Single stranded DNA re-synthesis at uncapped telomeres requires replication polymerases

PhD ThesisTelomeres are specialised DNA-protein structures capping or protecting the chromosome ends fromshortening,degradation and fusions. Telomere uncapping occurs when some of the proteins associated with telomeres lose their integrity. For example in yeast a point mutation in the gene encoding the telomere binding protein Cdc13called cdc13-1,leads to conditional telomere uncapping at temperatures above 26º C. In this thesis I have utilised the cdc13-1model system to study repair after telomere uncapping. De-protection of the telomere triggers resection of the AC rich strand in 5’ to 3’ direction and formation of single stranded DNA(ssDNA). Checkpoint proteins are readily recruited to the damage and halt the cell cycle. However nossDNA re-synthesishas been observed in cdc13-1cells with uncapped telomeres. Here I will show that the ssDNA damage in cdc13-1cells recruits polymerase α,εandδand the clamp PCNA but in normal circumstancesefficientrepair is not observed. Only when telomeres are recapped the ssDNA could be re-synthesisedand this depended on the polymerase δsubunit Pol32 but did not require the non-essential subunits Dpb3 and Dpb4from polymerase ε. Interestingly, ssDNA re-synthesisat uncapped telomeres could be stimulatedthroughmild osmotic pressureand required both polymerase δand ε. Furthermore mild osmotic pressurecould also rescue cells damaged with methyl methanesulfonate but not with UV light or hydroxyurea. My data suggeststhat single stranded DNA re-synthesis may bespecifically inhibited or compete with resection when telomeres are uncapped and that osmotic pressurestimulated re-synthesisby regulating polymerases α, εand/or δ.Laura Maringel

INFLUENCE OF CALCIUM SALT SOLUTIONS ON DIFFERENT CONCENTRATIONS OF THE PHYSICAL AND CHEMICAL QUALITY OF FERMENTED MILK PRODUCTS

В статті розглянуто вплив розчинів кальцій хлориду різної концентрації на основні фізико-хімічні показники якості знезжиреного домашнього коров’ячого молока. Експериментально було підтверджено, що основні показники якості молока змінюються продовж 3-х діб в залежності від наявності в пробі 0,3 %-ого і 3 %-ого розчинів CaCl2. В результаті експериментальних досліджень виявлено, що при додаванні 3 %-ого розчину кальцій хлориду уповільнюється скисання молока в 1,5 рази, в порівнянні з контролем. При збільшенні титруємої кислотності молока зменшується його активна кислотність. При додаванні водних розчинів солей кальцію спостерігається збільшення умовної в’язкості продукту.  Виявлено, що на 2-у добу досліджень найбільший вміст казеїну (у %) спостерігається при додаванні 3 %-ого розчину CaCl2 в порівнянні з контролем (5,15±1,42 %, 3,01±0,95 % відповідно).В статье рассмотрено влияние растворов кальций хлорида различной концентрации на основные физико-химические показатели качества обезжиреного домашнего коровьего молока. Экспериментально было подтверждено, что основные показатели качества молока изменяются течение 3-х суток в зависимости от наличия в пробе 0,3 %-ого и 3 %-ого растворов CaCl2. В результате экспериментальных исследований установлено, что при добавлении 3 %-ого раствора кальций хлорида замедляется скисание молока в 1,5 раза, по сравнению с контролем. При увеличении титруемой кислотности молока уменьшается его активная кислотность. При добавлении водных растворов солей кальция наблюдается увеличение вязкости продукта. Выявлено, что на вторые сутки исследований наибольшее содержание казеина (в %) наблюдается при добавлении 3%-ого раствора CaCl2 по сравнению с контролем (5,15±1,42 %, 3,01±0,95 % соответственно).In the article the influence of calcium chloride solutions on different concentrations on basic physical and chemical indicators of cow’s milk quality. It has been experimentally confirmed that the main indicators of milk quality change cont in 3 days depending on the presence in the sample  the 0,3 % and 3 % solution CaCl2. As a result of experimental studies found that the addition of 3 % solution of calcium chloride slows sour of milk in 1,5 times, compared with the control. With increasing acidity of milk tytrometicaly reduced its active acidity. When adding aqueous solutions of calcium, an increase the viscosity of the product, found thaton the second day the largest research casein content (in %) observed when adding 3 % solution CaCl2 compared with controls (5,15±1,42%, 3,01±0,95 %, respectively)

OBSTETRIC CARE IN THE HOME ENVIRONMENT FOR WOMEN WITH ONCOGYNECOLOGICAL DISEASE

Introduction: Every year, about 12 million patients with malignant tumors are diagnosed in the world, and every day about 20,000 people die from cancer. Although modern medical oncology has the means for early diagnosis and successful definitive treatment, this diagnosis causes panic and a sense ofdoom in the patient.Objective: To explore the opinion of women with oncological-gynecological diseases, regarding obstetric care in the home environment.Materials and methods: An anonymous survey was conducted for the period April 2022 - May 2023. 74 women with oncological and gynecological diseases hospitalized in the GynecologyDepartments of the Complex Oncology Center and MBAL - Shumen AD were surveyed.Results and discussion: Midwifery care in a home environment can provide greater patient comfort. They can be treated and cared for in their own home where they feel more relaxed and comfortable.Conclusions: Home obstetric care can be an effective way to manage onco-gynecological diseases and improve patients' quality of life. To ensure that patients' medical needs are met and that they receive appropriate midwifery support care

Role of the patronage midwives in strengthening maternal and child health

Introduction: patronage activity is based on the methodology developed in accordance with the Law on Child Protection and its Implementing Regulations, the Health Act, the Hospitals Act, Regulation №26 from 14.06.2007g. providing obstetric care to women without health insurance, Ordinance №1 from 08.02.2011g. Professional activities that nurses, midwives, associate medical professionals and health assistants may perform assignment or independently.Objective: To investigate the credibility of pregnant women and mothers of the Center for Maternal and Child Health gr. Shumen to the activities of patronage midwife.Materials and methods: An anonymous survey during February and March 2016 were interviewed 60 pregnant women and mothers with children up to three years by the Centre for Maternal and Child Health gr. Shumen (municipalities of Novi Pazar, Kaolinovo and Nicolas Kozlevo) . The questionnaire contains 15 questions developed specifically for this purpose.Results and discussion: Patients express confidence in the information (91.7%) they receive from midwives and prefer this communication because of its proximity that feel to it. Respondents expect the midwife to look a certain way and if it does not meet those expectations, they react with disbelief and to its professionalism. Rated demonstrations of respondents who receive nursing, nutrition (93.3%). The most important qualities that must have been the midwife responsibility (83.3%), politeness, which would lead to better communication (75 %) and tact (69.3%). Among the qualities you would not want to own midwife, respondents indicated lack of compassion, indifference and lack of professionalism.= The midwife is what their behavior should submit serenity, calmness›s good nature, to support mothers and pregnant. In this sense, the analysis of the survey results can be used as guidelines for potential development of personal qualities and professional skills of the midwife to win the confidence of pregnant women and mothers.Conclusions: The professional skills of the midwife is impossible to use without specific personal qualities of her. They have enormous significance and impact on the quality of the consultation process. Undoubtedly, the success of the consulting midwife builds on the qualities of the individual, supported by the best professional skills

Identification of nucleotide variation of growth hormone gene in rabbit populations reared in Bulgaria

[EN] Five rabbit populations of New Zealand White (NZW), Californian (CAL), crossbred NZW×GW and two generations of the synthetic population – SPF1 and SPF2 reared in Bulgaria were included in the present study with the aim of detecting the genetic variability of the growth hormone encoding gene (GH) via polymerase chain reaction with the restriction fragment length polymorphism analysis and direct sequencing. The targeted region of the rabbit GH gene was amplified and a fragment of a total of 231 bp was obtained in all studied populations. Allele identification was determined after enzymatic digestion, where two fragments of 62 and 169 bp correspond to allele C and an undigested fragment of 231 bp corresponds to allele T. Two additional bands of 107 and 124 bp evidenced A/G genetic polymorphism in the rabbit GH gene. Thirtyeight percent of the studied rabbits were carriers of the double mutation (C/T+A/G) in the same locus as the studied GH gene. The sequence analysis revealed two nucleotide substitutions – g.111C>T and g.156A>G in the non-coding region between the regulatory TATA box and 5’ UTR region, and a novel g.255G>A genetic variant in intron 1 of GH gene. The A>G transition was most frequent (40.57%), compared to the other ones, G>A (28.57%) and C>T (10.80%), respectively. The most frequent genotype in the NZW population was homozygous TT (0.93), with a prevalence of the T allele (0.97) over allele C (0.03) for g.111C>T SNP site. The distribution of the allele and genotype frequencies at the sites g.156A>G and g.255G>A in this rabbit group was identical, with the highest value of 0.93 for alleles A and G, respectively. The rabbit populations CAL and NZW×GW showed equal frequencies of the prevalent T allele (0.83) and for homozygous TT genotype (0.67) according to g.111C>T SNP. The highest values were obtained for the allele А (0.83) and for homozygous AA genotype (0.67) at c.33A>G SNP in these rabbit groups. The highest values (0.67, 0.60 and 0.80) for the heterozygous genotypes at g.111C>T, g.156A>G and g.255G>A SNPs, respectively, were detected among the SPF2 rabbit population, compared to the both homozygous genotypes. The results obtained in the present research indicates a significant degree of genetic variability of the studied polymorphic GH locus in the SPF2 rabbit group.Gencheva, DG.; Koynarski, TV.; Dafova, V.; Tanchev, SG. (2021). Identification of nucleotide variation of growth hormone gene in rabbit populations reared in Bulgaria. World Rabbit Science. 29(1):19-29. https://doi.org/10.4995/wrs.2021.12693OJS1929291Abdel-Kafy E., Hussein B., Abdel-Ghany S., El-Din A., Badawi Y. 2015. Single nucleotide polymorphisms in growth hormone gene are associated with some performance traits in rabbit. Int. J. Biol. Pharm. Allied Sci., 4: 490-504.Abdel-Kafy, E., Darwish, S., Elkhishin, D. 2016. Correlating single nucleotide polymorphisms in the myostatin gene with performance traits in rabbit. World Rabbit Sci., 24: 213-221. https://doi.org/10.4995/wrs.2016.4026Amalianingsih T., Brahmantiyo B. 2014. The variability of growth hormone gene associated with ultrasound imaging of longissimus dorsi muscle and perirenal fat in rabbits. Media Peternakan, 37: 1-7. https://doi.org/10.5398/medpet.2014.37.1.1Dimitrova I., Dimitrov T., Teneva A., Tzvetkova H. 2008. Rabbit production in Bulgaria. Biotechnol. Anim. Husbandry, 24: 149-154. https://doi.org/10.2298/BAH0802149DEl-Sabrout K., Aggag S. A. 2017. Associations between single nucleotide polymorphisms in multiple candidate genes and body weight in rabbits. Vet. World, 10: 136. https://doi.org/10.14202/vetworld.2017.136-139El-Sabrout K., Aggag S., de Souza Jr J.B.F. 2019. Some recent applications of rabbit biotechnology - a review. Anim. Biotechnol., 1-5. https://doi.org/10.1080/10495398.2018.1539005Fontanesi L., Dall'Olio S., Spaccapaniccia E., Scotti E., Fornasini D., Frabetti A., Russo V. 2012. A single nucleotide polymorphism in the rabbit growth hormone (GH1) gene is associated with market weight in a commercial rabbit population. Livest. Sci., 147: 84-88. https://doi.org/10.1016/j.livsci.2012.04.006Fontanesi L., Tazzoli M., Scotti E., Russo V. 2008. Analysis of candidate genes for meat production traits in domestic rabbit breeds. In Proc.: 9th World Rabbit Congress, June 10-13, 2008, Verona, Italy, 79-84.Gencheva D., Georgieva S., Velikov K., Koynarski T., Tanchev S. 2017. Single nucleotide polymorphism of the Growth Hormone Receptor (GHR ) encoding gene in Oryctolagus cuniculus. J. BioSci. Biotechnol., 6: 197-201.Heracle BioSoft. 2013. DNA Sequence Assembler v.4. https://www.DnaBaser.comHristova D.G., Tanchev S.G., Velikov K.P., Gonchev P.G., Georgieva S.J. 2018. Single nucleotide polymorphism of the growth hormone (GH ) encoding gene in inbred and outbred domestic rabbits. World Rabbit Sci., 26: 49-55. https://doi.org/10.4995/wrs.2018.7211Hristova D., Tanchev S., Velikov K., Gonchev P., Georgieva S. 2017. Rabbit growth hormone and myostatin gene polymorphisms. J. Agr. Res., 2: 000133. https://doi.org/10.23880/OAJAR-16000133Hussein B., Abdel-Kafy E.M., Abdel-Ghany S.M., Gamal A.Y., Badawi Y.M. 2015. Single nucleotide polymorphism in growth hormone gene are associated with some performance traits in rabbit. Int. J. Biol. Pharm. Allied Sci., 4: 490-504.Kumar S., Stecher G., Tamura K. 2016. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol. Biol. Evol., 33: 1870-1874. https://doi.org/10.1093/molbev/msw054Migdal L., Palka S., Kmiecik M., Derewicka O. 2019. Association of polymorphisms in the GH and GHR genes with growth and carcass traits in rabbits (Oryctolagus cuniculus). Czech J. Anim. Sci., 64: 255-264. https://doi.org/10.17221/27/2019-CJASNei M. 1973. Analysis of gene diversity in subdivided populations. Proc. Nat. Acad. Sci. USA, 70: 3321-3323. https://doi.org/10.1073/pnas.70.12.3321Sahwan F.M., El-Sheik, A.I., Sharaf, M.M., El-Nahas, A.F. 2014. Genetic polymorphism in growth hormone receptor gene (GHR) and its relationship with growth trait in pure and hybrid rabbit breeds. Alexandria J. Vet. Sci., 43: 45-51. https://doi.org/10.5455/ajvs.165197Stothard P. 2006. Sequence Extractor. https://www.bioinformatics. org/seqext/Tamura K., Nei M. 1993. Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol. Biol. Evol., 10: 512-526. https://doi.org/10.1093/oxfordjournals.molbev.a040023Thompson J.D., Higgins D.G., Gibson T.J. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res., 22: 4673-4680. https://doi.org/10.1093/nar/22.22.4673Wallis C., Wallis M. 1995. Cloning and characterisation of the rabbit growth hormone-encoding gene. Gene, 163: 253-256. https://doi.org/10.1016/0378-1119(95)00429-AYeh F., Yong R. 1999. POPGENE version 1.31 (02.04.2011). Microsoft based Freeware for Population Genetic Analysis. University of Alberta, Edmonton, Canada, http://www.ualberta.ca/~fyeh/fyehZaghloul A. R.; Khalil, M.H.; Iraqi, M. M.; Ramadan, Sh. and EL Nagar, A. G. 2019. Crossbreeding effects and polymorphic associations of genotypes of GH gene with growth traits in rabbits. Egyptian Journal of Rabbit Science, 29: 149-169. https://doi.org/10.21608/ejrs.2019.81100Zhang W.X., Zhang G.W., Peng J., Lai S.J. 2012. The polymorphism of GHR gene associated with the growth and carcass traits in three rabbit breeds. In Proc.: 10th World Rabbit Congress, Sharm El-Sheikh, Egypt, 75-78

Single stranded DNA re-synthesis at uncapped telomeres requires replication polymerases

PhD ThesisTelomeres are specialised DNA-protein structures capping or protecting the chromosome ends fromshortening,degradation and fusions. Telomere uncapping occurs when some of the proteins associated with telomeres lose their integrity. For example in yeast a point mutation in the gene encoding the telomere binding protein Cdc13called cdc13-1,leads to conditional telomere uncapping at temperatures above 26º C. In this thesis I have utilised the cdc13-1model system to study repair after telomere uncapping. De-protection of the telomere triggers resection of the AC rich strand in 5’ to 3’ direction and formation of single stranded DNA(ssDNA). Checkpoint proteins are readily recruited to the damage and halt the cell cycle. However nossDNA re-synthesishas been observed in cdc13-1cells with uncapped telomeres. Here I will show that the ssDNA damage in cdc13-1cells recruits polymerase α,εandδand the clamp PCNA but in normal circumstancesefficientrepair is not observed. Only when telomeres are recapped the ssDNA could be re-synthesisedand this depended on the polymerase δsubunit Pol32 but did not require the non-essential subunits Dpb3 and Dpb4from polymerase ε. Interestingly, ssDNA re-synthesisat uncapped telomeres could be stimulatedthroughmild osmotic pressureand required both polymerase δand ε. Furthermore mild osmotic pressurecould also rescue cells damaged with methyl methanesulfonate but not with UV light or hydroxyurea. My data suggeststhat single stranded DNA re-synthesis may bespecifically inhibited or compete with resection when telomeres are uncapped and that osmotic pressurestimulated re-synthesisby regulating polymerases α, εand/or δ.Laura Maringel

Need for providing health information midwives - opinion of expert from Family consultation center in Shoumen

For the prevention of abandonment and neglect of children to change attitudes in communities at risk and supporting families are created and operate family-counseling centers (existence SCC). In the district of Shumen are created three family-counseling centers: Shumen, Novi Pazar, Veliki Preslav exported workplace - affiliate in Varbitsa Municipality. Existence SCC represent innovative cross-sector service that combines a set of integrated social, health, educational services, measures and activities for prevention and support of children and families at risk with dominant mobile component to cover the most vulnerable groups living in separate quarters or isolated settlements in the municipality and region. Services and activities of existence SCC associated with search and targeting pregnant women and mothers at risk to the health system, which in the opinion of the experts (75%) is an important part of their business. Counseling and support for pregnant women takes place in the `Pregnancy at risk` aimed at developing parental capacity and increase health literacy in communities at risk.The attitude of specialists existence SCC accept midwife as part of the team is most convincing in smaller cities: Veliki Preslav (75%) and Novi Pazar (71%) and less in the big city Shumen (64%)

Crystal structure of all-cis-2,4,6-trihydroxycyclohexane- 1,3,5-triaminium chloride sulfate, C6H18ClN3O7S

C6H18ClN3O7S, trigonal, P31c (no. 159), a = 8.3990(14) Å, b = 8.3990(14)∘ , c = 9.6208(17) Å, V = 587.76(17) Å3 , Z = 2, Rgt(F) = 0.0497, wRref(F2 ) = 0.1404, T = 300.15 K