess formation of which is associated with the enzyme aromatase, which converts testosterone into estradiol. Despite the availability of laboratory determination of the level of estradiol, the result may not always coincide with the clinical picture, one of the reasons is the difference in the methods for determining the level of estradiol in each laboratory, there is no single standard or «reference¼ today. This article describes the evolution of TRT, and also focuses on the variability of estradiol levels from laboratory to laboratory, and explains in detail why the assessment of estradiol over time should be carried out in only one laboratory. A systematic literature search was carried out in the databases Medline, Scopus, Web of Science and Elibrary, CyberLeninka.The purpose of our review was the need to engage the attention of specialists to the problem of hyperestrogenism, the not always justified prescription of TRT in patients with obesity, hypogonadism and hyperestrogenism, as well as to the problem of laboratory diagnosis of hyperestrogenism, which is acute not only in our country, but all over the world.

[A role of fructose in urinary stone formation].

Obesity is one of the actual problems of modern medicine. The comorbid conditions in patients with obesity and metabolic syndrome have no less important value, attracting the attention of scientists and clinicians. Renal stone disease is one these diseases. We are interested in its development, which is directly related to the effect of excess amounts of such a monosaccharide, as fructose. For quite a long time, this carbohydrate was considered to have only positive properties, including its role in carbohydrate metabolism in general, and, in particular, in diabetes mellitus. Over the years, a pathological effect of excess fructose was detected. Since fructose was used to replace glucose, and accordingly, its amount in food, and especially in diabetic foods, was growing. In addition, the industrial production of corn-based fructose has become much less expensive than the production of sucrose, which has also contributed to the increased use of this monosaccharide in the food industry. In this article, the mechanisms leading to the development of pathologies which are associated with the excessive use of fructose, increase in the level of uric acid with subsequent formation of uric acid stones, are reviewed. Thus, the study of the effect of fructose on the pathogenesis of renal stone disease is interesting and extremely relevant, since for many specialists, it is animal protein that is associated with the development of kidney stone disease and the role of fructose is not only unobvious, but is also a revelation.

The transforming growth factor-Beta signaling pathway involvement in cardiovascular lesions in mucopolysaccharidosis-I.

Mucopolysaccharidoses (MPS) are a group of genetic disorders due to deficiency of lysosomal enzymes resulting in impaired glycosaminoglycan metabolism. All types of MPS can present with cardiovascular manifestation, although MPS-I, II, and VI seem to have more severe involvement than the other types. Enzyme replacement therapy (ERT) is available for MPS-I, II, and VI. Cardiovascular changes including hypertrophic cardiomyopathy, thickened valvular lesions, and coronary artery lesions often poorly respond to ERT and are well known as leading causes of death in patients with MPS-I. The mechanisms to cause these changes in MPS-I have not been well characterized. Immunohistopathological studies were conducted on the cardiac specimens from a patient with MPS-I who died due to sudden cardiac failure. Phosphorylated Smad2 staining showed hyperactive transforming growth factor-beta (TGF-ß) signals in the intimal layer with myointimal proliferation causing stenosis in the coronary arteries as well as in the thickened endocardium and in the myocardial cells. TGF-ß is involved in the pathogenesis of cardiovascular diseases including hypertrophic cardiomyopathy and vascular atherosclerosis. The primary mechanisms to cause hyperactive TGF-ß signals in MPS-I are unknown. The similar mechanisms leading to hyperactive TGF-ß signals may exist in the other types of MPS. The findings of TGF-ß hyperactivity in the cardiovascular lesions in a patient with MPS-I may lead to a new therapeutic approach. Further studies are warranted to evaluate the effectiveness of the medications that suppress TGF-ß signals, such as losartan, in preventing or improving cardiaovascular lesions in patients with MPS.

Postmortem studies on a patient with mucopolysaccharidosis type I: histopathological findings after one year of enzyme replacement therapy.

Deficiency of lysosomal α-L-iduronidase results in systemic accumulation of glycosaminoglycans (GAGs). Cardiac lesions due to accumulation of GAGs include hypertrophic cardiomyopathy, valvular insufficiency/stenosis, and coronary artery stenosis due to intimal proliferation. Cardiac dysfunction is one of the most common causes of death in patients with mucopolysaccharidosis type I (MPS I). Enzyme replacement therapy (ERT) with laronidase has shown clear effects in reduction of hepatomegaly and it has been unclear whether ERT could improve or prevent the cardiac lesions. Postmortem findings in a 3 1/2-year-old boy diagnosed with MPS I at age 2 years are described. He received ERT with laronidase at 100 U/kg/week for one year. He suddenly developed cardiorespiratory failure and died the next day after C2-3 spinal surgery for instability. Postmortem examination showed hypertrophic cardiomyopathy, severe aortic valve and mitral valve thickening with shortened chordae, and endocardial fibroelastosis. Histology of the cardiac tissue revealed increased perivascular and interstitial connective tissue in the myocardium and intimal thickening causing stenosis in the cardiac vessels. Electron-microscopic (EM) studies of the thickened endocardium revealed numerous histiocytes with enlarged lysosomes. EM examination of the liver and the cardiac muscle revealed no accumulation of GAGs. ERT with laronidase showed clear effects in removing GAGs from the liver and the cardiac muscle. However, it did not show a clear effect on the thickened endocardium, myocardial perivascular and interstitial connective tissue or intimal thickening in the epicardial vessels.

[Regulatory genes of garden pea (Pisum sativum L.) controlling the development of nitrogen-fixing nodules and arbuscular mycorrhiza: a review of basic and applied aspects].

The review sums up the long experience of the authors and other researchers in studying the genetic system of garden pea (Pisum sativum L.), which controls sthe development of nitrogen-fixing symbiosis and arbuscular mycorrhiza. A justified phenotypic classification of pea mutants is presented. Progress in identifying and cloning symbiotic genes is adequately reflected. The feasibility of using double inoculation as a means of increasing the plant productivity is demonstrated, in which the potential of a tripartite symbiotic system (pea plants-root nodule bacteria-arbuscular mycorrhiza) is mobilized.

[Prevention and treatment of postoperative nausea and vomiting (results of a multicenter study)]. / Profilaktika i lechenie posleoperatsionnoi toshnoty i rvoty (analiz rezul'tatov mul'titsentrovogo issledovaniia).

Described in the paper are the results of a multicenter study that was conducted at 11 health-care facilities of the Russian Federation and that was based on an analysis of 623 medical records of adult patients operated on with general anesthesia of different variations. There were 154 (24.7%) males and 469 (75.3%) females, aged 16 to 90. The study denoted the need in and possibility of preventing and arresting the PONV; it also outlined those forms of ondansetron (Zofran) that can be used for the purpose, they are: parenteral and lingual pills and rectal suppositories. An assumption was put forward on the significance of PONV conductive factors and on the most effective forms of ondansetron from among the mentioned above. An understanding, resulting from the study, on that PONV is of the multietiology nature and on that its probability increases in proportion to a compatibility of provoking factors, like sex, surgery technique as well as duration of anesthesia and its variation is of importance. The efficiency of medicamental prevention of PONV by a blocker of HT3-receptors, i.e. by ondansetron (Zofran), is to a great extent predetermined by that if the form and, probably, dose of the drug is in line with a combination of the above factors, which needs further research.

[Pharmacoeconomic assessment of prevention of the postoperative nausea and vomiting syndrome in plastic (esthetic and reconstructive) surgery]. / Farmakoékonomicheskaia otsenka profilaktiki sindroma posleoperatsionnoi toshnoty i rvoty v plasticheskoi (ésteticheskoi i rekonstruktivnoi) khirurgii.

A comparative "cost-efficiency" pharmacoeconomic analysis was made for 5 different anti-emitting drugs, i.e. for "Navoban", "Zofran" (intravenous administration, 8 mg; and suppositories, 16 mg) and for "Metipred" (methylprednisolone) used in aesthetic and reconstructive surgeries of 86 patients. All the drugs were found to be effective for the prevention of PONV. Clinically, the efficiency decreased in the following order: "Novoban" and "Zofran" suppositories 16 mg, "Metiored" and "Zofran" intravenous administration 8 mg. As for "cost-efficiency", preference should be given to "Metipred".

Infection as a cause of death in the extremely-low-birth-weight infant.

OBJECTIVE: To determine the causes of neonatal death for extremely-low-birth-weight (ELBW) infants. METHODS: All liveborn infants below 1000 g birth weight born from 1994 to 1998 who died and were autopsied were included. Maternal and infant characteristics, placental histology, autopsy material and culture results were obtained. RESULTS: A total of 263 ELBW infants were born alive, 104 (40%) died and 44 (42%) were autopsied. Placentas were available for 41 (93%). Infection was the leading cause of death in the autopsied babies (25/44; 57%). Sixteen (64%) of these deaths occurred within the first 48 h and were classified as being due to congenital infections. Twenty-two of 41 (54%) placentas showed evidence of infection. Infection as a cause of death peaked at 22 weeks. Other causes of death were lethal anomalies (20%), respiratory distress and its complications (9%) and immaturity, intraventricular hemorrhage and other conditions (14%). CONCLUSION: Congenital infection is the leading cause of death in ELBW infants.

Meconium-stained amniotic fluid and neonatal morbidity in near-term and term deliveries with acute histologic chorioamnionitis and/or funisitis.

OBJECTIVE: To determine the incidence of meconium-stained amniotic fluid (MSAF) and neonatal morbidity in near-term and term deliveries with histologic acute chorioamnionitis and/or funisitis compared to those with normal placental histology. STUDY DESIGN: In a retrospective case-control design, we compared the incidence of MSAF and neonatal outcome in 45 cases of acute histologic chorioamnionitis and/or funisitis with 89 cases of normal placental histology. We reviewed the obstetric and neonatal records for perinatal complications and neonatal morbidity. RESULTS: Mean birthweights (3372+/-473 vs 3287+/-518 g) were similar in infants born to mothers with histologic chorioamnionitis and/or funisitis compared to infants born to mothers with normal placental histology. The incidence of MSAF was significantly higher in the group with acute chorioamnionitis/funisitis (p<0.05). Similarly, the incidence of admissions to newborn intensive care unit, respiratory distress, meconium aspiration syndrome, and presumed sepsis was also significantly higher (p<0.05) in this group. CONCLUSION: The incidence of MSAF and neonatal morbidity is higher in the presence of acute inflammation of placental membranes. The presence of meconium in the amniotic fluid should alert the physician to the potential for infection and increased neonatal morbidity.

[Nodulation as a model for studying differentiation in higher plants]. / Kluben'koobrazovanie kak model' dlia izucheniia differentsirovki u vysshikh rastenii.

The stages of the legume-rhizobial symbiosis and nodule structure in various legume plants are briefly reviewed. Modern data on the mechanisms involved in the control of nodule initiation and morphogenesis are considered.

[Characteristics of certain symbiotic pea mutants by traits related to hormonal status]. / Kharakteristikia nekotorykh simbioticheskikh mutantov gorokha po priznakam, sviazannym s gormonal'nym statusom.

Association between traits for hormonal status and nodulation in the mutants of pea Pisum sativum L. with abnormal nodulation and original forms was analyzed. The sensitivity of plant tissues to exogenous phytohormones and changes in the concentration of the major auxin, indolyl-3-acetic acid, in plant roots during interaction with rhizobia were examined. Association between Nod(++)-phenotype and auxin balance was revealed: the supernodulating mutants were more sensitive to auxin treatment than the parental cultivars. Mutations in the sym8 gene, in contrast to those in the sym5 gene, had no effect on plant hormonal status. The level of indolyl-3-acetic acid during interaction with rhizobia depended on the time after inoculation and plant genotype. The mutations affecting nodulation were suggested to change auxin level in roots.

Fetal membrane histology in preterm premature rupture of membranes: comparison to controls, and between antibiotic and placebo treatment. The National Institute of Child Health and Human Development Maternal Fetal Medicine Units Network, Bethesda, MD, USA.

The objectives of this study were to test the hypotheses that antibiotic therapy will alter the histologic appearance of fetal membranes in preterm premature rupture of membranes (pPROM), and that the membrane histology will demonstrate distinct differences between term and preterm rupture of membranes. We also wished to test interobserver variability of pathologists. Placental membranes were sampled from 268 women participating in a randomized placebo-controlled trial of antibiotic therapy for pPROM at 24-32 weeks of gestation (cases) and from 4 control groups who were not in the randomized trial: (1) preterm labor without pPROM (n = 21), (2) term labor (n = 65), (3) term PROM (n = 21), and (4) term cesarean section (n = 27). The cases and controls were scored for 40 histologic features by pathologists blinded to the identity of each sample (case or control). pPROM histology of samples from patients receiving antibiotics and those receiving placebo was compared using a chi-squared test and with control groups using logistic regression. There were no histological differences between pPROM cases treated with antibiotic and those receiving placebo, nor with respect to duration of membrane rupture greater or less than 48 h. Concordance among pathologists was low for features other than acute inflammation. Logistic regression analysis controlled for race and pathologist, and demonstrated that all of the control groups had significantly fewer common markers of acute inflammation when compared with the pPROM cases. This study suggests that histopathologic evidence of infection is seen more frequently with pPROM than in preterm or term controls. The histologic features used in this study cannot be used to determine the effectiveness of antibiotic therapy.

Causes of death in the extremely low birth weight infant.

OBJECTIVE: To present primary and secondary causes of death confirmed by autopsy for the extremely low birth weight infant. METHODS: A total of 111 infants weighing between 300 and 1000 g at birth who died and were autopsied at our hospital during the 4-year period 1990-1993 were retrospectively reviewed. Clinical, pathologic, and laboratory data were retrieved including results of placental examinations and autopsy cultures. Primary and secondary causes of death were assigned by the authors. RESULTS: Infection was the most common primary cause (56/111) followed by respiratory distress syndrome/bronchopulmonary dysplasia (24/111) and congenital defect (15/111). Immaturity as an only cause appeared almost exclusively in infants weighing <500 g at birth. Infection was significantly underdiagnosed clinically with most of these deaths attributed to immaturity or respiratory distress syndrome. In only 1 case was intraventricular hemorrhage considered the primary cause of death although it was present as a secondary cause in 19/111. Infections were divided into congenital (30/56) and acquired (26/56) by time of death. The congenital infections (