To study and analyze the effect of perioperative comprehensive nursing intervention on transcatheter arterial chemoembolization (TACE) in patients with primary hepatic carcinoma (PHC). n = 55) and an observation group (n = 55) by random number sorting according to odd and even numbers. Patients in the control group received conventional nursing interventions and those in the observation group received comprehensive nursing interventions. The two groups were compared in terms of surgical status, quality of life (QoL), and nursing satisfaction.  One hundred and ten patients with PHC diagnosed in our hospital from May 2019 to January 2022 were randomly selected and divided into a control group (P < 0.05) shorter than those in the control group; the observation group had significantly (P < 0.05) higher scores of quality of life, including somatic function, emotional function, role function, social function, and cognitive function than the control group; chemotherapy adverse reactions including fever, abdominal pain, urinary retention, and gastrointestinal reactions in the observation group were significantly less than those in the control group (P < 0.05); the total incidence of complications in the observation group was significantly (P < 0.05) lower than that in the control group. The total satisfaction with nursing care in the observation group was significantly (P < 0.05) higher than that in the control group.  The operation time, postoperative bed rest time, and hospital stay in the observation group were significantly ( The perioperative application of comprehensive nursing intervention in TACE for patients with PHC aids in the smooth operation, improves patients' QoL, lowers the risk of chemotherapy reactions and complications, and enhances patient satisfaction and nursing quality. These advantages justify a wider perioperative application of comprehensive nursing intervention in TACE clinical practice. Primary hepatic carcinoma (PHC) refers to a malignant tumor occurring in hepatocytes or intrahepatic cholangiocytes . Recent PHC treatment is defined by the convergence of several approaches and disciplines in clinical practice, with a focus on interdisciplinary comprehensive care and individualized treatment. The main treatment approaches include hepatectomy, liver transplantation, local ablation, transcatheter arterial chemoembolization (TACE), radiotherapy, and systemic drug therapy. TACE is an interventional therapy that primarily involves femoral artery intubation and sending anticancer drugs or embolic agents into the hepatic artery , 9. ThisPrevious research findings reveal that TACE can not only lengthen patients' lives and control disease progression but also pave the way for secondary surgical treatment . Howevern = 55) and the observation group (n = 55) according to odd and even numbers through random number sequencing. Patients in the control group were treated with routine nursing intervention, while patients in the observation group received comprehensive nursing intervention. Their families and themselves were informed of the study and signed the informed consent form voluntarily. The experiment has been approved by the ethics committee of our hospital .A total of 110 patients who were diagnosed with PHC and received TACE in our hospital from May 2019 to January 2022 were randomly selected, including 81 males and 29 females, aged between 43 and 75 years, with a mean age of 60.34 ± 9.84 years. All patients were allocated to the control group (  Inclusion criteria: (1) patients who met the diagnostic criteria for PHC; (2) patients who received TACE treatment; and (3) patients with no distant metastasis found in clinical examinations.  Exclusion criteria: (2) patients with severe heart, lung, liver, and renal insufficiency; (2) patients with mental illness or unconsciousness; and (3) patients with complications such as severe jaundice or abnormal coagulation function.Patients in both groups were treated with TACE after being diagnosed and admitted to the hospital . They rePatients in the control group received routine nursing intervention, including observation of illness, improvement of preoperative examination, medication guidance, routine dressing change at the puncture site, bandaging, and prevention of infection.Before the operation, patients were informed of relevant information to allow them to fully and correctly understand the etiology, clinical manifestations, treatment methods, and prognosis of cancer. Any discomfort and complications that may occur after operation were explained in detail. According to the individual situation of patients, appropriate psychological care and health guidance were formulated, and a relationship of mutual trust with patients was established for patient encouragement and compliance as well as the elimination of negative emotions such as anxiety, fear, depression, and sadness that often appear in the treatment process. Preoperative guidance and preparation of surgical instruments and drugs were conducted, followed by a detailed physical examination and allergy tests. Patients were reminded to fast 4 hours before the operation.Intraoperative: patients cooperated with physicians to complete the operation, with the indexes of patients normally checked and the internal conditions of patients monitored. Patients were informed of the chemotherapy process promptly. Physicians could remind patients to be mentally prepared before injection chemotherapy, which may cause obvious discomfort. They could also tell patients that the operation was successful and praise the patient's compliance when it was approaching the end of the operation. Various conditions should be handled reasonably appropriately: pain care, attention to control, respiratory control, and position adjustment to relieve the patient's pain. Patients in severe pain may be given analgesics to relieve severe pain with the consent of the physician's diagnosis. Nursing for complications and adverse reactions was performed with prevention in advance by ventilating and maintaining a warm temperature in the ward, frequently changing sheets and clothes, closely monitoring the condition, and timely handling the side effects and complications of chemotherapy to different degrees. Diet nursing was followed according to the individual physiological and dietary characteristics of patients, with reasonable and healthy daily diet plans formulated, and the diet plans focused on light foods with high calorie, high cellulose, and high protein, boosting the patients' body resistance and immunity. Daily life nursing intervention was conducted through formulating activity plans according to the actual situation of patients after operation, such as encouraging patients to get out of bed for postoperative recovery, paying attention to sleep quality, guiding patients to rest more, drinking water properly, sleeping well, carefully following the doctor's instructions on medication, and regularly checking liver function and blood circulation .Patients in the observation group were treated with comprehensive nursing intervention. Specific treatment included the following:Detailed recording and comparison of the two groups, including the duration of surgery, postoperative bed rest, and length of hospital stay.QoL rating scale (SF-36) was used to evaluate patients' quality of life, which was divided into five dimensions: somatic function, emotional function, role function, social function, and cognitive function. The total score of each dimension was 100 points. The higher the score after evaluation, the better the patient's quality of life.Chemotherapy adverse reactions, including fever, abdominal pain, urinary retention, and gastrointestinal reactions, were recorded in detail and compared between the two groups.We also recorded and compared the complications of the two groups, including renal failure, tumor rupture, hemorrhage of the digestive tract, and puncture point bleeding. The total incidence of complications was calculated and compared between the two groups. Complication rate = number of people with complications/total number × 100%Using the Questionnaire of Nursing Satisfaction created by our hospital , we set four options of satisfaction  to understand the satisfaction of the two groups of patients and determine which one of the two groups of treatment methods was more effective through the results.t-tests were performed for enumeration data (n (%)) and measurement data (s), respectively. The difference was statistically significant if P < 0.05.The GraphPad Prism 8 software was used to process images, and data processing was conducted with the SPSS 22.0 software. Chi-square tests and P > 0.05) difference in general data between the two groups, but the two groups were comparable. See In the control group, there were 55 cases, including 41 males and 14 females, aged 43–75 years, with a mean age of 61.25 ± 9.24 years. The course of the disease was 0.5–7.1 years, with a mean course of disease of 3.53 ± 1.78 years. The diameter of the carcinoma was 5–13 cm, with a mean diameter of 7.82 ± 1.71 cm. There were 55 cases in the observation group, including 40 males and 15 females, aged 43–75 years, with a mean age of 59.42 ± 9.72 years. The course of the disease ranged from 0.8 to 7.5 years, with a mean course of disease of 3.68 ± 1.74 years. The carcinoma diameter ranged from 5 cm to 13 cm, with a mean diameter of 7.68 ± 1.84 cm. There was no significant (P < 0.05) shorter than those in the control group . See The operation time, postoperative bed rest time, and hospital stay in the observation group  were significantly (P < 0.05) higher scores of QoL, including somatic function, emotional function, role function, social function, and cognitive function  than the control group . See The observation group had significantly (P < 0.05) less than those in the control group (39 (70.91%), 32 (58.18%), 28 (50.91%), 31 (56.36%)). See Chemotherapy adverse reactions including fever, abdominal pain, urinary retention, and gastrointestinal reactions (12 (21.82%), 15 (27.27%), 8 (14.55%), 10 (18.18%)) in the observation group were significantly (P < 0.05) lower than that in the control group (32.73%). See There were 0 (0.00%) case of renal failure, 0 (0.00%) case of tumor rupture, 1 (1.82%) case of hemorrhage of the digestive tract, and 2 (3.64%) cases of puncture point bleeding in the observation group, while 5 (9.09%) cases of renal failure, 0 (0.00%) case of tumor rupture, 7 (12.73%) cases of hemorrhage of the digestive tract, and 6 (10.91%) cases of puncture point bleeding in the control group. The total incidence of complications in the observation group (5.45%) was significantly (P < 0.05) higher than that in the control group (72.73%). See The results of questionnaire survey showed that 15 (27.27%) patients in the control group were very satisfied, 25 (45.45%) were satisfied, 11 (20.00%) were not very satisfied, and 4 (7.27%) were dissatisfied. In the observation group, 26 (47.27%) patients were very satisfied, 28 (50.91%) were satisfied, 1 (1.82%) was not very satisfied, and 0 (0.00%) was dissatisfied. The total satisfaction with nursing care in the observation group (98.18%) was significantly (Primary liver cancer has become the fourth most common malignancy and the second leading cause of death in China, posing a serious threat to human safety and health . The etiTACE is a combination of medical imaging and clinical treatment that has become one of the most important treatments for the treatment of PHC, as it can help reduce tumor volume and allow for a second operation. Although liver resection or transplantation provides better results than other therapies in the local control of HCC, survival rates are unsatisfactory, particularly for large tumors, and it remains controversial whether HCC angiogenesis is enhanced after TACE even in patients undergoing radical resection . It is fPrevious studies have revealed that TACE for patients with PHC has noticeable adverse effects, such as digestive system reactions, that affect postoperative recovery and treatment effectiveness . Based oTACE has been shown to have positive efficacy. However, clinical controversy remains due to the necrosis of normal liver segments or lobe tissue in patients after occlusion of the liver donor vessels and the toxic side effects of chemotherapeutic drugs and embolic agents. The patient's postoperative recovery and overall treatment outcome are affected by a variety of adverse effects or complications. The application of comprehensive nursing intervention in the perioperative period requires postoperative nursing of complications and adverse reactions, which can be prevented in advance by ventilating and keeping warm in wards as well as changing sheets and clothes frequently. In the case of adverse reactions, medical staff will monitor the situation closely so that chemotherapy-related side effects and complications of varying degrees of severity can be managed promptly. This approach is effective in reducing the risk of chemotherapy reactions and complications for patients, as well as preventing patients from being bedridden, hospitalized for observation, or treated for adverse reactions and complications. The results also demonstrated that the observation group had significantly higher scores of QoL and total satisfaction with nursing care than the control group.The physical pain, organ damage, and negative emotions associated with PHC may increase the risk or exacerbate the extent of adverse reactions. Also, due to the technical aspects of TACE and the physical condition of the patient, one or more adverse chemotherapy reactions and complications often occur after the procedure, leading to a variety of negative emotions and a significant deterioration in the patient's quality of life. However, comprehensive nursing interventions are required throughout the perioperative period, which means that care should be provided before, during, and after the procedure. Pain care alleviates the patient's suffering. Dietary care improves the patient's immune system. Daily life nursing intervention encourages patients to get out of bed for postoperative recovery, guides patients to rest more, drink enough water, and sleep well. Patients are instructed to carefully follow the doctor's advice on medications and regularly check liver function and blood circulation. It not only satisfies patients' needs for love and belonging, eliminates negative emotions, and increases patients' confidence and compliance in treatment, but it also improves their QoL, contributing to a considerable increase in patient satisfaction. This is consistent with the findings of Li et al., who found that perioperative comprehensive nursing could improve patients' psychosocial and self-care abilities, followed by a higher QoL .To sum up, the perioperative application of comprehensive nursing intervention in TACE for patients with PHC aids in the smooth operation, improves patients' QoL, lowers the risk of chemotherapy resections and complications, and enhances patient satisfaction and nursing quality. These advantages justify a wider perioperative application of comprehensive nursing intervention in TACE clinical practice.


Chemical analysis was performed on sediment samples collected in two sampling sessions (July and October) from Podu Iloaiei Dam Lake, one of the most important water resources used for aquaculture in north-eastern Romania. The concentration of 15 trace elements (TEs), 8 refractory elements (REs), and 15 rare earth elements (REEs)—determined using inductively coupled plasma mass spectrometry—showed variability largely dependent of the sampling points and collection time. Manganese was the most abundant TE, V and Cr were the most abundant REs, while Ce was one of the most abundant REEs. The cerium negative anomaly and Gd positive anomaly were observed in the Chondrite-normalized distributions. In October, the Ce anomaly showed significant negative correlation with Mn, emphasizing the water body oxidation potential. The identified positive Gd anomaly was most likely associated with the use of Gd-chelating agents in magnetic resonance imaging in Iasi, the largest medical hub in north-eastern Romania. Principal component analysis extracted three factors explaining 96.0% of the observed variance, i.e., rock weathering, leaching from soil surface, contributions from urban stormwater and atmospheric deposition (50.9%), pedological contributions (23.7%), and mixed anthropogenic sources . The evaluation of pollution indices highlighted low and moderate degrees of contamination for most of the elements and a considerable degree of contamination for Cd. Assigned Cd sources included fertilizers and pesticides used in the near agricultural areas or the high traffic road located near the lake. Since contamination of aquatic ecosystems with harmful elements is a human health concern, further monitoring of specific vectors in the food chain of the investigated dam lake will be of the utmost importance. The nature of element sources in sediments, combined with their different mobility, persistence, and toxicity, are critical factors in the assessment of their environmental impact . DetermiIn urbanized areas, several heavy metals at trace levels are assigned as major pollutants. The direct discharge of industrial effluents, domestic wastewater into the environment, and use of agricultural chemicals has led to elevated heavy metal concentrations in urban soils and surface water bodies ,4,5. In The extensive use of relatively new technologies is a major source of rare earth elements (REEs) in the environment, and the emission of REEs into natural waters is increasing . The mos4+ and Eu2+ are of high interest [3+ to Ce4+ may occur [Although, in aqueous environments, the REEs chemistry involves almost exclusively the trivalent oxidation state, stable compounds formed by Ceinterest ,18, as tinterest ,13,19. Iay occur , with thay occur ,18. Bothay occur ,13,21. Gay occur ,22,23.The spatial distribution characteristics of metals in sediment may indirectly give an indication of the pollution sources localization. Multivariate statistics techniques  are powerful tools for pollution source assessments ,24. ThesgeoI), and enrichment factor (EF) [Sediment contamination across a wide range of environments is reported in several studies ,2,3,31. tor (EF) ,33,34,35tor (EF) ,37. In ttor (EF) ,39. Constor (EF) .To our knowledge, limited information on lake sediment quality in north-eastern Romania is available. Strungaru et al.  studied The present study aimed both to evaluate the distribution of potentially toxic heavy metals and rare earth elements and to assess the anthropogenic impact on lake sediments from Podu Iloaiei Dam Lake. The investigated lake is located on the Bahluet River from the Bahlui river basin and is considered as one of the most anthropized hydrological basins of the country in north-eastern Romania. The evaluation of possible contamination of the lake water and sediment is of the utmost importance since it is an important water resource for fish farming in the region. The objectives of this study were: (1) to investigate the spatial distribution characteristics of 15 trace elements (TEs), 8 refractory elements (REs), and 15 rare earth elements (REEs) in Podu Iloaiei Dam Lake sediments in north-eastern Romania; (2) to evaluate element sources in sediments and driving environmental processes using multivariate statistical methods; (3) to assess the pollution and to identify the harmful potential of these elements in the investigated area. The results obtained in this study may provide a reliable scientific database for the estimation of the local ecological status and implementation of pollution remediation strategies.The Podu Iloaiei Dam Lake is located on the Bahluet River from the Bahlui hydrographic basin, north-eastern Romania. More geographical details of the hydrographic area are elsewhere reported . The surIn the Prut–Barlad hydrographic area, including the Bahlui hydrographic basin, agricultural (71%), forest (14%), and residential (12%) areas are the dominant forms of land use. According to the estimate of the National Institute of Statistics, only a small area is occupied by surface waters (3%) .A total number of 28 sediment samples were collected from Podu Iloaiei Dam Lake within s two session measurement, on 16 July 2017 and 5 October 2017, and the distribution of the sediment sampling sites is presented in 2O2) was used as the oxidizing agent, as it is one of the strongest available [2O2 reagent proved to be suitable for complete sample dissolution.The solid-state dissolution procedure and inductive coupled plasma-mass spectrometry (ICP-MS) analyses were applied for the metal elemental analysis in the sediment samples. Sodium peroxide  was added to each sample. The samples were further mixed with the oxidizing agent, followed by transfer to the porcelain crucibles lined with aluminum foil. Thermal processing was performed at 460 °C. After cooling, the sintering mix was dissolved in HNO3 3% solution prepared in deionized water and stored in closed polypropylene vials until analysis.The sediment samples were dried slowly and, subsequently, were finely grinded using an agate mortar and pestle. Around 0.1 g of sediment sample was weighed, and 50 µL of internal standard solution , refractory elements , and rare earth elements  in the sediment samples were determined by the ICP-MS technique using the ICP-MS 7700X system .® Indium ICP standard from Merck KGaA  was used as an internal standard. Certipur® ICP multi-element standard solution XXI for MS  was used for TE quantification, Specpure® refractory metals plasma standard solution  was used for RE quantification, and TraceCERT® rare earth element mix for ICP  was used for REE quantification. Nitric acid 65% Suprapur®  was used for HNO3 3% preparation. Ultrapure water obtained from an Advantage A10 water purification system  was used for standards and sample dilution.Analyte quantification was performed by an internal standard calibration procedure. CertipurSpecial emphasis was placed on carefully controlling the entire array of preparatory and analysis steps in order to prevent the possible contributions brought by sample loss or contamination. For quality control assurance, blanks and control samples were frequently analyzed.The geostatistical approach was used in the present work to comprehensively describe the variation of elements concentration in the sediment of Podu Iloaiei Dam Lake. For the quantified elements, analysis of the geospatial distribution was used in order to discriminate between potential contributions of natural or anthropogenic sources. Google Earth Pro and ArcMap (included in the ArcGIS 10.2 package) programs were used for geospatial distribution analyses. For data processing inverse distance weighting (IDW), interpolation was used.Pearson correlation analysis and principal component analysis (PCA) were used to identify the possible sources of elements in sediment samples from Podu Iloaiei Dam Lake. Varimax normalized PCA was used to determine the association of the different variables by reducing the dimensionality of the dataset. The concentrations of the quantified elements from sediment samples were used as variables for the PCA. The validity of the PCA was checked with the Kaiser-Meyer-Olkin (KMO) statistic and Bartlett’s test using SPSS (version 22.0). The PCA was performed to determine possible chemical associations that could explain TE, RE, and REE behaviors and the relationships between them.p < 0.05.The Kolmogorov-Smirnov test, carried out in OriginLabPro 2022, was performed to check for possible differences between the sampling sessions. Differences were considered of significance at the level of The correlation matrix, including Pearson correlation coefficient values, pie charts, pollution indices heat map representations, and the boxplots of the potentially harmful elements, were graphically represented in OriginLabPro 2022.The normalization to Chondrite  and the N represents the normalized concentrations to the UCC values [The Ce and Gd anomalies were estimated as identified in the literature with Equation (1) for Ce/Ce* ,21,48,49C values .BSI) was evaluated using the formula presented in Equation (3) [REEs. The lREE, mREE, and hREE factors were determined using Equations (4)–(6), respectively. The concentration of REEs was normalized to UCC, and Ce was excluded from the lREE factor calculation due to anomalous behavior.The bell-shape index (tion (3) , with thCF), pollution load index (PLI), geoaccumulation index (geoI), and enrichment factor (EF) were estimated in the present work in order to assess the Podu Iloaiei Dam Lake sediment contamination with TEs and REs. The equation and the criteria for assessing the pollution indices are presented in EF, Mn was used as a reference element to determine the anthropogenic element enrichment.The contamination factor  from  were useThe potential of the contaminants to cause adverse effects on aquatic organisms was determined by comparing the concentrations of Cr, Zn, As, Pb, and Cd with the Sediment Quality Guideline (SQG) values for freshwater sediments.−1) values for the TEs, REs, and REEs determined in the sediment samples collected in July and October 2017 from Podu Iloaiei Dam Lake. REE concentration values in the water and the water-soluble fraction from sediments  are presented in −1) compared to October (1540 µg g−1). For the ∑RE concentrations, the difference was almost insignificant, with 276 µg g−1 in July versus 262 µg g−1 in October. While variation of the emission sources between the sampling sessions could be responsible for TE abundances, more stable sources could explain the RE behavior. For the ∑REEs, an opposite trend could be observed with a higher average concentration in October. However, more detailed aspects regarding the determined concentrations of REEs and potential sources will be discussed in In the sediment samples, the average of ∑TE concentrations was higher in July (1755 µg g−1 in July and 124 µg g−1 in October) were consistent with the values reported by Sandu et al. [−1 in July and 94.6 µg g−1 in October (−1 in summer season and 80.97 ± 29.74 µg g−1 in autumn season) [Regarding the TE concentrations in both sampling sessions, the highest levels were found for Mn, followed by Ba, Zn, and Rb, an observation which may indicate a natural accumulation process . Zinc cou et al.  for stre October , which a season) .−1 for July and 1.35–2.06 μg g−1 for October) compared with sediment from Lake Balaton, Hungary (0.1–0.7 μg g−1) reported by Nguyen et al. [−1) and Mn (160–760 μg g−1), similar concentrations for Co (1.7–17 μg g−1) and Zn (13–150 μg g−1), and higher concentrations for Pb (2.4–160 μg g−1) were presented. By comparison with a remote location, Bory Tucholskie National Park, Poland [The present study reports significantly higher concentrations for Cd , REs , Ce, LREEs* (LREEs without Ce), and HREEs spatial distributions in the sediment of Podu Iloaiei Dam Lake for July and October are presented in As observed from The profile of the spatial distribution of Mn, Ba, V, Cr, Mo, HREEs, and LREEs may indicate a displacement of the sediment from the input water area (in the west side) especially in July.Similar patterns were observed in July for the distributions of As, Cd, and Pb. In October, As seemed to have a more pronounced contribution from the river water source, while Cd and Pb seemed to be associated with contributions from the road and railway traffic. Cadmium, Pb, and other elements’ distribution in soil samples collected from nearby roads were also highlighted in recent studies ,58. HighWastewater discharge and traffic might be important contributors for Co and Zn in both sampling sessions. Additionally, the feeding area presented higher concentrations of these elements.Moreover, it is worth mentioning that V, Cr, and Mo presented higher concentrations in the feeding zone in both sampling sessions, with the higher concentration area being broader in October compared to July.4+ [A complementary spatial distribution of Mn and Ce was observed. This indicated that the oxidative dissolution of Mn from the sediment particles can generate Ce enrichment from Ce4+ ,60. The Element concentration variability was higher in July compared to October, with the coefficients of variation from 10 to 33% in July (19% in average) and from 5 to 26% in October (10% in average). This variability indicated that the urbanization, accumulation, and distribution of the potential toxic elements in lake sediments may lead to an increased spatial heterogeneity. For almost all REEs, the coefficient of variation values were higher in July (10 to 18%) compared to October (5 to 8%). These aspects indicated a homogeneous spatial distribution for these elements, except for Ce (with 33% in July and 26% in October). Differences in the input water quality, the weathering and leaching from soil surface, the local biochemical processes in the area with low water levels, and direct anthropogenic pressure could be important factors controlling the variability in the chemical composition of sediment samples from July. In October, the increase of the minimum concentration led to a reduced variability.For measured REEs, the variations observed between sampling sessions may be related to the input water source, which may change the pH and salinity of the lake, seasonal changes in water redox conditions, and also due to an enhancement of the adsorption–desorption reactions and REEs complexation processes .The distribution of Chondrite- and UCC-normalized REE concentrations for all sediment samples is presented in 3+ oxidation to Ce4+) on the surface of the water in October than in July. Under these circumstances, the sedimentation of Ce4+ due to scavenging by Fe-Mn oxide particles [The negative Ce anomaly (Ce/Ce* < 1) was determined for all investigated samples, with values ranging from 0.33 to 0.78 in July and from 0.37 to 0.92 in October. The individual values of the Ce anomaly are included in articles  may induarticles ,20,61,624+ is possible, followed by Mn-containing particle scavenging, which finally leads to a diminished Ce anomaly in the sediment due to sedimentation.Considering that the Mn concentration in sediment samples was fairly similar for both sampling sessions, it is assumed that, in October, the variation of the Ce anomaly was mainly due to the more pronounced oxidation potential of the water body. Only then, the formation of Ce2+ [2+ in enzymes [A positive Gd anomaly was obtained for both July and October sediment samples. The values obtained for the Gd anomaly varied from 1.15 to 1.24 in July and from 1.18 to 1.28 in October. The individual values are presented in 2+ ,63 or ma enzymes .BSI) was used to numerically evaluate the element enrichment in the investigated dam lake sediment. The individual values of the BSI are presented in BSI values ranging from 1.21 to 1.28 in July (average of 1.25) and from 1.19 to 1.28 in October (average of 1.25) suggested stability in terms of the sampling points and sampling sessions. The enrichment of mREEs  associated with phosphate minerals [As presented in minerals  was mostminerals ,66,67 buminerals  or the uminerals , P-fertip < 0.05) were observed for As with Mn (0.60) and Pb (0.79) in July. In October, the correlations extended to Co (0.66), Zn (0.59), and some REEs . Cadmium was significantly correlated (p < 0.05) with Co , Zn , Bi , and Pb  in both sampling sessions , V , Cr , and Mo . Negative significant correlations were observed between Mn and Zr (r = −0.68), Hf (r = −0.68), and Ce (r = −0.69) in October. These observations suggested that the reductive dissolution of Mn from the sediment can generate Ce, Hf, and Zr enrichment because these elements are redox condition sensitive [The correlations between the investigated elements (Pearson correlation coefficients) are presented in sessions . Signifi6), Zn 0., and somith Mn 0. and Pb , except for Ce in July. For October, a different behavior was observed, with a clear separation between LREEs and HREEs. p < 0.05), except for Ce, and between HREEs , except for Gd and Tb (p < 0.05). In both sampling sessions, Nb and Ta elements were highly correlated with most of the REEs.High values of Pearson coefficients were obtained between REEs , except for Gd and Tb, which presented strong loadings in Factor 2 among the LREEs . A slight distancing of Ce from the Factor 2 elements can be observed in CFs of Zn, Ga, As, and Pb showed moderate contamination for all sampling points, with the highest values obtained for S-8, S-9, S-10, S-11 and S-12, corresponding to the feeding area. Natural weathering of minerals is also a known source for Zn [The e for Zn . The usee for Zn  on agricThe PLI values obtained for the S-1 and S-2 sampling points  located geoI) are shown in geoI values indicate that the sediment was uncontaminated (geoI ≤ 0) for most of the investigated TEs and Res. None to moderate contamination (geoI = 0 ÷ 1) with Zn, Ga, As, and Ta was observed. For Cd, element moderate contamination (geoI = 1 ÷ 2) was found at 10 sampling locations, confirming the contributions from anthropogenic pollution sources. Only at the S-4, S-7, S-13, and S-14 sampling locations, the geoI values for Cd were below 1. The highest geoI values for Zn and Ga were found at S-3. Sediments at the S-9, S-10, and S-11 locations were hinted as uncontaminated to moderately contaminated with Ga, As, and Ta. In specific agricultural activities related to crops of sunflower, wheat, corn, and rapeseed, fruit-bearing orchards in the area, the use of agrochemicals  generally used at the European level can be responsible for As pollution, as suggested by Defarge et al. [The results obtained with the geoaccumulation index  for the S-3, S-8, S-9, and S-11 sampling points and minor anthropogenic modification (1.5 < EF < 3) for the other ten sampling points from Podu Iloaiei Dam Lake. The obtained results confirmed the anthropogenic source of Cd, with emissions associated mainly with fuel used for boats, railways, and road traffic.The influence of anthropogenic sources on the elements in the sediment samples was also assessed by determining the The potentially harmful element  concentration in the sediment samples of Podu Iloaiei Dam Lake, along with threshold effect concentration (TEC) and probable effect concentration (PEC) for each element, are shown in Among the investigated elements, Cd and Pb showed concentrations that were mainly below TEC in all samples. The obtained results indicated that these elements are very unlikely to pose a threat to the organisms living in the bottom of the aquatic system  in the iThe concentrations of Zn, As, Cd, and Pb did not exceed the PEC values for any of the investigated samples. Values higher than the PEC thresholds were identified for Cr in several samples (five sampling points in July and one in October), and such behavior may hint that Cr is one of the most important elements that could lead to toxicity in the investigated area. The sources of Cr pollution in Podu Iloaiei Dam Lake may be related to domestic wastewater from villages without centralized sewerage and fuel combustion associated with road traffic, both sources being located in the northern part of the lake.High variabilities of TE and RE concentrations in the Podu Iloaiei Dam Lake sediments were identified. For both TEs and REs, indication of their main geogenic origin was obtained. The concentrations of Pb, As, Cd, and Zn were mainly related to anthropogenic sources.mREEs , evaluated by BSI, indicated shared sources such as phosphates that may have both natural and anthropogenic origins. The statistical evaluation of redox-sensitive elements, such as Ce and Mn, and the Ce anomaly highlighted a higher oxidizing potential of the aqueous environment in October compared to July. A positive Gd anomaly was observed for the Chondrite-normalized concentration in both the July and October sampling sessions, which indicated micropollution and may be related to the use of Gd-chelating agents in MRI, a medical investigation technique with a growing use in the Iasi medical hub.The REEs had low variability, except for Ce. Normalized distributions of REEs and the observation of an enrichment of The assessment of the pollution indices emphasized that the sediment samples were highly enriched in Cd and contaminated with elements such as Zn, Ga, As, Pb, V, Cr, Nb, and Ta, related to anthropogenic sources. Sediment quality guidelines indicated that Cr posed a considerable threat to the aquatic biota.The present study brought high contribution to understanding the specific sources of REEs, TEs, and REs in the aquatic system used for intensive fish farming, from north-eastern Romania. The use of agrochemicals , road and railway traffic, and domestic wastewater discharge were found to be the major contributors to the potential harmful element contents in the Podu Iloaiei Dam Lake sediment.A close monitorization of the Bahluet River, which is the main water input on the investigated Podu Iloaiei Lake, and even extending the investigation on the Bahlui hydrographic basin may be necessary in order to fully understand the evolution of metal pollutants and micropollutants.


Glucomannan not only reduced the total cholesterol (TC)  and low-density lipoprotein (LDL) levels  compared with the control group, but also reduced the fasting blood glucose (FBG) , 2 h postprandial blood glucose (P2hBG) , fasting insulin (FINS) , and serum fructosamine (SFRA) levels . Our analysis indicates that glucomannan is an effective nutritional intervention for type II diabetes.The hypoglycemic and lipid-lowering effects of glucomannan are widely known, and it is a potential effective treatment for type II diabetes. In this study, we evaluated the effects of glucomannan supplementation on blood-lipid-related indicators, blood-glucose-related indicators, blood pressure (BP), and body weight (BW) in patients suffering from type II diabetes. We searched databases including PubMed, Cochrane, the comprehensive biomedical research database (Embase), Web of Science, and China National Knowledge Infrastructure (CNKI) for literature on glucomannan and type II diabetes. Six randomized controlled trials (RCTs) were eligible ( Glucomannan is a type of soluble hemicellulose. It has clear hypoglycemic and lipid-lowering effects, and its mechanism may be related to its rheological properties . As a soAmorphophallus konjac is a perennial herb of Amorphophallus Blume of Araceae [ Araceae . Its bul Araceae . Konjac  Araceae . Konjac  Araceae .Glucomannan is a high-quality dietary fiber and a safe food additive . GlucomaThe main difference between type Ι and type II diabetes is the presence of insulin resistance . The IntThe English and Chinese databases, including Embase, PubMed, Cochrane, Web of Science, and CNKI, were searched for relevant studies among the initial studies up to the date of 31 December 2021. We used the MeSH terms “Glucomannan” and “Diabetes Mellitus” in the retrieval process. The complete search combinations used in the search on Web of Science were “TS = (Glucomannan OR Konjac glucomannan OR Konjac mannan)” and “TS = (Diabetes Mellitus OR Diabetes OR Diabetic)”. All language and publication types were included in the search.Two independent researchers closely investigated each word of the title and each sentence of the abstract. We traced and downloaded the full text of the RCTs that meet these search standards. Two researchers analyzed the experiments used for detailed analysis and data extraction, with differences resolved through mutual consultation or discussion with a third independent author. The inclusion criterion was RCTs of glucomannan in patients with type II diabetes, and the exclusion criteria were cell and animal experiments, patents, conference speeches, retrospective analyses, nonrandomized controlled studies, reviews, and meta-analyses.The extracted basic data are included in 2 exceeded 50%, which we used for the heterogeneity analysis, and conducted sensitivity analysis to analyze the robustness and reliability of the merge results. When the I2 value did not exceed 50%, we used the fixed effects model; p values less than 0.05 were considered statistically significant. We assessed the risk of publication bias using funnel plots derived from the Cochrane bias risk assessment tool. We evaluated the quality of the six RCTs using seven criteria, i.e., random sequence generation, allocation concealment, patient and personnel blindness, outcome evaluator blindness, incomplete outcome data, selective reporting, and other biases not covered above. Deviation risk included three types, namely, “low risk”, “high risk”, and “uncertain risk”. The evaluation was based on the Cochrane manual.Review Manager 5.3 software  was used for the meta-analysis. The standardized mean difference (SMD) and 95% confidence interval were the basic analysis parameters of the continuous variables. We used a random effects model to determine the value when Ip < 0.05 indicated significant publication bias.Stata/MP 17.0 software  was used to perform Egger’s test to assess publication bias; We collected 198 studies from the Web of Science, 92 from Embase, 49 from PubMed, 25 from the Cochrane Library, 21 from CNKI, and 1 additional record from another source, with a total of 386 studies. After reviewing the titles and abstracts, we removed 126 duplicate articles and 247 unrelated articles, leaving 13 that met the requirements for searching their full texts. The full texts of three articles were not available to us. We read the remaining 10 articles and considered 4 of these to be RCTs in a unqualifying sense. Finally, there was a total of six RCTs of glucomannan on type II diabetes in the meta-analysis. The flow chart of the study is shown in Among the six RCTs included in the meta-analysis, two were from China, two were from Canada, one was from Thailand, and one was from Taiwan ,23,24,251.Triglyceride (TG)p = 0.81). The heterogeneity among the included RCTs was significantly different   A.2.Total Cholesterol (TC)p = 0.001). The heterogeneity among the included RCTs was significantly different   B.3.High-Density Lipoprotein (HDL)p = 0.15) and the heterogeneity  between the included RCTs p < 0.0001). The heterogeneity among the included RCTs was significantly different   D.1.Fasting Blood Glucose (FBG)p = 0.0002). The heterogeneity among the included RCTs was significantly different   A.2.Two-hour Postprandial Blood Glucose (P2hBG)p = 0.003). The heterogeneity among the included RCTs was significantly different   B.3.Fasting Insulin (FINS)p = 0.004). The heterogeneity among the included RCTs was not significantly different   C.4.Serum Fructosamine (SFRA)p < 0.0001). The heterogeneity among the included RCTs was not significantly different   D.1.Body Weight (BW)p = 0.40). The heterogeneity among the studies was not significantly different   A.2.Systolic Blood Pressure (SBP)p = 0.14). The heterogeneity among the studies was significantly different   B.3.Diastolic Blood Pressure (DBP)p = 0.12). The heterogeneity among the studies was not significantly different   C.p = 0.030).We used Egger’s test to determine the risk of publication bias. The results of the risk of publication bias are presented in the In this meta-analysis, which included six RCTs, we fully evaluated the effects of glucomannan on patients with type II diabetes, with multidimensional indicators of diabetes, such as blood lipid levels, glycemic index, BP, and BW. Our analysis results showed that glucomannan had significant reduction effects on TC, LDL, FBG, P2hBG, FINS, and SFRA. Glucomannan might have reduction effects on HDL, BW, SBP, and DBP, but the differences were not statistically significant. Glucomannan had no reducing effect on TG.As shown in As shown in As shown in 2 did not exceed 50%, which indicated that heterogeneity was low. On the contrary, when I2 exceeded 50% we used the random effects model. For indicators using the random effects model, we conducted sensitivity analysis to explore the reasons of heterogeneity. After removing each study in turn, merged results of the remaining studies did not have significant differences compared with the original results, indicating that the original results were not likely to change. In our meta-analysis, we used the fixed effects model when IIn this meta-analysis, the six RCTs were mainly performed in Asian countries and Canada in North America. The main researchers were mostly Asians, which might be related to the fact that konjac is mainly produced in Asia. The geographical and climatic environment in some parts of Asia is suitable for its growth, and the consumption of konjac has a long history in Asia. The main effective component of konjac is glucomannan. Specific doses of glucomannan were used in the six RCTs, making them more accurately studied than in usual food interventional research. The six RCTs were published between 1999 and 2007. No standard RCT results have been published in the past decade, but this might not hinder the authenticity and reliability of the results of this study. Glucomannan was a research hotspot for some time, and it may become a research hotspot again in the future.The research population mainly comprised adults with type II diabetes and included slightly more women than men, totaling 440 people. The daily doses of the glucomannan supplement in the experimental group ranged from 1.2 g to 15 g, and the duration of the experiment ranged from 3 to 16 weeks. The placebo or wheat bran was used in the control group. As the main component of konjac, glucomannan did not cause a definite adverse reaction during the study. However, to determine the safety and effectiveness of a drug ingredient, studies need a longer experimental time.Asian countries, represented by China, have a long history of consuming konjac food. Its effect of reducing weight was known before, but it had not been verified by scientific experiments before modern science confirmed its effective ingredients. Amorphophallus konjac is a natural food in which glucomannan, its main effective ingredient, can be naturally derived. The RCTs with the highest levels of evidence can directly and accurately verify glucomannan’s clinical effects. The effects of glucomannan on obesity, metabolic syndrome, and constipation have been studied. We carried out our study on the effects of glucomannan supplementation on type II diabetes in this research.The treatments for type II diabetes mainly include diet control, exercise regimens, oral medication, insulin therapy, and the recent trend of surgery. Considering the poor compliance with diet and exercise therapy, the side effects of long-term oral medication and insulin therapy, and severe trauma after surgical treatment, glucomannan exhibits certain advantages as a natural product for nutritional intervention.Glucomannan solution can also be dehydrated and properly treated to form edible and naturally degradable membrane materials . As a naBased on our literature search for this meta-analysis, we comprehensively evaluated for the first time the effects of glucomannan supplementation on various indicators in patients with type II diabetes. However, we failed to avoid several limitations: (a) Due to the limited number of large clinical trials, we included only six eligible RCTs, and there were only two studies including data of SFRA, SBP, and DBP. Most RCTs had a small sample size, so the results may not have been significant. (b) The effectiveness and safety of the long-term use of glucomannan and dose of glucomannan were not verified, nor was the daily dose of glucomannan. (c) The RCTs were conducted only in Asia and Canada, so our study lacked research from Europe and Africa. These limitations need to be overcome in future studies. Even with the above limitations, our findings clearly lend support to the suggestion that the supplementation of glucomannan has an ameliorative effect on type II diabetes mellitus.Our results indicate that glucomannan supplementation has significant reducing effects on the TC, LDL, FBG, P2hBG, FINS, and SFRA of type II diabetic patients. However, the beneficial effects of glucomannan on TG, HDL, BW, SBP, and DBP were not statistically significant. More RCTs with larger sample sizes, combined drugs, and longer experimental times are needed to further determine the effects of glucomannan on various indicators of type II diabetes patients. Perhaps glucomannan can then be developed into a highly effective natural product, thus benefiting more type II diabetes patients.


Trachoma is the leading infectious disease that leads to blindness worldwide, especially in developing countries. Though Ethiopia had targeted a trachoma elimination program by 2020, the problem worsens, particularly in the Amhara Region. Even though sustained intervention measures are undertaken across the region, it is unclear why trachoma is still a significant public health problem. So, this study assessed the prevalence of active trachoma and associated factors among 1–9 years of age children from model and non-model kebeles in Dangila district Amhara Region, Northwest Ethiopia.th September 2019 to 29th October 2019. A multistage stratified random sampling technique was used to reach 704 children from model and non-model kebeles. Samples were allocated proportionally to model and non-model kebeles. A structured and pretested data collection tool and observational checklist was used to manage the necessary data. Data were coded and entered in Epidata version 4.6, and further analysis was done using SPSS version 20 software. Bivariable and multivariable logistic regression analysis was employed to identify factors associated with active trachoma. Adjusted Odds Ratios (AOR), p-value, and respected Confidence Interval (CI) were used to report the findings.A community-based comparative cross-sectional study was conducted from 20Seven hundred four children were included in this study, with a response rate of 97.8%. The overall prevalence of active trachoma was 6% . The prevalence of active trachoma among non-model and model Kebele was not significantly different. Still, the prevalence of active trachoma among children from model Kebele were  relatively lower compared with non-model kebeles, . Moreover, not using latrine , fly-eye contact , presence of sleep in eyes , presence of ocular discharge , presence of nasal discharges  and washing faces with soap  were found significantly associated with the prevalence of active trachoma among children 1–9 years old.The prevalence of active trachoma in the model and non-model kebeles was high and did not show a statistical difference. Attention to be given to latrine utilization, washing face with soap, and other personal hygiene activities. Trachoma is a disease of the eye caused by infection with the bacteria Chlamydia Trachomatis , 2. BlinGlobally, around 50 million people are infected with trachoma, and 3 to 10 million people are becoming blind due to the infection . The annSeveral factors contribute to the occurrence of active trachoma. Evidence identified that secretions around the eyes that attract flies, scarcity of water for personal hygiene, limited access and utilization of latrines, overcrowded living conditions, close contact, and sleeping in the same bed with infected persons increased trachoma prevalence –16. The SAFE -based interventions have been implemented in endemic countries, including Ethiopia , 20. Duerd most populous and covers a large geographical area among the twelve districts at Awi-zonal administration. Danglia district (woreda) is a part of the Amhara region and Awi zone and is located in the western part of Ethiopia. Furthermore, it is found 78 km away from Bahir Dar regional city in the Southeast direction. The district had six health centers and thirty-one health posts or kebeles. Of these, 15 kebeles were verified as model kebele on the district transformation agenda by model kebele criteria. The rest 16 are non-model kebeles [A comparative cross-sectional study was conducted in Dangila district from October to November 2019. Dangila district is the 3 kebeles . The dis kebeles .All children whose age was found in the range of 1–9 years old in the Dangila district were the source population. Children aged 1–9 years living in the model and non-model kebeles were included in this study. On the contrary, temporarily residing children were excluded.The outcome variable showed signs of active trachoma among children aged 1 to 9 years old. The explanatory variables were age and sex of the child, residence, parental education, wealth index, availability of latrine facility, type of latrine, latrine utilization, household size, source of water, accessibility of water, the quantity of daily water used, presence of feces near the compound, waste disposal sites, solid waste disposal methods, availability of livestock, frequency of face washing, using soap for washing, discharge on the eye, facial cleanness and presence of flies on a child’s face, knowledge of mothers about trachoma and number of under ten years children.Active trachoma: The presence of Trachomatous inflammation–follicular or Trachomatous inflammation- intense [ intense .Trachomatous inflammation -follicular (TF): The presence of five or more follicles in the upper tarsal conjunctiva, each with at least 0.5 mm diameter in size [ in size .Trachomatous inflammation- intense (TI):- is pronounced inflammatory thickening of the upper tarsal conjunctiva that obscures more than half of the standard deep vessels [ vessels .Knowledge: assessed by developing trachoma transmission, prevention and control measures related questions. If the mother answered ≥ 80% of knowledge-related questions, then the mother classified as knowledgeable 50%-79% was reasonably knowledgeable. Less than 50% were classified as less knowledgeable [edgeable .Model kebeles: where the residents meet the following interrelated criteria as ≥80% of residents are model households, ≥85% of residents attended facility-based delivery, ≥85% of residents owned improved latrines, and ≥85% of schools become models [Non-model kebeles:—The low performance of the five interrelated criteria, namely; model HH (≤80%), Facility delivery (≤85%), improved latrine ownership (≤85%), and model schools (≤85%)as well as for urban kebeles it includes model youth centers performance (≤85) [livery ≤8%, improvThe sample size was determined by using the second objective using factors significantly associated with active trachoma by considering the following assumptions: 95% confidence level, 80% power of the study, two comparison groups population ratio 1:1, considering the availability of flies on the face children as significant determinants of trachoma and the proportion in the exposed group was 45.9%, the proportion in the unexposed group was 29% with an adjusted odds ratio (AOR) of 1.98 and design effect 2 . The calMultistage stratified sampling followed by a simple random sampling technique was used to select kebeles and households. Stratification of the kebeles was done based on their model and non-model kebele status. Of all kebeles in the district, 35% (5 kebeles in the model and five kebeles in the non-model) were randomly selected. Those kebeles are Chara-01, Dubi, Dimsa, and Affessa from graduated model kebeles; on the other way, Gumdri, Abadra, Misrak Zelesa, Gissa Balegiziabher, and Dengeshita were selected randomly from non-model kebele strata. Population proportion allocation was proposed to determine the required sample size for each randomly selected kebele.th interval value was used. Every 8th and 5th interval was used to allocate the sample size to model and non-model kebeles, respectively. Before starting the sampling, pen spiring was conducted to indicate the starting location of the village. Finally, one child was selected using the lottery method when two or more children live in 1–9 years old per a single household. Finally, 720 households were selected using a systematic sampling technique.In a total of 2087 and 3049 households, at least one children were aged 1–9 years old in model and non-model kebele, respectively. Households with children aged 1–9 years old were selected using a systematic random sampling technique. To determine the interval of Households in selected Kebele, the KData were collected using a pretested structured questionnaire, observational checklists, and physical examination. The data collection tool was developed after reviewing other available literature , 26–30. The questionnaire was pretested using 5% of the total sample in another kebele which was not included in the study, and the necessary amendments were done after the pre-test.After the data collection, the information was reviewed, and incomplete questionnaires were returned to the data collectors for revisiting the respondent and completion. Data were coded and entered in Epidata version 4.6 and exported to SPSS version 20 for analysis. Descriptive statistics were used to describe data. Principal Component Analysis(PCA) generated a wealth index and divided households into poor, medium, and high economic groups.The sampling adequacy for PCA was assessed using Kaiser-Meyer-Olkin (KMO) statistics. The KMO measures the adequacy of sampling thoroughly and the sampling adequacy for each variable that indicates the proportion of variance the variables might be caused. Sampling adequacy set at communality value > 0.5 with P-value > 0.05 and complex structure factor  greater than one was considered . BivariaDebre Markos University assessed ethics. An approval letter was obtained from the College of Health Sciences Ethical Review Committee. Moreover, the support letter was secured from the study setting, Dangila district Administrative and Health offices. The parents or guardians were informed about the study’s purpose, and verbal informed consent was obtained before data collection. Mothers or caregivers were also informed that they had the full right to discontinue participating in the study. Confidentiality was ensured by omitting the name and ID of the participants. A separated, secured, and conducive interview setting was selected. Each respondent was assured that their information was confidential and used only for research. Trachoma-infected children were referred to the nearest health facility.A total of 704 children in the age range of 1 to 9 years completed their responses, providing a response rate of 97.8%. Slightly more than half (55%) of the respondents were from the model kebeles. The mean (± standard deviation (SD)) age of children was 4.8 (±2.3) years and 4.0 (±2.2) years old from model and non-model kebeles, respectively. The proportion of female children who participated in this study was 185 (52.6%) from the model and 182 (51.7%) from non-model kebeles. The majority, 347 (98.6%) of the households, have two children, and 345 (98%) of them were from model and non-model kebeles. Regarding household economic status, 140 (39.8%) of households in the model and 95 (27%) were in the high financial group based on assets owned .2).Nearly all households had sanitation facilities in their compound. For instance, 97.8% and 96% of model and non-model kebele households had latrine access. Of them, 98.6% from model kebele and 72.7% from non-model kebele properly utilize latrine. However, human feces were detected near 3.7% and 50.8% of households among model and non-model kebeles, respectively (Table 3).Of 704 children, 669 (95%) had a habit of face washing. In contrast, only fifteen 15 (4.3%) in model and 20 (5.6%) in non-model kebeles had no tradition of washing their face regularly. There is a difference between children’s model and non-model kebeles’ hygiene behavior, especially facial hygiene. Similarly, 55.4%, 19.9%, and 29.7% of the children from the model kebele were observed washing their faces once, twice and more than twice a day, respectively, using soap. On the contrary, 73.5%, 17.5%, and 9% of the children from the non-model kebele were presented with washing their faces once, twice and more than twice a day, respectively, using soap . The prevalence found to be 4.5%  in the model and 7.7%  in the non-model kebeles .During the bivariate logistic regression, latrine utilization, residence, animal waste, solid waste disposal pit, ocular discharge, nasal discharge, flies on the face, fly-eye contact, sleep on the eye, antibiotics medication, and washing faces by soap were found to be a p-value less than or equal to 0.25 and selected as a candidate for the final multivariable analysis. But, the other variables, such as socio-economic variables with a p-value > 0.25, are not selected for multivariable analysis. During the multivariable logistic regression analysis, variables like the experience of fly-eye contact, the presence of sleep in the eye, the presence of discharge on the eye, face washing with soap, and nasal discharge were significantly associated with the prevalence of active trachoma.The odds of having active trachoma among children (1–9) from households who did not utilize latrine was 4.3 times higher than children from families using latrine . In the same manner, children who had fly-eye contact were three times  higher than children who had no fly-eye contact. Having an ocular and nasal discharge were 2.8 and 2.7 times higher to develop active trachoma than their counterparts, as well as children’s who had a sleep (collection of crusted secretions in the canthus of the eyes) were 2.5 times more likely to develop active trachoma. Lastly, the odds of having active trachoma among children (1–9) years old who had an experience of face washing with soap were 77.9% lower than their counterparts  .In Ethiopia, trachoma is the leading infectious disease that causes blindness and death . The curFurthermore, the prevalence of active trachoma in non-model and model and kebele differs at 7.7%  and 4.5% , respectively, the difference was not statistically significant. This consistent finding might be because the two study settings have had the same awareness and knowledge about trachoma signs, transmission mechanisms, and prevention methods. About 99.4% of mothers who live in the model had information about trachoma, while approximately the exact figure (98.9%) of mothers residing in non-model kebele had information about trachoma. Similarly, 38.9% and 32.9% of children’s mothers who live in the model and non-model households had good knowledge about trachoma. Besides this, the availability of latrines in both kebeles is nearly similar. Moreover, the similarity may be due to children’s hygiene practices such as face washing and soap use in both kebeles.The difference in model and non-model kebeles did not persist after multivariable analysis, indicating that both areas require continued implementation of trachoma control interventions to increase the prevalence under the elimination target of 5%. The lower-level maternal education might also explain the difference. The educational status of mothers has played a significant role in reducing trachoma prevalence as educated mothers had positive attention in caring for and teaching their children about trachoma prevention mechanisms. On the other hand, it may be due to the study conducted at the district discontinued MDA supplementation four years ago.After completing five annual MDA rounds, the pooled prevalence was higher than the 5% criteria set under the WHO trachoma elimination program and impact surveys in five districts in the Amhara region , 39. ThiThe pooled prevalence was higher than the 5% criteria set under the WHO trachoma elimination program; the prevalence of active trachoma in non-model kebele was higher . These mIn this study analysis, different factors have an association with active trachoma. Latrine utilization, washing faces with soap, fly-eye contact experience, sleep in the eyes, discharge on the eye and nose were variables found to be significantly associated with the prevalence of active trachoma among children (1-9y rears) old. The probability of being infected with active trachoma was higher in children 1–9 years old who had ocular and nasal discharges than those who had not. The finding is consistent with previous studies in Gambia and Tanzania , southerThis study also revealed that children (1–9) who had fly-eye contact were more likely to risk active trachoma than their counterparts. This finding is supported by the previous studies conducted in Ankober district and rural Ethiopia , 46. ThiNot using a latrine can be considered an indicator of hygienic behavior. Children (1–9 years old) who lived in households that did not use latrines were more likely to be affected by active trachoma. This finding aligns with a study conducted in the Gonji kolela, Baso-Liben, and Ankober districts, 47, 49.This study also revealed that using soap for face washing is significantly associated with reducing active trachoma prevalence. This finding is consistent with the previous studies in Leku town and the Baso-Liben district , 47. TheIn this study, the prevalence of active trachoma among children 1–9 years old was 6% and had not shown statistically significant variation between model and non-model kebeles. The prevalence of active trachoma among children 1–9 years old in the Dangila District was in line with the WHO-recommended trachoma elimination program. Latrine utilization, using soap, the experience of fly-eye contact, the presence of sleep, discharge on the eye and the nose were significantly associated with the prevalence of active trachoma among children (1–9 years old). Therefore, an intervention area needs to be improving personal hygiene-related activities such as washing children’s faces thoroughly with soap, removing dirt , and eye sleeping from their faces. Significant emphasis on the use of sanitation services, especially on latrine construction and use, is also required to eliminate open defecation.One of the strengths of this study is including large sample size. Moreover, this study investigated children in the 1–9 year age group in accordance with WHO guidelines. It is possible that the responses to the questionnaire might have social desirability bias and potential recall bias. However, many components such as environmental hygiene conditions, water source, and facial hygiene were directly observed. The other limitation of this study was the lack of qualitative components for triangulation.S1 File(DOCX)Click here for additional data file.S2 File(DOCX)Click here for additional data file.S1 Dataset(SAV)Click here for additional data file.


Quinate, neochlorogenic acid, and ouabagenin, taxezopidine K1, taxezopidine K2, and taxezopidine K3 in tobacco were influenced by the interaction of environmental factors and the genotype. Our results suggest that metabolomics based on UPLC-QTOF MS/MS could be used to analyze the ecological functions of biomarker metabolites and understand the mechanisms of plant adaption to the environment.Environmental factors affect plant metabolites, different climates, cultivation conditions, and biotic stresses and genotypes strongly affect their chemical composition and contents. Our aim is to examine the environmental and genetic interaction effects on tobacco metabolite composition. UPLC-QTOF MS/MS coupled with multivariate data analyses were applied for the metabolomics analysis of three tobacco cultivars from different planting regions in China. Principal component analysis (PCA) revealed that environmental factors have a greater effect on tobacco metabolism compared to genotypes. Twelve biomarkers were screened by orthogonal partial least squares discrimination analysis (OPLS-DA). Univariate analysis indicated that Malate, conjugated chlorogenic acid, chlorogenic acid, quercetin 3-rutinoside-7-glucoside, and unknown compound  Nicotiana tabacum) is an ideal model plant for molecular ecology studies, and the metabolites of tobacco are susceptible to environmental and genetic changes. As previously stated, different climates, cultivation conditions, biotic stresses and genotypes strongly affect their chemical composition and contents −). The assistant software packed in the MassLynx i-FIT algorithm was used to determine the elemental composition for the peak at m/z 771.1967. The result showed that there were two candidates and that the i-FIT value of the first one (C34H36N4O17) was low, whereas the second one (C33H40O21) had a higher i-FIT value with a low tolerance. Second, some databases, such as KEGG, METLIN and Chemspider, were used to search for MS and MSn information based on the accurate m/z and potential formula. Meanwhile, the MS/MS information  was obtained with collision energy from 10 to 60 V in the negative ion mode. The fragment ion at m/z 609.1401 was obtained through the loss of 163 (C6H11O5) from the ion at m/z 771.1967 ([M−H] −). The fragment ion  emerged at m/z 609.1401, 300.0269 and 271.0233 and were following fragments generated by rutin, which indicated that the structure of this fragment ion must include one rutin group and a hexose group. Therefore, the candidate from the database with neither a rutin group nor a hexose group was excluded. Finally, this candidate was tentatively identified as quercetin 3-rutinoside-7-glucoside, which has not previously been reported in tobacco but has been reported in tomato [Structural identification was conducted by a UPLC-QTOF MS/MS analysis platform based on its retention time, accurate n tomato . In addip ≤ 0.01) in different regions or cultivars. Based on multiple comparisons, the metabolites malate, conjugated chlorogenic acid, chlorogenic acid, quercetin 3-rutinoside-7-glucoside, and unknown compound 5 were statistically significant in different planting regions. However, no differences were found between the two cultivars from the same planting region. This showed that environmental factors mainly influenced the accumulation of these five metabolites in tobacco. Malate is involved in various metabolic pathways [The relative contents of the biomarkers in different planting regions and different cultivars are shown in pathways  and implpathways ,16. Sincpathways ,16. PolyIn our results, a significantly higher level of chlorogenic acid was observed in tobacco leaves from YJ, which had the longest sun exposure time and the lowest average temperature. The results suggest that chlorogenic acid increased with increasing latitudes, sun exposure time and decreasing temperature. Hårdh et al.  considerPhenolics can accumulate in the vacuoles of plants as glycosides. Flavonols and flavones, particularly kaempferol and quercetin, were conjugated to sugars, mainly glucose, rhamnose and rutinose. Rutin is formed by the conjugate connection of quercetin and rutinose. The rutin content was significantly different between XXY and samples in YJ (YJY and YJJ). Likewise, YJY was different from samples in XX (XXY and XXZ). The content of rutin in different cultivars of the same planting regions had no significant difference. In addition, XXZ and YJJ had no significant difference. It is possible that rutin was affected by both environmental factors and cultivars, but that the environmental factors still played a major role.Polyphenols were identified in this paper, and all of them were correlated with environmental changes. Coping with biotic and abiotic stress is the main biological function of polyphenols in plants , and the5 were only influenced by environmental factors independent of the genotype, whereas quinate, neochlorogenic acid, ouabagenin, taxezopidine K1, taxezopidine K2 and taxezopidine K3 in tobacco were influenced by the interaction of environmental factors and the genotype. The content of polyphenols reflects the mechanism of plant adaption to the environment. In agricultural practice, it is feasible to screen the cultivars that adapt better to the local environment according to the sensitivity of the cultivars and using the methods of metabolomics. The optimum mean daily temperature for growth is 20–30 °C; an atmospheric humidity of 80 to 85% and soil without high nitrogen levels is optimal. When grown commercially, tobacco requires a frost-free period of 90–120 days from transplanting to the last harvest of leaves. When grown as a crop, a dry period is required for the leaves to ripen and be harvested. Moreover, excessive rainfall causes thin, lightweight leaves. To develop its full aroma, sun-cured or oriental tobacco requires a relatively dry climate.The metabolites of tobacco leaves from different planting regions and cultivars were significantly different, and the influence of planting regions was greater than the cultivar influence. OPLS-DA selected twelve biomarkers, and seven of them were identified as polyphenols. Among these biomarkers, malate, conjugated chlorogenic acid, chlorogenic acid, quercetin 3-rutinoside-7-glucoside, and unknown compound


A nationwide diabetic retinopathy (DR) screening program has been established in Denmark since 2013. We aimed to perform an evaluation of adherence to DR screenings and to examine whether non-adherence was correlated to DR progression.The population consisted of a register-based cohort, who participated in the screening program from 2013 to 2018. We analyzed age, gender, marital status, DR level  and proliferative DR (PDR)), comorbidities and socioeconomic factors. The attendance pattern of patients was grouped as either timely (no delays > 33%), delayed (delays > 33%) or one-time attendance (unexplained).P < 0.05). Delays at previous screenings were associated with progression to severe NPDR or PDR (hazard ratio (HR) 2.27, 6.25 and 12.84 for 1, 2 and 3+ delays, respectively). Any given delay doubled the risk of progression (HR 2.28).We included 205,970 patients with 591,136 screenings. Rates of timely, delayed and one-time attendance were 53.0%, 35.5% and 11.5%, respectively. DR level at baseline was associated with delays  and one-time attendance  with relative risk ratios (RRR) of 1.68, 2.27, 3.14, 2.44 and 1.18, 2.07, 1.26, respectively (In a national cohort of 205,970 patients, almost half of the patients attended DR screening later than scheduled or dropped out after first screening episode. This was, in particular, true for patients with any levels of DR at baseline. DR progression in patients with delayed attendance, increased with the number of missed appointments. Diabetic retinopathy (DR) is a frequent complication of diabetes, and sight-threatening DR is among the leading causes of preventable blindness in the working-age population . AccordiDenmark has a national tax-funded screening program for DR. It is recommended that patients attend screening at either a practicing ophthalmologist or a hospital-based screening facility. Financial reimbursement is provided regardless of screening site and patients with proliferative DR (PDR) or diabetic macular edema are referred for treatment at the public hospital departments of ophthalmology. Denmark is divided geographically into five regions; the Capital Region of Denmark, Central Denmark Region, North Denmark Region, Region Zealand and Region of Southern Denmark . The regIn this study, we aimed to utilize the Danish registers to examine attendance patterns in the Danish nationwide DR screening program, to characterize timely, delayed and one-time attending patients, as well as explore the effects of delayed attendance on DR progression.In this retrospective nationwide cohort study, our population was defined by the data in The Danish Registry of Diabetic Retinopathy (DiaBase), which contains data of all patients who had attended DR screening at least once, from January 2, 2013, to December 30, 2018 . We inclWe utilized the Danish national registers where all data can be linked on an individualized level. This includes entire medical records, socioeconomic data and prescription medication usage.Diabase, which defined our population, contains data reported by the screening ophthalmologist, and the database has approximately 100,000 additions annually . From DiIn addition to DiaBase, we utilized the following registers:The Danish Civil Registry 1968) was used to link data across registries using an individual identification number (CPR number) given to all citizens in Denmark . We extr68 was usThe index date was defined as the first screening date, and delay was calculated according to the next recommended screening interval, as given by the screening physician. Patients were classified as having timely attendance if they were never delayed > 33% and did not miss any screenings during follow-up. Patients were classified as delayed if the actual date of the next screening was registered beyond 33% of the intended interval, e.g., a patient with a recommended interval of 90 days, would therefore be classified as delayed if the next screening date was more than 30 days after the planned screening date Fig. .Fig. 1TiOne-time attendance was defined as a patient only participating in screening once, with no follow-up appointments, without apparent reason. Patients with a scheduled next screening date beyond the observation period or who were referred for treatment (for DR or other eye-related illness) and therefore exited the screening program, as well as patients, who disappeared or died before their next screening, were censored at exit date and, thus, only included in the analyses in the periods where they could be clearly classified. DR progression was defined as a worsening in DR to either severe NPDR or PDR in either eye.2 test. Using a multinomial logistic regression model with relative risk ratio (RRR) calculations, we compared the characteristics of patients with delayed and one-time attendance to patients with timely attendance depending on various exposure variables. The model included a crude, semi-adjusted (age and gender) and fully adjusted multivariable analysis, adjusted for all statistically significant exposure variables from Table P-values < 0.05 and confidence intervals (CIs) not including 1.0 were considered statistically significant.Descriptive data on the population were reported in numerical format with percentages for all variables except age, which was reported in median and interquartile range. Statistical significance was calculated using the Chin = 205,970) consisted of 56.6% males, had a median age of 65.7 years , and 89.1% were of Danish lineage ), divorced or widowed ), having type 2 diabetes ), DR level 2–4 , 2.07 , 1.26 ) or CCI scores of 2 or 3 , 1.09 ) compared to patients attending screening at recommended intervals  was seen more often in patients with delayed attendance, and the number of delays was correlated to a significantly increased risk of progression. This is in accordance with a study from England , IrelandThe inclusion of a large nationwide cohort with a considerable amount of screenings, and detailed, validated register information on an individualized level, is a clear strength of this study. The addition of socioeconomic data ensured the completeness of the characterization of the study population.As our study focused on adherence to the screening program, and patients attending DR screening at least once, we did not address the issue of patients never attending screening, which might add another dimension. Due to the register-based nature of the study, the subjective reasons for non-adherence were not addressed. This would, however, be important for future reference, as an involvement of patients and a prioritization of their prerogative will be crucial in order to improve attendance.In conclusion, our study of non-adherence successfully added information on a population basis using a national cohort of patients in the Danish screening program of DR in Denmark. We highlighted younger age, divorce, presence of DR, competing illnesses and low income as the characteristics of patients with delayed and one-time attendance in the Danish screening program and showed twice the risk of progression to severe NPDR and PDR in patients with delayed attendance.


Acinetobacter, Brevundimonas, Dolosigranulum, Finegoldia, Haemophilus, Leptotrichia, Moraxella, Peptoniphilus, Pseudomonas, Rothia, and Staphylococcus were the only 11 bacterial genera that presented significant age-associated differences. Other bacterial genera such as Anaerococcus, Burkholderia, Campylobacter, Delftia, Prevotella, Neisseria, Propionibacterium, Streptococcus, Ralstonia, Sphingomonas, and Corynebacterium appeared in the population with a very high frequency, suggesting that their presence might be biologically relevant. Therefore, in contrast to other anatomical areas such as the gut, bacterial diversity in the nasopharynx of healthy subjects remains stable and resistant to perturbations throughout the whole life and in both sexes. Age-associated abundance changes were observed at phylum, family, and genus levels, as well as several sex-associated changes probably due to the different levels of sex hormones present in both sexes at certain ages. Our results provide a complete and valuable dataset that will be useful for future research aiming for studying the relationship between changes in the nasopharyngeal microbiome and susceptibility to or severity of multiple diseases.Functional or compositional perturbations of the microbiome can occur at different sites, of the body and this dysbiosis has been linked to various diseases. Changes in the nasopharyngeal microbiome are associated to patient’s susceptibility to multiple viral infections, supporting the idea that the nasopharynx may be playing an important role in health and disease. Most studies on the nasopharyngeal microbiome have focused on a specific period in the lifespan, such as infancy or the old age, or have other limitations such as low sample size. Therefore, detailed studies analyzing the age- and sex-associated changes in the nasopharyngeal microbiome of healthy people across their whole life are essential to understand the relevance of the nasopharynx in the pathogenesis of multiple diseases, particularly viral infections. One hundred twenty nasopharyngeal samples from healthy subjects of all ages and both sexes were analyzed by 16S rRNA sequencing. Nasopharyngeal bacterial alpha diversity did not vary in any case between age or sex groups. Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes were the predominant phyla in all the age groups, with several sex-associated.  Among the remaining challenges in biomedical sciences, one of the most important is to fully understand the effect of aging on human biological processes, health, and wellness. In recent years, solid evidence has been collected to support the idea that the microbial communities that inhabit the different anatomical areas of the human body could play a key role in these processes, and there has been much speculation about possible medical interventions ,3,4,5,6.The human upper respiratory tract that comprises the anterior nares, nasal cavity, sinuses, nasopharynx, Eustachian tube, middle ear cavity, oral cavity, oropharynx, and larynx is the major portal of entry for infectious droplet- or aerosol-transmitted microorganisms . Among tLarge cohort studies of human microbiome data with appropriate controls are particularly valuable, especially of all ages and both sexes, as these datasets are difficult to obtain due to multiple factors, including our long lifespans, heterogeneity in consent and other sample access issues, and because of socioeconomic confounds. Therefore, human studies have tended to focus on a specific component of the lifespan, such as infanthood, or studies of the elderly, rather than examining variation across an entire population. Knowledge about the relationships between changes in the nasopharyngeal microbiota and susceptibility to viral infections is a good example of this, since most studies have focused only on children . A crucial factor to consider in studies of variation across the lifespan is sex as a biological variable. For aging research, this includes the understanding that females and males may have different aging trajectories ,20,21, iHere, we analyze, for the first time, the diversity and relative abundance of the nasopharyngeal microbiota across the whole lifespan in 120 healthy individuals of all ages and both sexes, the taxonomic changes in the nasopharynx associated to age or sex, and the possible biological relevance of several taxa whose frequency of appearance in the population is high. We therefore provide a very comprehensive and valuable dataset that will be the base for future research aimed at identifying relationships between age- and sex-associated changes in nasopharyngeal microbiome and susceptibility to or severity of the diseases of interest.Staphylococcus (13.06%), Dolosigranulum (11.99%), Corynebacterium (10.18%) and Ralstonia (10.08%). The most abundant species were Dolosigranulum pigrum (24.55%), Ralstonia pickettii (19.02%), Corynebacterium pseudodiphtheriticum (4.87%) and Propionibacterium acnes (4.85%). We excluded one sample with an abnormally high proportion of Chlamydophila, which is an indication of an abnormal sampling or pathological disorder of this individual ‘C_A1_M8’. To reveal age-related progression of nasopharyngeal microbiota, we divided the samples into six age groups, each divided into females and males to be able to also study possible sex-associated differences  were obtained after quality control analyses and OTU filtering. The 16S rRNA sequences were binned into 128 families, 250 genera and 561 species. The most abundant families were Staphylococcaceae (12.14%), Burkholderiaceae (11.52%), Carnobacteriaceae (11.48%) and Corynebacteriaceae (9.47%). The most abundant genera were ferences . There wferences .We sought to determine the ways in which different samples were grouped according to their OTU composition. To that end, we applied nonmetric multidimensional scaling (NMDS), which is a powerful statistical tool that enables complex multivariate data sets to be visualized in a reduced number of dimensions, to determine the clustering patterns of samples according to their Bray–Curtis distances  . The anaThe fact that significant changes in bacterial diversity throughout life had previously been described in the well-studied gut microbiota of healthy individuals  promptedTo determine differences in nasopharyngeal taxa abundance among age and sex classes, we compared the nasopharyngeal microbiome of healthy women and men in the six different age groups established for this study . The firNext, we proceeded further in this study and moved to the family level, finding that 24 distinct bacterial families presented an average abundance of >1% in at least one of the age groups studied g. Our anStaphylococcus was the dominant genus in age group A1, Dolosigranulum in A2, Staphylococcus in A3 and A6, Ralstonia in A4, and Streptococcus in A5 (Dolosigranulum or Staphylococcus) that represented the majority of their nasopharyngeal microbiomes  in relative abundance between the distinct age groups in 11 bacterial genera: Acinetobacter, Brevundimonas, Dolosigranulum, Finegoldia, Haemophilus, Leptotrichia, Moraxella, Peptoniphilus, Pseudomonas, Rothia and Staphylococcus , it dramatically decreased in the group of individuals over 70 years old  bacterium b; or . Therefore, these results reveal that, in terms of relative abundance of bacterial genera, the nasopharyngeal microbiota of the youngest and oldest subjects is more different from that of the other age groups than that of any other age group. Among these age-associated changes, from a clinical perspective, it is particularly concerning that Dolosigranulum, which is an opportunistic pathogen that causes pneumonia in elderly patients . According to the exclusion criteria we established for this study, (1) individuals younger than 1 year of age were disqualified because the microbiome of infants is known to be highly fluctuating with age, and (2) subjects who were tested for SARS-CoV-2 infection because they had respiratory or any other kind of symptoms were also excluded to avoid the enrolment of individuals who could have any infection or disease that could alter their nasopharyngeal microbiota although they were not infected by SARS-CoV-2.To facilitate the study of age- and sex-associated changes in the nasopharyngeal microbiota throughout life, and to ensure that the sample size of all the age and sex groups were homogeneous, we decided on an experimental design that distributed the 120 nasopharyngeal samples that we could analyse into six age groups with 20 individuals each, of which 10 were females and the other 10 were males . For thi® XP Beads , and quantified using QubitTM dsDNA HS Assay Kit in a Qubit 3 fluorometer . Bacterial identification was performed by sequencing the 16S rRNA gene’s hypervariable regions. The 16S rRNA gene was amplified by multiplex PCR using Ion Torrent 16S Metagenomics kit , with two sets of primers, which targets regions V2, V4, and V8, and V3, V6–7, and V9, respectively. Amplification was carried out in a SimpliAmp thermal cycler  running the following program: denaturation at 95 °C for 10 min, followed by a 3-step cyclic stage consisting of 25 cycles of denaturation at 95 °C for 30 s, annealing at 58 °C for 30 s, and extension at 72 °C for 20 s; at the end of this stage, the program concludes with an additional extension period at 72 °C for 7 min and the reaction is stopped by cooling at 4 °C. The resulting amplicons were tested by electrophoresis through 2% agarose gels in tris-acetate-EDTA (TAE) buffer, purified with AMPure® XP Beads and quantified using the Ion Universal Library Quantitation Kit  in a QuantStudio 5 Real-Time PCR Instrument . The libraries were then pooled and clonally amplified onto Ion Sphere Particles (ISPs) by emulsion PCR in an Ion OneTouch™ 2 System (Ion Torrent) according to the manufacturer´s instructions. Sequencing of the amplicon libraries was carried on an Ion 530™ Kit (Ion Torrent) on an Ion S5™ System (Ion Torrent). After sequencing, the individual sequence reads were filtered by the Torrent Suite™ Software v5.12.1 to remove low quality and polyclonal sequences.A library was generated from each sample using the Ion Plus Fragment Library Kit (Ion Torrent), whereby each library is indexed by ligating Ion Xpress™ Barcode Adapters (Ion Torrent) to the amplicons. Libraries were purified with AMPure® 16S Reference Library v2013.1 and the Greengenes v13.5 database. For an OTU to be accepted as valid, at least ten reads with an alignment coverage of ≥90% between hit and query were required. Identifications were accepted at the genus and species level with sequence identity of >97% and >99%, respectively. Annotated OTUs were then exported for analysis with R (v.4.1.2) (https://www.R-project.org/), where data were converted to phyloseq object [https://bioconductor.org/packages/devel/bioc/vignettes/phyloseq/inst/doc/phyloseq-mixture-models.html#import-data-with-phyloseq-convert-to-deseq2, accessed on 10 January 2022). The raw abundance matrix was imported into phyloseq object (as specified in the documentation of phyloseq with DESeq2) and subsequently converted to DESeq2 object. Then, estimated size factors were used with the DESeq2 function to obtain the differential abundance. DESeq automatically searches for outliers and, if possible, replaces the outlier values estimating mean-dispersion relationship. If it is not possible to replace, p-values are replaced by NA. R (v.4.1.2) was also used to perform a non-metric multidimensional scaling (NMDS) analysis on Bray–Curtis dissimilarity measures among samples based on relative OTU abundances . The relative abundances of OTUs were also used to test for statistically significant differences among age and sex groups. Group OTU compositions were compared through the non-parametric statistical tool ANOSIM. The 90% confidence data ellipses for each of the age groups were plotted. Alpha diversity was estimated based on Chao1, Shannon, and Inverse-Simpson indices by using the phyloseq package. To test for statistically significant differences between pairwise groups in alpha diversity, the non-parametric Wilcoxon test was used. Frequency of appearance was obtained by calculating the percentage of individuals in each age group in which that taxon occurs. The bar plots aggregated by groups (age and/or gender) show the aggregated relative abundance (sum of relative abundances). Krona charts that aid in the estimation of relative abundances even within complex metagenomic classifications were generated as previously described [The obtained sequences were analyzed and annotated with the Ion Reporter 5.18.2.0 software  using the 16S rRNA Profiling workflow 5.18. Clustering into OTUs and taxonomic assignment were performed based on the Basic Local Alignment Search Tool (BLAST) using two reference libraries, MicroSEQq object  and abunq object  that useescribed . All theescribed .Our study shows that bacterial diversity in the nasopharynx of healthy subjects remains very stable and resistant to perturbations throughout the whole life and in both sexes. Age-associated changes in taxa abundance were observed at phylum, family, and genus levels, as well as several sex-associated changes probably due to the different levels of sex hormones present in both sexes at certain ages. We provide a complete and valuable dataset that will be useful for future research aiming for studying the relationship between changes in the nasopharyngeal microbiome and susceptibility to or severity of multiple diseases.