Background: In developed or developing countries, the most common cancer in women is breast cancer with a pick in 40–50 years in Asia. Herein, we compared the association between IHC with FISH in HER2-positive breast cancer patients and affection of trastuzumab on disease free survival and overall survival (OS).Subjects and Methods: Immunohistochemical (IHC) analysis of hormone receptors and HER2 was performed in 133 patients with breast cancer between 2003 and 2014. Patients were selected for Herceptin adjuvant treatment, according to IHC 3+ or FISH+. The specimens for pathology reports were fixed at 10% neutral-buffered formalin (pH=7.4) for 24 hours, then sliced into 4 μm sections.Results: The mean age of patients at diagnosis was 46.39 years , 100% female. Concordance rates between IHC and FISH were 31.1% for IHC 2+ and 84.1% for IHC 3+ (p<0.001). The 87 patients had age ≤50 years and 46 patients had >50 years. Of the 133 patients, 30 patients (22.6%) had metastasis and 72 (54.1%) had right involvement. Ninety three (69.9%) patients had lymph node invasion. 48 patients (36.1%) were treated with trastuzumab and 85 (63.9%) were treated without trsastuzumab. The 10-year survival rate was 70% and the mean survival was 49 months.Conclusion: We recommend clinicians that FISH analysis is as a predictor in breast cancer patients with IHC score 2+. In contrast, FISH analysis of IHC 3+ samples was no useful. Trastuzumab therapy is effective and tolerated for breast cancer with IHC 3+ and probably IHC 2+/FISH+. This study was approved by Ethics Committee of Kermanshah University of Medical Sciences, Kermanshah, Iran . Between 2003 and 2014, 133 breast cancer patients with HER2-neu positive in IHC test were referred to our clinic. The patients were categorized on the basis of IHC values as +2 or +3. Only patients with IHC 3+ and FISH+ were selected for herceptin adjuvant treatment and were centrally reviewed ER, PR and HER2 copy numbers. The magnitude of trastuzumab benefit was assessed using the Cox proportional hazards model for the DFS and the OS. The specimens for pathology reports were fixed at 10% neutral-buffered formalin (pH=7.4) for 24 hours, then sliced into 4-μm sections. Her-2 protein expression was measured using a commercial available S-P kit. FISH for Her-2 gene amplification was performed in laboratory of Taleghani Hospital using a commercial available double-color probe. Monoclonal antibodies against ER, PR, HER2, as well as IHC kit were purchased from Pars Azmon Co. Equivalent phosphate-buffered saline (PBS) was used as a negative control for primary antibodies.Statistical analysis The OS was calculated using Kaplan-Meier method and the DFS and the OS with log-rank test. The OS is defined as time from randomization until death from any cause or endpoint with minimum two years follow-up. Statistical analysis was performed with IBM SPSS software version 19 and the enumeration data were compared with Fisher’s exact test. P<0.05 was considered to indicate a statistically significant difference (95% CI). The DFS is defined as the time from treatment with trastuzumab until recurrence of tumor or death (for 3 years). In our study, the mean age of women with breast cancer at diagnosis was 46.39 ± 10.81 years  . Among 1The concordance rate is defined as the number of agreed to IHC 2+ and 3+ cases divided by the total number of IHC 2+ and 3+ cases. Also, the discordance rate is defined as the number of discrepant to IHC 2+ and 3+ cases (IHC 2+ or 3+ but Her-2 FISH negative) divided by the total number of IHC 2+ and 3+ cases. Concordance rates were 31.1% for IHC 2+ and 84.1% for IHC 3+. In other hand, discordance rates were 68.9% for IHC 2+ and 15.9% for IHC 3+  .Of 133 patients, 87 patients had age ≤50 years that 50 of them were ER positive, 43 were PR positive, 23 had IHC 2+ and 64 had IHC 3+. In the other hand, 46 patients had >50 years that 27 patients were ER positive, 30 were PR positive, 22 had IHC2+ and 24 had IHC3+(p>0.05) .Among 133 patients, 48 patients (36.1%) were treated with trastuzumab and 85 (63.9%) were treated without trastuzumab. Furthermore, in the group of 29 patients who had IHC positive that were treated with trastuzumab for 3 years, one patient died, however in the group of 29 patients who never received trastuzumab, 8 patient died.10 It is a leading cause of death in women. Adjuvant chemotherapy, commonly include alkylating agents and anthracyclines, improves survival rate in treated breast cancer.11 A study on 231 patients with breast cancer,12 showed that the mean age the patients was 45 years and also another study indicated that the mean age in the patients was 48.30 years.13 Breast cancer is the most common malignancy among women.14 Therefore, the need for accurate detection of the Her2 alteration has now become even more important, because therapeutic decisions for patients with breast cancer are increasingly dependent on this information.9,15 This study evaluated the results of IHC and FISH among 133 patients with HER2 positive. There was an association between IHC 3+ and FISH. However, there was a discordance rate between IHC 2+ and FISH. This was similar to another study9 that reported FISH-positive and-negative rate for IHC2+ were 36 and 64%, respectively; 91 and 9% for IHC3+, respectively. This was statistically significant (p<0.01). A few studies,16-18 showed an association between IHC 3+ and no association between IHC 2+ with FISH.In our study the mean age was 46.39 years and was similar to results of other studies. HER2, a proto-oncogene, also known as c-erbB-2 or HER2/neu, located on chromosome 17q21, is considered to be closely associated with the occurrence and development of breast cancer and also IHC detection has become essential to many malignancies and plays a key role in tumor diagnosis, treatment and prognostic assessment.19 and Park et al.20 showed an association between IHC 2+ and FISH. Therefore, an agreement between IHC and FISH is still controversial. The concordances in the cases of IHC 3+ are similar but there is a difference in the cases of IHC 2+. This difference may be due to the different sensitivity and specificity of the antibodies and probe used,19 or because of subjective interpretative variation in reporting, variable pre-analytical conditions, imperfect IHC methodology.18 ER and PR have important prognostic role in breast cancer and they also dictate the endocrine treatment, and important prognostic markers.14 Our study showed that in all of the patients, the positive rates of ER and PR were 54.8% and 61.6%, but results weren’t statistically significant (p>0.05). Her2+ reduces affection of treatment in the patients of ER and PR positive.Sui et al.14,21-23 revealed the relationship of expression between subtypes  with age. ER and PR-high expressions were observed in female who were 50 or younger,14,24 such as this study. HER2-high expression was in female with >50 years,14 but we showed that it was in female ≤50 years such as result of Shomaf et al.25 There were no significant differences between the subtypes regarding age in present study. Trastuzumab was a humanized monoclonal antibody directed against the HER2/neu oncoprotein and has the ability to inhibit tumor growth in breast cancer patients overexpressing HER2.26 The mean survival of 48 treated patients with trastuzumab and 85 patients without it were 27.8 and 20.8 months. The five-year survival rate was 74.71% for patients with IHC 2+ and IHC 3+ that is favorable in comparison with previously reported series.27 The 10-year survival rate for our patients with IHC 2+ and IHC 3+, was 70% with mean survival 49 months. These results showed that the OS in our patients is better than other studies. The three-year DFS between two subgroups was statistically significant (p=0.006). The DFS rate for patients treating with trastuzumab and without trastuzumab was 93.3% and 76.6%, respectively. Therefore, trastuzumab therapy improves the survival of HER2-positive breast cancer patients.Several studies We advise clinicians that FISH analysis is a predictive factor in breast cancer patients with IHC score 2+. In contrast, FISH analysis of score 3+ samples was not useful. Also, trastuzumab therapy is effective and tolerated for breast cancer with IHC 3+ and probably IHC 2+/FISH+.


Pleurotus eous strain P-31 on the growth and yield performance of pepper and tomato seedlings under greenhouse conditions. Sandy loam soil was combined with different percentages of SMC to obtain the following combinations  %. Lower concentrations SMC5, SMC10 and SMC15 promoted vegetative growth  of the two test plants. Tomato seedlings grown in SMC10 recorded the highest plant height (50.3 ± 7.2cm); leaf area (378.8 ± 1.2cm2); number of floral buds (51) and flowers (28) whereas SMC5 recorded the highest chlorophyll content 34.1 ± 0.9CCI though SMC15 recorded the highest number of leaves (8). Tomato seedlings grown in SMC30 produced both the maximum number of fruits (8) with corresponding high weight (34.2 ± 7.7g). Pepper seedlings grown in lower concentrations (SMC5–15) recorded the highest plant heights (29.8–30.8cm), chlorophyll content (20.3CCI) and leaf area (53.5–66.2 cm2). Although the different combinations of sandy loam soil and SMC did not significantly (p ≥ 0.05) affect the number of axillary branches developed; different combinations significantly (p ≤ 0.05) affected the number of floral bud, flower and fruit, weight of fruits formed and value of each of these increased with increasing percentage of SMC. Pepper seedlings grown on SMC30 recorded the maximum number of floral buds (32.0 ± 3.6), number of flowers (19.4 ± 1.3), number of fruits (10.8 ± 1.2) and weight of fruits (31.9 ± 3.4g). Tomato seedlings raised on SMC100 (spent mushroom compost only) and soil only did not significantly (p ≥ 0.05) differ from each other however, was statistically significant (p ≤ 0.05) from amended sandy loam soil by all criteria investigated. The study shows that SMC provide favourable soil conditioners for the cultivation of fruits, vegetables and foliage crops as it improved growth and yield of tomato and pepper seedlings.The objective of this study was to investigate the influence of spent mushroom compost of  Throughout the world, mushrooms are the most popular, delicious, nutritious and medicinal vegetables considered as the most promising for crop diversification. This is due their flavour, texture and richness in proteins, minerals, vitamins, low levels of calories with an appreciable amount of unsaturated fats. Mushrooms also accumulate secondary metabolites such as phenolic compounds, polypetides, terpenes, and steroids . Mushrooet al. 2014; Apart from using Spent Mushroom Compost as feeding material for vermicomposting, plant disease management, preparation of organic-mineral fertilizer and bioremediation of contaminated soils . The unsCapsicum annuum L. var. Legon 18) and Tomato (Solanum lycopersicon Fr. Lycopersicon esculentum Mill. var. Wosowoso) seeds were obtained from the Department of Crop Science, School of Agriculture, University of Ghana.This study was carried out in the greenhouse at the Department of Plant and Environmental Biology, University of Ghana Legon. Potted plastic buckets of dimensions 25 cm and 22 cm diameter were used to pot the test seedlings. Pepper  cultured in polyethylene bags on rice straw after a cropping period of 8 weeks. The bags were obtained from the Mycology Unit of the Food Research Institute of the Council for Scientific and Industrial Research, Ghana. Sandy loam soil used for this present study was collected from the University’s Farm in the Legon Botanical Gardens University of Ghana, Legon. The soil sample was taken to the Department of Soil Science, School of Agriculture for authentication, identification and elemental nutrient analysis.Spent mushroom compost (SMC) derived from mushroom (Capsicum annuum L. var. Legon 18) and Tomato (Solanum lycopersicon Mil. var. Wosowoso) were used as test plants because they are among the plants most preferred for cultivation in backyard gardens and commercial farms in Ghana. To study the effect of the spent mushroom compost (SMC) on vegetative growth of the two crops, a modified method of ; SMC5 (5% of SMC to 95% soil); SMC10 (10% of SMC to 90% soil); SMC15 (15% of SMC to 85% soil); SMC20 (20% of SMC to 80% soil); SMC25 (25% of SMC to 75% soil) and SMC30 (30% of SMC to 70% soil).Pepper  6.5 kg for each plastic bucket. Thoroughly mixed of SMC and sandy loam soil the growth medium in each plastic bucket was irrigated with 1 l running tap water. Three weeks old seedlings of either pepper or tomato from a wooden tray were transplanted into each plastic bucket. (5 seedlings per bucket). The potted plants in the plastic buckets were arranged in a randomised complete-block design (RCBD) with five replicates for each of the treatments afore-mentioned above. Five replicates plastic potting buckets of the same size and dimension each was filled with either 6.5 kg SMC, or soil medium served as controls for comparison purposes. Soil: SMC combinations/mixtures served as treated medium. Seedlings were watered every other day with equal volumes of 500 ml of tap water. The following parameters were recorded on weekly intervals: plant height, number of leaves, floral buds, flowers, fruit, axillary branches, leaf area, and chlorophyll content of leaves, stem girth and no. of fruits and weight of fruits.SMC in the following ratios SMCThis was determined after 8 weeks of transplanting seedlings using a chlorophyll meter (Optic Sciences CCM-200 plus Model USA) by attaching the chlorophyll meter knob to five randomly selected leaves of equal age and size and average chlorophyll recorded as Chlorophyll Content Index (CCI).2 was determined using a digital leaf area meter (LI-3000C) via attaching five randomly selected leaves of same age and size.The number of leaves formed was counted manually per week during the 12 weeks growing period. Leaf area in mmThis was estimated by using a meter rule placed firmly on the surface of substrate to determine the height of the plant in centimeters.TM, Radiometer Analytical A/S Copenhagen, Denmark).Moisture content of loamy soil and spent mushroom compost served as controls was determined using the conventional gravimetric method. 1 g of the sample was weighed in a can using a Digital Computer Scale . Drying was done in a hot oven  at 105°C for 3 h allowed to cool in a desiccator and was re-weighed. The cooled sample was returned to the oven for further drying and cooling until no further reduction in weight was obtained. The moisture was expressed as a percentage. The pH of the sandy loam soil and spent mushroom compost served as controls using a pHM92 pH meter  were dismembered into shoot and roots, placed in brown paper envelopes for each species before weighing each separately and placing them in an oven  at 100°C for 24 h and then re-weighed after cooling.4 dilutions. One millitre aliquots of the serially diluted 1 g per 100 ml of 0.1% peptone water was mixed with either 20 ml of Cooke’s medium or Dichloran Rose Bengal Chloramphenicol Agar (DRBC) in 9.0 cm sterile petri plates and the plates swirled to obtain uniform distribution of spores. There were three replicates at each dilution level. The plates were incubated at 28 ± 2°C for 7 days after the total number of colonies were calculated in log10 CFU/g sample. The species of fungi appearing were identified according to their colour, morphological cultural characteristics as outlined by This was determined using the decimal serial dilution technique up to 1:10Percentage crude protein, hemicellulose, cellulose and lignin were determined by the conventional Kjeldahl’s method  and JameThe following elements  were estimated in the soil and SMC using the conventional methods . 250 mg of powdered air dried sample was weighed into a beaker was then placed in ignition muffle furnace  for drying at 400°C for 24 h. Five millitres of hydrochloric acid (HCl) was added to the sample and the solution was dried again; subsequently 5 ml of nitric acid (HNO3) was added. After evaporation, the sample was diluted to 50 ml with water. Sodium (Na) and calcium (Ca) content of the ashed sample were determined by flame photometer. K, P, Mg, Cu, Zn, Mn, Fe, and Pb were determined by Unicam 929 Atomic Absorption Spectrophotometer (AAS) (Model PinAAcle 900T). Nitrogen was estimated using the micro Kjeldahl method .Data obtained were subjected to statistical analyses using the Statistical Package for Social Sciences (SPSS version 20 for Windows). Significant differences between vegetative parameters measured among plants in treated media (soil: SMC) and controls media were determined using Duncan’s Multiple Range Test (DMRT) at 5% probability.5–30) were significantly (p < 0.03) higher when compared to that of the control (i.e. those grown in loamy soil only) (10 was the highest (69.9 cm)  ; the hei69.9 cm) .p > 0.05) in the height of pepper plants grown in sandy loam soil amended with SMC5–25 after 8 weeks of growth. Growth of pepper seedlings in unamended loamy soil (control) was the lowest (18.2 cm) and was statistically (p < 0.05) different from data obtained in the sandy loam soil medium amended with SMC5–30  were also significantly (p ≤ 0.05) larger when compared to the values of those grown in the control planting medium  also had significantly (p ≤ 0.05) higher effect on the growth/expansion of leaf area in the tomato seedlings  significant. The differences in the stimulation of leaf production by SMC5–20 were initially indistinguishable after 2–6 weeks and remained so until 8 weeks at least for concentration of SMC5–20 (2 in 8 weeks as compared to seedlings grown in pots with 15 and 20% SMC (2 (near 3 times). The differences observed after 8 weeks were statistically significant (p ≤ 0.05). Maximum net photosynthetic area of the leaf was obtained in plants grown in 10% SMC and was similar to what was obtained for 15 and 20% SMC. Data on leaf area of pepper seedlings grown in soils amended with 5%–30% SMC is presented in p > 0.05) after 2–4 weeks but was significantly different (p ≤ 0.05) after 8 weeks. The largest leaf area was obtained in soil amended with 5% SMC which was statistically (p ≤ 0.05) different from the leaf areas of plants grown in soil amended with 15% SMC. As the SMC: soil ratio increased the leaf area declined (2 was recorded in plants growing in the unamended native soil.Leaf formation by pepper plants grown in both control medium (soil only) and loamy soil amended with SMC 8 weeks . The dif SMC5–20 . Leaf ar 20% SMC  with a ldeclined . Interesp ≤ 0.05) decreased with increasing levels of SMC in soil in the following descending order 5% >10% >15% >20% >25% >30%. However, the lowest chlorophyll content recorded in soil amended with SMC was still superior statistically (p ≤ 0.05) to that of the control (p < 0.05) (Chlorophyll content of tomato plants significantly ( control . Chlorop < 0.05) .100) . Thereafter increased in dry weight of shoot did not correspond with increased dry weight of root (SMC15–30) p ≤ 0.05) than that obtained in loamy soil only (The lowest dry matter of tomato plant was obtained in the unamended soil which recorded shoot weight of 90.3 ± 5.9 g and roots weight of 45.3 ± 2.9 g . There w100) . On the oil only  & 2.10 CFU/g sample; in the SMC the fungi population estimated in the two-media varied from 4.3–4.5 log10 CFU/g. There was therefore no statistically significance difference (p > 0.05) between the mycoflora population recorded on the two media  varied between 4.2–4.3 logwo media .Aspergillus, Cladosporium, Fusarium, Mucor, Penicillium, Rhizopus, Rhodotorula, Trichoderma, Scopulariopsis) other yeasts and Mycelia sterilia were encountered in both samples  (The use of two isolation media enabled a wider spectrum of fungi to be detected. Eighteen (18) different fungi belonging to 9 genera ( samples . Aspergiesidents . A. flav (8%–9%) .The soil sample did not contain Cu and Fe, although Zn, Mn, Pb, Ca, Mg, Na, P, K and N  were detp ≤ 0.05) increase plant height and number of fruits produced by cucumber  and broccoli . Their results revealed that SMC and peat or SMC could be used as seedling media for both kale and broccoli. Lentinula subnudus as soil conditioner for cowpea and tomato. Their findings revealed that composted spent mushroom substrate mixed with loamy soil produced greater vegetative growth and yields of both vegetables than loamy soil (controls). Studies by Zea mays) in Nigeria.SMC is currently disposed of as waste and constitutes an environmental waste problem in Ghana. However, there can be economic benefit from its use in agriculture as bio-fertiliser, if properly harnessed. P. eous contained the following: 90.8 ± 1.79% dry matter, 7.76 ± 0.48% crude protein, 32.34 ± 1.97% cellulose, 7.26 ± 1.94% lignin, 53.3 ± 6.0% NDF and 50.42 ± 4.64% ADF  although it did not approximate the best growth in height in 10% SMC after 8 weeks  between the performances of the plants in terms of height, number of leaves and leaf area produced per plant (p ≤ 0.05) from pepper seedlings grown on SMC5–25. Albeit it performed better than seedlings grown in sandy loam soil only. Growth of pepper plants in soil only medium (control) was lower by all criteria than plants grown in soil media amended with (5–30) % SMC  % SMC . Total cCapsicum annuum L. and Solanum tuberosum L. by Recent studies shows the influence of SMC from oyster and button mushroom on growth yield of Lactuca sativa L) and showed that it provided the most adequate conditions for the growth and development of crisp head lettuce seedlings. P. ostreatus to cultivate okra (Abelmoscus esculentus Moench) and found that the growth and yield of okra (fresh and dry weight of fruits and number of okra fingers) was significantly enhanced. Data from this study suggest that spent mushroom compost of P. eous used as a soil conditioner to a certain level can be used as a bio-fertiliser for the enhancement of growth and yield of tomato and pepper at least under greenhouse conditions in Ghana.P. eous contained high dry matter (90.8 ± 1.79%), protein (7.76 ± 0.48%), cellulose (32.34 ± 1.97%), lignin (7.2 ± 1.94%), NDF (53.3 ± 6.07%), and ADF (50.42 ± 4.64%) rich enough for utilisation by the resident fungi for lignolytic, cellulolytic, pectinolytic, and proteolytic metabolic activities “in situ”  was the best suitable medium to support optimum vegetative growth of both test plants. These nutrients acted in concert to promote growth of the two plants in the greenhouse at least to a certain level of concentration of soil with SMC. The pH of the soil and spent mushroom compost were within the pH ranged of 6.6 and 6.8 which are well pH’s suited for growth of both tomato and pepper. Therefore, application and use of SMC in cultivation of these vegetables for field trial is highly recommended.Current findings indicate that both tomato and pepper seedlings raised on different concentrations of SMC  % amended with sandy loam soil supported vegetative growth  and fruit yield. SMC


PALB2, CHEK2 and ATM make it difficult to estimate precisely associated cancer risks. Population-based family studies have provided evidence that at least some of these mutations are associated with breast cancer risk as high as those associated with rare BRCA2 mutations. We aimed to estimate the relative risks associated with specific rare variants in PALB2, CHEK2 and ATM via a multicentre case-control study.The rarity of mutations in PALB2 c.1592delT, c.2816T>G and c.3113G>A, CHEK2 c.349A>G, c.538C>T, c.715G>A, c.1036C>T, c.1312G>T, and c.1343T>G and ATM c.7271T>G. We assessed associations with breast cancer risk (42 671 cases and 42 164 controls), as well as prostate (22 301 cases and 22 320 controls) and ovarian (14 542 cases and 23 491 controls) cancer risk, for each variant.We genotyped 10 rare mutations using the custom iCOGS array: PALB2 c.1592delT OR 3.44 , PALB2 c.3113G>A OR 4.21  and ATM c.7271T>G OR 11.0 . We also found evidence of association with breast cancer risk for three variants in CHEK2, c.349A>G OR 2.26 (95% CI 1.29 to 3.95), c.1036C>T OR 5.06 (95% CI 1.09 to 23.5) and c.538C>T OR 1.33 (95% CI 1.05 to 1.67) (p≤0.017). Evidence for prostate cancer risk was observed for CHEK2 c.1343T>G OR 3.03  for African men and CHEK2 c.1312G>T OR 2.21  for European men. No evidence of association with ovarian cancer was found for any of these variants.For European women, strong evidence of association with breast cancer risk was observed for This report adds to accumulating evidence that at least some variants in these genes are associated with an increased risk of breast cancer that is clinically important. PALB2, CHEK2 and ATM.The rapid introduction of massive parallel sequencing (MPS) into clinical genetics services is enabling the screening of multiple breast cancer susceptibility genes in one assay at reduced cost for women who are at increased risk of breast (and other) cancer. These gene panels now typically include the so-called ‘moderate-risk’ breast cancer susceptibility genes, including PALB2 c.1592delT (a Finnish founder mutation), PALB2 c.3113G>A and ATM c.7271T>G. These mutations have been estimated to be associated with a 40% (95% CI 17% to 77%), 91% (95% CI 44% to 100%) and 52% (95% CI 28% to 80%) cumulative risk of breast cancer to the age of 70 years, respectively.BRCA2: 45% (95% CI 31% to 56%).Estimation of the age-specific cumulative risk (penetrance) of breast cancer associated with specific mutations in these three genes has been limited to those that have been observed more frequently, such as  95% CI 2% to 80% Case-control studies provide an alternative approach to estimating cancer risks associated with specific variants. This design can estimate the relative risk directly, without making assumptions about the modifying effects of other risk factors. However, because these variants are rare, such studies need to be extremely large to provide precise estimates.PALB2, CHEK2 and ATM relate to protein truncating and splice-junction variants.et alATM, which is comparable to that for truncating variants. Specifically, ATM c.7271C>G has been associated with a more substantial breast cancer risk in several studies.et al,CHEK2 from similarly designed studies were 6.18 (95% CI 1.76 to 21.8) for rare protein-truncating and splice-junction variants and 8.75 (95% CI 1.06 to 72.2) for evolutionarily unlikely missense substitutions.11The clearest evidence for association, and the most precise breast cancer risk estimates, for rare variants in PALB2, CHEK2 and ATM could be associated with increased risk of cancers other than breast cancer, as has been observed for BRCA1 and BRCA2 and both ovarian and prostate cancers.PALB2 carriers, there is little evidence to support or refute the existence of such associations, although a few individually striking pedigrees have been observed.18–20It is plausible that monoallelic mutations in PALB2, CHEK2 or ATM. These included three rare variants in PALB2: the protein truncating variants c.1592delT (p.Leu531Cysfs)ATM c.7271T>G .22In this study we selected rare genetic variants on the basis that they had been observed in breast cancer candidate gene case-control screening projects involving Participants were drawn from studies participating in three consortia as follows:Breast Cancer Association Consortium (BCAC), involving a total of 48 studies: 37 of women from populations with predominantly European ancestry (42 671 cases and 42 164 controls), 9 of Asian women (5795 cases and 6624 controls) and 2 of African-American women (1046 cases and 932 controls). All cases had invasive breast cancer. The majority of studies were population-based or hospital-based case-control studies, but some studies of European women oversampled cases with a family history or with bilateral disease (see online The 10.1136/jmedgenet-2016-103839.supp1Supplementary dataProstate Cancer Association Group to Investigate Cancer Associated Alterations in the Genome (PRACTICAL) involving a total of 26 studies: 25 included men with European ancestry (22 301 cases and 22 320 controls) and 3 included African-American men (623 cases and 569 controls). The majority of studies were population-based or hospital-based case-control studies (see online The Ovarian Cancer Association Consortium (OCAC), involving a total of 46 studies. Some studies were case-only and their data were combined with case-control studies from the same geographical region (leaving 36 study groupings). Of these groupings, 33 included women from populations with predominantly European ancestry (16 287 cases (14 542 with invasive disease) and 23 491 controls), 25 included Asian women (813 cases (720 with invasive disease) and 1574 controls), 17 included African-American women (186 cases (150 with invasive disease) and 200 controls) and 29 included women of other ethnic origin (893 cases (709 with invasive disease) and 864 controls). The majority of studies were population-based or hospital-based case-control studies ATM We selected for genotyping 13 rare mutations that had been observed in population-based case-control mutation screening studies. These variants were PALB2 variants c.2323C>T (p.Gln775*), c.3116delA (p.Asn1039IIefs) and c.3549C>G (p.Tyr1183*) were unable to be designed for measurement on the custom Illumina iSelect genotyping array and were not considered further . p Values were determined by the likelihood ratio test comparing models with and without carrier status as a covariate. We also applied conditional logistic regression, defining risk sets by study, and found that this made no difference to the OR estimates, CIs or p values to two significant figures; since model convergence was a problem for this latter regression analysis, all subsequent analyses were based on unconditional logistic regression. For the main analyses of breast cancer risk in European women, we also included as covariates the first six principal components, together with a seventh component specific to one study (Leuven Multidisciplinary Breast Centre (LMBC)) for which there was substantial inflation not accounted for by the components derived from the analysis of all studies. Addition of further principal components did not reduce inflation further. Data from all breast cancer studies were included to assess statistical significance. Data from cases selected for inclusion based on personal or family history of breast cancer were excluded in order to obtain unbiased OR estimates for the general population of white European women (leaving 37 039 cases and 38 260 controls from 32 studies). Multiple testing was adjusted for using the Benjamini-Hochberg procedure to control the false discovery rate, with a significance threshold of 0.05.PALB2 c.1592delT (Leu531Cysfs) was only observed in 35 cases and 6 controls, all from four studies from Sweden and Finland (Helsinki Breast Cancer Study (HEBCS), Kuopio Breast Cancer Project (KBCP), Oulu Breast Cancer Study (OBCS) and Karolinska Mammography Project for Risk Prediction Breast Cancer (pKARMA); see online −5); the OR estimate was 4.52 (95% CI 1.90 to 10.8) based on all studies and 3.44 (95% CI 1.39 to 8.52) based on unselected cases and controls  was identified in 44 cases and 8 controls from nine BCAC studies. Only one carrier of the variant was of non-European origin. Strong evidence of association with breast cancer risk was observed (p=6.9×10−8), with an estimated OR of 5.93 (95% CI 2.77 to 12.7) based on all studies and 4.21 (95% CI 1.85 to 9.61) based on unselected cases and controls. There was no evidence of a differential association by ER status (p=0.15).PALB2 variants (c.1592delT or c.3113G>A) was 3.85 (95% CI 2.09 to 7.09).Based on unselected cases, the estimated OR associated with carrying either of these PALB2 c.2816T>G (p.Leu939Trp) was identified in 150 cases and 145 controls and there was no evidence of association with risk of breast cancer. There was no evidence of association with risk of prostate or ovarian cancer for any of the three PALB2 variants (see ants see  and 4.CHEK2 c.349A>G (p.Arg117Gly) was identified in 44 cases and 18 controls in studies participating in BCAC; all of these women were of European origin. We found evidence of association with breast cancer (p=0.003), with little change in the OR after excluding selected cases (OR 2.03 (95% CI 1.10 to 3.73)).CHEK2 c.538C>T (p.Arg180Cys) was identified in 158 breast cancer cases and 142 controls in studies of white Europeans. Evidence of association with breast cancer risk (p=0.016) was observed, with an unbiased OR estimate of 1.34 (95% CI 1.06 to 1.70). A consistent OR estimate was observed for Asian women, based on 45 case and 45 control carriers (OR 1.16 (95% CI 0.75 to 1.76)).CHEK2 c.715G>A (p.Glu239Lys) mutations were identified in 15 cases and 9 controls, all European women participating in BCAC and no evidence of association with risk of breast cancer was observed (p=0.21).CHEK2 c.1036C>T (p.Arg346Cys) was identified in nine cases from seven studies and two controls from two different studies in BCAC (neither control carrier was from a study that had case carriers), all of European origin. We found evidence of association with breast cancer risk (p=0.017) with reduced OR estimate of 3.39 (95% CI 0.68 to 16.9) after excluding selected cases.CHEK2 variants (CHEK2 c.349A>G (p.Arg117Gly); c.538C>T (p.Arg180Cys); c.715G>A (p.Glu239Lys) and c.1036C>T (p.Arg346Cys)) were found to be associated with an increased risk of prostate or ovarian cancer (CHEK2 variant c.1312G>T (p.Asp438Tyr) was not associated with risk of breast cancer for European women (p=0.91). Variant c.1343T>G (p.Ile448Ser) was not observed in any breast cancer cases of European or Asian origin. It was detected in 48 cases and 29 controls of African origin, giving weak evidence of association ). CHEK2 c.1312G>T (p.Asp438Tyr) was identified in 23 cases and 11 controls from PRACTICAL, all European, providing evidence of association with prostate cancer risk ). CHEK2 c.1343T>G (p.Ile448Ser) was observed in 35 cases and 11 controls, all African, participating in PRACTICAL and was also associated with an increased risk of prostate cancer ). There was no evidence that these CHEK2 variants were associated with risk of ovarian cancer  was identified in 12 cases and 1 control in studies participating in BCAC, all of European origin, giving evidence of association with breast cancer risk (p=0.0012). The OR estimate based on unselected studies was 11.0 (95% CI 1.42 to 85.7). There was no evidence of association of this variant with prostate or ovarian cancer risk see  and 4.24Gly wasrare variants in so-called ‘moderate-risk’ genes are associated with an increased risk of breast cancer that is of clinical relevance.The present report adds to an accumulating body of evidence that at least some PALB2, CHEK2 and ATM, variants generated from these new testing platforms is not being used in clinical genetics services due to lack of reliable estimates of the cancer risk associated with individual variants, or groups of variants, in each gene. Previous analyses have been largely based on selected families, relying on data on the segregation of the variant. The present study is by far the largest to take a case-control approach. Consistent with previous reports,PALB2 c.3113G>A (p.Trp1038*), PALB2 c.1592delT (p.Leu531Cysfs) and ATM c.7271T>G  were found to be associated with substantially increased risk of breast cancer all with associated relative risk estimates of 3.44 or greater.These findings are presented at a time when detailed information about variants in these genes is becoming more readily available via the translation of diagnostic genetic testing from Sanger sequencing-based testing platforms to MPS platforms that test panels of genes in single assays.PALB2 variants (c.1592delT and c.3113G<A) were consistent with each other and with estimates based on segregation analysis.PALB2 c.2816T>G (p.Leu939Trp), with an upper 95% confidence limit excluding an OR >1.5 which is notable given the Align-Grantham Variation Granthan Deviation (Align-GVGD) score and the observed impact on protein function.30The estimates for the two loss-of-function ATM c.7271T>G  was also consistent with that found by segregation analysis.ATM c.7271T>G  could be due to the reduction in kinase activity  observed for ATM p.Val2424Gly,32The estimate for PALB2 c.3113G>A and 1.9 for c.1592delT and c.3113G>A combined.In contrast, we found no evidence of an association with risk of prostate or ovarian cancer with any of these three variants: however, the confidence limits were wide; based on the upper 95% confidence limit we could exclude an OR of >1.4 for prostate cancer for the loss-of-function CHEK2. Two of these  and c.1036C>T (p.Arg346Cys)) had evidence of a significant impact on the protein based on in silico prediction. We proposed these variants for inclusion in the iCOGS design as they had been identified in 3/1242 cases and 1/1089 controls and 3/1242 cases and 0/1089 controls, respectively, in a population-based case-control mutation screening study of CHEK2.et al, estimated an OR of 8.75 (95% CI 1.06 to 72.2) for variants with an Align-GVGD score C65 (based on nine cases and one control). The current analysis provides confirmatory evidence of this association in a much larger sample (OR 2.18 (95% CI 1.23 to 3.85)) including 40 unselected case and 18 control carriers. The evidence that CHEK2 is a breast cancer susceptibility gene is largely based on studies of protein truncating variants, in particular CHEK2 1100delC.We analysed six rare missense variants in PALB2 c.1592delT, c.3113G>A and ATM c.7271T>G because these associations had previously been reported; our aim was to more precisely estimate the associated relative risks. All three associations with breast cancer risk reported for CHEK2 variants remained statistically significant after adjusting for the other tests conducted in relation to breast cancer risk, but not after correcting for all tests for all cancers. Nevertheless, the findings for CHEK2 c.349A>G and c.1036C>T confirmed those reported previously, although collectively. The association observed with CHEK2 c.538C>T requires independent replication.The p values reported above have not been adjusted for multiple testing. This was not considered appropriate for the associations with breast cancer risk of et al,PALB2 c.1592delT; c.3113G>A ATM c.7271T>G) would place the carrier in a high-risk group, especially if other risk factors, such as a family history, are present. The high level of breast cancer risk associated with PALB2 c.1592delT and c.3113G>A reported here is consistent with the penetrance estimate reported for a group of loss-of-function mutations in PALB2Do this approach and new data have an impact on clinical recommendations for women and families carrying these rare genetic variants? Although age-specific cumulate risks for cancer are more informative for genetic counselling and clinical management of carriers, our study provides information that is relevant to clinical recommendations. As discussed in Easton The consistency of the relative risk estimates with those derived through family based studies supports the hypothesis that these variants combine multiplicatively with other genetic loci and familial risk factors; this information is critical for deriving comprehensive risk models. Even with very large sample sizes such as those studied here, however, it is still only possible to derive individual risk estimates for a limited set of variants, and even for these variants the estimates are still imprecise. This internationally collaborative approach also has limited capacity to improve risk estimates for rare variants that are only observed in specific populations. Inevitably, therefore, risk models will depend on combining data across multiple variants, using improved in silico predictions and potentially biochemical/functional evidence to synthesise these estimates efficiently. It will also be necessary develop counselling and patient management strategies that can accommodate a multifactorial approach to variant classification.10.1136/jmedgenet-2016-103839.supp2Supplementary Financial Support


Reliable data on causes of death form the basis for building evidence on health policy, planning, monitoring, and evaluation. In Ethiopia, the majority of deaths occur at home and civil registration systems are not yet functional. The main objective of verbal autopsy (VA) is to describe the causes of death at the community or population level where civil registration and death certification systems are weak and where most people die at home without having had contact with the health system.Causes of death were classified and prepared based on the International Classification of Diseases (ICD-10). The cause of a death was ascertained based on an interview with next of kin or other caregivers using a standardized questionnaire that draws information on signs, symptoms, medical history, and circumstances preceding death. The cause of death, or the sequence of causes that led to death, is assigned based on the data collected by the questionnaire. The complete VA questionnaires were given to two blinded physicians and reviewed independently. A third physician was assigned to review the case when disagreements in diagnosis arose.Communicable diseases (519 deaths [48.0%]), non-communicable diseases (377 deaths [34.8%]), and external causes (113 deaths [10.4%]) were the main causes of death between 2007 and 2013. Of communicable diseases, tuberculosis (207 deaths [19.7%]), HIV/AIDS (96 deaths [8.9%]) and meningitis (76 deaths [7.0%]) were the most common causes of death.Tuberculosis, HIV/AIDS, and meningitis were the most common causes of deaths among adults. Death due to non-communicable diseases showed an increasing trend. Increasing community awareness of infections and their interrelationships, tuberculosis case finding, effective local TB programs, successful treatment, and interventions for HIV are supremely important. Data on causes of death are important for mapping the geographical burden of disease and for health sector planning, including assessing programmatic needs, monitoring progress of interventions, and reassessing health priorities. Cause of death data are crucial for understanding the overall epidemiological profile of disease in a population , 2.It is reported that more than two-thirds of deaths in the world occur in developing countries but causes of death are not well documented, and vital registration systems are nonexistent or at rudimentary stages , 4. ManyEthiopia is one of the SSA countries with no functional vital registration system yet. Additionally, health service utilization is very poor, with total outpatient use of government health facilities estimated at 0.3 visits per person per year . Thus, dCognizant of the lack of information on basic demographic and health outcomes, the Dabat Health and Demographic Surveillance System (HDSS) was established, with the purpose of, among other things, knowing community-level causes of death. This study therefore aimed to assess the causes of adult deaths in the population covered by the Dabat HDSS using VA data between 2007 and 2013.The Dabat HDSS study site is located in a rural part of the Amhara regional state in northwest Ethiopia. It is 850 km northwest of Addis Ababa, the capital city of Ethiopia. The altitude of the district ranges from about 1000 to 2500 and above meters above sea level. The population consists mainly of subsistence farmers  method.The VA data presented have been collected since 2007. Verbal autopsy is a method used to ascertain the cause of a death based on an interview with next of kin or other caregivers. The VA questionnaire prepared by World Health Organization (WHO) and INDEPTH  was used –12.A paper form platform questionnaire was adapted and translated into Amharic, the national language of Ethiopia. The cause of death, or the sequence of causes that led to death, is assigned based on the data collected through a questionnaire and any other available information. Eighteen diploma-graduated and trained and resident full-time enumerators visited the household every month for events and reported all deaths to the Dabat HDSS database. VA interviews were carried out after 45 days of the date of death in respect of the local mourning time. VA was conducted using a standardized World Health Organization (WHO) questionnaire endorsed by international Network for Demographic Evaluation of Populations and Their Health (INDEPTH network) for all deaths occurring in the HDSS . For thiThe data collectors received training on questionnaires, recording, contacting close relatives, and data collection procedures. The training curriculum included sessions about individual symptoms and description in local language for easy recognition by the respondents. The VA interviewer was informed about new deaths by the resident enumerators and conduct verbal autopsy interviews. Five trained supervisors were involved in data collection and did regular supervision by rechecking the data collected by enumerators. The research team also did regular supervision to recheck the progress of data collection and sort out problems that may have been encountered by enumerators. A total of nine trained physicians  were involved in assigning the causes of deaths. International Classification of Diseases (ICD-10) was used to classify main, underlying, and contributing causes of death following independent assessment of the completed questionnaires by two or three physicians .2) test. Binary logistic regression model was used to identify the relationship between the outcome variables  and the independent variables. Variables in the bivariable analysis with a P-value of <0.2 were entered into the multivariable logistic regression analysis to control confounders. Adjusted odds ratio (AOR) with corresponding 95% confidence interval (CI) were estimated to show the strength of association. The model fitness was checked using the Hosmer and Lemeshow goodness of fit test. All statistical tests were two-sided and considered statistically significant at a p-value of 0.05.VA data entered using visual basic and exported to Stata version 12 for analysis. Frequencies, proportions, and summary statistics were used to describe the socio-demographic characteristics of the deceased individuals. Differences in causes of death among groups were reported using Pearson’s chi-square  of the deaths were among females. The median age of death was 60.1 . The majority of the deaths (687 [63.5%]) were at ages greater than 50 years old. About 784 (72.5%) and 801 (74%) of the deceased were uneducated and self-employed, respectively. About 899 (83%) of the deaths occurred at home and 726 (67.0%) of deaths happened in mid- to lowland rural areas (Table The highest number of deaths were registered in the years 2009 (201 deaths), 2010 (196 deaths), and 2012 (196 deaths). Of the total deaths during the period between 2007 and 2013, communicable diseases (48.0%); non-communicable diseases (NCDs) (34.8%); and external causes (10.4%) were the main causes of death in the study area. In this study, the proportion of total deaths caused by NCDs increased from 2007 to 2013. Though deaths from NCDs increased, deaths varied from year to year during the study period. Pregnancy-related causes of death were high in the year 2007  were in adults age 50+, and the next highest number, 273 (25.2%), were among ages 25–49 years old. Tuberculosis (139 [20.2%]), cerebrovascular diseases (55 [8.0%]), meningitis (40 [5.8%]) and HIV/AIDS (35 [5.1%]) were the leading specific causes of death for adults 50 years old and older. Tuberculosis (54 [19.8%]), HIV/AIDS (52 [19.0%]), meningitis (19 [7.0%]), and cerebrovascular diseases (3 [1.1%]) were the leading specific causes of death for those 25–49 years old. Meningitis (17 [13.9%]) and tuberculosis (14 [11.5%]) were the leading specific causes of deaths for those 15–24 years old. Women aged 50+ were killed by tuberculosis (76 deaths [36.7%]), meningitis (30 [50.8%]), and cerebrovascular diseases (14 [27.5%]). More men were killed by malaria/leishmaniasis (11 deaths [40.7%]), and accidental falls (11 [41.7%]) compared with counterparts (Table The majority of specific causes of death identified between 2007 and 2013 were tuberculosis (155 [20.8%]), HIV/AIDS (55 [7.4%]), and meningitis (51[6.9%]). These diseases were the top three specific causes of death from 2007 to 2011. In 2012, tuberculosis, cerebrovascular diseases, and congestive heart failure were the major specific causes of death. In 2013, tuberculosis, meningitis and cerebrovascular diseases were the major specific causes of deaths. Of the total deaths during the period of the study, 3.6% of deaths among females were due to meningitis, as were 3.3% of deaths among males. Of the total deaths, 122 (11.3%) were due to meningitis between 2007 and 2013. Of these, 76 (67.9%) occurred between 2011 and 2013, and 46 (41.0%) occurred between 2007 and 2010. The largest number of meningitis deaths (33 [29.5%]) were in the 65 and above age groups, followed by the age group between 15 and 34 (24 [21.4%]). Of the deaths due to meningitis, 67.1% occurred in rural residents , HIV/AIDS (55 [7.4%]), and meningitis (51 [6.9%]). These diseases, tuberculosis (24.2%), HIV/AIDS (9.3%), and meningitis (7.7%) were the top three specific causes of death in rural areas from 2007 to 2010. In urban areas, tuberculosis (52 [15.4%]), HIV/AIDS (41 [12.2%]), cerebrovascular diseases (26 [7.7%]), and meningitis (25 [7.4%]) were the major specific causes of death between 2007 and 2013 (Table Tuberculosis (34 [15.9%]), intestinal infectious diseases  (15 [7.0%]), HIV/AIDS (13 [6.1%]), malaria (11 [5.1%]), and meningitis (11 [5.1%]) made up the majority of specific causes of death among lowlands and midlands dwellers. In the highlands, the majority of specific causes of death during the study period were tuberculosis (173 [19.9%]), HIV/AIDS (83 [9.6%]), meningitis (65 [7.5%]), cerebrovascular disease (53 [6.1%]), and congestive heart failure (42 [4.8%]). Of the total deceased, 776 (67.1%) lived in highlands. Of these, 388 (46.6%) were women (Table HIV/AIDS (8 [12.9%]), meningitis (8 [12.9%]), tuberculosis (7 [11.3%]), and other maternal causes (6 [9.6%]) were the major specific causes of death among women aged 15–49 years. Tuberculosis (139 [20.2%]) and cerebrovascular diseases (55 [8.0%]) were the most common specific causes of death for women aged 50 and older.The variables sex, residence, age group, and education status were significantly associated after adjusting for possible confounding variables. Females were at 1.4 (95% CI: 1.1–1. 8) times greater risk of death due to communicable diseases compared with their male counterparts. The odds of death from communicable disease are 1.5 times higher among rural residents than urban (95% CI: 1.1–2.1). Age groups between 25 and 49 years are the highest-risk age group for communicable diseases. The risk of dying from communicable diseases for the age group 25–49 years was 1.6 times higher than in the 15–24 age group (95% CI:1.0–2.5). Those who completed high school or greater were at 3.4 times higher risk of death from communicable diseases compared with those who were not educated (95% CI: 1.1–10.1) . Older ages (50 and above) have a 1.8 (95% CI:1.1–3.0) times higher risk of death due to non-communicable diseases as compared to the 15–24 age group. However, the association with climatic zone was not statistically significant after adjusting for confounding. External causes of death were associated with sex, age group, and climatic zone. Males had a 4.5 times higher risk of death from external causes than females (95% CI: 2.8–7.4). The chances of death due to external causes were high in the 15–24 age group (3.9 [2.2–7.1]) and the 25–49 age group (2.6 [1.7–4.3]) compared to the 50+ group  times higher among rural residents than urban. With respect to climatic zone, highlanders had a 1.5 (95% CI: 1.0–2.3) times higher risk of death due to tuberculosis compared to other causes  were significantly associated with deaths due to non-communicable diseases. Increasing community awareness of infections and their interrelationships, intensifying tuberculosis case finding, implementing effective local TB programs, and providing successful treatment and increased interventions for HIV are of supreme importance.This study showed that the majority of the deaths among adults were from tuberculosis, HIV/AIDS, and meningitis. Deaths due to non-communicable diseases showed an increasing trend


Streptococcus mutans count in saliva among 12-year-old school children.To evaluate the effect of fluoride varnish on mutans count in saliva among 12-year-old school children. A total of 42 school-going children attending schools in Vadodara district, Gujarat, India, were divided into two groups. Group I was treated with fluoride varnish and group II received no treatment. Assessment of S. mutans was carried out at baseline and 3 to 6 months postfluoride varnish application. Friedman analysis of variance test and post hoc test were applied to detect statistically significant differences between baseline, 3 to 6 months of fluoride varnish application, and also between groups I and II.A field experiment was conducted to evaluate the effects of fluoride varnish on S. mutans value found in case group at baseline, 3 to 6 months was 31.23 ± 1.119, 9.27 ± 0.852, and 9.39 ± 0.908 × 104 colony-forming unit CFU/mL respectively. The difference in S. mutans count from baseline to 3 to 6 months was highly statistically significant (p = 0.000), but the difference from 3 to 6 months was not statistically significant (p = 0.142). In control group, the mean S. mutans value found at baseline, 3 to 6 months was 30.63 ± 1.436, 31.23 ± 1.351, and 31.40 ± 1.374 × 104 CFU/mL respectively. The differences between these values were not statistically significant (p = 0.11).The mean number of salivary S. mutans count in saliva was seen 3 to 6 months after fluoride varnish application.Statistically significant reduction in S. Streptococcus mutans Count in Saliva. Int J Clin Pediatr Dent 2017;10(1):62-66.Badjatia S, Badjatia RG, Thanveer K, Krishnan ACG. Effects of Fluoride Varnish on  Dental caries is still one of the most common diseases in the world.Streptococcus mutans, Lactobacillus spp., Veillonella spp., and Actinomyces spp. A variety of carbohydrates provide substrates for these organisms to grow on, and the waste products of their metabolism, acids, initiate the tooth decay process by dissolving tooth enamel.1Dental caries will not occur if the oral cavity is free of bacteria. These bacteria are organized into a yellowish film known as dental plaque on the surface of the teeth. Many types of bacteria are present in the mouth; the most caries-active appear to be S. mutans and its ability to ferment sucrose. It ferments sucrose to produce significant amounts of acid and extracellular polysaccharides (plaque).1 The most important understanding of caries process is that dental caries do not occur either in the absence of dental plaque or dietary fermentable carbohydrates. The S. mutans plays a significant role in the development of dental caries and is the chief pathogen.23Research on the bacteriology of dental caries has focused on the ubiquitous S. mutans is required for dental caries initiation, and it has been suggested that an S. mutans count higher than 105 colony-forming unit CFU/mL of saliva is related to higher caries risk. Hence, microbial monitoring has been considered as an alterna-five method for evaluating caries activity.4Oral colonization by 5 Fluoride-containing toothpastes (dentifrices), mouth rinses, gels, and varnishes are the modalities most commonly used at present, either alone or in combination.6 The use of fluoride varnish to prevent and control dental caries in children and adults is expanding in both public and private dental practice settings and in nondental settings that incorporate health risk assessments and counseling.7As dental caries has multifactorial etiology, preventive measures usually involve a combination of dietary counseling, oral hygiene measures, and fluoride application.8 The most important anticaries effect of fluoride is considered to result from its action on the tooth/plaque interface, through promotion of remineralization of early caries lesions and by reducing tooth enamel solubility.6 Fluoride at low concentration is bacteriostatic and at high concentration, it is bacteriocidal.7 A high fluoride concentration in the oral cavity might inhibit acid production by bacteria and may reduce the number of certain species.9Fluoride works primarily via topical mechanism inhibiting demineralization and enhancement of reminer-alization at the crystal surface and inhibition of bacterial enzymes.S. mutans, does not show conclusive results. Moreover, the effect of fluoride varnish over a period of time has not been studied.Limited study, which explored the effect of fluoride varnish on S. mutans count in saliva among 12-year-old schoolchildren.To evaluate the effect of fluoride varnish on S. mutans in saliva at baseline. To estimate the count of S. mutans in saliva 3 to 6 months after fluoride varnish application. To estimate the count of S. mutans count with that of 3 to 6 months postfluoride varnish application. To compare baseline salivary S. mutans count in saliva among 12-year-old schoolchildren of Vadodara district, Gujarat, India. The study protocol was reviewed and approved by the institutional ethics committee. Permission from respective authorities was taken prior to the study.A field experiment was conducted to evaluate the effects of fluoride varnish on 10 with salivary S. mutans count of ≥104 CFU/mL of saliva .Subjects having at least one carious tooth  Children who were caries free at baseline examination. Children whose parents refused to give informed consent. Children having history of antibiotics for past 3 to 4 weeks or were taking antibiotics. Children having history of fluoride treatment for past 6 months.All the 12-year-old schoolchildren were examined for the presence of caries , out of The selected samples were then divided equally into two groups:Group I: Subjects who received fluoride varnish application at baseline (study group)Group II: Subjects who did not receive any fluoride varnish application (control group)In the present study, data were collected by personal interview.S. mutans analysis to the microbiology laboratory.Saliva collection and microbial analysis  and 3 waStep 1: Teeth cleaned with the help of cotton and tweezersStep 2: Teeth dried with the help of chip blower.Step 3: A small amount of varnish (0.5 mL) will be dispensedStep 4: Varnish applied to the teeth with the help of applicator tipStep 5: Varnish was allowed to dry for 1 minute as per the instruction of manufacturer.These steps were performed quadrant-wise sequentially starting from the upper arch. Subjects were instructed not to rinse their mouth for 1 hour and not to eat for 4 hours .S. mutans count at baseline, 3 to 6 months, and Mann-Whitney test was used to compare S. mutans count between group I (case group) and group II (control group).The data were analyzed using Statistical Package for the Social Sciences version 11. Friedman analysis of variance (ANOVA) test was used to compare Data collected from the study subjects by personal interview reveals that all the subjects requited for the study had not undergone any antibiotic treatment in the past 6 months. Information regarding their oral hygiene practice, methods to clean their teeth, and their rinsing habits was also collected. Among the 42 subjects, 29 (69.0%) subjects cleaned their teeth once daily, 12 (28.6%) subjects cleaned their teeth twice daily, and 1 (2.4%) subject did not clean his/her teeth. About 24 (57.2%) subjects cleaned their teeth by using toothbrush, 10 (23.8%) used Datun, and 7 (16.6%) used finger. About 22 (52.4%) subjects cleaned their teeth by toothpaste, 12 (28.6%) by tooth powder, whereas 7 (16.6%) used other materials to clean their teeth. About 23 subjects (54.7%) rinsed their teeth once daily, 17 (40.5%) twice daily, and 2 (4.8%) after every meal. All the 42 (100%) subjects cleaned their teeth by using nonfluoridated toothpaste and had never received any fluoride treatment in the past 6 months.4 CFU/mL S. mutans.Saliva sample analysis at baseline revealed that all the subjects who participated in the study had more than 10S. mutans at baseline, 3 to 6 months after treatment with fluoride varnish in case group. The mean number of S. mutans value found at baseline, 3 to 6 months was 31.23 ± 1.119, 9.27 ± 0.852, and 9.39 ± 0.908 × 104 CFU/mL respectively. The difference between these value was highly statistically significant; p = 0.00 .post hoc comparison of S. mutans from baseline, 3 to 6 months after treatment with fluoride varnish in group I (case group). The difference in S. mutans count from baseline to 3 to 6 months was highly statistically significant , but the difference from 3 to 6 months was not statistically significant .S. mutans from baseline, 3 to 6 months in group II (control group). The mean S. mutans value found at baseline, 3 to 6 months was 30.63 ± 1.436, 31.23 ± 1.351, and 31.40 ± 1.374 × 104 CFU/mL respectively. The difference between these values was not statistically significant; p = 0.11 .post hoc comparison of S. mutans from baseline, 3 to 6 months after treatment with fluoride varnish in group II (control group). The difference in S. mutans count from baseline to 3 months, baseline to 6 months, and 3 to 6 months was not statistically significant .S. mutans from baseline, 3 to 6 months between group I (case group) and group II (control group). The S. mutans value at baseline was not statistically significant , but highly statistically significant at 3 to 6 months .S. mutans count in saliva at baseline, 3 to 6 months.This is a unique study showing the effect of fluoride varnish on S. mutans plays a major role. The S. mutans are generally considered to be the principal etiological agent of dental caries.Dental caries is a transmissible infectious disease in which S. mutans are most commonly associated with dental caries, several other microorganisms also have the ability to produce organic acids that induce demineralization of tooth structure and lead to clinically detectable caries.11A measure of caries activity and caries risk is the concentration of cariogenic bacteria within saliva. Although S. mutans. Now, microbial monitoring has been considered as an alternative method for evaluating current caries activity and future caries risk.11Caries activity is evaluated on the basis of data obtained from clinical examination and assessment of factors associated with the pathogenesis of the disease. These data can be collected by traditional visual inspection and probing or by some objective detection methods, which rely on the mineral changes as a basis for evaluation of caries activity and risk assessment. None of these methods aims at the estimation of the chief pathogen 6 Fluoride therapy has been the centerpiece of caries-preventive strategies since the introduction of water fluoridation schemes over 5 decades.12The prevention of dental caries in children and adolescents is generally regarded as a priority for dental services and considered more cost-effective than its treatment.13Fluoride varnish, which is also used as caries-preventive strategy, works by increasing the concentration of fluoride on outer surface of the teeth, thereby enhancing fluoride uptake during early stages of demineraliza-tion. Varnish hardens on the tooth as soon as it contacts saliva, allowing the high concentration of fluoride to be in contact with tooth enamel for an extended period of time (about 1-7 days). This is a much longer exposure as compared with other high doses of topical fluorides, such as gels or foams, which takes 10 to 15 minutes. The most important anticaries effect of fluoride results from its action on tooth and plaque interface, by promotion of remineralization of early caries lesions and reducing tooth enamel solubility.S. mutans count in saliva at baseline, 3 to 6 months.Hence, the present study was undertaken to compare the effect of fluoride varnish on S. mutans levels were higher than 104 CFU/mL.The subjects in the present study were accepted as a high-caries-risk group since they had at least one or more decayed tooth. The baseline salivary The age of the present study population is around 12 years. This age is appropriate because it conforms to one of the World Health Organization indexed age groups.S. mutans count of group I was 31.23 ± 1.119 × 104 CFU/mL. The mean value of S. mutans at 3 to 6 months after fluoride varnish application was 9.27 ± 0.852 and 9.39 ± 0.908 × 104 CFU/mL respectively.In the present study, it was found that the baseline mean S. mutans count at baseline, 3 to 6 months was 30.63 ± 1.436, 31.23 ± 1.351, and 31.40 ± 1.374 × 104 CFU/mL respectively.Whereas in group II, the mean S. mutans count in group I from baseline to 3 to 6 months was highly statistically significant , but the difference from 3 to 6 months was not statistically significant  and also the difference in group II from baseline to 3 months, baseline to 6 months, and 3 to 6 months was not statistically significant .The difference in S. mutans from baseline, 3 to 6 months were compared between groups I and II. The S. mutans value at baseline was not statistically significant , but was highly statistically significant at 3 to 6 months .In the present study, the mean values of 14 found that the fluoride varnish treatment had no significant effect on the plaque and salivary levels of S. mutans at baseline and 4, 10, and 21 days after treatment with fluoride varnish.Zickert and Emilson15 observed that the application of fluoride varnish causes a significant suppression of S. mutans compared with baseline, 3 to 6 months.Araujo et al16 found no statistically significant difference for S. mutans in dental plaque after treatment with fluoride varnish at 1 week and 1 month relative to baseline.Ekenbäck et alS. mutans count at baseline of at least 104 CFU/mL are included. Further studies with larger sample size and a placebo group can reveal the long-term effect of fluoride varnish on the S. mutans count in saliva of carious dentition.In the present study, children with at least one carious teeth and having S. mutans count in saliva as compared with baseline, 3 to 6 months in our present study. Thus, we can conclude that fluoride varnish can be applied on teeth for broad spectrum antimicrobial activity, as an effective agent against S. mutans and in caries reduction.Fluoride varnish had a statistically significant reduction in the


Purpose. To evaluate the efficacy of intravitreal conbercept (IVC) plus modified grid laser photocoagulation (MGP) versus IVC alone for treatment of diffuse diabetic macular edema (DDME). Methods. In this retrospective study, 51 DDME patients were treated with either IVC alone (IVC group) or IVC plus MGP (combined group) with 12 months of follow-up. The clinical records of those patients were reviewed. Results. 26 patients (31 eyes) received IVC alone and 25 patients (30 eyes) received combined therapy. At month 12, the mean best-corrected visual acuity (BCVA) letter score improvement was 9.1 ± 4.5 and 7.5 ± 4.2 in the IVC group and the combined group and the mean central retinal thickness (CRT) reduction was 145.1 ± 69.9 μm and 168.5 ± 53.6 μm, respectively. There was no statistically significant difference of improvement in BCVA (P = 0.164) and decrease in CRT (P = 0.149) between the two groups. The mean number of injections delivered was significantly higher (P < 0.001) in the IVC group (5.6 ± 0.8 per eye) than in the combined group (3.3 ± 1.2 per eye). Conclusions. IVC alone or combined with MGP appeared to be effective for treatment of DDME, achieving the similar clinical efficacy. Moreover, MGP helps to reduce the number of injections.  Diabetic retinopathy (DR) is the most frequent and severe ocular complication of diabetes mellitus, the leading cause of blindness in the working age population . DiabetiDME consists of two main forms: diffuse and focal . DiffuseThe Early Treatment Diabetic Retinopathy Study (ETDRS) has demonstrated the value of focal/grid laser treatment in preventing or reversing visual loss from DME . Its benVascular endothelial growth factor (VEGF) has been proved to be an angiogenic inducer and a vascular permeability factor, which increases the retinal vascular permeability. Recently, anti-VEGF agents  have been reported to be effective in reducing DME and improving the best-corrected visual acuity (BCVA) when injected intravitreally –15. AlthThe most recent anti-VEGF drug is conbercept, also named KH902 , which is a recombinant fusion protein containing the second immunoglobulin (Ig) domain of VEGF receptor 1 (VEGFR1) and the third and the fourth Ig domains of VEGFR2 and the Fc region of human IgG . Its affThus, this study was conducted to evaluate the efficacy of intravitreal conbercept (IVC) plus MGP versus IVC alone in the treatment of DDME in real-life clinical practice.This was a retrospective study performed at the Department of Ophthalmology of the Tongji Hospital affiliated to Tongji University . We reviewed the clinical records of 51 consecutive patients (61 eyes) with DDME treated with IVC alone or IVC plus MGP. All the patients received initial injection from 01.04.2014 to 31.03.2015 and had 12 months of follow-up. This study was approved by the Review Board/Ethics Committee of the Tongji Hospital and was conducted according to the ethical standards laid down in the 1964 Declaration of Helsinki. After discussion with the patient regarding the benefits, risks, and alternatives to treatment, informed consent was obtained from each participant.μm measured by OCT and best-corrected visual acuity (BCVA) letter score between 78 and 24 measured by the ETDRS protocol. Exclusion criteria included previous macular laser photocoagulation, macular traction as evidenced by fundus biomicroscopy and OCT, macular ischemia as diagnosed by FA, prior intraocular injection of anti-VEGF or steroids, history of glaucoma or ocular hypertension, presence of proliferative diabetic retinopathy (PDR) with high-risk characteristics, significant media opacity, and previous panretinal laser photocoagulation or intraocular surgery performed within the last 6 months. Patients with uncontrolled hypertension and recent thromboembolic events were also excluded.In this study, DDME was defined as having two or more disk areas of retinal thickening and involving part of the foveal avascular zone . The diaμm) and/or an increase of diffuse fluorescein leakage involving the center of the macula on FA. In the combined group, MGP was performed 3 weeks after the initial IVC injection. Supplemental laser treatments were performed at a minimum of 3 months from the previous laser treatment if OCT and/or FA confirmed the presence of residual or recurrent DDME.Patients in both groups were started on the intravitreal injection of 0.5 mg conbercept. The retreatment of IVC was performed whenever there was a recurrence of DDME. Recurrence was defined as a decrease of BCVA associated with OCT evidence of increasing macular thickness  was measured by Cirrus HD-OCT  utilizing six diagonal slow 6 mm radial line scans, with software version 4.0. The CRT of the 1 mm central retina was obtained using the macular thickness map for our calculations.Intravitreal conbercept injections were performed in the operating room under the complete aseptic conditions with topical anesthesia and insertion of a lid speculum. After disinfection, 0.05 mL of solution containing 0.5 mg of conbercept was injected intravitreally 3.5 to 4 mm posterior to the limbus with a 30-gauge needle through the superotemporal quadrant. After the treatment, patients were examined at day 1 and day 7 and each month thereafter. All eyes underwent an ophthalmic examination for anterior chamber (AC) reaction and intraocular pressure (IOP) rise. All ocular and systemic adverse events (AEs), including information on their relationship to study drug and procedure, were recorded at the visits.μm spots were applied to the parafoveal region, where there was retinal thickening up to and including the edge of the foveal avascular zone; the lesions were placed 100 μm apart. Then, 150 μm to 200 μm spots were placed approximately 200 μm apart and were applied to all remaining areas of retinal thickening and in all areas of capillary nonperfusion [Modified grid laser photocoagulation (MGP) was performed by one trained ophthalmologist with double-frequency Nd:YAG (532 nm) laser. After topical anesthesia and placement of a contact lens, two to three rows of 100 erfusion . Laser tPrimary outcome measures included the changes in BCVA and CRT. Secondary outcome measures were the mean number of injections needed during the 12-month study period. t-test was performed to compare the BCVA and the CRT to baseline values within each treatment group. The independent samples t-test was used to determine statistically significant differences between two groups as regards mean change in BCVA and CRT, as well as mean number of injections consumed. All statistical tests were 2-sided. A P value less than 0.05 was considered to be statistically significant.Statistical analysis was performed using SPSS software version 17.0 . Qualitative variables were stated using percentage, and quantitative data were reported as mean ± standard deviation (SD). Differences in categorical variables were assessed with the chi-square test. The paired samplesμm. The baseline ocular and systemic characteristics of each treatment group were compared and presented in We reviewed the clinical records of 51 consecutive patients (61 eyes) with DDME. All patients had 12 months of follow-up. 31 eyes of 26 patients were treated with IVC alone, and 30 eyes of 25 patients were treated with IVC plus MGP. The mean age of the participants was 61.4 ± 11.7 years; 49.0% were women, and 51.0% were men. A total of 94.1% of the participants had type 2 diabetes, and the mean duration of diabetes was 13.6 ± 4.7 years. The mean visual acuity letter score at baseline was 47.4 ± 10.2, and the mean CRT was 483.4 ± 95.7 P < 0.001) improvements in the BCVA letter score were observed for both the IVC group (7.5 ± 5.1) and the combined group (6.0 ± 3.9). These improvements were maintained throughout the 12 months of follow-up  and 7.5 ± 4.2 in the combined group (P < 0.001). There was no statistically significant difference of improvement in BCVA between both groups  in the IVC group (5.6 ± 0.8 per eye) than in the combined group (3.3 ± 1.2 per eye) throughout the study period. The numbers of the macular MGP procedures performed were 2.1 ± 0.6 in the combined group. In the IVC group, 3 eyes (9.7%) received 7 injections, 14 eyes (45.2%) received 6 injections, 12 eyes (38.7%) received 5 injections, and 2 eyes (6.4%) received only 4 injections over the 12 months of study period. In the combined group, 2 eyes (6.7%) received 6 injections, 3 eyes (10.0%) received 5 injections, 5 eyes (16.6%) received 4 injections, 15 eyes (50.0%) received 3 injections, 3 eyes (10.0%) received 2 injections, and 2 eyes (6.7%) received 1 injection.In our study, the mean number of injections consumed was significantly higher  in the IVC group and in 7 eyes (23.3%) in the combined group. Mild vitreous hemorrhage was reported in 1 eye (3.2%) in the IVC group. No systemic adverse events were observed in both groups throughout the study duration.P = 0.164) and decrease in CRT (P = 0.149) between two groups.This is a single-center retrospective study that evaluates the anatomical and functional outcomes during 12 months of follow-up in patients with DDME treated with either IVC or IVC plus MGP. Our results indicated that intravitreal injection of 0.5 mg conbercept with and without MGP provided fast decrease in CRT and improvement in BCVA at month 1, and these changes were maintained throughout 12 months of follow-up. At the end of the 12 months, there was no statistically significant difference of improvement in BCVA  combination treatment group with 6 months of follow-up. Bevacizumab plus MPC combination treatment could maintain visual acuity and reduce the recurrence of macular edema . In the Previous studies have demonstrated the efficacy of intravitreal injection of anti-VEGF drug in patients with DME –15. HoweP < 0.001) in the combined group (3.3 ± 1.2 per eye) than in the IVC group (5.6 ± 0.8 per eye) throughout the study period, which meant that combining IVC with MGP could decrease the rate of recurrence of DDME, the frequency of repeated IVC injections, and the possible adverse effects associated with intravitreal injections. This matches with the results of the study conducted by Solaiman et al. [P < 0.05) in the combined group (2.36 per eye) than in the IVB group (3.27 per eye).In our study, we also found that the mean number of injections was significantly lower  are some factors that contribute to chronic capillary damage and increased permeability . Anti-VELimitations of our study include that it is nonrandomized, retrospective, and single-centered, which precludes any estimation of the efficacy or safety of IVC with or without MGP. A larger size sample and a longer follow-up period would be ideal to verify and confirm the results of the present study, to select the correct timing for MGP after IVC, and also to establish parameters for selection of patients who are most suitable for undergoing combined therapy. Moreover, there was no standardized adverse event form to collect the safety data in our study. Despite these limitations, our results are very promising and suggest the possible needs for the further investigations.In conclusion, this preliminary study showed that IVC injections both with and without MGP are effective in the treatment of DDME, achieving the similar clinical efficacy. The additional MGP reduces the number of the repeated intravitreal injections, which not only decreases the possible adverse effects of the intravitreal injections, but also minimizes the financial cost. Evaluation in a multicenter randomized controlled clinical trial with longer follow-up is needed to evaluate the safety and efficacy of IVC plus MGP versus IVC alone for treatment of DDME.


Smoking Cessation Clinics (SCCs) involve the use of cognitive behavior therapy and effective medications provided by specialists. Our objective was to report socio-demographic data, investigative services provided, and determine the smoking cessation success.Data from all hospitals affiliated with the Beyoglu Public Hospitals Union were obtained between January 1, 2015 and December 31, 2015. Data from Sisli Hamidiye Etfal Training and Research Hospital were reviewed via retrospective chart review in the same period. Frequency and average values were determined using statistical software. In the evaluation of related factors, chi-square and student t tests were applied; p ≤ 0.05 was considered statistically significant.The mean admission age was 38.72 ± 12.20 years . Women tended to be older than men at the time of admission but men started smoking younger than women. Less than the high school educated subjects started smoking at early ages. The smoking cessation rate was 39.3% (n = 219) when treated with medication and behavioral therapy.About 48% smokers stopped smoking after treatment with medications and behaviour therapy. Most of the smokers were between 30-50 years of age. More Smoking Cessation Clinics should be established to allow access to more highly educated patients to smoking cessation resources. Smoking addiction is a serious public health problem worldwide. The World Health Organization (WHO) defines smoking as “the world’s fastest-spreading and longest-running epidemic”.3The first legal regulation in the fight against tobacco in Turkey was made in 1996. In 2004, the WHO Framework Convention on Tobacco Control was put into practice. In 2007, the Provincial Tobacco Control Boards were established in 81 provinces so that the implementation of the National Tobacco Control Program as a whole, including follow-up activities, could be implemented in the provinces.Epidemiological data show that 70% of smokers want to quit smoking, and 46% have attempted to quit at least once.6In Turkey services are provided for nicotine dependence reduction as well as for smoking cessation. Components of the system that help smokers quit include brief referrals and withdrawal assistance lines as well as smoking cessation clinics involving the use of cognitive behavior therapy and effective medications provided by experienced specialists.7The smoking cessation clinic (SCC) at our hospital was previously run by both psychiatry and family medicine departments until 2015 but has subsequently shifted to the family medicine clinic. Our objective was to report socio-demographic data, investigate services provided, and determine the smoking cessation success of our SCC at the Sisli Hamidiye Etfal Training and Research Hospital.This retrospective descriptive study was performed on 4.119 smokers admitted between January 2015 and December 2015 to SCCs of hospitals of Beyoglu Public Hospitals Union (BPHU) in Istanbul. There was no need for ethical approval because the study was retrospective so that’s why permission was taken from BPHU General Secretariat only.Hospitals which are member of BPHU and have common electronic database in addition to SCCs were included in this study: Sisli Hamidiye Etfal Training and Research Hospital, Okmeydani Training and Research Hospital, Gazi Osman Pasa Taksim Training and Research Hospital; Balta Harbor Bone Diseases Training and Research Hospital, and Eyup State Hospital. Required data were collected from SCCs and BPHU General Secretariat database. Sisli Hamidiye Etfal Training and Research Hospital (SHETRH) SCC data were reviewed via retrospective chart review in the same period because it was our main SCC. All records in that period were included in the study Frequency and average values were determined using statistical software. In the evaluation of related factors, chi-square and student t tests were applied; p ≤ 0.05 was considered statistically significant.In 2015, a total of 4.119 patients were referred to the SCC affiliated with the General Secretariat of BPHU. There were 2.443 (59.3%) male patients and 1.676 (40.7%) female patients. The minimum age was 13, the maximum age was 95, and the mean age was 39.73 ± 12.39 years. The mean age of female patients was 41.22 ± 11.76 years, and the mean age of male patients was 38.71 ± 12.72 years.The most frequent admission was in the age group of 35–49 . Males were most frequently in the 20–34 year age group, while females were most frequently in the 35-rs and 26 of these (53.06%) were female.The mean number of admissions per patient to SCCs was 1.55 ± 0.98 times/year. When the responsible physicians of SBPs were evaluated according to specialty, the distribution of patients were 2355 (57.2%) family physicians, 1424 (34.6%) pulmonologists, and 340 (8.3%) psychiatrists.When the distribution of applicants to these centers was examined, most admissions were made to Sisli Hamidiye Etfal Training and Research Hospital . A significant number of patients  were admitted to family medicine’s SCC. One thousand-forty-five (61.5%) of these patients were male, and 655 (38.5%) were female. The minimum admission age was 13 years, the maximum was 94 years, and the mean age was 38.72 ± 12.20 years. Twenty-one (21.2%) patients under the age of 18 applied. The average age of males was 37.70 ± 12.42 years, while the average age of females was 40.34 ± 11.66 years. The most frequent admission was in the 20–34 age group . Table-IIAccording to family medicine-related SCC data from SHETRH, 524 applicants were female (38.7%) and 834 were male (61.3%). The mean age was 39.35 ± 12.17 years . The mean age of females was 41.18 ± 11.44 years , while it was 38.20 ± 12.47 years  in males. According to age group, most smokers were in the age group of 35–49 years. Females were more likely to be in the 35–49 year age group  and males in the 20–34 year age group . There was a significant relationship between gender and age (p = 0.000). Women tended to be older than men at the time of admission.While 788 (51.2%) smokers admitted to SHETRH were high school educated or above, 582 (42.8%) received less education than high school. Sixteen (1.2%) patients were illiterate. The education level of males was found to be significantly higher than females (p = 0.000). We found that the level of education decreased significantly with increasing age at admission (p = 0.000).While 810 (59.1%) of those who came to the SCC were single, at least one person smoked in the family were 98.8% (n = 1.344). There was no significant relationship between marital status and number of smokers in the family (p = 0.783).When the age of smoking initiation was questioned, the mean age was 17.40±5.20 years . According to gender, smoking initiation began in females at 18.38 ± 5.75 years, while in males it was found to be 16.79 ± 4.72 years, and this was statistically significant (p = 0.000). When the educational status  and the age at which the person started to smoke were compared, it was found that the age at which smoking began was higher in the high school and above educationed subjects (p = 0.000).The mean pack/year among applicants to SCCs was 21.99 ± 13.70 . When the relationship between gender and number of cigarettes (pack/year) was examined, the pack/year average of females was significantly higher than the pack/year average of males (p = 0.046). When the relationship between educational status  and the number of cigarettes smoked (pack/year) was examined, the number of cigarettes smoked (pack/year) was lower in high school and above education (p = 0.000).About 44.9% (n = 1423) of the patients who had medication and cognitive behavioral treatment were quit smoking when all hospitals belonging to the BPHU General Secretariat were taken into consideration, and patients who were not discharged were not evaluated. The smoking cessation rate was 39.3% (n = 219) when treated with medication and behavioral therapy.The frequency of tobacco product use among individuals 15 years and older in Turkey is 27.1%. This value corresponds to 14.8 million people. Monso et al found Male gender and older age to be effective in smoking cessation.SCC admissions are most frequently patients in the 25–44 year age group according to a study from USA.Females who applied to the SHEEAH’s SCC were found to be significantly older than males (p = 0.000). At the same time, the age at which males started to smoke was less than that of females (p = 0.000). Similarly, in a study published in 2012, the average age of onset of cigarette smoking was 17 (6–37) years, and the mean age of onset of smoking in females was 18.09 ± 5.14 years and 16.09 ± 4.43 years in males.According to one study, 58.7% of cigarette smokers are smoking before 18 years of age, the legal age at which one can purchase cigarettes.According to 2010 data, smoking is lower in females with lower educational levels, and smoking rates also increased as education level increased.Smoking cessation treatment consists of behavioral therapy and motivation as well as pharmacological treatment.16According to statistical data of our country, 13.6% of those who attempted to quit smoking used pharmacologic treatment. 8% of those who attempted to quit were given counseling, and most (73.4%) attempted to quit without any help. 9The smoking cessation rate was 39.3% (n = 219) when treated with medication and behavioral therapy. In Turkey, smoking cessation drugs were provided by the SCC free of charge to the patient. Free drug treatment may increase individual motivation to quit smoking.Smoking cessation treatment is a special treatment consists of cognitive behavioral therapy and motivation as well as pharmacological treatment. Smoking Cessation Clinics are the most important centers for smoking cessation and prevention from cigarette related diseases. The widespread use of SCCs is important both in terms of individual and social wellbeing. For this reason, necessary certification trainings should be organized, and new SCCs should be opened to allow access of more highly educated patients to smoking cessation resources.MTE conducted this research and is responsible and accountable for the accuracy and integrity of this work.GZO helped in collection of data and its interpretation.MHB reviewed the manuscript.DT helped in data analysis.