Morphological and Molecular Studies of the Rapa Whelk, Rapana venosa (Neogastropoda, Muricidae), from Odesa Bay

The gastropod Rapana venosa (Valenciennes, 1846) is a successful worldwide invader occurring in the Black Sea. The aim of this study is to overview specific population features of this mollusk from Odesa Bay through integrative systematic approach by means of morphological and molecular research. For this purpose, the mollusks were collected from the Black Sea and examined using morphological methods: traditional morphometry, which employs linear parameters of shells (height, width, whorl height, whorl width, height of the last whorl) and shell weight, and geometric morphometrics of the shell shape data. For a molecular genetic test, the COI gene region was used. Among all conchological variability, the two morphotypes were defined: the first has a “broad” shape — shells have a thick and durable last whorl and a low spire, and the second one — “extended” shape: shells are relatively slender with an elongated high-conical spire. According to the geometric morphometric data, R. venosa has statistically significant differences between defined morphotypes (F = 4.12, p = 0.001); however, the shapes in males and females are not significantly different (F = 1.13, p = 0.318). No genetic diversity, neither novel haplotypes were revealed by the molecular analysis: in Odesa Bay, the haplotype occurring also in other regions of invasion across the world is present.

Joubert's syndrome as a cause of cholestasis in children

Literature review and a case report of Joubert syndrome in a preschool child are presented. This syndrome is accompanied by chronic intrahepatic cholestasis, characteristic facial dysmorphia, congenital malformations of the urinary system, eyes pathology, confirmed by a molecular genetic test of TMEM67 gene mutations. Diagnostic aspects of this pathology have been reviewed. The clinical case description. Boy K was admitted to the pediatric hepatology department with respiratory and renal failure, decompensated acidosis. On admission, complaints included shortness of breath, severe itching, pale and dry skin, poor appetite and delayed physical development. On examination, dysembryogenic stigmas: large head circumference, elongated face, protruding forehead, high rounded eyebrows, deeply set eyes, hypertelorism, antimongoloid incision of the eyes, low auricles. The skin was pale, dry with the scratch marks on the arms and legs flexor surfaces. Ankles, wrists skin lichenification, bruising and hyperpigmentation was observed. Moderate palmar erythema, spider veins on the abdomen, watch-glass nails were present. The abdomen was enlarged due to hepatosplenomegaly. A clinical and paraclinical examination was conducted, the patient was consulted by related specialists. Based on the obtained data and molecular genetic examination, the diagnosis was made. Conclusion. Joubert's syndrome may have a clinical presentation similar to that in cholestatic liver disease with typical clinical and laboratory symptoms. The molecular genetic testing is an important workup component in children with liver pathology combined with damage to other organs and systems. Successful treatment is possible providing a multidisciplinary approach used. The study was conducted by the principles of the Declaration of Helsinki. The research protocol was approved by the Local Ethics Committee of the abovementioned institution. Informed consent of the child's parents was obtained for the research. The authors declare no conflict of interest. Keywords: Joubert syndrome, cholestasis, children.

Basic characteristics and features of the molecular genetic test systems designed for non-invasive diagnostics and prognosis of prostate cancer and bladder cancer

Improving the laboratory diagnosis of prostate cancer and bladder cancer are still an actual problem in modern urologic oncology. Test systems for DNA or RNA alterations that occurred during carcinogenesis and associated with the malignant tumor and the prognosis of disease have been actively developed in recent years. Here we reviewed the data published mainly in the last 5 years about the molecular genetic kits for diagnosis (Progensa, SelectMDx, ExoDx Prostate Test, Prosta-Test, Confirm MDx) and assessment of prognosis (Prolaris, Decipher, Oncotype DX) in patients with prostate cancer, discussed their sensitivity and specificity. The characteristics of analogous kits and panels for bladder cancer (UroVysion, CertNDx Bladder Cancer Assay, UroSEEK, mutations in the FGFR3 and TERT genes, and the Cxbladder Monitor/Detect/Triage kit's line) were systematized. Particularly we focused on the description of the patient cohorts for whom kits mentioned above have greater diagnostic accuracy, described limitations of these test systems in consequence both a methodological and registration aspects, and their use in combination with other tumor markers. This review is aimed at oncologists, urologists, laboratory geneticists and specialists in related professions.

CREATION OF A MOLECULAR GENETIC TEST SYSTEM FOR EARLY DIAGNOSTICS AND EVALUATION OF EFFECTIVENESS OF TREATMENT OF INFLAMMATORY PERIODONTAL DISEASES

Inflammatory periodontal diseases represent a serious dental and general medical problem due to the high prevalence among the adult population, the presence of clinical forms leading to the destruction of the dentition and tooth loss, insufficient treatment effectiveness and the frequency of relapse, including in connection with the formation of biofilms. A molecular genetic test system has been developed to evaluate the content of periodontopathogenic microorganisms Porphyromonas gingivalis, Treponema denticola, Streptococcus oralis, Streptococcus sanguis and Streptococcus sobrinus in the contents of periodontal pockets. The analytical characteristics of the test system were determined, and testing was carried out on clinical samples of patients with chronic generalized periodontitis of moderate severity.The constructed diagnostic kit allowed us to conduct a comparative analysis of the effectiveness of various types of treatment of inflammatory periodontal diseases based on quantitative data on the content of bacteria in the contents of periodontal pockets.

Hypophosphatasia in children. Three faces of one disease

Hypophosphatasia (HPP) is a rare multisystem inherited metabolic disorder caused by mutations in ALPL gene that encodes tissue nonspecific alkaline phosphatase responsible for bone mineralization. HPP is characterized by impaired bone mineralization, skeletal abnormalities, and systemic manifestations which result in significant morbidity and mortality. Clinical presentations of HPP vary greatly. Early (perinatal and infantile) HPP is characterized by the most severe symptoms, i.e., respiratory and neurological disorders are of crucial importance being the leading causes of death. Progressive skeletal impairment, rickets-like deformities, reduced mobility, and severe disability are typical of childhood-onset HPP. The biochemical hallmark of HPP is low alkaline phosphatase (ALP) activity. HPP diagnosis is verified by clinical symptoms in combination with persistently low ALP activity (adjusted for age and sex). Molecular genetic test to identify ALPL gene mutation is performed as needed. Three case reports addresses authors’ experience with the diagnosis and treatment for HPP.Keywords: hypophosphatasia, case series, alkaline phosphatase, impaired bone mineralization, asfotase alfa.For citation: Boykov S.A., Chernyak I.Yu., Shatokhina N.S. et al. Hypophosphatasia in children. Three faces of one disease. Russian Journal of Woman and Child Health. 2020;3(2):136–141. DOI: 10.32364/2618-8430-2020-3-2-136-141.

Measuring the chronology of the translational process of molecular genetic discoveries

Background The process of technology validation and transfer of new molecular diagnostic tests towards the clinic faces challenges and needs to be improved. There is no empirical measure of the chronology and pace of technology transfer of molecular genetic discoveries. Methods We studied these for 29 molecular genetic test discoveries in order to (1) provide estimates of the timeframe between discovery of a clinical application and complete clinical implementation, and (2) compare the trajectories between different new tests to identify common patterns. We identified 11 publicly available “timestamps” for the technology transfer process ranging from discovery of the marker to use in a clinical setting. For each test selected, we searched public databases to identify available timestamps and dates. We plotted and compared trajectories of individual tests, including chronology. Results We show that there is much variability in the chronology of transfer between biomarkers. The median time between discovery of the marker and availability of the clinical test was 9.5 years (minimum 1). There was a median time of 18 years between test discovery and FDA approval (minimum 7 years), and it took a median of 17 years between discovery and the availability of a certified reference material for the 10 assays that have one (minimum 9 years). Conclusions We conclude that new molecular genetic tests take significant time between discovery and clinical implementation, and that further work is needed to pinpoint key factors, including policy and organization factors, that may allow for improving and streamlining this process.

DIFFICULTIES OF TUBERCULOSIS DETECTION BY A LOCAL PHISICIAN IN THE PERIOD OF THE EPIDEMIC OF INFLUENZA (CLINICAL CASE)

Purposeof issue is to show difficulties of detecting tuberculosis the general physician during epidemic of acute respiratory viral infection and flu, to show efficiency of modern methods of mycobacterial and HIV evaluation.Materials and methods. Clinical case from general physician’s practice was discussed. All methods from primary healthcare were used.Results. Possibilities of tuberculosis diagnostics by general physician during epidemic were shown. Tuberculosis was held under mask of flu. While carrying out diagnostic minimum negative results have been received. As a result of plane radiography dissemination was found. After analysis of social risk factors, analysis for HIV and «Diaskin-test» were made. Sputum also was sent for mycobacterium evaluation to antituberculosis center laboratory. Positive «Diaskintest» (8 mm), positive Gene-expert molecular-genetic test and BACTEC fluid growth were found. Diagnosis of tuberculosis was stated and patient was sent to specialized center.Conclusion. During flu epidemic alertness of general practitioners about tuberculosis is needed especially in social risk groups. It is recommended to enlarge diagnostic minimum using molecular genetic methods. These actions can improve rate of tuberculosis proving.

Визначення поліморфних варіантів фрагментів ДНК збудників хламідійних інфекцій сільськогосподарських тварин

Under the currently existing classification, adopted at the Second European Symposium «Animal Chlamydioses and Zoonotic Implications (EMAC-2)», Chlamydia pathogens of animals and humans are intracellular gram-negative bacteria belonging to Chlamydiales order, Chlamydiaseae family, Chlamydia genus. The above mentioned genus includes 11 species: C. abortus, C. avium, C. caviae, C. felis, C. gallinacea, S. muridarum, C. pecorum, C. pneumoniae, C. psittaci, C. suis and C. trachomatis, 10 of them being pathogenic for animals and Chlamydia trachomatis being exclusively human Chlamydiosis agent. Development of highly sensitive and specific molecular genetic test systems for indication and species differentiation of the said Chlamydia genus bacteria will permit to reliably study various aspects of chlamydial infection. Aim of the study was to perform bioinformatiс alignment of different genes’ nucleotide sequences to determine polymorphic DNA regions of Chlamydia genus bacteria, which are the basis for designing oligonucleotide primers for molecular genetic test systems capable of differentiating pathogens, causing disease in cattle and small ruminants, horses and pigs, by species. Bioinformatic study has been performed in 411 primary nucleotide DNA sequences of genes encoding 16S rRNA, RNase P RNA and MOMP of four chlamydial infections agents of farm animals (C. abortus, C. pecorum, C. pneumoniea, C. suis) obtained from the international electronic databases «GenBank» and «PubMed». Meanwhile, it was determined that the gene encoding the main outer membrane protein (MOMP) of Chlamydia possesses the highest variability level, making 97.1%. Using the method of aligning the primary sequences of the said gene, by means of «MEGA4» and «MEGA7» software, polymorphic fragments of nucleotide DNA sequences have been determined for C. abortus, C. pecorum, C. pneumoniea and C. suis. Specificity testing of polymorphic fragments, determined for each of the four chlamydia species with nucleotide sequences of microorganisms, both opportunistic and other infections pathogens, was carried out using «Blast» online software applications. Polymorphic fragments of the gene encoding MOMP of the above Chlamydia genus bacteria will be used to design oligonucleotide primers of test systems for indication and specific differentiation of farm animals’ chlamydial infections agents in polymerase chain reaction.

De novo SCN1A géndeletio terápiarezisztens Dravet-szindrómában

Severe myoclonic epilepsy in infancy (Dravet’s syndrome) is a very rare form of epilepsy. Mutations of SCN1A gene encoding voltage-gated sodium channel alpha-1 subunit are major causes of the autosomal dominant disorder. Most cases are associated with a de novo point mutation, but some patients have copy number variations. The protein encoded by the SCN1A gene plays a role in the generation and propagation of action potentials. Loss of function caused by the majority of gene mutations leads to hyperexcitability of the neuronal network that finally results in the formation of the epileptic seizures. Molecular genetic test for copy number variations of SCN1A gene is available in the department of the authors since 2013 besides sequencing analysis of the whole gene. This article presents the case of a 7-year-old patient with two years of recorded patient history outside of the author’s department. Molecular genetic test, which detected a de novo SCN1A gene deletion in heterozygous form, revealed SCN1A gene associated monogenic epileptic syndrome being in the genetic background of therapy-resistant seizures. Orv. Hetil., 2015, 156(49), 2009–2012.