Family planning is one of the four pillars of safe motherhood initiative to reduce maternal death in developing countries. Despite progress in contraceptive use, unmet needs are wide open and fertility remains high. Ethiopia have a higher fertility rate which contributes to maternal and child health destitution, putting pressure on the already weak health system. This study examined individual and community-level factors associated with contraceptive use in Ethiopia.Data from Ethiopian Demographic and Health Survey 2016 were used to identify individual and community level associated factors among reproductive-age women. Non-pregnant, fecund and sexually active women aged 15–49 were included. Six hundred forty-two communities and 6854 women were involved from this two-stage cluster sampled data. The analysis was done using two-level mixed-effects logistic regression to determine fixed effects of individual and community-level factors and random intercept of between characteristics.From the total eligible women for contraceptive use 2393 (34.9%) of them were users. Injectables were the commonest of all contraceptive methods. Various individual-level variables were associated with contraceptive use. Household wealth index, women’s age, number of living children, husband’s occupation, ever experience of a terminated pregnancy, current working status of the women, number of births in the last 3 years, and hearing of FP messages through different media were significantly associated individual-level variables after adjusting other factors. Community characteristics like region, place of residence, religion, and community-level wealth were the factors associated with contraceptive use.Both individual and community-level characteristics were significant predictors of use of contraceptives in Ethiopian women. Besides the individual-level factors, interventions should also consider community-level associates. In the current growth rate of the world population, 1.13%, about 80 million people will be added per year . With thThe use of modern contraceptives ranges from 3.6% in the Somali region to 56.3% in Addis Ababa with a national prevalence of 27.3% in 2011 , 9. The Modern contraceptives in Ethiopia are available in every health facility free of cost and pushing to increase utilization. The use of contraceptives in Ethiopia usually focused on short-acting methods like birth control pills and injectable, which have a high chance of discontinuation , 12. FeaThe number of children, women’s current age, age at first marriage, education, religious affiliation, media exposure about family planning, wealth index, occupation, husband’s occupation, place of residence, husbands approval couple’s discussion, are the main determinant factors for utilization of modern contraceptives among married women of reproductive age , 14, 15.Several community-level factors were found to be associated with contraceptive use elsewhere. Access to family panning in nearby store or health facility, female autonomy, female education, religion, place of residence, proportions of polygynous marriages and community level media exposure were community-level determinants in Nigeria , 17. AveRepresentative studies on the use and determinants of modern contraceptives in Ethiopia are scares. Besides deficiency of fertile evidence, small inconclusive studies in different areas without considering community-level factors put different dissimilar, mutually exclusive determinant factors, lacking to draw relevant conclusions in national level.Generally, even if previous studies have given important clues to policymakers, programmers, and other stakeholders, they mainly lack consistency and representatives to be used for national-level policy making and programming by concerned bodies. Thus, the present study aims to determine multilevel determinants of modern contraceptive use among married women of the reproductive age group in Ethiopia using the 2016 Ethiopian Demographic Health Survey (EDHS 2016) data.Ethiopia is the second populous country in Africa next to Nigeria with a population of more than one hundred million. Administratively, Ethiopia is divided into nine geographical regions  and two administrative cities, Addis Ababa and Diredawa. Ethiopia shares the boundaries in the North with Eritrea, in the South with Kenya and Somalia, in the West with South Sudan and North Sudan, in the East with Djibouti and Somalia. The survey was a population-based cross-sectional study conducted from January 18, 2016, to June 27, 2016, across the country , and only 14.2% of them had attended secondary education or higher. By contrast, their husband had a better education with a decreased number of no formal education males 2981(43.5%), again 28.3% of them had secondary education or higher. More than two-thirds of women 4696 (68.5%) had either one or two under-five children, while less the one-fourth of the total 1446(21.1) had no under-five children. Majority of the women 4674(68.2%) were not working in the time of the survey, this figure is bigger when we consider the husband’s status of 681(9.9%), indicating disparities in work. Summarized in Table Exposure to mass media including radio, television, magazine/newspaper, at least for one of the media, remains below half of the study population. Only 41% of the women had exposure to medias 12 months preceding the survey. Media exposure for family planning was also low with 2090(30.5%) of mothers heard through radio or TV or newspaper in the last few months before the survey. Health insurance coverage was extremely low as well. More than 95% of the women were no insured. While 1 in 10 mothers experienced a terminated pregnancy, smoking in reproductive age fecund Ethiopian women was low (1%). The mean age of mothers during the survey was 29 years, the median value for under-five children in the household during the survey was one child and the mean value for the number of total living children in the household preceding to the 2016 EDHS were three.n = 642).From the total 645 EDHS clusters, 642 clusters had married, non-pregnant reproductive age women eligible for contraceptive use and included in this study. Below one-third of clusters were from urban areas. Three hundred eighty-two (59.5%) of clusters had lower Muslim proportions at the community level. Comparing with the median educational level of the country, women in 306 (55.8%) clusters had lower education. Almost half of the clusters had lower employment status, and media exposure about family planning was poor even at community-level. Description of community-level variables was presented in Table A two-level mixed-effect logistic regression was used to analyze the effect of women’s individual characteristics and community-level factors in determining women’s use of contraceptives. As depicted in the empty model 40% of the total variance in the odds of contraceptive use was accounted by between cluster variation of characteristics. The between cluster variability declined over successive models from 40% in the empty model into 23% in individual-level only model, 16% in community-level factors only model and 14% in the combined model. Thus, the combined model of individual-level and community-level factors were preferred for predicting women’s contraceptive use.In the model I only individual-level variables were added. The result signified that wealth index, women’s age, number of living children, husband’s occupation, ever experience of a terminated pregnancy, current working status of the women, number of births in the last 3 years, and hearing of family planning messages through different media were significantly associated variables with contraceptive use.The ICC in Model I indicated that 23% of the variation in contraceptive use was attributable to difference across the communities. From PCV in Tables The number of living children affects women’s use of contraceptive at the individual level. Women with 1–4 children have higher odds of using contraceptive OR = 3.6 than women with no living children. Even the odds were highest in women having living children between five and eight 4.0(95% CI 2.8–5.5), although the odds have slightly decreased for women having more than 8 children there was a significant difference with those no living children at all 3.2. Wealth index was another factor influencing women’s use of contraceptive, as wealth index increases from poor to rich the odds of using contraceptive was also increased. Compared to the poor the middle and the rich had higher odds 1.4.Among several husband’s characteristics that affect women’s use of contraceptive methods, one was partner’s occupation. Different occupations had different odds for women’s utilization, professional/technical/managerial and Clerical/sales/services/skilled labor positions of the husband gave highest of odds 1.9, while farming/unskilled followed with odds of 1.6. working women during the time of the survey had higher odds with 1.3, and experience of terminated pregnancy had increased the use of contraceptives in currently in-union women. Women’s individual level determinant factors were summarized in Table To examine if the characteristics of the cluster affect women’s contraceptive use, regardless of women’s individual characteristics, we analyze community-level attributes. In Model II only community-level variables were added. Cluster characteristics like region, place of residence, religion, and community-level poverty were the factors associated with contraceptive use. The ICC in this model implied that difference between communities account for about 16% of the variation in women’s contraceptive use. The PCV also showed that 88% of the variation in contraceptive use between communities was explained by community level characteristics. As a cluster factor region was a strong predictor of contraceptive use. Considering Addis Ababa, the capital of the country as a reference, dwelling in Amhara region had the highest odds OR = 2.7 (95% CI 1.8–4.1) of contraceptive use. On the contrary Somali region had the lowest odds for contraceptive use OR = 0.1 compared with the people of the capital.Compared with urban women, rural women had lower odds for contraceptive use OR = 0.7, and community-level poverty decreased odds. Wealthy clusters had higher odds than poor clusters, regardless of household wealth index OR = 1.7. Religious affiliations had an effect on contraceptive utilization, being in a Muslim dominant cluster had lower odds of utilizations OR = 0.6 after controlling other individual and community level factors. Controlling other individual and community level factors, hearing information about family planning on radio and other information sources in the last few months didn’t contribute for utilization P value > 0.05 in all cases.Family planning is one of the four pillars of safe motherhood initiative to reduce maternal death in developing countries. Contraceptives prevent unwanted and unplanned pregnancies, as well as complications, arising from these conditions. Increasing contraceptive use in developing countries has cut maternal death by 40% in past 20 years, a further 30% maternal deaths could be averted by fulfilling the unmet needs of family planning , 22. BesIn the current EDHS, about 34.9% of married women used modern contraceptive methods. This figure was much higher than previous percentages of EDHS data. In 2000’s and 2005’s DHS data contraceptive use was 8 and 14% respectively , furtherHousehold wealth index positively affects contraceptive use in the current study, as found previously –33. In EWomen age had also effects on contraceptive use, where younger women had higher odds than older women in child bearing age. Although the denominators are different, similar scenarios have occurred in previous studies , 43. TheHusbands have a determinant role in contraceptive use of their wife, and approval by the husband gives the women higher chance of contraceptive use . If husbAt the community level, there was lower odds of contraceptive use in Muslim dominant clusters. Similarly, women in Muslim communities of Asian countries usually had more children, desire for additional children and if they desire no more children, less likely to use contraceptive methods than the non-Muslims . Urban wCommunity level women employment and community level poverty were also significant cluster characteristics affecting contraceptive use. These factors were significant in different studies –57. Econ. The study used the most recent and representative nationwide surveys. The study also, applied a multilevel analysis to accommodate the hierarchical nature of the Ethiopian Demographic Health Surveys data. Having the above strengths, the study has also some limitations: Since the data used in this study was cross-sectional data, which limits the conclusions about the causality of the factors on the dependent variable. Moreover, recall bias and not using sampling weight for the analysis were the limitations of the study.This study has some strengthsIn nutshell, this study indicated both individual and community level factors can influence the use of contraception by women. Wealth index, women’s age, number of living children, husband’s occupation, ever experience of a terminated pregnancy, current working status of the women, number of births in the last 3 years, and hearing of FP messages through different media were significantly associated individual-level factors. Cluster characteristics like region, place of residence, religion, and community-level poverty were the factors associated with contraceptive use. Individual and community characteristics were significant predictors of use of modern contraceptives in Ethiopia and thus, these factors should be considered in programming for family planning in the country. Both individual and community-level characteristics should be priority areas in future measures to be taken in policies regarding contraceptive use in Ethiopia.


Second malignant neoplasm is one of the most devastating late effects of childhood cancers. This study aimed to evaluate the incidence and survival outcomes of patients developing second malignant neoplasms (SMNs) after surviving childhood cancer in Korea.Medical data of childhood cancer patients diagnosed between 1993 and 2012 were obtained from the Korea Central Cancer Registry. The risk of developing SMNs was calculated using standardized incidence ratio (SIR), excess absolute risk (EAR), and cumulative risk. Kaplan-Meier survival curves were estimated, stratified by SMN status.A total of 28,405 childhood cancer patients were diagnosed in the study period, and 337 (1.2%) developed SMN. The total follow-up period was 197,359 person-years at risk (PYR), with a median follow-up duration of 5.6 years. Overall SIR was 20.0, which was 23.2 in women, and 17.6 in men. The overall EAR was 16.4 per 10,000 PYR. The most common types of SMNs, in order of incidence, were other malignant epithelial neoplasms, leukemia, and soft tissue sarcomas. The cumulative incidence of developing SMNs was 0.7% at 5 years, 1.2% at 10 years, and 2% at 15 years. After primary cancer diagnosis, the 10-year overall survival rate of patients with SMNs was 65.1%, which was lower than the 73.4% in patients without SMN. After SMN diagnosis, the 10-year overall survival rate was 55.8%.Through this registry-based study of 5.6 years of follow up, childhood cancer survivors were found to be at 20-fold higher risk of developing a malignant neoplasm compared to the general population. The majority of malignant neoplasms are malignant epithelial neoplasms, leukemia, and soft tissue sarcomas. Continued surveillance for assessing long-term risks, and guidance for appropriate long-term follow up of childhood cancer survivors, are needed. Over the past two decades in Korea, the incidence of childhood cancer has increased substantially. A recent nationwide childhood cancer study in Korea showed that the age-standardized incidence rate for all cancers among children aged 0 to 14 years was 134.9 per million children in 1999–2011 [On the other hand, the increase of childhood cancer survivors has resulted in a substantial increase in the incidence of second malignant neoplasms (SMNs) after childhood cancer . The occA proper surveillance program for childhood cancer patients is needed to detect other malignant diseases and provide more preemptive management. Research has consistently demonstrated that the incidence of neoplasms in patients who have previously had cancer is higher than that in the general population , 8–11. EAccording to previous studies, it has been reported that risk of SMN increases with follow-up time, and that cancer treatment determines the risk of secondary cancer –15. The The total number of childhood cancer survivors in Korea is estimated to be approximately 30,000; however, recent statistics for SMNs in Korea are not yet available. A previous retrospective multicenter study involving 11 centers in Korea reported that 102 cases of second malignant neoplasm after childhood cancer were treated between 1998 and 2011 . HoweverThus, in the present study, we aimed to evaluate the incidence and survival outcomes of SMNs that occurred in childhood cancer survivors in Korea, through analysis of national registry data.Registry-based data on childhood cancer patients (aged 0–19 years old) diagnosed between 1993 and 2012 were obtained from the Korea National Cancer Incidence Database (KNCIDB) of the Korea Central Cancer Registry (KCCR). The KCCR is a population-based cancer registry that was begun in 1980 by the Korean Ministry of Health and Welfare, and which has covered the entire Korean population since 1999. The KNCIDB includes information about sex, age, date of first diagnosis, primary tumor site, and method of diagnosis. Detailed information about KCCR data has been reported previously . MortaliWe considered that primary malignancy treatment did not affect a second cancer diagnosis that was established within 6 months of the first cancer diagnosis. Therefore, patients who developed a second malignancy within the first 6 months of follow-up were excluded from the study . As a rerd edition [http://kosis.kr) was used as the general reference population. To measure the overall burden of SMN, excess absolute risks (EARs) of SMN were calculated by dividing the difference between the observed number and the expected number by the PYR, and were presented as numbers per 10,000 PYR. The number of PYR was defined from 6 months after the date of the first childhood cancer diagnosis to the date of death or the end of this study, whichever occurred first. In addition, we calculated the cumulative incidence of second primary cancer, which was defined from the time of first cancer diagnosis.Childhood cancer patients in this study were classified according to the diagnosis of the SMN, using the 12 main diagnostic groups of the International Classification of Childhood Cancers, 3 edition . SMNs in edition . The expP-values <0.05. All calculations of SIR and EAR were performed using the SEER*Stat software . The cumulative incidence rates and survival curves were generated, and log-rank tests were performed in SAS  and Stata (http://www.stata.com/).Kaplan-Meier survival analyses were done for primary cancer patients and SMN patients. Survival differences between primary cancer patients and SMN patients were estimated using a log-rank test. All statistical tests were two-sided and considered statistically significant for Malignant neoplasm was diagnosed in 28,405 patients aged 0–19 years, in 15,452 men and 12,953 women, who contributed a total of 197,359 PYR over the 20-year study period (1993–2012). Among these patients, 340 cases of subsequent malignant neoplasms were observed in 337 patients, including 3 patients who developed third cancers .The median age at initial diagnosis was 11 years old and the median follow-up period was 5.6 years. The most common type of SMN was ‘other malignant epithelial neoplasms and malignant melanomas’ (Group XI), diagnosed in 111 cases. This group is composed of adrenocortical carcinomas, thyroid carcinomas, nasopharyngeal carcinomas, malignant melanomas, skin carcinomas, and other and unspecified carcinomas. Thyroid carcinoma  was the most common type of neoplasm in this group .Following ‘other malignant epithelial neoplasms and malignant melanomas’, the second, third, fourth, and fifth most commonly-observed types of SMNs were ‘leukemias, myeloproliferative diseases, and myelodysplastic diseases’ , ‘soft tissue and other extraosseous sarcomas’ , ‘malignant bone tumors’ , and ‘central nervous system (CNS) and miscellaneous intracranial and intraspinal neoplasms’ , respectively. The overall SIR was 20.0 . The folThe SIR was higher in women  compared to men , but the difference was not statistically significant .According to the primary malignant neoplasm types, ‘soft tissue and other extraosseous sarcomas’ showed highest SIR (51.61) for developing SMN. . PatientOn comparing between the incidence of primary and secondary cancers types, all 5 retinoblastoma patients with SMN developed secondary “malignant bone tumors”. In addition, “leukemias, myeloproliferative diseases, and myelodysplastic diseases” was the most common type of SMN after primary “malignant bone tumor”  .Almost half  of SMNs occurred within 5 years of diagnosis of the primary cancer, while 93 cases (27.4%) occurred 5–10 years after diagnosis of primary cancer, and 86 cases (25.3%) occurred more than 10 years after diagnosis of primary cancer . AccordiThe overall EAR was 16.4 per 10,000 PYR . EAR wasThe median latency between first and second cancers was 5.3 years, which was shortest for secondary ‘lymphomas and reticuloendothelial neoplasms’ (1.3 years) and ‘neuroblastoma and other peripheral nervous cell tumors’ (1.3 years). On the contrary, ‘carcinomas of breast’ (11.9 years) and ‘thyroid carcinomas’ (10 years) had the longest median latency. The median age at SMN was 16 years, while the development of hepatic tumors occurred at the oldest age (25 years) . The cumThe 5-year overall survival rates were 76.2% for patients with only one primary cancer and 80.5% for patients with second malignant neoplasms. The 10-year overall survival rate of patients with SMN was 65.1%, lower than the 73.4% overall survival rate of patients without SMN, but statistically non-significant . The 5-yThrough this study, we concluded that risk of SMN in childhood was 20-fold greater in childhood cancer survivors than the risk of developing a malignant neoplasm in the general population. SIR and EAR of SMN among the childhood cancer survivors in this cohort were 20.0 and 16.4, respectively. When compared to previous reports from various nations, SIR in this study was higher than in others, which reported values between 5.0–8.0 , 21–23. When divided according to primary malignant neoplasm, breast cancer showed a particularly high SIR in this study. A high proportion of breast cancer cases occurred after a diagnosis of malignant bone tumor, soft tissue sarcoma, germ cell tumor, or other malignant epithelial neoplasm (ICCC-3 group XI). The median age at diagnosis of breast cancer in Korean women is approximately 10 years younger than that of women in the United States, and this difference may be associated with different environmental and genetic backgrounds between ethnicities . In addiIn our study, the risk of SMN was higher in females than in males, concordant with previous studies , 11, 22.TP53 mutation or RB1 mutation in osteosarcoma. Newly discovered mutations have been reported in various types of sarcomas [In this study, the primary cancer with highest SMN risk was soft tissue sarcoma and malignant bone tumor. Due to lack of data about treatment in our data set, it is impossible to identify the relationship between the treatment and results in SMN. However, previous studies have shown a possible relationship between treatment and SMNs in sarcoma patients. The high incidence of SMN in these groups could be partially related to treatment, including alkylators, anthracyclines, or epipodophyllotoxins . Radiatisarcomas . ExpansiSeveral studies showed high incidence of SMN after treatment of Hodgkin lymphoma , 22. HowAccording to this study, secondary leukemia or lymphoma mostly occurred within 5 years of the primary diagnosis, and carcinoma of thyroid or breast was found after 5 years of latency. Based on these results, a routine blood test is needed for at least 5 years after cancer treatment to screen for secondary hematologic malignancy. Beyond 5 years, surveillance of thyroid and breast cancers with physical examinations or imaging studies would be beneficial. In short, our results indicate the need of clinical follow up for more than 5 years and emphasizing continued surveillance to patients is of particular importance. As many cancer survivors do not realize the health risks that are related to their childhood cancer, many do not undergo the regular medical follow-up that is recommended for secondary cancer screening .In this study, the 10-year overall survival rate of patients with and without SMN were 65.1% and 73.4%, respectively. Although there are several long-term studies of childhood cancer survivors, only a few studies reported the overall survival of patients who developed SMN after childhood cancer. A recent Japanese study showed overall survival of survivors who developed secondary cancer, before and after 5 years from primary cancer diagnosis, to be 30% and 62%, respectively .This study has several limitations. First, as data was extracted from a population-based cancer registry database, it lacks detailed information about individual treatment methods, such as radiotherapy, chemotherapy and surgical therapy. Several studies have reported that secondary cancer risk is increased by the treatment received for the first cancer , 21, witIn spite of these limitations, this study retains its significance, as many studies concerning SMNs in childhood cancer do not provide the survival data. Furthermore, this study is valuable as the first Korean, nationwide, population-based study of SMN after childhood cancer. This study will be helpful for planning cancer control program strategies such as screening, surveillance, and treatment to decrease the incidence of SMNs and improve childhood SMN survival rates in Korea.Our findings suggest that among childhood cancer survivors, the risk of SMN is increased 20-fold when compared with the risk for the general population. Due to this increased risk, continued surveillance of SMN is recommended for childhood cancer survivors. Additionally, attempts should be made to reduce treatment intensity, and to select proper treatment modalities. Further studies with longer follow up time periods are needed to clarify the pattern of SMN, to reveal the etiology and risk factors for SMN in childhood primary-cancer survivors, and to provide an advanced follow up guide for childhood cancer survivors.


Chickpea flour was found to be nutritionally superior compared to refined wheat flour, and conventional processing of the flour was found not to affect its content of protein, fats, carbohydrates, and phosphorus, while total dietary and crude fibers were significantly increased. The fatty acid profile was minimally affected, while magnesium and potassium were reduced. The sensory test conducted among panelists (n = 60) showed that the incorporation of processed chickpea flour into the dough of mankoushe zaatar with ratios of 30% and 50% provided an end-product with better taste and overall acceptability compared to the regular mankoushe. Hence, conventionally processed chickpea flour can be used as a fortifier to improve the nutritional quality of bakery products without negatively affecting their sensory properties.Chickpea flour is known to have good nutritional values. Nevertheless, it is commonly made from ground grains, and characterized by an “off-flavor”. Processing of chickpea grains before flour formation reduces the intensity of the off-flavor. Therefore, two experiments were conducted: first to examine the effect of conventional processing  on the nutritional composition of the chickpea flour; and second, to investigate the impact of processed chickpea flour incorporation with different ratios on the sensory properties of  Pulses are important crops cultivated around the world because of their nutritional value. They play an essential role in human nutrition, particularly in the dietary pattern of low-income individuals in developing and underdeveloped countries. Pulses are good sources of protein, carbohydrates, minerals, and vitamins. These grain legumes are characterized by high dietary fiber content, oligosaccharides, slowly digestible starch, and resistant starch ; thus, tWorldwide, chickpeas received attention because of their physiological benefits. Extensive efforts have been conducted to increase the intake of these grain legumes by integrating them as flours into food products: bread ,8, spaghmankoushe zaatar. It is a widely consumed pastry similar to pizza, in which the dough base is topped with mixed herbs, known as zaatar. In this study, chickpea grains were conventionally processed , and the impact of processing on the chemical composition of the flour was also examined.Thus, the aim of this study was to examine the effect of processed chickpea flour incorporation with different ratios on the sensory properties of the traditional Lebanese pastry Cicer arietinum L.), Kabuli type, were purchased from the local Lebanese market. Chickpea grains were hand-sorted to remove foreign materials. Some of the grain was kept raw, and some was soaked in distilled water in the ratio of 1:10 (w/v) for 17 hours, after which the water was discarded and fresh tap water was added to the chickpeas in the ratio of 1:10 (w/v). Grains were boiled at 100 °C for 1 hour in a double-jacketed kettle, drained, then dried in an industrial air drier  for 8 hours at 38–40 °C. The two types of chickpea, raw and treated grains, were ground into powder form with an electrical, multifunction swing type portable grinder , sifted through a 60-mesh stainless steel sieve (250 µm particle size) to obtain a fine flour. Each kind of flour was packed in a vacuum form into nylon bags for later use. Refined wheat flour (as control) was obtained from Crown Flour Mills, Beirut, Lebanon. Chickpea grains  Offshore, Beirut, Lebanon. The additional ingredients  were all purchased from the local market. Three samples were prepared for the proximate fatty acids and macrominerals analysis. Sample 1 contained refined wheat flour as a control, sample 2 contained raw chickpea flour, and sample 3 contained processed chickpea flour.For the topping, The proximate composition analysis was carried out on the control flour, the raw chickpea flour, and the processed chickpea flour, according to the standard methods of the Association of Official Analytical Chemists  . MoisturFatty acid (FA) composition was measured in the three flour samples. Palmitic, palmitoleic, stearic, oleic, linoleic, and linolenic FA were determined according to the International Organization for Standardization (ISO) (ISO 12966-4:2015) . The com3 and 2 mL of H2O2. Then 0.5 g of the sample was diluted in 50 mL. Some tubes were spiked with 1 ppm of P, Mg, and K each. One blank was spiked with 5 ppm of P, Mg, and K. Minerals were determined via inductively coupled plasma-atomic emission spectrometry (ICP-AES) [The macromineral content of phosphorus (P), magnesium (Mg), and potassium (K) was determined in the three different samples. All samples were digested with 4 mL of NHOICP-AES) . All theMankoushe is a dough-based pastry topped with mixed herbs, known as zaatar. zaatar is composed of ground oregano leaves, sesame seeds, sumac spices, and salt, mixed in the following percentages, respectively: 35.86%, 43.04%, 17.93%, and 3.15%. In the present study, three types of mankoushe were made in which the base was modified and the topping was maintained: regular mankoushe with 100% refined wheat flour, as a control; “chickpea-30” mankoushe, containing dough made up of a mixture of refined wheat/processed chickpea flour (70/30); and “chickpea-50” mankoushe, in which the dough was made up with a mixture of refined wheat/processed chickpea flour (50/50). Afterward and upon use, sunflower oil was added to the topping mixture based on the following percentages: 31.70% topping mixture and 68.30% oil. As for the dough, the three types were prepared under controlled conditions, according to the formulation given in mankoushe were baked in an electric oven  for 6 min at 300 °C. Food samples were prepared just before the sensory test.The base is usually made of refined wheat flour, and n = 60), aged between 20 and 40 years (mean ± standard deviation (SD) of 24.15 ± 3.8). Subjects were completely blinded to the three mankoushe samples. The test samples, assigned with three number codes, were presented randomly to the panelists and served at room temperature. Four sensory characteristics: texture, color, taste, and overall acceptability were evaluated using a nine-point hedonic scale, with a score of 1 (Dislike Extremely), 5 (Neither Like nor Dislike), or 9 (Like Extremely) [The study was approved by the Ethics Committee at the Doctoral School of Science and Technology (DSST), and all subjects were asked to sign a consent form before participation. Participants were recruited through an advertisement posted at the DSST, Lebanese University. A sensory analysis (Preference Test) was performed under optimal conditions . For the purpose of maximizing participant homogeneity, the study was conducted among untrained university females (tremely) . p < 0.05 level.Data were analyzed using Statistical Package for Social Sciences . Food composition results are expressed as the mean value ± standard deviation of three separate determinations, and data were evaluated using two-way ANOVA and Tukey’s post-hoc test. Sensory parameters were analyzed by comparing the means using one-way ANOVA followed by Tukey’s post-hoc test. In all analysis, significant differences were determined at the p < 0.001) in both chickpea flours compared to the refined wheat flour. However, moisture and carbohydrate contents were detected to be considerably lower. Chickpea flours appeared to be of higher nutritional value compared to refined wheat flour in terms of proteins, fat, ash, and fibers. These findings were in line with those reported by Hefnawy et al. [The proximate compositions of flours are presented in y et al. .p < 0.001) in processed (1.85%) compared to the raw (2.9%) chickpea flour. Total dietary fibers (15.75%), and crude fibers (3.7%) were modestly, but significantly (p < 0.001) increased in processed chickpea flour compared to raw chickpea flour, in which total dietary fibers and crude fibers were 14.70% and 1.63%, respectively. It appears that conventional processing of chickpea grains ultimately increased the fiber content of the flour. Fibers are the edible parts of pulses that are resistant to digestion and absorption in the human small intestine, with complete or partial fermentation in the large intestine. Fibers are of high importance since they have beneficial physiological effects, including laxation, blood glucose, and cholesterol attenuation [Between the two types of chickpea flour, proteins, fat, and total carbohydrates appeared to be similar. However, ash content was significantly lower  in palmitic acid between refined wheat (21.96%) and both chickpea flours (about 12%). While the amount of saturated FA stearic acid (2.72%) was lower in wheat compared to raw and processed chickpea flours (about 4%), the linolenic, linoleic, oleic, and palmitoleic acid contents were similar between flours . The fatty acids profile  shows a In the current study, conventional processing had minimal effects on the FAs, except for stearic. Nevertheless, the impact of processing on the FA profile of chickpea flour is not well documented, and varied affects have been reported .Furthermore, findings show that the main FAs found in raw and processed chickpea flours is the linoleic acid (about 58%) followed by the oleic acid (23%). Linoleic acid has numerous beneficial impacts, including improved blood lipid profile, improved insulin sensitivity, lower incidence of type 2 diabetes, and anti-arrhythmic effects . Oleic ap < 0.001), while P was minimally affected (9.26%). Mineral losses are likely to be attributed to their leakage into the cooking water or to the conventional processing steps. Soaking, cooking, drying, and milling are critical processes in flour production. Nonetheless, these steps are known to affect the concentrations of inorganic elements in grains [The phosphorus (P), magnesium (Mg), and potassium (K) content of chickpea flour  was highn grains .P, Mg, and K are essential minerals to human health. They have been suggested to be potentially protective against obesity and metabolic disorders ,16,17. PChickpea flours (raw and processed) in the present study differed in their mineral content from the ones reported earlier ,19. The mankoushe—regular, chickpea-30, and chickpea-50—are presented in mankoushe with regard to texture and color. As for the taste, chickpea-50 mankoushe was significantly accepted (p = 0.026) compared to regular. Regarding the overall acceptability, a significantly higher score (p = 0.005) was observed between the regular and the two types of chickpea mankoushe. However, the chickpea-30 and chickpea-50 varieties were similar in all sensory attributes.The sensory acceptability results of the three types of mankoushe significantly. The sensory enhancement seen in this study could also be attributed to Maillard reactions [Processing is well-known to enhance the flavor of pulses . In the eactions . It has eactions .The use of raw chickpea flour as an ingredient in bread generated inconsistent data. Chickpea bread with an incorporation rate of 30% showed either similar  or lowermankoushe at different ratios enhances the sensory qualities of the baked product. Therefore, it is a promising fortifier to improve the nutritional properties of wheat-based products. Raw and processed chickpea flours appear to be of a high nutritional value compared to refined wheat flour, even though conventional processing of chickpea grains affects the chemical composition of the flour. Nevertheless, processed chickpea flour is still considered to be a good source of protein, fibers, minerals, and healthy fats. Processed chickpea flour is suitable as an ingredient for bakery products, since its incorporation into the dough of


To determine the prevalence of hepatic steatosis (HS) in children andadolescents with cystic fibrosis (CF) and associate it with nutritionalstatus. Cross-sectional study with children and adolescents with CF diagnosis.Weight and height were used to calculate the body mass index (BMI) andsubsequent classification of the nutritional status. The midarmcircumference (MAC), triceps skinfold thickness (TSF) and midarm musclecircumference (MAMC) were used to evaluate body composition. Abdominalultrasonography was performed for diagnosis of HS. The statistical testsused were Student’s t test, Mann-Whitney test and chi-square test withsignificance level of 5%. 50 patients with CF were evaluated, 18 (36%) were diagnosed with HS (GroupA) and 32 (64%) without HS (Group B). The mean age of Group A was 13,2±4,9years old and Group B 11,7±4,9; for BMI, the value for Group A was 18,0±4,1and Group B was 15,7±3,8; the TSF of Group A was 8,4±3,5 mm and Group B was7,0±2,5 mm. For these variables, there was no significant difference betweenthe groups. The mean of MAC and MAMC differed significantly between thegroups, being higher in the HS group, with p values of 0,047 and 0,043. The frequency of HS in patients with CF is high and it is not related tomalnutrition, according to the parameters of BMI, TSF and MAMC. The valuesof MAC and MAMC indicated a greater reserve of muscle mass in patients withHS. Recently, attention has been given to hepatic involvement in cystic fibrosis (CF), asthis is one of the main causes of death, respiratory failure and complicationsrelated to lung transplantation.There are several reasons that hamper understanding the actual prevalence of liverdisease in CF: there is no unanimously accepted definition of criteria for diagnosisof liver disease in CF, most patients are asymptomatic, and highly sensitive andnon-invasive tests are scarce. The term hepatic disease in CF is non-specific andhas been used in several studies to describe a broad spectrum of hepatobiliarydiseases such as: neonatal cholestasis, elevated liver enzymes, imagingabnormalities including liver parenchymal heterogeneity upon ultrasonography, portalhypertension, liver failure and histological abnormalities such as fibrosis,cirrhosis, and hepatic steatosis (HS).HS is reported in a case series with frequency of 20-60%.In this context, this study aimed to describe the prevalence of HS found by abdominalultrasonography in children and adolescents with CF and associate it withnutritional status.This was a cross-sectional study carried out with 50 children and adolescents agedbetween 2 and 19 years of age, with diagnosis of CF established by two sodium andchloride dosages in sweat above 60 mEq/L.UniversidadeEstadual de Campinas (UNICAMP), Campinas, SP, Brazil, in 2016, andwhose caregivers signed the free informed consent form. Exclusion criteria were:patients on hepatotoxic drugs with elevated aminotransferases, severe dyslipidemia,or other liver diseases that may occur with HS .Inclusion criteria were: patients assisted at the Cystic Fibrosis Outpatient Clinicof Clinical Hospital of the School of Medical Sciences, 2), and BMI was classified according to the WorldHealth Organization (WHO) growth curves.Weight, height, midarm circumference (MAC) and triceps skinfold thickness (TSF) weremeasured by the anthropometric techniques recommended by Lohman. A Lange skinfoldmeasure was used to measuring TSF, and a Sanny tape was used to measure MAC. Thebody mass index (BMI)/age was calculated using the Quetelet index wascalculated, according to The percentage of adequacy was measured according to Patients with values ≤90% were classified as malnourished, while eutrophic patientswere those whose percentile resulted >90. Patients with presenting values>110% were considered as having excessive body fat only for TSF.Abdominal ultrasound is the most commonly used imaging method to identify HS becauseit is relatively low-cost, non-invasive, easy to apply and available in mostservices. The sensitivity of the method was 89%, and specificity was 93%.Pancreatic insufficiency was considered present in patients using pancreatic enzymesand/or presenting with steatorrhea. Diabetes mellitus was considered according tothe classification by the Brazilian Guidelines for the Diagnosis and Treatment ofCystic Fibrosis.The method for selection of participants was convenience sampling; the 85 patientscared for in 2016 were invited to participate and 50 of them met the inclusion andexclusion criteria. Patients were divided into two groups according to ultrasoundresults:Group A: patients with CF and HS.Group B: patients with CF and without HS.Data were analyzed in the IBM Statistical Package for the Social Sciences (SPSS)software version 20.0. The Kolmogorov-Smirnov test was applied to evaluate thedistribution of quantitative variables in Gaussian curve. The descriptive analysisof continuous variables included calculation of means and respective standarddeviations for variables with normal distribution, and calculation of medians and25th and 75th percentiles for variables that did not adhere to the normality test.Categorical variables were expressed in percentage values. Student’s t-test was usedto compare the variables with normal distribution according to the presence of HS;for variables without normal distribution, the Mann-Whitney test was applied. Thechi-square test was used to investigate the association between HS and categorizedanthropometric indicators. As some variables were allocated in more than twocategories, the verification of local association between categories was analyzed bycalculating adjusted residuals. Adjusted residual values greater than 1.96 wouldindicate a statistically significant association between both categories.This study was approved by the Research Ethics Committee of the School of MedicalSciences of UNICAMP, protocol 494,781.Fifty patients from the Cystic Fibrosis Outpatient Clinic of UNICAMP ClinicalHospital were evaluated, being 23 (46%) females and 27 (54%) males aged between 2and 19 years of age (12.2±4.9); there were four children and 14 adolescents in GroupA, 13 children and 19 adolescents in Group B, considering the classification forchildren aged <10 years.Eighteen patients (36%) were diagnosed with HS by abdominal ultrasonography (Group A)and 32 (64%) were diagnosed as not having HS (Group B). When evaluating theassociation between HS and nutritional status, the variables BMI and TSF did notdiffer significantly between groups, meaning no association between HS andmalnutrition. MAC and MAMC were significantly different between groups, with thehighest difference in the HS group, indicating greater muscle mass .From all patients evaluated, 48 (96%) had pancreatic insufficiency and seven (14%)had diabetes mellitus and no significant association with presence or absence of HS.These data show that the mean BMI of the HS group (18.0±4.1) was higher whencompared to the non-HS group (15.7±3.8), but with no significant statisticaldifference; 15/18 patients (83.3%) with HS were considered eutrophic . When adThere is a broad spectrum of hepatic involvement in CF including HS, withmalnutrition as one of the causes involved. In the present study, we report a highfrequency of HS, but not related to malnutrition. The exact mechanism of liverdisease in CF is not well-known. The primary alteration is known to involve agenetic defect of the cystic fibrosis transmembrane conductance regulator (CFTR) ofbile epithelial cells, leading to the production of thick biliary secretion,evolving with biliary ductal obstruction and resulting in the development offibrosis and biliary cirrhosis.The causes of HS secondary to CF have not yet been fully clarified. The pathogenesismay be related to malnutrition, deficiencies of essential fatty acids, carnitine,choline, oxidative stress, and insulin resistance, and not only to a CFTR gene.When evaluating the relationship between HS and BMI/age, 16.7% of patients presentedmalnutrition, but no significant association with the non-HS group, since 31.2% ofpatients without HS were malnourished. TSF showed adipose tissue depletion in bothgroups, but without statistical difference, unlike MAC and MAMC, which were shownsignificantly different between groups, demonstrating that in the presence of HS thepatient maintains a better reserve of lean mass and, consequently, higher MAC.Isolated MAC analysis does not allow to affirm that there is an increase in leanmass, but by analyzing low TSF values, it can be concluded that MAC values reflecthigher muscle reserves. This result should be confirmed with larger samples andother methods that evaluate muscle mass.In the presented series, 13/50 patients (26%) presented malnutrition when classifiedby BMI/age, regardless of HS. When the mean percentile was evaluated, it was belowthe recommended (percentile 31.52). The Cystic Fibrosis FoundationAccording to the publication of 2014 by the European Cystic Fibrosis Society,The Brazilian Registry of Cystic Fibrosis (REBRAFC),A study conducted in Rio Grande do Sul with 82 patients aged 7 months to 16 yearsshowed that 26.8% of the sample was malnourished by BMI <10 percentile as cutofffor malnutrition.,Nutritional status in our study was assessed according to WHO standards. Otherstudies have used the recommendations by the international consensuses of CF,comparing divergent studies, since there is a reduction in the number of individualsconsidered eutrophic and an increase in cases of malnutrition. This was identifiedin another study.Malnutrition is multifactorial in CF, including poor dietary intake, increased dailyenergy requirement and is associated with poor nutrient digestion. Patientsdiagnosed in newborn screening programs benefit from early intervention, which isassociated with positive nutritional status.One of the limitations of this study was not evaluating body composition byelectrical bioimpedance, to better interpret anthropometric measures, which showedgreater reserve of lean mass in HS group. Another limitation was the lack of asurvey on eating habits to identify unbalanced diet and use of dietary supplementsthat could interfere with nutritional status. Another limitation was the smallsample, composed by subjects selected by convenience sampling.In conclusion, the frequency of HS is high in patients with CF and it is notassociated with malnutrition, according to the parameters of BMI, TSF and MAMC. MACand MAMC values indicated greater reserve of muscle mass in patients with HS.


Fatty acids (FAs) of milk fat are considered to be important nutritional components of the diets of a significant portion of the human population and substantially affect human health. With regard to dairy farming, the FA profile is also seen as an important factor in the technological quality of raw milk. In this sense, making targeted modifications to the FA profile has the potential to significantly contribute to the production of dairy products with higher added value. Thus, FAs also have economic importance. Current developments in analytical methods and their increasing efficiency enable the study of FA profiles not only for scientific purposes but also in terms of practical technological applications. It is important to study the sources of variability of FAs in milk, which include population genetics, type of farming, and targeted animal nutrition. It is equally important to study the health and technological impacts of FAs. This review summarizes current knowledge in the field regarding sources of FA variability, including the impact of factors such as: animal nutrition, seasonal feed changes, type of animal farming , genetic parameters (influence of breed), animal individuality, lactation, and milk yield. Potential practical applications  of FA profile information are also reviewed. Fatty acids (FAs) in milk fat are considered to be important nutritional components of the diets of a substantial part of the human population. According to scientific knowledge, they can also affect human health. In the past, FAs have been regarded as having negative impacts on human health; however, in the last ten years this notion has been greatly reassessed. Currently, the impact of milk fat on human health is thought of in a much more positive way than it was in previous periods ,4,5,6,7.In the case of dairy farming, the FA profile is also seen as an important factor in the technological quality of raw milk. Therefore, the FA profile has the potential to significantly contribute to the production of dairy products with higher added value. As such, FAs also have an economic importance. Current developments in analytical methods and their increasing efficiency enable the study of milk FA profiles not only for scientific purposes but also in terms of practical technological applications. It is important to study the sources of FA variability in milk, which include population genetics, farming management, and targeted animal nutrition . It is eEfforts to carry out practical improvements of milk FA profiles to benefit consumers are usually driven by two reasons: (1) from a nutritional point of view, a lower proportion of saturated FAs (SFA) and a higher proportion of unsaturated FAs (UFA), especially polyunsaturated FAs (PUFA) n-3, is desirable; and (2) from a usability point of view, higher proportions of UFA are preferred . However, there are also problems associated with having high UFA content in milk fat, including its lower stability and the accompanying phenomena such as oxidation and possible sensory changes.c9, t11; isomer of conjugated linoleic acid (CLA)) content in milk fat.Making desirable changes to the FA profile requires a thorough knowledge of the various factors that influence milk fat composition. It is also important to know the extent to which these relevant factors are involved in influencing the FA profile. Some factors affecting the FA profile of milk , lactation stage (days in milk), and diet) have been described previously ,11,12,132 > 0.5), as well as very good models for trans isomers of UFA (TFA) (R2 > 0.6) have been utilized for this purpose [In-depth knowledge of these factors can also be used to predict milk FA profile. This can be carried out effectively for bulk milk samples based on information about farm practices . Good prediction models for SFA and PUFA . Therefore, a more effective analytical process is necessary. Over the past twenty years, there have been significant advances in practical hardware and software development for effective routine indirect analyses of milk FAs ,35,36,37c9 in oven-dried milk, approximate for PUFA, C18:0, vaccenic  and RA, and poor for linoleic acid , alfa-linolenic  acid, PUFA n-6, and PUFA n-3. Quantification was more accurate for oven-dried milk, but good results were also obtained for SFA, MUFA, UFA, and C18:1 c9 in liquid milk . Lower result reliability  was obtained for TFA and RA .Near infrared spectroscopy has been used and evaluated for successful estimation of milk FA profiles . Resultsc9, C18:1 c9, SFA, and MUFA in milk . In all cases [p < 0.001); whereas, they were lower for TFA and PUFA . This means that 50.3% and 88.2% of the variability in routine values of SFA and UFA were explainable by variations in reference values. These indirect routine analytical methods are now opening the door to more intensive and effective research studies and quantifications of the sources of variability in milk FA profiles. Nevertheless, it is technically more effective to use mid infrared spectroscopy with Fourier transformation (MIR-FT) in a flow design apparatus for liquid milk ,35,37,38The amount of milk fat and its composition depends mainly on two processes: lipid metabolism in the rumen and lipid metabolism in the mammary gland. Furthermore, FAs released from body reserves during negative energy balance at early lactation also contribute to the final composition of milk fat . Metaboltrans isomers (bacteria firmly attached to feed particles), or branched-chain FAs (bacteria loosely attached to feed particles) [c9, VA, RA, and MUFA compared to silage-based feeding [In many countries, intensive production of milk relies on two feeding strategies: year-round indoor feeding based on preserved feed or seasonal feeding based on grazing during summer combined with indoor feeding during winter. Preserved feed is represented mainly by corn, grass, legume, grass-legume silages or a combination of the above, all supplemented with concentrates. The spectrum of plants used for feeding purposes depends largely on local soil and climatic conditions. Furthermore, the proportion of forage in the diet of dairy cows can range from 50 to 90 percent of dry matter. Diet composition is the main factor that can cause shifts in the microbial diversity of the rumen  with subrticles) . Recent rticles) , oil suprticles) , changesrticles) , or a swrticles) , as also feeding .Of the types of silage mentioned in According to the literature ,51, the c9, RA and cis-MUFA, and decreases the proportion of SFA compared to silage-based feeding. Of the preserved feedstuffs, the most suitable seem to be either legume silage or mixed silage [From the perspective of enhancing the HFA/C18 ratio see , grazingd silage .c9 (20.3%) and ALA (6.8%), rapeseed products, in general, are a good source of C18:1 c9 (more than 50%) with lower amounts of LA (20.6%) and ALA (8.9%) [c9 and RA and by decreasing C14:0 and C16:0  [c9, VA, and RA.Using oilseeds in dairy cow diets is a common nutritional strategy used to improve the FA profile of milk fat. Their effect on milk FA composition has been recently reviewed in many studies ,52,53. SA (8.9%) . As docuilseeds) . Based oAs the most important factor affecting the variability of milk FA profile is dairy cow nutrition, comparison of organic (OS) and conventional (CS) farming systems seems to be very important. OS is characterized by grazing, with a reduced amount of conserved forage (but a higher ratio of this type of feed than concentrates) and a small proportion of grain concentrates compared to CS. The main reason for the differences between organic and conventional milk is predominantly in terms of nutritionally desirable components, rather than a genotypic effect ,60.In terms of nutrition, comparing milk fat of organic and conventional milk is interesting. Statistically significant differences between OS and CS were only found in the proportions of PUFA and TFA. Both groups of FAs were found to have higher proportions in organic milk ,61 than In extensive herds (low-input non-organic farms) with mostly pasture-based feeding (94% of dry matter in feed ration) and no conserved forage, the increase in RA, ALA and MUFA was even more pronounced than in OS and CS where pasture was combined with conserved feeding . On the c9, LA, and ALA, were more abundant in organic milk compared to conventional milk in the relevant study [p < 0.05). In contrast, C16:0 and C18:0 were 10% higher in conventional milk (p < 0.05). SFAs were 4% higher in conventional milk; whereas, UFAs were 9% higher in organic milk (p < 0.05). PUFAs, including the essential FAs n-6 and n-3, were 25% higher in organic milk. Furthermore, the nutritionally desirable FA parameters indicated by the n-6/n-3 and PUFA/MUFA ratios were 33% and 25% higher in organic milk than in conventional milk, respectively. In general accordance with the above-mentioned results, concentrates, corn (or other) silage, hay, and straw decreased the nutritionally desirable FAs such as MUFA, PUFA, PUFA n-3, ALA, and n-6/n-3 ratio; whereas, these feeds increased SFA, PUFA n-6, and LA [Similarly, C18:1 nt study . In part, and LA .Results for MUFA and PUFA show greater consistency, and higher proportions of VA, RA, ALA, and eicosapentaenoic acid  in organic milk have been reported independent of the country of origin .de novo and preformed FAs [It is interesting that organic milk usually yields more nutritionally favorable FA profiles  than conventional milk ,61,71. Trmed FAs  or precurmed FAs . Also, hrmed FAs ,76,77.In general, up to 44% of milk fat can originate from a cow’s diet. Therefore, management of animal nutrition during lactation is considered to be critical for producing a high quality FA profile in milk fat, more so than any other factors such as cow breed and genotype, age, health, and aspects of lactation ,62,65,76de novo synthesis in mammary glands; ruminal biohydrogenation; body fat mobilization) [Lactation stage, along with energy balance of dairy cows, has an impact on the FA profile of cow’s milk. Changes in milk FA composition during lactation, particularly at the beginning of lactation, originate from altered activity in pathways of FA derivation  ,79,80,81ization) . Therefoization) , approxiization) , or evenization) . Mammaryization) . Body reization)  through ization) ,88. Thisization) ,90,91 asization) . NEFA meization) ,93,94,95de novo synthesis of FAs through the inhibition of acetyl coenzyme A carboxylase [c9, decrease in proportion until week 12 pp [Thus, the beginning of lactation is the most demanding period in terms of energy status and also herd health management ,86,96,97ek 12 pp ,81,100 . Three FAs that are referred to as HFA  are loweactation ,81. RA iactation . Accordiactation ,116.In terms of parity, primiparous cows have a nutritionally more desirable FA composition, with a lower proportion of SFA and a higher proportion of UFA and RA ,118.Use of genetic knowledge to alter milk fat is possible only with these prerequisites: (1) genotypic and phenotypic variation; and (2) accurate estimation of genetic parameters . Fat content is much more variable in comparison to other milk constituents . High variability was found not only in fat content, but also in the proportion of FAs and their groups ,119,120.Finally, knowledge of genetic parameters is important for the design of animal breeding programs and for prediction of selection responses . EvaluatThe first studies on heritability of FA profiles began in the 1970s ,128. TheDe novo synthesized FAs are influenced mainly by animal factors ; whereas, pre-formed FAs are influenced by feed factors [Generally, heritability depends on FA saturation and on carbon numbers of the FAs . As such factors . However factors ,125 have factors . Therefo−1 of milk vs. g·100 g−1 of FA vs. g·100 g−1 of fat); (2) the methods used for FA analysis (reference GC vs. routine MIR-FT); (3) the models used for estimation of heritability ; and (4) the database structure. These database structures are determined by the relative genetic and environmental variances because each have a different design given by the number of herds  or by the number of samples available for analysis [As shown in analysis ,132,133.analysis  and the analysis .Some studies ,122,126 Heritability estimates for FAs are usually higher for FAs expressed in 100 g of milk or in g per day, rather than in 100 g of fat or 100 g of FA ,134. ThiOverall, results for genetic correlations indicate that cows are genetically predisposed to produce more SFA when fat content increases . On the FAs were negatively correlated with milk yield and protein yield, but positively correlated with fat yield, fat content, and protein content . Very sic9, which is an indicator of body fat mobilization (0.42 at 5 days in milk and −0.40 at 230 days in milk).It has been noted  that corSimilar to heritability estimates, genetic correlations vary depending on parity , with thIt should be noted that it is also possible to apply genetic information in attempts to make changes to fat composition, even though UFA expressed a strong negative phenotypic correlation and a weak genetic correlation .ACSS2, FABP3), synthesis and desaturation of FAs , triacylglycerol synthesis , regulation of transcription , and others [A large number of genes participate in milk fat biosynthesis . Despited others . PolymorDGAT1 (K232A) and SCD1 (A293V) genes have been thoroughly studied [DGAT1 is lower (3–15%) than the variance explained by the SCD1 (6–52%) polymorphism [In relation to FA profile, polymorphisms of the  studied ,145,146.morphism .Cattle breed may affect the FA profile of milk ,150,151;−1 of FAs) as follows (for the same order of breeds as listed in the previous sentence): 25.0–39.9; 20.9–22.5; 20.9–29.7; 21.5–24.8; and 16.1–22.0. The same figures for PUFA were: 3.3–4.9; 3.2–3.5; 2.8–4.7; 2.5–4.3; and 2.6–3.2.Some papers have described significant milk FA profile differences between cattle breeds ,150,151.c9, C17:1 t9, LA, ALA, C20:1 c9, C20:4 n-6, PUFA, and UFA, as compared to Holstein-Friesian (HF) milk. Additionally, Simmental milk had a lower amount of C15:0, C18:0, C20:0, C22:0, and CLA. Compared to HF milk, the PUFA/SFA and UFA/SFA ratios in the Simmental milk were significantly higher; whereas, the thrombogenic index and the LA/ALA ratio were significantly lower [In a recent study from an organic farm, Simmental milk had, in the total content of FAs, a significantly higher content of C12:0, C16:1 ly lower .Some minor local breeds have also been studied with regard to milk FA profile . The PolIn general, it is possible to state that local cattle breeds tend to have a better milk fat composition with respect to desirable FAs than breeds that are more efficient in terms of dairy yield. However, these breeds are more often kept under extensive conditions (more often OS), which is also related to a typical variation in dairy cow diet—the proportion of ingredients in the feeding ration is more in favor of forage (fresh or preserved) compared to grain concentrates than under intensive conditions. There have not yet been enough experiments on breed differences in milk FA profile that have been carried out on cows with the same nutritional conditions . Of course then, it is possible that a portion of these breed FA differences is primarily defined by dietary sources.The variability of FAs in bovine milk due to genetic factors and FA profile differences among cattle breeds have been well-studied ,150,172.Because it is difficult to evaluate the nutritional and also technological value of milk fat from the content of individual FAs, some ratios or indices have been proposed. First, these indices were used primarily to evaluate the negative effect of C12:0, C14:0, and C16:0 on human health. Formulas for calculating the atherogenic (AI) and thrombogenic (TI) indices as well as a modified calculation of S/P were proposed by Ulbricht and Southgate :AI =  is calculated according to Santos-Silva et al. :HH =  and can be calculated on the basis of product/substrate [The activity of Δ9-desaturase is essential in the process of UFA synthesis. This enzyme catalyzes the introduction of a C18:1 c9 ,176. Thaubstrate ; substraubstrate , or prodc9/(C14:0 + C14:1 c9) ratio has been suggested as the best indicator for Δ9-desaturase activity [c9 is likely to be the product of Δ9-desaturase activity [Although it is possible to calculate DI in different ways, it should be noted that the C14:1 activity  because activity :DI = 100c9, which has been shown to be the most accurate indicator of butter hardness [All the abovementioned indices together with PUFA/SFA and n-6/n-3 PUFA ratios are widely used to evaluate the nutritional value of milk fat. It is supposed that milk fat with high AI and TI values may be more likely to contribute to the development of atherosclerosis or coronary thrombosis in humans; whereas, milk with high HPI and HH ratios may have a protective effect against cardiovascular diseases . Althoughardness ). The oxs, e.g., , the invAs presented in c9 [As expected, DI was influenced by the type of forage because of greater differences in the concentration of C18:0 and C18:1 c9 . The effc9 ,32,33.Of the possible forages, the best SI of milk fat was found after feeding corn and grass silages, while the highest value was calculated for pasture-based feeding systems. For oilseeds, an increased SI value and thus softer butter fat was produced after feeding rapeseed products, which has also been documented by e.g., .p < 0.05). All relationships of MUFAs to milk indicators were insignificant (p > 0.05). Relationships between PUFAs and milk indicators were narrower: fat ; lactose ; milk alcohol stability ; titration acidity ; cheese curd quality ; milk fermentationability ; streptococci count in yoghurt ; and total count of noble bacteria in yoghurt . Relationships of CLA to selected milk indicators were as follows: fat ; lactose ; alcohol stability ; and cheese curd quality . Thus, higher CLA levels were associated with higher fat and lower lactose content, as well as lower alcohol stability. Biological variability in milk metabolic indicators  can also be explained, in part, by the relationships among physiological, technological, and health milk indicators. Relationships between FAs and their groups and selected milk indicators (bulk milk samples) were studied in Czech Fleckvieh and Holstein cows . The onlc9, were not significantly related to either lactation sum or daily production parameters, or to the content of basic components of milk with the exception of C16:0 (30.93 ± 4.81% in milk fat), which has a negative relationship to daily milk (−0.4) and protein production (−0.35). This FA has also been positively associated with fat content (0.44) and negatively associated with lactose content (−0.31). CLA was negatively correlated with daily fat production (−0.41), fat content (−0.27), and fat/protein index (−0.42). In order to better understand and interpret milk FA profiles, knowledge of the relationships between major FAs and their groups and other milk indicators is also important.Correlation coefficients (>0.3) for the summarized values of lactation, which were calculated in regular milk recording were observed  at MUFA In addition to the above mentioned milk indicators, there are also documented associations between the milk FA profile and metabolic disorders such as ketosis or between the milk FA profile and reproduction performance ,136. Morc9 in the liver as well as C16:1 c9 and C18:0 in adipose tissue. Energy-protein supplementation significantly affected the content of most FAs in blood  and liver , but it did not affect the profile of adipose tissue in cows. Therefore, it is necessary to consider these effects when developing methods to control the production of animal raw materials for the food industry.Last but not least, dairy cow nutrition substantially influences the profiles of other body tissues, especially body liquids, in addition to the milk FA profile. Accordingly, an analysis of the percentage (by weight) of FAs in different body tissues  was doneMilk and dairy products play an important role in human nutrition because they provide not only essential nutrients such as high-quality proteins, fat, lactose and minerals, but also various physiologically active compounds such as vitamins, bioactive peptides, and antioxidants ,188,189.Milk and some dairy products (butter) have been criticized for the unfavorable FA profile in milk fat. Indeed, bovine milk fat contains on average about 70% SFA, 25% MUFA, and 5% PUFA ; whereasThe SFAs that play the greatest role in a negative view of milk fat , have often been associated with having adverse effects on indicators of cardiovascular risk . This iAt present, there is strong research interest in the nutritional quality and health benefits of food, which is supported by public awareness and an on-going desire to improve our quality of life. Consequently, targeted modification of the FA profile of milk fat is desirable. Developments in analytical methods have played an important role in increasing the efficiency and feasibility of studies on the sources of FA variability. In recent decades, many studies have been devoted to improving milk FA composition by increasing the amount of FA with beneficial effects on human health and appropriate technological properties. Accordingly, knowledge of the important factors affecting milk FA composition and their relationships, including physiological aspects such as rumen fermentation, fat synthesis in mammary gland tissue, and energy status of animals, is essential from both a research and practical point of view. This review has therefore focused on the main sources of FA variability. Breed, animal genetics, metabolic and lactational effects, management, and other factors were mentioned; nevertheless, feeding strategy is undoubtedly considered to be the most efficient way to modify milk FA composition. Some factors are well known—e.g., role of diet and stage of lactation—while others, such as polymorphism, are only beginning to be understood. The possibility of using milk FA profiles as indicators to predict animal health or even methane production levels is currently under active investigation. Further research on sources of FA variability is important for finding effective ways of improving the health benefits and technological quality of milk products through modifications in the FA profile. This will most likely be achieved by targeted changes to feeding and breeding strategies.


PSMB5, PRMT5, ACTB, PDE3A, NPC1, FSCN1, NR5A2, IQCG, LHX8, and DMRT1 were identified in these window regions for these five semen traits. Our findings provided a solid basis for further research into genetic mechanisms underlying semen quality traits, which may contribute to their accurate genomic prediction in Chinese Holstein population.Efficient production of high-quality semen is a crucial trait in the dairy cattle breeding due to the widespread use of artificial insemination. However, the genetic architecture  underlying such semen quality traits remains unclear. In this study, we performed genome-wide association studies to identify genes associated with five semen quality traits in Chinese Holstein population, including ejaculate volume, progressive sperm motility, sperm concentration, number of sperm, and number of progressive motile sperm. Our dataset consisted of 2,218 Holstein bulls in China with full pedigree information, representing 12 artificial insemination centers, with 1,508 genotyped using the Illumina BovineSNP50 BeadChip. We used a weighted single-step genome-wide association method with 10 adjacent Single nucleotide polymorphisms (SNPs) as sliding windows, which can make use of individuals without genotypes. We considered the top 10 genomic regions in terms of their explained genomic variants as candidate window regions for each trait. In total, we detected 36 window regions related to one or multiple semen traits across 19 chromosomes. Promising candidate genes of  The fertility of dairy cattle has decreased over recent decades due to highly intensive selection for milk production . Male feSemen traits such as progressive sperm motility (SM) and ejaculate volume (VE) are complex and affected by genetic factors  as well In our previous study, a single-SNP GWAS was performed for five semen traits in a population of 692 animals . Due to In order to study the genetic basis of the semen traits, a weighted single-step GWAS (WssGWAS) was performed in a larger Chinese Holstein bull population. Recent studies have shown that WssGWAS can improve precision in estimating SNP effects and perform better than traditional methods under certain conditions, including with small sample sizes with both genotypes and phenotypes where traits are regulated by loci with large effects . It alsoAll procedures pertaining to the handling of experimental animals were conducted in accordance with and approved by the Animal Welfare Committee of China Agricultural University (Permit Number: DK996). All efforts were made to minimize discomfort and suffering.9 spermatozoa per ml. Number of sperms (NSP) per ejaculate was calculated by multiplying VE and SC, and the number of motile sperm (NMSP) was obtained by multiplying NSP and SM. After removal of the error data , we yielded phenotypic records of 527,207 records of VE, 527,231 records of SC, 521,150 records of MS and NMSP, and 522,906 records of NSP. The means, standard deviations, and minimum and maximum values of these traits are shown in A total of 2218 Holstein bulls born between 1996 and 2016 were represented in semen samples and phenotypes, from 12 artificial insemination (AI) centers across China where routine semen testing has been carried out since 1995. Distributions of bulls’ birth year and location are given in DNA was extracted from 1508 semen samples using a standard phenol–chloroform method. Illumina BovineSNP50 BeadChip versions 1 and 2  were used to genotype these individuals. For filling the missing genotypes, genotype imputation was carried out using BEAGLE software version 3.3.1 , and 52,The complete pedigrees of all individuals included in this study comprised a total of 4,237 animals, which had 497 sires, 1,512 dams, and 2,695 bulls. The number of bulls with phenotypes was 2,218, and the number of bulls with both phenotypes and genotypes was 1,508.P value to evaluate the importance of SNPs, and the other is a Bayesian method that utilizes all SNPs simultaneously as random effect and output SNPs effect to evaluate their importance ; ais the vector of additive genetic effects with X, Z, and W are the design matrices of β, a, and p, respectively. I is an identity matrix, and H–1 is created as follows: A22 is the numerator relationship matrix for genotyped animals , D is a diagonal matrix of weights for SNP variances , and q is a weighting factor. The weighting factor can be derived based on SNP frequencies  to investigate the gene function based on orthologous genes of human.We also used the web of genecards . On BTA 8, a window region of 43.78–44.28 Mb was shared by NMSP and NSP, with five genes located there. Of these, DMRT1 was a cell-identifying gene in the testis cord and seminiferous tubules, according to LifeMap Discovery (https://discovery.lifemapsc.com/in-vivo-development/testis). On BTA 24, the window region 33.25–33.69 Mb was shared by VE, NMSP, and NSP, and nine genes were located there. Of these, the expression of NPC1 was related to the testis interstitium according to LifeMap Discovery (https://discovery.lifemapsc.com/in-vivo-development/testis). In total, we identified 36 window regions across 19 chromosomes, out of which 8 window regions were overlapped by at least two semen traits. On BTA 5, a window region of 113.47–113.87 Mb was shared by VE, NMSP, and NSP, and 12 genes were located in this region. Out of these 12 genes, P = 0.001), which is likely to be related to semen traits. Six genes were in this GO term, including CYB5R3, SLCO1B3, NPC1, SLCO1A2, SLCO1C1, and NR5A2. It is worth noting that the DMRT1 gene was in the GO term of male germ cell proliferation (GO: 000217) and spermatogenesis (GO: 0007283), indicating that DMRT1 is a promising candidate gene.The GO enrichment analysis of the 10 top regions for the five semen traits revealed 11 significant GO terms, including biological process, cellular component, and molecular function. The details of the significant GO terms are shown in Based on analysis above, we concluded 10 promising candidate genes for these five semen traits Table 3.In WssGWAS, different weights were given to SNPs according to their importance, and linkage disequilibrium made the consecutive SNP window method more effective at detecting Quantitative Trait Loci (QTL) compared with a single-SNP method . As a rePDE3A and SLCO1C1  on BAT 1 was detected in our study, which was also identified in GWAS by IQCG knockout mice can show total immobility and severe malformation of spermatozoa, due to disorganization of the sperm flagellum axoneme . LHX8, preferentially expressed in testicular germ cells  between NMSP and NSP were confirmed in our previous study , and ourrm cells , may plarm cells .NPC1 and NR5A2 were included in the GO term of steroid metabolic process (GO: 0008202) (P = 0.001), which is the most important to semen traits due to their spermatogenesis function being regulated by steroid hormones, the most important of which is testosterone. NPC intracellular cholesterol transporter 1 (NPC1) on BTA 24 was located in an overlapping window region among VE, NMSP, and NSP. Lack of a functional NPC1 protein can cause multiple defects in mouse sperm and result in sterility , expressed from NR5A2, is produced in human steroidogenic tissues and activates transcription of genes encoding steroidogenic enzymes  and spermatogenesis (GO: 0007283). DMRT1 is located in an overlapping window region between NMSP and NSP, and is required for spermatogonial stem cell replenishment and maintenance in mice  on BTA 25 was identified as a promising candidate gene for MS. ACTB is expressed in sperm and is distributed in the acrosomal and postacrosomal regions of ejaculated spermatozoa, where it is potentially involved in membrane changes during the acrosome reaction with important implications to sperm function (B-actin (ACTB) gene regulates the process from spermatogenesis to fertilization . All efforts were made to minimize discomfort and suffering.SZ and JL designed and supervised the study. HY and CZ contributed to genomic DNA extraction and conducted GWAS analysis, HY and CZ contributed to population construction and statistical analysis with help from JL and DS. SS contributed to genomic DNA extraction, phenotypes collection, and sample collection. HY drafted the manuscript, which was critically remarked by LF, JL, DS, and SZ. All authors read and approved the final manuscript.This work is supported by the 863 project (2013AA102504); the National Science and Technology Programs of China (2011BAD28B02); National Key Technologies R&D Program (2012BAD12B01); Beijing Dairy Industry Innovation Team, China Agricultural Research System (CARS-37); and Xinjiang Province Key Technology Integration and Demonstration Program (201230116). We are deeply grateful to all donors who participated in this program.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.The reviewer GM declared a past co-authorship with one of the authors LF to the handling editor.


The advent of RNA-sequencing (RNA-Seq) technologies has markedly improved our knowledge and expanded the compendium of small non-coding RNAs, most of which derive from the processing of longer RNA precursors. In this review article, we will present a nonexhaustive list of referenced small non-coding RNAs (ncRNAs) derived from eukaryotic ribosomal RNA (rRNA), called rRNA fragments (rRFs). We will focus on the rRFs that are experimentally verified, and discuss their origin, length, structure, biogenesis, association with known regulatory proteins, and potential role(s) as regulator of gene expression. This relatively new class of ncRNAs remained poorly investigated and underappreciated until recently, due mainly to the a priori exclusion of rRNA sequences—because of their overabundance—from RNA-Seq datasets. The situation surrounding rRFs resembles that of microRNAs (miRNAs), which used to be readily discarded from further analyses, for more than five decades, because no one could believe that RNA of such a short length could bear biological significance. As if we had not yet learned our lesson not to restrain our investigative, scientific mind from challenging widely accepted beliefs or dogmas, and from looking for the hidden treasures in the most unexpected places. The ribosomes are ribonucleoprotein (RNP) complex assemblies required for the translation of all proteins ,2. DurinMammalian ribosomal DNA (rDNA) genes code for rRNAs that are generally comprised of several hundreds of transcription units organized in tandem repeats and clustered on a number of chromosomal loci ,8,9. ForEach ribosome is composed of rRNA—constituting its functional core—and ribosomal proteins. In eukaryotes, ribosomes consist of four rRNAs and approximately 80 ribosomal proteins arranged into two subunits: 60S and 40S . During Nevertheless, it has been shown that distinct ribosomal genes (rDNA) are expressed at different stages of development, leading to the incorporation of alternative forms of rRNA with heterogeneous sequences into ribosomes ,28. It iSince the discovery of the first small silencing RNA in 1993 , a notewThese studies range from the small RNAs like the snoRNA   (~80%), mRNAs (~5%), and transfer ribonucleic acid (tRNA) (~15%) are the most abundant RNA molecules. Despite their relatively high enrichment and potential function, the small ribosomal RNA-derived fragments (rRFs) are usually removed as a by-product of RNA degradation from RNA-sequencing (RNA-seq) or small RNA-seq analyses ,57. SimiHowever, even when rRNA sequences are included in the analyses, some rRFs may be missed because standard approaches use a reference transcriptome that does not include all the possible variations from rDNA sequences. To detect rRFs that differ from the rDNA sequences of reference, the challenge will be to annotate all repeat variability and existing SNPs (single-nucleotide polymorphisms) . NotablyNevertheless, over the past few years, scientists have begun investigating the existence, role, and function of specific small rRFs, which will be the topic of this review article. Whereas rRNA plays a role in ribosome heterogeneity, rRFs may be involved in the control of translation, albeit not excluding other important biological functions. Here, we will discuss about the discovery, biogenesis, protein-binding capacity, and function of small ncRNAs derived from rRNA. The 28S rRNA is the longest rRNA and forms the large subunit (LSU) of eukaryotic cytoplasmic ribosomes . The matAmblyomma testudinarium model and in human cell lines . Th. Th40]. Finally, the longest rRF originating from the 28S rRNA, as reported among other rRFs in an RNA-seq study of Zebrafish development, measures 80 nt ,28. ThisNeurospora crassa) are approximately 20–21 nt in length and form a hook structure [Previously described qiRNAs  reported by Locati and al.  (ZebrafiShort hairpin RNA (shRNA)-induced depletion of the 28S rRF3, in the H1299 cell line, significantly increased cell apoptosis and inhibited cell proliferation . MoreoveQDE-2-interacting small RNAs have been shown to be required for the DDR and repair pathway in rice ,67. In aStudies involving Argonaute (Ago) protein immunoprecipitation, followed by high-throughput sequencing, on various species, including Arabidopsis and Drosophila models and human cell lines, revealed that rRFs co-immunoprecipitated with Ago1 and Ago2 ,73,74,75Also similar to miRNAs, phasiRNAs encoded by PHAS play important regulatory roles by targeting protein coding transcripts in plant species . GeneralThe biogenesis pathway leading to the formation of the 18S rRNA is different from that of the 5.8S and 28S rRNAs ,78; rRFsThe most abundant maternal-type rRF detected during Zebrafish developpement  comes frInterestingly, a group of clustered noncanonical miRNAs derive from pre-rRNA , and thrThe 130-nt rRF ncRNA derived from the 18S rRNA, as described above, has a secondary structure with a stem and a complex hinge with three smaller hairpins . In factIt has been reported  that botThe existence of rRNAs and of rRFs suggest dual molecular functions ,65,83,94Finally, these clustered miRNAs were differentially expressed in different tissues, suggesting that rRNA processing into rRFs may be placed under specific spatiotemporal control . ExpressThe existence of two forms of 5.8S rRNA, with 7 or 8 different nt at their 5′ end, is widely described in eukaryotes ,35,36,96RNA-seq data, obtained from developing Zebrafish , unveilePiper nigrum77, originating from the 5′ end of the 5.8SL rRNA and representing the largest subset of rRFs of 23 nt [Because the rRFs mentioned above are not the only ones originating from the 5.8S RNA in eukaryotes; some forms are shorter and more abundant ,102,103,The majority of the 20-nt long fragments deriving from the mature sequences of tRNAs, rRNAs, snoRNAs, and small nuclear RNAs (snRNAs), are produced, in a specific cleavage pattern, from the 5′ or 3′ end ,104. TheInterestingly, three regions of 5.8S rRNA can form helices by base pairing with the 28S rRNA as we can see in the secondary structure diagrams found in Andreas M. et al., supplementary data . The lonA 5′ consensus 5.8S rRF sequence of 22 nt was found in all seed plants (Spermatophyte plants) . The majA new small ncRNA, mapping to the 5′ region of rRFs and called cosRNAs (clustered organellar short RNA fragments), were recently identified (Plasmodium) . These rA large group of piRNAs and endogenous small interfering RNAs (endo-siRNAs), produced upon rRNA processing, have been identified as “unconventional” small ncRNAs with regulatory functions in mouse . These rThe 5.8S rRNA participates to ribosome translocation and thus exerts an essential role in protein synthesis ,119. TheArabidopsis thaliana 5.8S rRNA predicts a secondary structure composed of hairpins and of a noncanonical miRNA-like short hairpin precursor, to which the second most abundant class of 5.8S rRFs could be mapped [In silico analysis of e mapped . Locati e mapped  discovere mapped . This pre mapped ,124,125.e mapped . Once thConcerning expression patterns, the phenomenon of 5′–3′-specific processing is observed across all major classes of ncRNAs, except mRNAs . MoreoveArabidopsis thaliana and Oryza sativa revealed an association between 5.8S rRF5 variants and Ago complexes [RNA-seq analysis of Ago co-immunoprecipitation experiments ,127,128 omplexes , such asomplexes . Specifiomplexes . Interesomplexes . Nonetheless, the Ago-rRF interaction could have another explanation: would it be possible for rRFs to be the targets of a miRNA? For instance, a study identified chromatin-binding sites for Ago2 throughout the 45S region of the human rRNA gene . This woOn one hand, this interaction could be dependent on Dicer, as the cross-linking of Ago2 to the rRNA was reduced to background levels in the absence of Dicer . MoreoveTogether, these results suggest that rRFs may exert important functions in fundamental mechanisms through their association with specific proteins, as each rRF may exert a specialized function depending on its incorporation into specific RNP complexes. Deep sequencing analysis of small RNAs that emanate from the highly repetitive rDNA arrays of Drosophila revealed the existence of small RNAs deriving from internal transcribed spacer (ITS) of rRNA . The autFurthermore, in the filamentous fungus model Neurospora, numerous qiRNAs derived from the external and internal transcribed spacer regions  have been described . They arAccording to a study conducted by Son et al. , it is nLike mature rRNA, the ITS1 and ITS2 sequence length increased during evolution . These lTaken together, the studies discussed in this review article demonstrate that the 28S, 18S, and 5.8S rRNAs, and even the ITS1 and ITS2, produce one or more small rRFs. These rRFs are present at various, yet significant, levels in different cell types or organs, and during development, like in the often described Zebrafish development model. The generation of these small rRFs does not appear to result from random degradation of the associated mature rRNAs. Moreover, the degradation rate of mature cytoplasmic rRNAs is generally beyond detection under normal conditions , as the Whereas the notion of degradation products is the first to come to mind when considering small rRFs, it was shown recently that tRNAs and rRNAs undergo stress-induced cleavage to produce stable rRFs products, and that this mechanism is conserved from yeast to human cells ,114,116.It is likely that these rRFs are not generated in a random manner, but by a highly regulated cleavage and maturation process. Knowing that they derive from a highly expressed rRNA precursor, the rRF generation could be under strong selective pressure. It may be that the biogenesis of a given rRF is guided by RNA motifs and the recruitment of specific RNA binding proteins ,67. Moreover, some specific cleavages have been observed, especially for rRFs coming from the 3′ and 5′ ends of rRNAs . In someMoreover, the same locus in two different organisms could generate two different rRFs involved in different functions . That isA comparable mechanism could be imagined for the rRFs deriving from mature 18S, 28S, and 5.8S rRNAs, but, so far, there is little known about the sequence variability of these rRNAs . In both cases, these rRFs could derive from mature and functional rRNAs that would be differentially processed depending on the environmental condition. Another possibility is that they would be cleaved from certain rRNA pseudogenes that are not matured and incorporated into ribosomes, but instead serve as precursors for rRFs.Finally, when looking at the targets of these rRFs, there is evidence that some of these act like miRNAs and target mRNAs to prevent their expression by inhibiting their translation or leading them to degradation ,86,129.