The treatment of the diaphyseal bones defects using the method of induced membrane (preliminary study)

Department of Orthopaedics and Traumatology, Nicolae Testemitanu State University of Medicine and Pharmacy, Chisinau, Republic of Moldova, The 6th International Medical Congress for Students and Young Doctors, May 12-14, 2016Introduction: The management of segmental long-bone defects is a challenge. The literature has described many techniques, but each is fraught with specific difficulties. Masquelet’s technique of induced membrane is now a reference surgical procedure for the treatment of complex lesions requiring bone regeneration. The concept of induced membrane was introduced by Alain-Charles Masquelet in 1986. The Masquelet method consists in formation of an induced membrane by a foreign body which has secretory properties, influencing positive on the regeneration and strengthening of the cancellous bone grafts. Aim: to investigate the morphological properties and characteristics of induced membrane which was modeled in an experimental group of rabbits in order to asses and to optimize the effectiveness of the Masquelet method in the clinic. Materials and methods: Experimental work was done using a group of rabbits (n=10) with the weight 5,5±0,5kg and the age 5 months. The investigation had 3 steps. The first step of the study consisted in creating the bone defect, filling it up with an antibiotic-impregnated cement spacer and stabilizing it with a plate. The second step of the study was 21 days later, consisting in incision of the induced membrane, removing the spacer and filling up the space with cancellous bone chips collected from iliac crest. At this stage we sacrificed 5 rabbits in order to perform the histological and morphological examination. At the sixth week we switched to the third step – ablation of metal construction and the radiological control exam. At this stage we sacrificed 5 rabbits to study the morphological aspect of the healed bone. Results: The histo-morphological examination performed at the 21 days demonstrated the presence of an inflamator process characterized by neutrophilic, eosinophilic elements and regeneration’s elements – fibroblasts. Also, it was determined a pseudo synovial metaplasia and a villous hyperplasia with formation of an synovial epithelium on the internal face of the induced membrane. The histo-morphological exam performed at the 6 weeks has demonstrated the continuation of the neoformating process and of the bone modelation, the regeneration process prevailed over the inflammatory one. The morphological aspect was formed by agglomerations of fibroblasts, myoblasts and collagen and numerous vascular buds, that promote a good neoangiogenesis and osteogenesis of the bone. Conclusion: The morphological study demonstrated an intense process of cell proliferation and differentiation, which highlights the biological role of induced membrane by foreign body with secretion of the osteoinductive factors, promoting the vascularization and corticalization of the bone. The Masquelet method is an effective method that allows getting the consolidation of the bone in case of critical size bone loss

Multiple plasmocytoma - a rare case of three atypical presentations

Introduction: The solitary plasmocytoma represents less than 5% of all plasma cell neoplasms. The multiple plasmocytoma is 20 times rarer than solitary plasmocytoma. Progression to multiple myeloma is common. Methods: Case study Results: We report a very rare case of multiple plasmocytoma developed in the bone and two atypical sites - renal and gastric. The patient was treated with radiotherapy, surgery and chemotherapy. The case is presented due to its rarity. Conclusion: The high risk of a progression towards a multiple myeloma justifies a comprehensive initial assessment and regular monitoring of all plasmocytomas. The management of a patient with multiple plasmocytomas will be determined by the sensitivity and the site of the tumor

Stem cell niche: location, structure and function

Department of histology, cytology and embryology, State University of Medicine and Pharmacy "Nicolae Testemiteanu" Chișinău, Republic of Moldova, Congresul consacrat aniversării a 75-a de la fondarea Universității de Stat de Medicină și Farmacie „Nicolae Testemițanu” din Republica Moldova, Ziua internațională a științei pentru pace și dezvoltareIntroduction: Stem cell niche is a specific histological structure capable of regulation tissue proliferation and proper functioning. The stem cell niche typically has a spatial organization that provides anatomical and functional interactions. These interactions are mutual and dynamic. Purpose: To study the location, structure and function of the stem cell niche by analyzing bibliographic sources. Material and methods: To realize the research, we reviewed materials from Medscape and PubMed, 2006-2020 yy. We specified the localization of the niche, the types of stem cells and the possible. Results: The niche is basic unit of tissue physiology., integrates signals of proliferation, regeneration, differentiation and migration of stem cells. The niche is formed by the ensemble of stromal cells and factors they produce, including adhesive signals, soluble factors and matrix proteins. Conclusions: Stem cell niche is the microenviroment that maintains stem cell homeostasis

Hipertensiunea pulmonară primară

Este prezentat un caz (bărbat, 26 de ani) de hipertensiune pulmonară severă, complicată cu suferinţa ventriculară dreaptă, clasa funcţional 2/3 NYHA. Investigaţiile efectuate au fost neinvazive şi invazive. Diagnosticul clinic de suferinţă al ventriculului drept era susţinut electrocardiografic; radiografia toracică era compatibilă cu diagnosticul de HP. Singura metoda noninvazivă care a stabilit cu certitudine diagnosticul de HP a fost examenul ecocardiografic - era esenţial pentru diagnosticarea şi aprecierea severităţii hipertensiunii pulmonare. Scintigrafia pulmonară de perfuzie a exclus embolismul pulmonar. Au fost excluse multiple cauze cardiace care pot determina creşterea presiunii sistolice in artera pulmonară (hipertensiunea pulmonară secundară). Cateterismul cardiac a confirmat hipertensiunea pulmonară şi tipul de hipertensiune. Evoluţia imediată ar putea fi relativ bună, prognosticul tardiv este nefavorabil, dar tratamentul medicamentos este limitat. Complicaţiile în viitor depind direct de accentuarea hipertensiunii pulmonare şi de agravarea insuficienţei ventriculare drepte

Severe axonal neuropathy is a late manifestation of SPG11

Complex hereditary spastic paraplegia (HSP) is a clinically heterogeneous group of disorders usually inherited in an autosomal recessive manner. In the past, complex recessive spastic paraplegias have been frequently associated with SPG11 mutations but also with defects in SPG15, SPG7 and a handful of other rare genes. Pleiotropy exists in HSP genes, exemplified in the recent association of SPG11 mutations with CMT2. In this study, we performed whole exome sequence analysis and identified two siblings with novel compound heterozygous frameshift SPG11 mutations. The mutations segregated with disease were not present in control databases and analysis of skin fibroblast derived mRNA indicated that the SPG11 truncated mRNA species were not degraded significantly by non-sense mediated mRNA decay. These siblings had severe early-onset spastic paraplegia but later in their disease developed severe axonal neuropathy, neuropathic pain and blue/black foot discolouration likely caused by a combination of the severe neuropathy with autonomic dysfunction and peripheral oedema. We also identified a similar late-onset axonal neuropathy in a Cypriot SPG11 family. Although neuropathy is occasionally present in SPG11, in our SPG11 patients reported here it was particularly severe, highlighting the association of axonal neuropathy with SPG11 and the late manifestation of axonal peripheral nerve damage

Morphological evaluation of the different methods used for protection of colonic anastomosis

First Department of Surgery “N. Anestiadi” and Laboratory of Hepato-Pancreato-Biliary Surgery, Medical University "N.Testemitanu", Chisinau , RMIntroduction: Despite the performances of modern medicine, especially of colorectal surgery, anastomotic leakage remains one of the most dangerous postoperative complications, without significant trend of decreasing. Morbidity and mortality increase considerably after the development of an anastomotic leakage. Anastomotic leakage presents an important problem of public health with major socio-economic impact and can be considered one of the quality indicators of specialized surgical centers' activity. There are multiple studies running in order to create and assess the efficacy of colonic anastomosis local protection methods. Aim of study was morphological evaluation of the methods used for local protection of anastomotic zone and their influence on the anastomosis healing. Materials and methods: Sixty three rats were divided in three groups: colonic anastomosis was performed and topical latex tissue adhesive was applied in the group I (n=21); colonic anastomosis with local application of collagen patch in the group II; colonic anastomosis without local protection in the group III. Results: Anastomotic leakage was not determined in the group I vs the group III, where were detected 5 cases of anastomotic leakage. According to the present study’s data in the group I was determined early diminution exudativ-detersiv process’ activity vs groups II and III (p<0.01). Latex tissue adhesive has positive influence on the processes of neoangiogenesis and fibrilogenesis in the anastomotic zone on the 14th POD vs the group II and III (p<0.05). According to ours data latex tissue adhesive has considerable compatibility with colonic tissue that represents the absence of giant like „foreign bodies” symplasts and insignificant immunologic reaction of large bowel. Aggressive bacterial colonization in this group has contributed for appearance of anastomotic leakage, formation of abscesses and granulomatous processes like „foreign bodies”. Mentioned processes considerable have complicated synchronous evolution of neoangiogenesis and fibrilogenesis in the anastomotic zone, resulted in decreasing of the primary healing, appearance of anastomotic deformations and expression of the adhesion process vs anastomosis from the groups I and III. Conclusion: Using of latex tissue adhesive for local protection of colonic anastomosis improves anastomotic healing, processes of neoangiogenesis and fibrilogenesis. Using of collagen patch for local protection of colonic anastomosis doesn’t have any advantages and provokes delaying of regeneratory processes and persisting of an inflammatory process

Familial epilepsy – clinical-epidemiological characteristics and next-generation sequencing in the Republic of Moldova’s population

Background: Although several theories are implicated in the origin of epilepsy, its cause is still unknown in about 50% of cases. To associate a gene with epilepsy for the first time, families with multiple affected members are needed. The aim of our study is carrying out a clinical-genetic study of multiplex families from the Republic of Moldova, for estimating the genetic biomarkers and establishing their weight in epileptogenesis. Material and methods: An epidemiological, descriptive study (2018 – 2023) started with lancing a National Epilepsy Registry for multiplex families. Whole Exome Sequencing (WES) was performed on the first 11 families. Preliminary statistical methods were applied. Results: Our National registry counts now 74 families including 186 members. First 11 families’ WES results showed that the most involved chromosomes with candidate epileptogenic variants are the 1, 2, 3, 4, 7, 12, and 17. Top affected genes are the AUTS2, ATXN1, KCNMA1, IRF2BPL, SUFU, CENPE, SACS, EDC3, RYR2, ANKRD11, PTPRD, CHL1, MYH1, CC2D2A, LIAS, TBCD and AARS. From all the detected variants, 20.3% were classified as deleterious and probably pathogenic, 38.9% were marked as tolerated and benign and 22.8% were variants of unknown significance (VUS). Conclusions: Our results represent an absolute novelty for our country, such studies having been never previously performed. Subjects continue to be recruited and the National Register of presumed genetic epilepsy is constantly being updated

Înțelegerea caracteristicilor genetice ale familiilor multiplex cu epilepsie din Moldova folosind secvențierea întregului exom

Laboratory of Neurobiology and Medical Genetics, Department of Neurology no. 2, Nicolae Testemitanu SUMP, Department of Neuromuscular Diseases, Institute of Neurology, UCLBackground. Although several theories have been proposed to explain the origin of epilepsy, its cause is still unknown in about half of cases. In most cases, the link between a gene and the condition is not yet clear and studying multiple affected members of a family is needed. Objective of the study. To estimate the genetic biomarkers of multiplex epilepsy families from the Republic of Moldova and their role in epileptogenesis. Material and Methods. Whole Exome Sequencing (WES) was performed on the first 11 epilepsy families from a newly started National Epilepsy Registry. It was followed by a descriptive analysis of the data. Results. Our National registry counts now 74 families including 186 members. WES results of the first 11 Moldovan multiplex epilepsy families revealed that the most prevalent epileptogenic variants are those involving the 1, 2, 3, 4, 7, 12, and 17 chromosomes. Top genes affected by candidate variants include AUTS2, ATXN1, KCNMA1, IRF2BPL, SUFU, CENPE, SACS, EDC3, RYR2, ANKRD11, PTPRD, CHL1, MYH1, CC2D2A, LIAS, TBCD and AARS. From the detected variants, almost 23 % were classified as of unknown significance (VUS), 20% were identified as deleterious and probably pathogenic by two known predictors (SIFT and Polyphen), and 39% are known as tolerated and benign. Conclusion. The preliminary results of our studies are truly revolutionary, as they represent an absolute novelty for the country and the eastern “genetically virgin” territories.Introducere. Deși au fost propuse mai multe teorii pentru a explica originea epilepsiei, cauza sa rămâne necunoscută în aproximativ jumătate dintre cazuri. În majoritatea acestora, legătura dintre genă și afecțiune nu este clară și se impune studierea mai multor membri afectați ai unei familii. Scopul lucrării. De a estima biomarkerii genetici ai familiilor multiplex cu epilepsie din Republica Moldova și rolul lor în epileptogeneză. Material și Metode. Secvențierea întregului exom (WES) a fost efectuată în cadrul primelor 11 familii cu epilepsie dintr-un Registru Național nou constituit. Ulterior, a fost efectuată o analiză statistică descriptivă a datelor obținute. Rezultate. Registrul nostru național numără acum 74 de familii și 186 de membri. Rezultatele WES ale primelor 11 familii cu epilepsie multiplex au arătat că cele mai răspândite variante epileptogene sunt cele care implică cromozomii 1, 2, 3, 4, 7, 12 și 17. Genele de top afectate de variantele candidate include: AUTS2, ATXN1, KCNMA1, IRF2BPL, SUFU, CENPE, SACS, EDC3, RYR2, ANKRD11, PTPRD, CHL1, MYH1, CC2D2A, LIAS, TBCD și AARS. Dintre variantele detectate, doi predictori de patogenitate cunoscuți (SIFT și Polyphen) au clasificat aproape 23% ca având o semnificație necunoscută (VUS), 20% ca dăunătoare și probabil patogene, iar 39% ca fiind tolerate și benigne. Concluzii. Rezultatele preliminare ale studiilor noastre sunt cu adevărat revoluționare, întrucât reprezintă o noutate absolută atât pentru țară, cât și pentru teritoriile estice, considerate „virgine genetic”

Aplicabilitatea testării genetice de nouă generație în epilepsie prin prisma secvențierii întregului exom

Background. Epilepsy affects around 1% of the general population. With strong genetic contributions >50% of cases still remain undiagnosed. Recently, novel genetic testing lead to massive gene discovery, epilepsy included. Objective of the study. Our review highlights the progress in the field of epilepsy genetics and discusses how the genetic architecture of common epilepsies is starting to be unraveled. Material and Methods. Relevant studies were searched using key-words: epilepsy genetics, next-generation sequencing (NGS), whole exome sequencing (WES) – in the PubMed and Google Scholar databases. A study was included if at least 2 of the key-words matched. Results. Since the 1995 finding of CHRNA4 mutation, more than 500 genes were estimated to play a role in epilepsy. To date, WES’s diagnostic rate varies from 12,5% in focal epilepsies to 33% in overall cohorts. The remaining undiagnosed forms are likely multifactorial. However, NGS introduces new challenges, yet to be resolved. Conclusion. Over the past decade, WES studies have increasingly been used to uncover the role of the coding genetic material in the human genome, being a prerequisite for personalized treatment approaches and reducing the epilepsy patient’s “diagnostic odyssey”. Introducere. Epilepsia afectează aproximativ 1% din populația generală. Cu contribuții genetice marcante, >50% din cazuri rămân încă nediagnosticate. Recent, testarea genetică de nouă generație a dus la descoperirea masivă de noi gene, inclusiv în epilepsie. Scopul lucrării. Review-ul nostru evidențiază progresul în domeniul geneticii epilepsiei și discută modul în care arhitectura genetică a epilepsiilor comune începe treptat să fie dezvăluită. Material și Metode. Studii relevante au fost căutate folosind cuvintele-cheie: genetica epilepsiei, secvențierea de nouă generație (NGS), secvențierea întregului exom (WES) - în bazele de date PubMed și Google Scholar. Un studiu a fost inclus dacă au corespuns cel puțin 2 dintre cuvintele-cheie. Rezultate. De la constatarea în 1995 a mutației în gena CHRNA4, peste 500 de gene au fost descrise având un rol în epilepsie. Până în prezent, rata diagnosticului în WES variază de la 12, 5% în epilepsiile focale la 33% în cohortele generale. Cazurile rămase nediagnosticate sunt probabil multifactoriale. Cu toate acestea, NGS lansează noi provocări, care încă necesită să fie rezolvate. Concluzii. În ultimul deceniu, WES a fost utilizată tot mai mult pentru a descoperi rolul secvențelor codificatoare din genomul uman, fiind o premisă indispensabilă pentru abordarea personalizată și reducerea „odiseei diagnostice” a pacientului cu epilepsie