Citrus sinensis Osbeck cv. Newhall) and examined the effects of soil REE on internal fruit quality in Xinfeng County, Jiangxi province, China. Path analysis showed that soil REE, pH, cation exchange capacity (CEC), and Fe oxide (Feox) significantly affected pulp REE concentrations. A Freundlich-type prediction model for pulp REE was established: log[REEpulp] = -1.036 + 0.272 log[REEsoil] - 0.056 pH - 0.360 log[CEC] + 0.370 log[Feox] . From the prediction model, it was inferred that even when soil REE and Feox were as high as 1038 mg kg-1 and 96.4 g kg-1, respectively, and pH and CEC were as low as 3.75 and 5.08 cmol kg-1, respectively, pulp REE concentrations were much lower than the food limit standard. Additionally, soil REE levels were significantly correlated with selected fruit quality indicators, including titratable acidity , total soluble solids  and vitamin C . Generally, under routine methods of water and fertilization management, the cultivation of navel oranges in rare earth ore areas of south China with soil REE ranging from 38.6 to 546 mg kg-1 had improved in internal fruit quality.The effects of soil rare earth element (REE) on navel orange quality and safety in rare earth ore areas have gained great attention. This study investigated the transfer characteristics of REE from soil to navel orange pulp ( The rare earth element (REE) comprise a group of 16 elements with very similar chemical and physical properties, including the lanthanides Z = 57–71) and the element yttrium Y, Z = 39) . The lan9 . The l7–71 and With the application of REE fertilizers, positive effects on the growth, yield and quality of numerous crops  have been observed for pot or field experiments in many countries, including the United States, the United Kingdom and China ,6. HowevIn addition, establishing empirical regression models is also an important method for determining whether the REE in crop production comply with the food safety limit. Numerous prediction models for the uptake of heavy metals by grains and vegetables have been developed –18. Thes3) digestion has been widely accepted in the environmental sciences as providing a good estimate of the maximal element availability for plant uptake  and the corresponding predicted log[REEpulp] is shown in pulp], log[HREEpulp] and log[REEpulp] were within the 95% prediction intervals, indicating that the prediction models derived in the present study display good accuracy. Moreover, the root-mean-square error (RMSE) values were 0.14, 0.18 and 0.14 for the LREE, HREE and REE prediction models, respectively , TSS  and Vc , indicating that navel orange internal quality improves with higher total REE present in the soil in the investigated area.The contents of TA, TSS and Vc in the navel orange pulp ranged from 0.4% to 0.9%, 10.2% to 16.3% and 31.1 to 69.2 mg 100 gthe soil . SignifiGenerally, the content of REE in the main organs of the navel orange decreased in the following order: root > leaf > peel > pulp ,22. The When comparing the TFs of LREE and HREE, most of the TF values for the HREE are higher than those for the LREE in navel orange pulp. One possible reason for this result is that although the content of the LREE in soils is markedly higher than that of the HREE, plants have the capacity to self-adjust, resulting in the translocation of only small amounts of these metals to the aerial parts of plants . Anotherox were the most important factors for explaining the variability in REE content in navel orange pulp in this study. The negative effect of pH on REE content in navel orange pulp was consistent with the general expectation that REE exhibit higher bioavailability at lower soil pH values; as the soil pH increases, the REE are more likely to precipitate and less likely to be released from soils [The results of the path analysis were consistent with the stepwise multiple regression equation, including that the soil REE content, pH, CEC and Feom soils . CEC is om soils . Previouom soils . Thus, aom soils , which eom soils ,39. One ox are sufficiently wide to represent the soil properties and soil REE contents in the navel orange production location of Xinfeng. Back-calculation from these models may not be accurate for soils in which the common variables  are outside the boundaries for which these models were derived [The prediction models developed in this study can be used to obtain reliable predictions of REE contents in navel orange pulp and therefore to assess the potential risk to humans. In addition, these models can also assist in the management of navel orange orchard soils to ensure the safety of fruits. For example, according to the prediction model, when cultivated on soil with high REE content, pulp REE content can be decreased by increasing soil pH. In this study, the ranges of REE content, pH, CEC and Fe derived . Becauseox. In the literature, the maximum soil REE content in south China is reported to be 1038 mg kg-1 [ox were determined from this investigation, not from the literature, due to the large variations in these values observed in the literature. These values were determined to be 3.75, 5.08 cmol kg-1, and 96.4 g kg-1, respectively. When these values were used in equation 14, a navel orange pulp REE content of 1.13 mg kg-1 dry weight was calculated. When converting the REE dry-pulp-weight content into the REO fresh-pulp-weight content with a water content of 87% and a conversion factor of 0.86 [-1 fresh weight, which is much lower than the food safety limit in China (0.7 mg kg-1 fresh weight) . Theref-1 is beneficial for improving the internal fruit quality, and the REE food safety limit is not exceeded in the pulp of these navel oranges.In addition, REE in soil have positive effects on the internal quality of navel oranges, increasing both the TA and TSS contents in navel orange pulp. The levels of TA and TSS represent the contents of organic acid and sugar in fruit, respectively, and they are key components in the perception of sour and sweet . Therefoox were significant variables affecting pulp REE concentrations. The total REE contents in soils were safe for planting navel oranges in rare earth ore area of south China. Even when total soil REE content was as high as 1038 mg kg-1, the navel orange was still safe enough for consumption. Under routine methods of water and fertilization management, internal fruit quality of navel orange increased with the increase of soil REE in the study area.The present results showed that soil REE content, pH, CEC, and Fe


Especially, the performances of TiS and CdF2 can be comparable to that of the most popular commercial ZnS at high temperature. Finally, we propose possible ranges of infrared transmission for halogen, chalcogen and nitrogen compounds respectively to guide further exploration. The present strategy to explore IR window materials can significantly speed up the new development progress. The same idea can be used for other material rapid searching towards special functions and applications.Long-wave infrared (8–12 μm) transmitting materials play critical roles in space science and electronic science. However, the paradox between their mechanical strength and infrared transmitting performance seriously prohibits their applications in harsh external environment. From the experimental view, searching a good window material compatible with both properties is a vast trail-and-error engineering project, which is not readily achieved efficiently. In this work, we propose a very simple and efficient method to explore potential infrared window materials with suitable mechanical property by first-principles gene-like searching. Two hundred and fifty-three potential materials are evaluated to find their bulk modulus  and phonon vibrational frequency . Seven new potential candidates are selected, namely TiSe, TiS, MgS, CdF Infrared radiation (IR) is with longer wavelengths than those of visible light, extending from the red edge of the visible spectrum at 700 nm to 1 mm. It has been broadly used in industrial, scientific, medical and military fields. Besides the extensively focused IR absorption material, IR transmitting material is another critical component for sensing and detecting system12425675An ideal IR-window material usually possesses a high (and wide) transmittance, and a good mechanical strength including high durability, hardness, chemical stability, wind/rain-erosion resistance and thermal shock resistance22, HgF2, CdO, and SrO which have not been used before. Especially, the performances of TiS and CdF2 are comparable to that of the most popular commercial ZnS at high temperature. The relations between the infrared transmission and the mechanical property are also discussed.Many researchers have expected to improve and reinforce the mechanical performance of the well-known IR transmitting materials, for example, a thin film with fine mechanical performance is deposited on a IR window material1101315161819Bv to predict mechanical strength2123et al.233–105 cm−1. Yet, the 2-phonon region in the range of 101–103 cm−1 also cannot be neglected. This is the region including the overtones of the fundamental vibrations (e.g. 2TO and 2LO) and the combined bands 10c is the light speed. In other words, the wavelength at Bvis almost the same as the experimental counterparts except a slight difference. The average discrepancy to the experimental data can be as low as −4.8%. On the other hand, the optical −1 as the criterionBv and First of all, before the large-scale searching, two key descriptors for the optical and the mechanical performance, which are readily calculated, are defined and compared with the experimental data from the current popular transmitting materials. One descriptor is chosen as the Voigt-type bulk modulus AxBy (A for cation and B for anion) are listed according to the periodic table. The simple binary compositions with x:y = 1:1, 1:2, 1:3, 2:1, 3:1, 2:3, 3:2, which is easy for future experimental production, are adopted. Secondly, the popular crystal structures for each composition are used. Thirdly, the two descriptors discussed above are calculated. Bv and the Bv. This can be readily understood by the behavior of the simple harmonic vibration,k is the force constant, which reflects the bonding strength in a solid and is positively related to Bv. m is the reduced mass2In the next step, the candidates with both the fine optical and mechanical performance for LWIR windows are explored. The procedures are as follow: firstly, the combinations of 2, with its CAL slightly below the midline (dot line) in the LWP region, has a consistent experimental ETL of 9.4 μm slightly below the midline (10 μm) of 8~12 μmTo evaluate the results, a standard to choose the most possible candidates is defined. Considering the average discrepancy (20.5%) between the ETL and the CAL/FAL by the benchmark test above, the concerned LWIR window (8~12 μm) is accordingly modified to (9.6~14.5 μm), as shown in the inset including LWP  and LWF  transmitting window regions. The bulk modulus ≥72 GPa is chosen according to the mechanical performance of the popular ZnS midline  μm of 8~Bv are used as criteria, it seems substantial materials are available. However, the zero-band-gap material is not suitable for windows due to the feasible IR absorption by their intrinsic free carriers generated at the elevated temperature. Thus, a further filtration is necessary through band structure evaluation for the candidates in LWP and LWF cover regions. Here, to enable a more reasonable prediction of metallic or insulating characteristics, the GGA+U method is applied to the transition-metal compound which has no band gap in a normal GGA calculation28293031AxBy materials, the orange squares highlight the potential candidates which are suitable for LWIR transmission, namely, TiSe, TiS, MgS, CdF2, HgF2, CdO and SrO . In fact, except TiSe, other candidate shows a larger band gap (>1.7 eV) with further adding 25% ExHF in a HSE calculation. According to a rule-of-thumb by HiltonIf only the 2, HgF2, CdO, and SrO are displayed in Stability is another important factor for the application of the new potential candidates. The crystal structures, phonon dispersion and band structures of TiSe, TiS, MgS, CdFNVT simulation, the snapshots of each crystal structure for TiSe, TiS, MgS, CdF2, HgF2, CdO and SrO are shown in 2 are seriously distorted after the annealing . Especially, for TiSe, the bond length fluctuates significantly. On the other hand, TiS, MgS, CdF2, CdO and SrO have the good thermal stability due to the stable bond-length evolution. After further consideration of chemical activity2 in the air. Thus, TiS and CdF2 should be the most possible candidates for LWIR transmitting windows at high temperature. SrO, MgS and CdO may also work if they are insulated from water and air.To further examine the applicability of these IR candidates in harsh external environment such as at the very high temperature, molecular dynamic simulations are carried out for these compounds. High-temperature molecular dynamics (HTMD) methods are widely used to test the thermal stability and behavior in materials1Last but not least, we try to summarize the “gene” behaviour of the materials towards the IR window application and tell the possible design rule. Bv, well describe the properties for IR window materials. Especially, in the benchmark test, the theoretical simulations are well consistent with experiments. From two hundred and fifty-three binary compounds with AxBy form, seven new potential candidates of TiSe, TiS, MgS, CdF2, HgF2, CdO, and SrO are finally decided. Especially, the performances of TiS and CdF2 can be comparable to the most popular ZnS at high temperature and in chemical stability. The intrinsic “gene” behaviour of the IR material is also studied to facilitate the future search for IR window materials. Here, we stress that the development of a practical IR window is in fact a big engineering project. To meet the requirement, people must carefully control the defects during the growth of material because defects or unintentional impurities can always lead to IR absorptions due to the local state inside the band gap or the free carrier supplement at band edge. Also because some large scale defects  can scatter IR seriously, thus worsen the transmission. On the other hand, some cover layers are also required to protect the IR window materials from the erosion by wind and rain. At this stage, we still have not considered the problems mentioned above, however the present strategy at least tells people which candidate should be focused on for IR window materials. We believe the same idea can also be used in searching other materials toward a special performance with the help of material gene-like exploration.In this work, a simple and efficient method has been proposed to explore the potential LWIR window materials with both the suitable mechanical and optical performances by a first-principles gene-like searching. The two descriptors, namely AxBy binary compounds, based on first-principles calculations. The theoretical investigations employ the density functional theory36Ab-initio Simulations Package (VASP)38AxBy  in inorganic crystal structure database (ICSD). Here, the phonon dispersion calculation uses the supercell method by phonopy codeBv (Voigt-type) for cubic, hexagonal, or tetragonal lattice is calculated from the elastic constant (Cij) by Voigt-Reuss-Hill approximationsNVT ensemble is usedPhonon dispersion and elastic constants are obtained to estimate the long-wave absorption limit How to cite this article: Du, J.-R. et al. Exploring long-wave infrared transmitting materials with xAyB form: First-principles gene-like studies. Sci. Rep.6, 21912; doi: 10.1038/srep21912 (2016).


Given the high prevalence of polycystic ovary syndrome (PCOS), and that a lifestyle is recognized effective in development of many diseases, this study aimed to compare lifestyle of women with PCOS and healthy women. Nor are there sufficient studies on the difference between lifestyle of these people with that of healthy people. Furthermore, studies show that changes in lifestyle improve this disease. This descriptive-comparative study was conducted on 65 women with PCOS and 65 healthy women of 18 to 45 years old who presented to hospitals affiliated to Shahid Beheshti University of Medical Sciences in 2013. The subjects were selected using multi stage random sampling method. The data were collected using questionnaires for diet, International Physical Activity Questionnaire, and unhealthy behaviors and were analyzed in SPSS v. 17, using descriptive statistics, Man-Whitney, independent t, Chi-square and logistic regression tests. The results showed there was a significant relationship between PCOS and inappropriate diet (p=0.009), low physical activity (p=0.009), but no relationship was observed between PCOS and unhealthy behaviors. Given the results obtained, training and awareness raising is necessary for women and girls especially about appropriate diet and regular physical activity. Polycystic ovary syndrome (PCOS) is the most prevalent endocrine disorder in women at reproductive ages, and the most common cause of hyperandrogenism and hirsutism. The prevalence of PCOS varies from 2.2% to 26% in different countries and has been reported as 7.1% based on National Institutes of Health (NIH) criteria, as 11.7% based on Androgen Excess Society (AES) criteria, and as 14.6% based on Rotterdam criteria in Iran . This sy3, and confirmation of the gynecologist in the center. Healthy women were selected from women visiting the same hospitals, but without signs of the disease according to Rotterdam criteria. Sample size was determined using the below formula with CI 95% as 65 people in each group.This descriptive-comparative study was conducted on 65 women with PCOS and 65 healthy women presenting to hospitals affiliated to Shahid Beheshti University of Medical Sciences in 2013. Inclusion criteria are: age between 18 and 45 years old, Iranian and Persian speaking, ability to read and write, no known medical conditions according to self-report or using patients’ records including diabetes, hypertension, thyroid disorders, hyperprolactinemia, Cushing’s syndrome, adrenal hyperplasia, no antidepressant medication, no hormone therapy, no oral contraceptive, no glucocorticoids, no anti-obesity medication, no special diet, and not being a professional athlete. Women were selected based on Rotterdam criteria (at least two out of three criteria) including amenorrhea, oligomenorrhea, clinical or lab evidence of hyperandrogenism including hirsutism, alopecia, acne or increased serum levels of androgens, ultrasound evidence of PCOS including more than 12 follicles in each ovary with a diameter of 2-9 mm or increased volume of ovary to more than 10 cm2σ2/ (μ1-μ2)2n= 2 (Zα + Zβ)  (1)In this equation, type I error, test power, and effect size in estimation of differences in lifestyle were determined in both groups as 5%, 80%, and 0.5, respectively.1 and unhealthy behavior questionnaires, tape measure and scale. Demographics questionnaire included age, height, weight, BMI, education level, marital status, if married: husband’s education, monthly income, menstruation history such as menarche, interval between menses, obstetrics history like parity, number of miscarriages, number of preterm births, and use of assisted fertility methods. The researcher-made diet questionnaire comprised 28 items with 0 to 112 points, where higher score showed more appropriate diet. There were 16 items for positive diet and 12 items for negative diet. Total score was measured by considering from 4 points for daily to 0 for never regarding the positive items and vice versa for the negative items. The total score was converted to 0 to 100 for statistical analysis, and 0-33, 34-66 and 67-100 were considered for inappropriate, quite appropriate and appropriate diet, respectively. Content validity was used to validate the questionnaire. Its reliability was measured as 0.80 using Cronbach’s alpha. IPAQ was developed in 1998 in Geneva by WHO and CDC for age groups of 15-69. It has 27 items that report physical activity based on MET-min.week. One MET equals the amount of energy consumed in one minute of rest. IPAQ classifies people based on MET into 3 groups of low activity (<600 MET), average activity (600-3000 MET) and high activity (>3000 MET). This questionnaire is standard and its validity has been confirmed using content validity in previous studies .2 in the PCOS and the healthy groups, respectively. There was no significant difference between the two groups. There was no difference between the two groups in terms of subjects’ education, their husbands’ education, subjects’ occupation, their husbands’ occupation, monthly income and marital status. , and the number of miscarriages (p=0.142). Nonetheless, there was a significant difference between the two groups in terms of parity (p=0.004) and the number of preterm births (p=0.049).As can be seen in The analysis of the data showed a significant relationship between PCOS and inappropriate diet (p=0.009), low physical activity (p=0.009). In recent years, researchers have attended to factors involved in incidence or severity of PCOS. Nutritional habits are one dimension of lifestyle which affects people’s physical health (Few studies have been conducted on lifestyle and nutritional habits of people with PCOS . ApproprThere was no significant relationship between the mean score of unhealthy behaviors between the two groups. Several studies have proved that smoking reduces estrogen levels and increase androgen levels in women at reproductive age. Therefore, smoking increases metabolic syndrome and hyperandrogenism in women with PCOS (The results showed that there is a difference between lifestyle of people with PCOS and those without in several domains. The need to attend to nutrition and physical activity in women and girls is clear.


Trichiurus lepturus. The ichthyoplankton was identified via canonical correspondence analysis to three assemblages: an estuary assemblage dominated by Chaeturichthys stigmatias, a coastal assemblage dominated by Engraulis japonicus and Stolephorus commersonii, and an offshore assemblage dominated by Trichiurus lepturus. Regarding environmental factors in the Yangtze Estuary, suspended matter and surface seawater salinity were the main factors influencing the distributions of the different assemblages, while sediment from the Yangtze River during the rainy season and chlorophyll a were the principle drivers of the annual variances in the distribution of ichthyoplankton assemblages. Our aims in this study were to provide detailed characterizations of the ichthyoplankton assemblage in the autumns of seven years, examine the long-term dynamics of autumn ichthyoplankton assemblages, and evaluate the influence of environmental factors on the spatial distribution and inter-annual variations of ichthyoplankton assemblages associated with the Yangtze Estuary.This study investigated the response of the ichthyoplankton community to environmental changes in the Yangtze Estuary using canonical correspondence analysis. Ichthyoplankton community and environmental data were recorded during the autumns of 1998, 2000, 2002, 2003, 2004, 2007 and 2009. Among the ichthyoplankton, the dominant larval and juvenile families were the Engraulidae, Gobiidae and Salangidae, and the most common eggs were from  The Yangtze Estuary, an area of complex and varied environmental conditions, is the link between the Yangtze River and the East China Sea . Sea curIchthyoplankton, including fish eggs, larvae and juveniles, is the basis for the sustainable utilization of fishery resources , with suTo date, investigations into ichthyoplankton communities in Yangtze Estuary have mainly concentrated on species composition, fauna distribution and seasonal variations i.e., , discusser mouth .Biological and oceanographic data were collected during seven fishery-evaluation cruises in seven autumns. Biological samples were collected by surface towing a larva net  equipped with a flow meter. At each station, the net was towed at a depth of ∼0.5 m from the surface for 10 min against the tidal flow, at a towing speed of approximately 2–3 knots. After completion of each tow, the nets were washed and the samples fixed and preserved in 5% buffered formaldehyde–seawater solution.a (Chla), were determined according to the GB/T 12763-2007, Specification for Marine Monitoring (http://www.cjh.com.cn/pages/nsgb.html).Geographical locations of sampling stations were provided by GPS, version Magellan 315. At each station, environmental variables, including depth (D), salinity (S), temperature (T), transparency (Trans), dissolved oxygen (DO), pH, total phosphorus (TP), total nitrogen (TN), total suspended matter (TSM), chemical oxygen demand (COD), and chlorophyll nitoring . Becausenitoring  and the Ichthyoplankton were identified in the laboratory to the lowest possible taxonomic level. Numerical density for each species was standardized to catch per unit effort (CPUE) as abundance per 10 tow units.The dominant species were determined using the Index of Relative Importance (IRI) developed by N∗100% and F∗100% are the relative abundance and frequency of occurrence, respectively. The IRI of the dominant species should be greater than 100.x + 1) transformed to reduce the dominant effect of some species (P < 0.05) analysis simulation and the forward selection option within CANOCO , which is utilized to visualize and describe the relationship between fish species and environmental variables . Only species that occurred in >1% of the catches, based on all species, were included in the analysis. Species abundance data were log10 , TSM, S, TSedi from the Yangtze River during the rainy season, Chla and PH of surface seawater were the most active environmental parameters affecting ichthyoplankton. Total variation due to TSM and TSedi was higher than other environmental factors. The patterns of D, TN, TP, TSM and S were highly correlated with the first CCA axis, and this CCA1 axis represented a spatial gradient from the Yangtze River to the sea. There was a high correlation between the second axis and TSedi, i.e., sediment from the Yangtze River during the rainy season, and Chla; the CCA2 axis represented a temporal gradient , representing a decrease in TSM from the river to the Offshore with increasing salinity, depth and transparency. There was a positive correlation between TSedi and Chla, both impacting on changes in the annual ichthyoplankton assemblage. There were combined effects of environmental factors in the CCA biplot of ichthyoplankton species. The resulting hierarchical classification yielded both species and station groups.Three species groups were revealed by CCA :Chaeturichthys stigmatias, Salanx prognathous, Pseudolaubuca engraulis and Pseudorasbora parva), distributed in an area of low salinity and high suspended matter;Estuary assemblage: This group included four species , distributed in brackish water;Coastal assemblage: This group included seven species , distributed on top-left of the CCA biplot and correlated with salinity.Offshore assemblage: This group included seven species  was the most abundant species, followed by Salanx prognathous (22.05%). Salinity in this area was 2.34 ± 1.1‰, and the total suspended matter was 121.66 ± 61.54 mg/L.I. Estuary area consisting of five sites: 35, 36, 37, 38 and 39, distributed in the river system. In this group, Salanx prognathous (29.81%), Stolephorus commersonii (23.40%) and Engraulis japonicus (20.06%). Salinity in this area was 19.41 ± 8.17‰ and the total suspended matter were 134.77 ± 240.10 mg/L, the highest of three groups.II. Coastal area consisting of 16 sites: 9–11, 15–18, 21–24, 28–31 and 40, distributed in water near the mouth of the Yangtze Estuary. The dominant species in this group included Trichiurus lepturus (58.90%) and Engraulis japonicus (10.68%) were the dominant species. Salinity in this area was 19.41 ± 8.17‰ (the highest of three groups), and the total suspended matter was 7.78 ± 13.99 mg/L (the lowest of three groups).III. Offshore area consisting of 19 sites: 1–8, 12–14, 19, 20, 25–27 and 32–34, located in offshore areas outside the Yangtze Estuary. In all, environmental conditions were significantly different among the three stations groups, and the distributions of ichthyoplankton varied with environmental factors from the estuary to the offshore area.S. commersonii assemblage group remained unchanged compared to the results in springs, although the seawater temperature within the estuary water may have changed between the springs and autumns (As a result of the interaction between biological and abiotic factors, in this study, coastal and offshore ichthyoplankton were dominant in the Yangtze Estuary during the November sampling periods. Compared with the characteristics of the ichthyoplankton assemblage within the Yangtze Estuary in springs , species autumns . Moreove autumns , but theSeasonal patterns of abundance of ichthyoplankton are mainly linked to reproductive strategies of adult population and phases of their life cycles, which, in turn are often associated with oceanographic and meteorological features . Most ofa.The ability of larval and juvenile fish to survive the pelagic phase and migrate to a suitable adult habitat may be dependent on their ability to regulate their dispersal or migration ; larvae a were the major factors affecting changes in the annual ichthyoplankton assemblage. Other environmental factors, such as temperature, nutrients, depth, dissolved oxygen and chemical oxygen demand, had no significant impact on ichthyoplankton.CCA analysis was utilized to reflect the correlations between community groups and environmental factors. The ichthyoplankton assemblage was divided into three groups: an estuary, coastal and Offshore assemblage. Compared with the ichthyoplankton assemblage pattern in spring after 2004 , there wIn estuaries, there are significant gradients in salinity and turbidity . The spaThe salinity was negatively correlated with suspended matter , which wFreshwater inflow is the critical factor determining abiotic and biotic variability , and hasa also influenced changes in the annual ichthyoplankton assemblage, and was positively correlated with sediment entering the Yangtze Estuary. Chlorophyll a is a relative accurate reflection of the standing crop of phytoplankton; the greater the phytoplankton biomass, the higher the primary productivity (a and nutrients from freshwater entering the estuary. When nutrient levels were high, chlorophyll a levels in the estuary were also high, and food resources, which are necessary for the development and growth for larvae and juveniles, were abundant. Rich food resources also increased survival and successful foraging rates. From the standpoint of trophodynamics, high chlorophyll a levels provided a food indicator for recruitment in the early life-stage of fish species (a had a major impact on the distribution of three freshwater fish, Stolephorus commersonii, Salanx ariskensis and Engraulis japonicus. a and the distribution of eggs, larvae and juveniles of E. japonicus. Moreover, a study on the distribution and variation in chlorophyll a demonstrated the relationship between phytoplankton and the environment and changes in environmental conditions (Chlorophyll uctivity . There i species . Figure nditions .PH is another important environmental factor influencing the distribution of an ichthyoplankton assemblage. There are three species groups and three station areas revealed by CCA based on the biological and environmental data in the seven autumns in the Yangtze Estuary which was consistent with the results of the springs and other estuarine systems.SPM and S were the most important environmental factors influencing the ichthyoplankton assemblage pattern. However, TSedi from the Yangtze River during the rainy season and Chla were the major factors affecting changes in the annual ichthyoplankton assemblage. Other environmental factors, such as T, nutrients, D, DO and COD, had no significant impact on ichthyoplankton.10.7717/peerj.1922/supp-1Supplemental Information 1Click here for additional data file.10.7717/peerj.1922/supp-2Data S1Click here for additional data file.10.7717/peerj.1922/supp-3Data S2Click here for additional data file.


Puccinia coronata f. sp. avenae) and powdery mildew (Blumeria graminis f. sp. avenae) are among the most important constraints for the oat crop. Breeding for resistance is one of the most effective, economical, and environmentally friendly means to control these diseases. The purpose of this work was to identify elite alleles for rust and powdery mildew resistance in oat by association mapping to aid selection of resistant plants. To this aim, 177 oat accessions including white and red oat cultivars and landraces were evaluated for disease resistance and further genotyped with 31 simple sequence repeat and 15,000 Diversity Arrays Technology (DArT) markers to reveal association with disease resistance traits. After data curation, 1712 polymorphic markers were considered for association analysis. Principal component analysis and a Bayesian clustering approach were applied to infer population structure. Five different general and mixed linear models accounting for population structure and/or kinship corrections and two different statistical tests were carried out to reduce false positive. Five markers, two of them highly significant in all models tested were associated with rust resistance. No strong association between any marker and powdery mildew resistance at the seedling stage was identified. However, one DArT sequence, oPt-5014, was strongly associated with powdery mildew resistance in adult plants. Overall, the markers showing the strongest association in this study provide ideal candidates for further studies and future inclusion in strategies of marker-assisted selection.Diseases caused by crown rust ( Avena sativa L. including both white and red oat (formerly A. byzantina K. Koch) is the main cultivated oat. Several pathogenic fungi can infect oats and drastically reduce its yield including biotrophic pathogens such as powdery mildew  and crown rust (Puccinia coronata f. sp. avenae Eriks). These fungi have very efficient spreading mechanisms, hampering their control by crop management measures such as rotation and the use of resistant varieties is one of the best control alternatives  associated with important agronomic traits using a number of experimental bi-parental oat populations. Examples include vernalization response, flowering, and heading date , qualitysistance , powderysistance , and crosistance . Howeversistance .Association analysis is an alternative approach that overcomes many of the limitations of conventional QTL mapping and has received increasing attention from plant geneticists during the last few years  followinIn recent years, genome wide association studies (GWAS) have identified marker-trait associations for a range of agronomic traits in many crops including maize, rice, sorghum, and foxtail millet . HoweverA. sativa accessions (110 white and 31 red oats) kindly provided by the “Centro de Recursos Fitogenéticos,” INIA, Madrid, Spain, and 36 commercial varieties supplied by the Andalusian Network of Agriculture Experimentation (RAEA) was used. Oat cultivars studied were: Ac1, Acebeda, Adamo, Aintree, Alcudia, Anchuela, Araceli, Brawi, Caleche, Cannele, Chambord, Chappline, Charming, Cobeña, Condor, Cory, Edelprinz, Flega, Fringante, Fuwi, Hammel, Kankan, Kantora, Karmela, Kassandra, Kazmina, Mirabel, Mojacar, Norly, Orblanche, Pallini, Patones, Prevision, Primula, Rappidena, and Saia. Details of the origin of all accessions and of their genetic relationship have been previously reported in For this study, a germplasm collection of landraces consisting of 141 -2 s-1 photon flux density supplied by high-output white fluorescent tubes.Seedlings were grown in 0.5 L pots filled with peat:sand (3:1) in a growth chamber with 20°C, 65% relative humidity (RH) and under 12 h dark/12 h light with 250 μmol mFirst leaves from 40 12-days-old seedlings were harvested, pooled together, and DNA extracted according to the method stipulated by Diversity Arrays P/L, Canberra, ACT, Australia and described by To remove possible errors and redundancies in markers that may cause false associations in GWAS, data cleaning was performed according to k-value were run. Then, the mean estimate across runs of the log posterior probability of the data for a given k, were plotted to enable the determination of the k-value of the population. As this point is known to be difficult to determine, the Δk, related to the second order rates of change of the likelihood function with respect to k, was also used  isolate Co-04, previously multiplied on the susceptible cultivar Araceli, was used. The virulence of this isolate on an oat differential set collection has been described in -2 (checked by counts made from glass slides laid adjacent to leaves). After inoculation, plants were incubated for 9.5 h in darkness at 100% RH and 18°C, and thereafter at 20°C under a 12 h photoperiod with 250 μmol m-2 s-1 photon flux density. Infection frequency (IF) was determined as previously described in The -2 with one isolate of B. graminis f. sp. avenae race five maintained on seedlings of oat cv. Selma, in a spore proof glasshouse. After inoculation, plants were maintained in the growth chamber for 8 days before assessment of the percentage area covered by powdery mildew on the inoculated leaf. Disease scores were converted into relative values, expressed as the percentage of the susceptible reference cultivar Selma and referred to as the relative disease severity ]) to normalize data and stabilize variances throughout the data range, and subjected to analysis of variance using SPSS software, after which residual plots were inspected to confirm that data conformed to normality. Significance of differences between means was determined by contrast analysis (Scheffe’s). The percentage of variation of each trait explained by the structure was computed through multiple linear regression of the phenotypes on the percentages of admixture using R .p-value for each marker, which is the probability of a greater F-value under the null hypothesis that the polymorphic site is independent of phenotype. All models were assessed for their ability to control for type I error by plotting the distribution of the p-values for the markers, where uniformly distributed p-values indicate proper control for type I errors  to further characterize the most significant markers identified.As many of the DArT markers used here have been previously sequenced , the NCBOf the initial 15,000 DArT markers assessed, 1,587 showed polymorphism in the oat collection. In addition 499 SSR alleles were also polymorphic. From the total 2086 polymorphic markers, 11 markers that showed a call rate lower than 80% and 56 markers that showed a MAF < 0.01 were removed. A total of 476 redundant markers were also merged in 169 groups representing these markers. Following data curation a total of 1,712 markers were used for association purposes in the oat collection of 177 white and red commercial varieties and landraces.Figure 1). Indeed, the correlation between SSR and DArT+SSR results was high with a correlation coefficient of 0.84 (p < 0.001). However, slight modifications of the genotype-cluster assignation and the corresponding percentage of admixture were observed. According to both analyses ∼30% of the accessions showed less than 80% of membership for a particular cluster. The differences of genotype-cluster assignation were always related to these accessions and when they were discarded from the analysis the correlation coefficient increased up to 1. According to STRUCTURE, subpopulation 1 showed the highest degree of admixture with 75.6% of the genotypes with less than 80% of membership to this subpopulation followed by subpopulations 3 and 4 with 18% of genotypes with less than 80% of membership in these groups. Subpopulation 2 in which only 15% of genotypes showed less than 80% membership to the corresponding subpopulation was the subpopulation with lowest admixture.A previous genetic diversity study of the oat collection with only SSR markers revealed a structure of four subpopulations . In the Figure 2). Cluster analysis was implemented on the first four principal components cumulatively explaining ∼50% of the variation with 23.3, 13.8, 8.11, and 4.5% for each of the components, respectively. Although separation between clusters was clear, some accessions were not part of the clusters but formed links between them . The number of lines in each cluster ranged from 33 to 64. The first cluster included mainly the white commercial varieties, cluster 2 the red oats, cluster 3 the white oat landraces characteristic to high altitude locations, and cluster 4 white oat landraces more adapted to low altitude locations.Multivariate analysis based on PCA also revealed a separation of four subpopulations which indicates a high consistency of the data . These relationships between clusters were in agreement with those depicted by the PCA scatter plot . These results suggest that clustering was also efficient in separating the oat types for the germplasm used in this study.As previously stated, PCA showed a separation between clusters but also a clear pair-wise relationship between clusters. Quantitative results for genetic distance (according to Nei’s parameter) between clusters are shown in r2, and the probability . A total of 507,042 pairs of markers showed a significant LD value with an average p = 0.004. From these, 277,920 pairs of markers showed an r2 < 0.1 chosen here as nominal level, according to the studies performed by Identification of disequilibrium between markers is highly useful since it may condition the strength of the association study. Since physical map distances between markers were not available, LD was represented by the disequilibrium matrix visualizing the linear arrangement of LD between polymorphic sites, represented by Figure 4). Means of the 177 accessions assigned to the four subpopulations for the different traits, excluding the admixed accessions were compared (Table 2). Significant differences between subpopulations were observed for all traits. Thus, subpopulation 4 had a significantly lower RIF after rust inoculation than the others (p < 0.005) and showed a high resistant response. Subpopulation 2 had lower RDS to powdery mildew than the others . Powdery mildew was the trait most affected by population structure although the proportion of variance explained by population structure remained under 4% (Table 3).Both traits followed a normal distribution with accessions ranging from highly resistant to highly susceptible . This last model showed almost a perfect fit between the observed and expected p-values except for few of the values, which is the characteristic of a model that sufficiently accounts for the number of false positives. Three additional markers, AM30-178, AME176-3, and oPt-15665 were significantly associated in all GLM models including those accounting for population structure  but not in the MLM models (Table 4).As expected, considerably fewer markers showed a significant association with rust resistance when applying a correction accounting for the population structure than when using GLM alone . The low ability of the GLM alone to account for false positives was confirmed when the distribution of observed e Figure . The dist Figure . Five map-values  and since it was not highlighted by the other models it was not considered to be strongly associated. Indeed, distribution of the p-values in the GLM model corrected with Q indicated also a relative over-abundance of low p-values. Interestingly, the reverse tendency was observed in the MLM with p-values moving down the expected values, indicating a scarcity of low p-values and indicating that the GLM+PCA or GLM+Q+PCA were the best fitted models. This indicated the importance of testing the models in order to help in the selection of the most robust markers.A significant reduction of associated markers with powdery mildew resistance in oat at seedling stage was also observed after correcting for the population structure than when using GLM alone. The low efficiency of the GLM alone in this data set was confirmed by the low fit of the observed and expected distribution of the s Figure . Since t2r < 0.1 .In order to find markers associated with powdery mildew resistance we took advantage of a previous detailed evaluation for powdery mildew adult plant resistance performed in a subpopulation of this collection . In this Figures . AssociaTable 6 show the most significant matches found with BLAST searches (BLASTX) against public databases for associated DArT markers. For most DArT markers, no significant match to specific genic sequences was identified with most significant hits corresponding to repetitive sequences and retrotransposons . However, moderately significant matches to disease resistance genes, including the wheat rust resistance locus Lr21, were seen for a DArT marker flanking MAMA5, oPt-14345  oats. Indeed, in a study by As a first step for the association study, population structure was inferred since it has great implications on the design and analysis of GWAS. The different approaches used here indicated moderate population structure within the germplasm collection evaluated. Thus, four oat groups could be detected albeit they presented a certain degree of admixture according to STRUCTURE software with up to 30% of accessions having less than 80% membership to a determinate group. This was also observed following PCA with several accessions covering “gaps” between clusters. Interestingly the group comprising the commercial varieties showed the highest degree of admixture, most likely due to a sharing of common ancestors in their genealogy as reported by Despite the relative moderate structure of this oat population, it contains a reasonably high genetic diversity, showing ∼10% of polymorphic markers as in the original oat panel . This is2r to 0.15 have been reported to occur within 500 bp  would explain, on average 20% of QTL variance. This is not very different from other self-pollinated cereals such as barley in which LD decay is expected to occur over relatively long map distances compared with allogamous cereals such as maize for which reduction of n 500 bp . Our worThe molecular marker data set in combination with phenotype evaluation was used to examine linkage-related marker-trait associations. Separating the role of population structure and genetic linkage as causes for marker-trait association remains the greatest challenge in association analysis . The fivArabidopsis mutant, atg2-2, has been reported to have enhanced resistance to powdery mildew , mapped onto rice chromosome 9. MAMA5 is reported by Pc38 that cluster with Pc62 and Pc63 . According to stachyon , a powdesome 5BL  and a loand Pc63  also mapand Pc63  together3 (Prq1b . AME176 n mapped . Finallyn mapped  onto chrOverall, the markers showing the strongest association in this study provide ideal candidates for further studies and future inclusion in strategies of MAS.GM-B, NR, and JS-M carried out most of the experimental work and data analysis. DR contributed to the disease resistance aspects. TL, CH, and LM contributed to the genetic aspects. EP designed experiments, and contributed to the interpretation of results and writing of the manuscript. NR, DR, TL, CH, and LM also contributed to critical reading and writing.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Background: Knowledge of the epidemiology of traumatic brain injury (TBI) is required both to prevent this disorder and to develop effective care and rehabilitation approaches for patients.Objective: The aim of this article is to find solutions to decrease the incidence of TBI and offer recommendations for their prevention.Material and methods: We analyzed epidemiological studies on TBI by performing a systematic review of literature, using information reported by different centers, collecting data on demographics, showing characteristics of TBI including incidence, identification of risk groups on differences in age, gender, geographical variation, severity and mortality.Results: Studies suggest that the incidence of TBI is between 18 and 250 per 100,000 persons per year. Men and people living in social and economical deprived areas, usually young adults and the elderly are high-risk groups for TBI.Discussion: Prevention remains the “key point” in medicine and especially for TBI, saving the patient from unnecessary often-harsh sufferance.Conclusions: Most public epidemiological data showed that TBI is a major cause of mortality and disability. The effort to understand TBI and the available strategies to treat this lesion, in order to improve clinical outcomes after TBI, may be based on an increase in research on the epidemiology of TBI. A coordinated strategy to evaluate this public health problem in Romania would first of all rely on a related advanced monitoring system, to provide precise information about the epidemiology, clinical and paraclinical data, but concerning the social and economic connected consequences, too. Abbreviations: CNS = central nervous system, ED = emergency department, EU = European Union, FTE = Full Time Employees, GCS = Glasgow Coma Scale, TBI = traumatic brain injury, US = United States, WHO = World Health Organization.  This article approaches a number of aspects related to the epidemiology of traumatic brain injury (TBI) as it became a worldwide public health problem, which can lead to long-term disability, which presents by far consequences that should not be neglected, with personal “destiny fractures” and substantial socio-economic costs for the affected people and their relatives, but also for the community. Therefore, there is a great need for more effective ways to prevent TBI. 1,2]. Therefore, patients with consequent disabilities, required for treatment and rehabilitation, should always be in the center of extended and integrated, coherent approaches, in order to eventually prevent long term medical and psychological complications as well as the socio-professional, family and/ or economic effects.Knowledge about the epidemiology of TBI is necessary in order to prevent this disorder, to improve public health programs and to efficiently implement protocols of managing it, being able to provide effective medical care and rehabilitation services for patients, in view of obtaining better outcomes and reducing not only physical disability but also cognitive and emotional sequels that tend to evolve in relation to the severity of trauma [3]. Other confusing variables in determining the epidemiology of TBI include the use of different definitions that may not clearly characterize the type of lesion, as well as the appearance of conflicting data within the results of diagnostic imaging used at separate times [3].Analyzing the epidemiological studies on TBI during an extended period, we found that while collecting data, several problems could occur, including the fact that many patients with moderate TBI do not present to the hospital and those who do, can be discharged from the emergency units without appropriate documentation. Additionally, deaths following severe TBI that result after accidents or during transport to the hospital cannot be fully accounted for collecting data as necessary in related epidemiological studies .The pressing need to advance research in TBI is obvious, especially when considering the predictions of the World Health Organization (WHO) that TBI will become the third leading cause of death and disability in the world, by 2020 [6].TBI is the leading cause of death in the first forty years of life, and the number of years lost due to accidents is far greater from cardiovascular or neoplastic disease [7]. The first place in the etiological structure of acute brain lesions is held by TBI, followed by ischemic or hemorrhagic stroke, brain tumors, diffuse cerebral hypoxia after resuscitation [Incidence8]. The general incidence of TBI in developed countries is frequently stated to be 200 of 100.000 population/ year, including only patients admitted to hospitals , while in 1995, the mean annual incidence rate of hospitalized survivors after TBI was 99 per 100.000 population  [9].Later, in the 1990’s, the incidence of TBI in the US was estimated at 200/ 100,000 population, meaning 500,000 new cases in a population of 250 million inhabitants .More recently, in 2003, there were over 1 million emergency department (ED) presentations due to head trauma (423 of 100.000), thus resulting in an increase of them [12]. Within the last decade (2001–2010), rates of TBI-related ED visits increased by 70%. Therefore, in 2010, in the US approximately 2.5 million people sustained a TBI, representing an important increasing rate: to 823/ 100,000 [13]. TBI generates about 1 million presentations to the emergency units per year in the European Union (EU). About 235 per 100,000 inhabitants/ year are frequently hospitalized, the incidence including both inpatients and persons who died after TBI [14]. According to the preliminary results of an epidemiological research conducted in the neurosurgery departments in Romania, in 1997, by the group “Neurotrauma” of the Romanian Society of Neurosurgery, TBI incidence varied between 25-95%, being the leading cause of death [Incidence by ageFig. 2) [8]. Comparing the incidence of TBI in adolescents/ young adults (15-30 years) age group, among different geographical areas, it has been concluded that in the US it reached values between 154 to 415 per 100,000, in France it achieved 535 per 100.000, and in Australia it attained 240 per 100.000 persons. While the incidence among adults in US was 93 per 100.000 persons, in Australia it reached 35 per 100.000, whereas in France it reached 190 of 100.000. Regarding the incidence in geriatric population, in US it was about 173 of 100.000, in France was nearly 275 of 100.000 and in Australia, it was 100 of 100.000 and . Regarding the mechanism of injury in the US, motor vehicle accidents - the leading cause of TBI in the general population - were responsible for 50% of all TBI.3]. The very young people, also frequently suffer TBIs due to falls [3]. Overall, findings suggest that the overall annual rates of fall-related TBI deaths increase dramatically with increasing age [16]. Falls - the second leading cause of TBI in the US - were accounting for about 20-30% of all TBIs. In elderly, over 65 years, falls are the most common causes of TBI [Fig. 4) [17]. More accurately, the annual average percentage of the presentations in the emergency room, of the hospitalizations and deaths regarding the causes of injuries due to TBI in the US, between 1995 and 2001, were found in a study as it follows: 28% from falls, 20% by motor vehicle accidents, 19% by blows, 11% assaults, 9% unknown, 7% other, 3% no motor vehicle accidents (cyclist), 2% other transport, 1% suicide , and a leading cause of TBI among persons aged 25-34 years, being more prevalent in African Americans than in Caucasians [8].Sports and recreational injuries were responsible for 10% of all TBI [18]. A Romanian study showed that during 2001 and 2005, 333 TBI patients, with the mean age around 40 years, needing rehabilitation, were transferred to Physical & Rehabilitation Medicine Clinic Division of “Bagdasar-Arseni” Clinical Emergency Hospital, Bucharest, the etiology of head trauma being the following: car accidents – 194 cases (58%), falls from height – 63 cases (19%), aggression - 44 cases (13%), other causes - 32 patients (10%) [Incidence by severity8], an exponential increase of mortality being correlated with the decrease in GCS score [19]. The Glasgow Coma Scale (GCS), described by Teasdale and Jennet in 1974, is the widespread used clinical classification for TBI severity, also having prognostic value [20].Classically, the GCS is stratified into 3 layers of severity by score frames, as it follows: severe (8 points or less), moderate (9-12p) or mild (13-15 p) [19]. Yet, a Romanian study asserted that a score over 7 shows a 90% probability of a favorable evolution or moderate neurological deficit. A score below 7 suggests a considerable increase in mortality or persistent vegetative state, the risk being 60-90% when GCS score is 3 [8].Accordingly, about 80% of TBI are found to be mild, 10% moderate and 10% severe [8].However, as described in another related epidemiological article, in the US, between 58% and 73% are mild, between 8% and 25% are moderate and between 6% and 8% are severe [3] Dawodu ST.Approximately 1.74 million people suffer minor TBIs causing temporary disability for at least one day [13].In the EU, the ratio between mild/ moderate/ severe TBI is 22/ 1.5/ 1 [Prevalence21]. The prevalence in the US is estimated to be of around 5 million people, i.e. 1-2% of the population have a disability related to TBI, while in Europe, 7,7 million population surviving after brain trauma, suffer from disabilities [3], the survivors are rather difficult to be systematically followed up on long term.Prevalence of TBI is not well documented very probably because most of the cases are not hospitalized [22].Estimations based on existing disabilities, showed that 2.5-6,500,000 Americans are living with TBI sequels, which often need long term complex management and care. As a short related complementary digression, it should be noted that in the US, 2.8 is considered the necessary number of full time employees (FTE) for such a patient: most often for months, sometimes for life [3]. The number of patients who require extended hospitalization and rehabilitation is a different measurement of the prevalence. In this respect, there are disparities between different parts of the world; these may be allocated to methods for checking various cases of TBI and in the way of referring to the cause of the lesion [23]. As prevalence refers to a long-term related status, precise data regarding TBI are helpful, including for improving rehabilitation programs [Mortality and morbidityFig. 5) [8]. TBI is the major cause of death injury induced. In the US, TBI represents about 40% of all deaths due to acute injury and the leading cause of mortality among people younger than 45 years. To note that, mortality due to TBI in the US was between 10 and 35 per 100.000 persons but decreasing across most etiologies and that death occurring outside hospital was of about 17 per 100,000 people, compared to that of hospitalized patients, which was estimated at about 6 per 100,000 people  [Financial cost24]. These injuries have both short-term and long-term effects on individuals, their kin and society; therefore, their overall cost is huge.Aside from the human suffering, which is incalculable, the direct and indirect costs are enormous, being appreciated, in the US, as amounting to $9- $10 billion spent on acute TBI and rehabilitation services annually. TBI may cost an additional amount of $1 billion per year by lost wages and/ or income taxes, and increased public assistance . In the context of many medicines that alter both the attention and promptness of reactions, persons in treatment, with potential risk may be repeatedly warned of the need for temporarily giving up driving.Globally, alcohol and/ or drugs intake are an important cause of death, also in relation to traffic accidents, so, a careful monitoring of the blood concentration in persons involved in road events and a good implementation of regulations regarding driving under the influence of alcohol and drugs must be rigorous  refers to the protective role of a very well known accessory device to be warred by different types of bikers. More precisely, it determines a 50,2% decrease in motor vehicle–related deaths “over 18 years”, after the introduction of the mandatory helmet law in Taiwan (passed in June 1997). It must be noted that in Romania wearing a helmet on motorcycle has been mandatory for almost 40 years.An interesting article  but not only, and also a significant multimodal challenge for the community: medically, socially, economically, educationally, advocationally and regulatory. According to the WHO and most public epidemiological data, lesions from TBI are a leading cause of death and respectively, for the survivors and their relatives, huge global burden below age 60 years [Continuous research/ documentation on such primary preventable events, education by appropriate means and implementation of appropriate legislation remain basic necessary prophylactic measures.Conflict of interests The authors declare no conflict of interests.


Objectives: Oral submucous fibrosis (OSMF) is the most prevalent precancerous condition in India. Low levels of lipids serves as a marker and prognostic indicator in the early detection of oral precancerous and cancerous states. In spite of the high prevalence and its potential to undergo malignant transformation, this condition has not widely been investigated with respect to the serum lipid levels. In the present study, an attempt was made to analyze the complete serum lipid profile, total cholesterol (TC), triglycerides (TG), high density lipoprotein (HDL) cholesterol, low density lipoprotein (LDL) cholesterol and very low density lipoprotein (VLDL) cholesterol in OSMF and controls.Material and Methods: The study was conducted in 45 clinically and histopathologically diagnosed cases of OSMF and 45 age and sex matched controls. The complete lipid profile including TC, TG, HDL cholesterol, LDL cholesterol and VLDL cholesterol was analyzed.Results: The serum lipid levels were significantly lower in the patients with OSMF than in the controls. When the values were compared between different disease stages, the maximum reduction of lipids was evident for stage 3 OSMF. From the present results, it is evident that the level of serum lipids decreases with progression of the disease. Conclusions: From these findings, it appears that the decrease in the lipid levels may be considered as a useful marker in the early diagnosis of oral premalignant condition like OSMF. Key words:Oral submucous fibrosis, lipids, premalignant condition. Oral submucous fibrosis [OSMF] is a chronic debilitating disease and a premalignant condition of the oral cavity . Oral caet al. (Over the years, the incidence of OSMF has increased in various parts of the Indian subcontinent . Murti eet al.  demonstret al. . Early det al. .In recent years, emphasis has been placed on detecting molecular markers from body fluid, such as saliva, urine and others, for detecting cancer, predicting prognosis, and monitoring disease progression. The idea of screening and following patients with malignancy by blood-based tests is appealing from several points of view including its ease, economic advantage, non-invasiveness, and possibility of repeated sampling .Usefulness of variations in tissue ⁄ blood cholesterol levels in diagnosis and treatment of various diseases has been studied by several workers. Although, its prime role in pathogenesis of coronary heart diseases has been consistently found, researchers have reported an association of plasma ⁄ serum lipids and lipoproteins with different cancers -10. An aet al. (Rose et al.  reportedet al. . Only a et al. . HoweverIn spite of the high prevalence and its potential to undergo malignant transformation, OSMF has not widely been investigated with respect to the serum lipid levels. Moreover, to our knowledge, literature pertaining to serum lipid levels in relation to OSMF and its clinical grading is rare. In the present study, an attempt was made to analyze the complete serum lipid profile in OSMF patients. Further, in this study the comparison in the serum lipid profile with grading in OSMF was done.et al. (The study was conducted in 45 clinically and histopathologically diagnosed cases of OSMF who attended the Department Of Oral Medicine and Radiology, Institute Of Dental Sciences, Bareilly, Uttar Pradesh-India. All the patients underwent recording of detailed history, and clinical diagnosis was made on the presence of characteristic features like vesicles, ulceration, mucosal blanching, burning, stiffness of oral mucosa, presence of characteristic fibrous bands and progressive inability to open the mouth. Also, histopathological examination was carried out in all the cases following incisional biopsy from the most affected area of the buccal mucosa. OSMF was divided clinically and functionally into three stages according to the criteria reported by Haider et al. , based oClinical Staging:• Stage 1: Faucial bands only• Stage 2: Faucial and buccal bands• Stage 3: Faucial, buccal and labial bandsFunctional Staging:• Stage 1: Mouth opening >20 mm• Stage 2: Mouth opening 11-19 mm• Stage 3: Mouth opening <10 mm45 healthy individuals, matched for age and sex, who consecutively presented at the same department for various other complaints, and had no adverse habits or oral lesions, or any other major illness in recent past, were included as controls. Informed consent was obtained from patients in both the case and control groups. The study was approved by the institutional ethical committee.Exclusion criteria were patients with systemic diseases/conditions that may be associated with alterations in the serum level of lipid profile like obese subjects weighing more than 20% above their ideal weight, known cases of uncontrolled diabetes mellitus, thyroid disorder, cardiac patients, liver dysfunction, malabsorption syndrome or those refusing for biopsy procedure and patients who had undergone or was on treatment for OSMF.The subjects of the study were as follows:• Group I [Clinical and functional stage 1 OSMF]: 15 patients• Group II [Clinical and functional stage 2 OSMF]: 15 patients • Group III [Clinical and functional stage 3 OSMF]: 15 patients• Group IV [Controls]: 45 patientsAll those patients who fulfill the above mentioned criteria were included in the study.- Estimation of lipid profileet al and Fossati et al for triglycerides and Phosphotungstic Acid method for HDL.First, the details of each patient, including medical history, were taken. After a clinical diagnosis of OSMF had been made, the purpose and procedure was explained to each patient, who then provided informed consent to participate. The patient was then asked to attend the next morning with a minimum of 12 hour of fasting for blood examination for complete lipid profile. Upon returning, the patient was asked to sit comfortably in a dental chair in a reclining position. A tourniquet was then applied above the right cubital fossa, and 3 ml of fas-ting blood was collected from the most prominent vein under aseptic conditions and was stored in test tubes. After the blood had coagulated, the test tube containing the blood was subjected to centrifugation for about 4-5 min at 2500 rpm. The test tube was then removed from the centrifuge, and the serum layer was pipetted into a vial, which was then stored in a refrigerator under protection from light. The serum lipid profile was analyzed using the kit Erba Mannheim  as per instructions provided by the manufacture, using semi-auto analyser. Complete lipid profile was checked in each case. TC, TG, HDL choles-terol, LDL cholesterol and VLDL cholesterol were monitored using Modified Roeschlau’s method for cholesterol, method of Wako and the modifications by McGowan VLDL and LDL levels were calculated using Friedwald’s formula:• VLDL= Triglycerides/5• LDL= Total Cholesterol – [VLDL+HDL]- Statistical analysisAll the variables from the study were statistically analyzed using the Statistical Package for the Social Sciences program [SPSS version 13.0]. Analysis of variance [ANOVA] was used to assess the statistical significance of differences between the stages of OSMF and the control group. Student’s t-test was used to compare serum lipid profile in all the three case groups with those in the control group.The present study comprises of 90 subjects, of whom 64 were males and 26 were females. Group IV comprised of 45 age and sex matched healthy individuals without any contributory habits or premalignant lesions/conditions. Group I, II, III comprised of 11, 10, 12 males and 4, 5, 3 females, respectively. The mean age of the patients in group I, II, III and IV was 25.13 years, 27.60 years, 34.53 years and 29.09 years respectively with most of the patients in the second and third decades of life. All of the patients had been taking areca nut in one form or another. The mean serum TC, serum HDL cholesterol, serum LDL, serum VLDL and serum TG levels in oral submucous fibrosis group were 151.07 mg/dL, 37.80 mg/dL, 95.15 mg/dL, 18.11 mg/dL and 94.18 mg/dL respectively. However, in the control group the corresponding values were 191.42 mg/dL, 51.36 mg/dL, 113.64 mg/dL, 26.43 mg/dL and 132.16 mg/dL respectively.  A statisSimilarly, in different stages of OSMF i.e stage 1, stage 2 and stage 3, these values were 174.20 mg/dL, 152.27 mg/dL and 126.73 mg/dL for TC; 42.47 mg/dL, 38.20 mg/dL and 32.73 mg/dL for HDL; 106.04 mg/dL, 97.27 mg/dL and 82.15 mg/dL for LDL; 25.69 mg/dL, 16.80 mg/dL and 11.85 mg/dL for VLDL; and 128.47 mg/dL, 92.80 mg/dL and 61.27 mg/dL for TC, respectively. These values were found to decrease progressively as one moved from control group to that of OSMF cases. It also shows that as the disease progresses i.e from OSMF stage 1 to stage 3, there is a gradual fall in lipid profile levels .In inter-group and intra-group comparisons of serum lipid, for all variables p-value using ANOVA showed that four groups differ significantly that is non-homogeneity for the means of four groups .For pair-wise comparison a Post-hoc Tukey test was conducted. It showed that for TC all pairwise differences were statistically significant. For HDL all pairwise difference was statistically significant except for Group I & II and Group II & III. For LDL all pairwise differences were statistically significant except for Group I & IV and Group I & II. For both VLDL and TG all pairwise differences were statistically significant except for Group I & IV and Group II & III .Oral submucous fibrosis is a chronic, insidious, disabling precancerous condition of the oral cavity . It has In this study, an attempt was made to analyze serum lipid levels in patients with OSMF in comparison with normal healthy individuals, thereby providing a marker and prognostic indicator in the early detection of oral cancer. An attempt was also made to compare these levels among different stages of the disease.The mean age of our study patients was 29.09 years. Most of our patients were in the second and third decades of life. Among our 45 OSMF patients, 32 were males and 13 were females. All the patients in our study had been chewing areca nut in some form. Areca nut chewing has been identified as the most important etiological factor of OSMF. A statistically significant reduction [P<0.001] was noted in serum TC, serum TG, serum HDL, serum LDL and serum VLDL between the control group and OSMF cases. Similarly, in different stages of OSMF i.e stage 1, stage 2 and stage 3, these values were found to decrease progressively as one moved from control group to that of OSMF cases. It also shows that as the disease progresses i.e from OSMF stage 1 to stage 3, there is a gradual fall in lipid profile levels. The reduced levels of serum lipid profile in OSMF patients may be a conse-quence of disease, probably mediated by the greater utilization of lipids for new membrane biogenesis.Tilakaratne et al.  reportedet al. (et al. (The results of the present study clearly shows that the serum TC, HDL, LDL, VLDL and TG were significantly reduced in OSMF patients group when compared with the control group. Patel et al.  showed t (et al.  reportedThe results of the present study show the evidence of an inverse relationship between serum lipid profile and OSMF. From these findings, it appears that the decrease in the lipid levels may be considered as a useful marker in the early diagnosis of oral premalignant condition like OSMF. Lipid profile can be added on to other tests as an additional indicator and can serve as another evaluating parameter to denote initial changes occurring during carcinogenesis. However, this will also have to be confirmed by using a larger sample size.


This disease is associated with approximately 3.4 million illnesses and 681,316deaths, particularly in Africa. Salmonella is a major cause of bloodstream infectionsworldwide, and HIV-infected persons and malaria-infected and malnourished childrenare at increased risk for the disease. We conducted a systematic literature review toobtain age group–specific, population-based invasive nontyphoidalSalmonella (iNTS) incidence data. Data were categorized by HIVand malaria prevalence and then extrapolated by using 2010 population data. Thecase-fatality ratio (CFR) was determined by expert opinion consensus. We estimatedthat 3.4 (range 2.1–6.5) million cases of iNTS disease occur annually . Africa, whereinfants, young children, and young adults are most affected, had the highestincidence  and number ofcases (1.9 [range 1.3–2.9] million cases). An iNTS CFR of 20% yielded 681,316 deaths annually. iNTS disease is a major cause ofillness and death globally, particularly in Africa. Improved understanding of theepidemiology of iNTS is needed.Nontyphoidal  Salmonella (NTS) disease, a major cause of diarrheal diseaseglobally, is estimated to cause 93 million enteric infections and 155,000 diarrheal deathseach year  and standardsecondary search terms  inthe following databases: PubMed, System for Information on Grey Literature in Europe,World Health Organization library, Food and Agriculture Organization of the UnitedNations, and Bath Information and Data Services. Each primary search term wascombined with all secondary search terms by using the AND operator. The time periodfor the search was January 1990–December 2012.We conducted a systematic review for iNTS incidence data by following guidelines ofthe Preferred Reporting Items for Systematic Reviews and Meta-Analyses  librarian during thedevelopment of the searches. Two teams of reviewers, each led by a co-author (T.T.A.or J.A.C.) and assisted by S.H. or D.B. (see Acknowledgments), processed thecitations for relevant publications in 3 stages: title, abstract, and full-textreview. If there was a disagreement during any stage, a tie-breaker from the otherteam (T.T.A. or J.A.C.) decided if the publication was appropriate. We includedforeign language articles that had at least an English translation of the title forthe first stage of review. If a citation met the requirement at this stage, we lookedfor the English translation of the abstract for the second and subsequent stages.We sought to obtain incidence data for all age groups. For studies identified throughthe systematic review that did not have incidence data for all age groups, we createdincidence profile curves to extrapolate the available incidence data to other agegroups in that particular population by using proportion curves that were calculatedfrom 2 sources that had complete case counts for each age group : US FoodPlasmodiumfalciparum transmission. Countries were classified as having low (0%),moderate (0.1%–10.0%), or high (>10.0%) proportions of their populations atrisk for malaria. Each of the iNTS incidence data points from the systematic review were assigned within the matrix,depending on the HIV and malaria epidemiology in the country source of the data. If amatrix cell had >1 data source after the iNTS incidence data were assigned, themean incidence from all sources was calculated as the reference incidence for thatmatrix cell. A minimum to maximum range was also identified by using the respectivevalues within a specific age group. If a matrix cell had only 1 incidence reference,the minimum and maximum incidences were calculated by using the median to mean ratioof cell A  due to iNTS. In addition, we solicited expert opinions in person at the8th International Conference on Typhoid Fever and Other Invasive Salmonelloses inDhaka, Bangladesh, 2013, and by email to develop a consensus CFR. We asked experts toprovide a single value, taking into account variations by co-infections, geography,age, infection rate, strain type, and study setting . The consensus CFR was then applied to the estimated number of iNTScases and the minimum and maximum range to estimate iNTS deaths and minimum andmaximum range. We also varied the CFR to understand the effect of the consensus CFRon global iNTS deaths. To understand the effect of the consensus CFR, we performed asensitivity analysis by applying the range of CFRs found in the systematicreview.We identified 9,739 unique citations, 2,029 (20.8%) of which were fromnon–English language journals. We excluded 9,011 by title screening alone. Of theremaining 728 citations, 700 were excluded by abstract screening. Of the remaining 28potential eligible citations with relevant abstracts, 10 were eligible for full textreview ; Table 1We found substantial geographic variation in age-specific incidence . OverallWe estimate 3.4 (range 2.1–6.5) million cases of iNTS disease each year and anoverall incidence of 49 (range 30–94) cases/100,000 population . Africa A range of potential CFRs, from 3% .We estimate that 3.4 (range 2.1–6.5) million cases of iNTS disease occurred in2010, with a CFR of 20%, for an overall incidence of 49 (range 30–94)cases/100,000 population and 681,316  deaths. Forcomparison, globally there were ≈1.2 million malaria-associated deaths and 1.5million HIV-associated deaths in 2010  occurred, accounting for more than half ofglobal cases. The region with the second highest number of cases in 2010 was Europe, but the number was substantially lower thanthat for Africa. The high number of cases in Europe is mainly driven by cases in Russia,Ukraine, and Estonia, 3 eastern European countries with large populations and relativelyhigher iNTS incidence profiles compared with those for countries in western Europe. Ourfindings, together with contemporary longitudinal descriptive literature, provideadditional evidence that a largely underappreciated epidemic of iNTS has been occurringin Africa, driven in part by co-infection with HIV or malaria. Antimicrobial drugresistance emerging across the continent is likely to influence the incidence of iNTSand associated deaths  is now common among iNTS strains inAfrica 41 in  2. Antim42,43 in  2. DecliOur results demonstrate substantial differences in the ratio of enteric to invasivedisease by geographic area. Whereas the incidence of enteric disease is highest in Asiaand Oceania, the incidence of invasive disease is highest in Africa, where the incidenceratio of invasive to enteric disease was 1:1 , which sOur study has several limitations. First, our rigorous systematic search forpopulation-based incidence data in the literature resulted in a limited number ofeligible sources. Despite this scarcity of eligible studies, the existing data are ofhigh quality and, we believe, representative of the settings from where they werecollected. Second, we did not consider other host risk factors, such as malnutrition andsickle cell disease. Excluding the effect of these conditions might result inunderestimation of iNTS incidence rates and the number of cases and deaths. Third, weassumed that the country-level prevalence for the HIV- and malaria-infected populationat risk is uniform, even though there is likely considerable subnational variation forthese measures. Fourth, we did not take into account the declines in malaria worldwidein the past decade, nor did we account for changes in HIV seroprevalence and progresswith provision of care and treatment globally. We used the most contemporary HIV andmalaria data available. However, iNTS incidence data used in these estimations may havebeen collected under different HIV seroprevalence and malaria population at-riskconditions than those observed in 2010. In addition, the incidence curve for the low HIVand high malaria grouping had an artificially high peak in the 35- to 39-year-old agegroup, which was due to the assumption we made that, across the rows in our incidencereference grid, the middle cell is the average of the 2 cells on each side. Because weknow that malaria is predominantly associated with iNTS disease among young children, webelieve this artifact is a result of our assumption. However, because there were only 2countries in this group (Comoros and Madagascar), and we believe that any potentialoverestimation of incidence in the 35- to 39-year-old age group in these countries willbe negligible. In addition, HIV and malaria also contribute differently as risk factorsin different ages in a given population. In populations with high HIV seroprevalence,the risk factors are highest among young adults and persons in younger age groups; inhigh malaria areas, the persons at highest risk for iNTS are those <5 years ofage.In the absence of a standard CFR for iNTS, we relied on expert opinion to estimate themost likely value. Our estimate of deaths from iNTS disease was high compared with theestimated number of deaths associated with HIV, malaria, and protein energymalnutrition. It is likely that many iNTS-associated deaths are currently counted asdeaths resulting from these underlying conditions. This high estimate might have beenbiased by expert opinions from hospital-based studies and experiences, especially inestablished research programs that have better diagnostics and appropriate antimicrobialdrug treatments and a higher level of care than in other settings.Salmonella disease burden globally, which is urgently needed to setthe scientific and policy agenda. We hope that our estimate will be refined in thefuture by incorporating new population-based surveillance data, improved estimates ofthe CFR, and more sophisticated approaches to extrapolation and modeling. We havedemonstrated that iNTS disease is a major cause of illness and death globally,particularly in Africa. Improved understanding of the epidemiology of iNTS is needed tounderpin effective efforts for prevention, control, and improved patient management.Despite these limitations, we provide a baseline estimate of invasive nontyphoidalCountry profile assignments for invasive nonthyphoidal Salmonella disease. Additional references for main text.


In spermatogenesis, spermatogonial cells differentiate to the haploid gametes. It has been shown that spermatogenesis can be done at in vitro condition. In vitro spermatogenesis may provide an open window to treat male infertility.The aim of this study was to evaluate the effects of a novel scaffold containing human serum albumin (HSA)/tri calcium phosphate nanoparticles (TCP NPs) on the mouse spermatogonial cell line (SCL).oC water for 30 min. In the next step, each scaffold was cut (2×2mm), placed into sterile well of microplate, and then incubated for 1, 2, and 3 days at 37oC with mouse SCL. After incubation, the cytotoxicity of the scaffolds was evaluated by different tests including 3--2, 5-diphenyl-tetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) assay, vital staining, and cell counting. On the other hand, the release of TCP NPs and HSA from the scaffolds was measured. First, TCP NPs were synthesized by reaction of calcium nitrate and diammonium phosphate at pH 13. Then, serial concentrations of TCP NPs were separately added to 500 mg/mL HSA, and incubated in the 100Based on microscopic observation, the size of cavities for all scaffolds was near 200-500 µm, and the size of TCP NPs was near 50-100 nm. All toxicity tests showed that the increase of TCP concentration in the scaffold did not affect mouse SCL. It means that the percentage of cell viability, LDH release, vital cells, and cell quantity was 85%, 105%, 90%, and 110%, respectively. But, the increase of incubation time led to increase of LDH release (up to 115%) and cell count (up to 115%). Also, little decrease of cell viability and vital cells was seen when incubation time was increased. Here, no release of TCP NPs and HSA was seen after increase of TCP concentration and incubation time. It can be concluded that the increase of TCP concentration in HSA/ TCP NPs scaffold does not lead to cytotoxicity. On the other hand, the increase of incubation time leads to increase of mouse SCL cell death. In this study, it was found that TCP NPs and HSA could not release from the scaffolds. In future, both proliferation and differentiation of mouse SCL on HSA/TCP NPs scaffold must be checked over more wide incubation times. Spermatogenesis is a developmental process in which spermatogonial cells differentiate to the haploid gametes , 2. Extret al used carbon nanotubes (CNTs) scaffold for spermatogonial cell. Based on their microscopic analysis, the cells maintained their shape and function on the scaffold for at least 21 days  and tri calcium phosphate nanoparticles (TCP NPs). HSA protein and TCP NPs were used because they are non-toxic and highly biocompatible. Also, many cells can uptake HSA by receptor-mediated endocytosis . ImportaMaterials3 were sourced from Zyst Fannavar Shargh Company (ZFS Co.), Yazd, Iran. Alamar blue reagent, LDH kit, and CellTiter-Glo Luminescent reagent were purchased from Invitrogen, UK.This study was a lab-trial study. It was done in Pajoohesh lab, at summer 2014. Calcium nitrates, diammonium phosphate, HSA, MTT, trypsin containing 5% EDTA, RPMI1640, fetal calf serum (FCS) were provided from Sigma-Aldrich Chemical Co, . Isopropanol (70% v/v), trypan blue, and HNOSynthesis of TCP NPs3)2) and 12 gr of diammonium phosphate ((NH3)2HPO4) were dissolved in 525 mL and 375 mL of distilled water (DW), respectively. Then, 25 mL of calcium nitrate were added to 25 mL of diammonium phosphate solution, adjusted to pH 13, and incubated for 6 hr at room temperature. After incubation, the synthesized product was rinsed with DW, and heated at 100oC for 30 min. Dried TCP NPs were hardly ball-milled for one hr, and then characterized by scanning electron microscopy (SEM)  and dynamic light scattering (DLS)   .Synthesis of HSA/TCP NPs scaffoldoC water and incubated for 30 min. After incubation, HSA/TCP NPs scaffold was freezen at -20oC, and then held in the 37oC water for 30 min. In this study, four HSA/TCP NPs scaffolds were prepared which their final concentration of TCP NPs was 10, 5, 2.5, and 1.2 mg/mL, respectively. Then, to measure the diameter of the scaffold cavities, they were cut, and observed by an optical microscopy (400x) and by a SEM. In this study, each type of scaffold was cut (2×2 mm), and placed into sterile well of ELISA microplate. For characterization of scaffold cavities, the scaffolds were cut by microtome, stained by Giemsa, and observed by optical microscopy   was separately added to 4 mL of 500 mg/mL HSA, and hardly mixed for 1 min straightaway, HSA/TCP NPs mixture was held in the 100n, Iran) .The preparation of mouse SCL suspensionoC. After initial incubation, the cells were washed and then transferred to culture flask containing fresh RPMI 1640-10% FCS, and incubated for 14 days at 37oC. Since the cells had been adhered to the culture flask, one mL of 100 mg/mL trypsin containing 5% EDTA was added and shaked for 15 min, in order to release of the adhered cells. Then, the cells were rinsed three times with RPMI1640-10% FCS, and the cell suspension was prepared (approximately 10000 cells/mL)  and antibiotic was added to cells, and incubated for 4 days at 37ells/mL) .MTT assayoC. Then, 50 µL of treated cells and 10 µL MTT were added and incubated at 37oC for 3 hr. In the next step, 100 µL of isopropanol (70%) was added to each well, and optical density (OD) of each well was read by ELISA reader  at 490 nm. Finally, the cell viability was measured. The cells which were not exposed to any scaffolds, considered as negative control  was separately added and incubated 5 min at room temperature. After incubation, the mean OD of each sample was read at 340 nm , and then divided to mean OD of control . In the negative control, the cells were not incubated with the scaffolds .Vital stainingoC. Finally, the quantity of live cells (not-stained cells) was recorded by optical microscopy, and the percentage of live cells was calculated. In negative control, the cells were not treated with the scaffolds  were separately incubated for 1, 2, and 3 days. Then, 10 µL of 10% trypan blue was added to 10 µL of treated cells, and incubated for 30 min at 25caffolds .The cell countFirst, each scaffold piece was separately incubated for 1, 2, and 3 days with 100 µL of cell suspension. After incubation, 10 µL of the cell supernatant was held on the Hemocytometer , the quantity of the cells was counted by optical microscopy, and then divided to the cell count of control . In the negative control, the cells were not treated with the scaffolds .The release of TCP NPs and HSAoC. After incubation, 50 µL of cell suspension was incubated with 50 µL of HNO3 for 24 hr, and then centrifuged at 10000 rpm for 15 min. Final, the quantity of Ca2+ ions was measured by atomic adsorption spectroscopy, and normalized to control. To measure HSA release, after incubation of the cell suspension with scaffold pieces, the cell suspension was centrifuged at 10000 for 15 min. Then, the supernatant OD of each sample was read at 280 nm , and then normalized to control. The cells which were not exposed to any scaffolds, considered as negative control  was used to evaluate the significant differences. In this test, p<0.05 was considered as significant difference.Characterization of TCP NPs and scaffoldsBased on the observation of scaffold slices by optical microscopy, the size of cavities for all scaffolds was near 200-500 µm . Since oThe cytotoxicity resultsThe results of MTT assay, LDH assay, vital staining, and cell counting are shown in the The release resultsThe release of TCP NPs and HSA is shown in the In this study, HSA/TCP NPs scaffold was synthesized, and its cytotoxic effects on the mouse SCL were investigated. Here, different tests including MTT assay, LDH assay, vital staining, and cell counting were used to show the effects of HSA/TCP NPs scaffold on mouse SCL. The main logics of this study were: 1) designing of a new scaffold based on natural components, 2) evaluating of the scaffold on the mouse SCL, and 3) using of HSA/TCP NPs scaffold in future for male infertility. The results of all tests revealed two important findings: The first important finding was that the increase of TCP NPs in the scaffold did not lead to change of cytotoxicity. The authors explain that the result is due to powerful binding of TCP NPs and HSA in the scaffold. This phenomenon is confirmed by the release test. As mentioned, no release of TCP NPs and HSA was observed after increase of concentration and incubation time. The authors think that the higher concentrations than 10 mg/mL must be tested in the future study. The reason of usage of these concentrations is addressed to initial concentration of TCP NPs, i.e., we could not prepare the suspension of TCP NPs higher than 100 mg/mL because of rapid precipitation. Supervising, increase of TCP concentration not only did not affect the result of cytotoxicity and release tests, but also did not change the strength (solidity) appearance.The second important finding was that the increase of incubation time led to few increase of LDH release and cell proliferation, and few decrease of cell viability and vital cells. Overall, these results showed that the more incubation time, the higher cytotoxic effects. It seems that the bad environmental and nutritional conditions are the reason of the higher cytotoxic effects when SCL incubated more times with scaffolds. Note, this pattern is common in the cell culture, and is not a supervising result , 11. In et al used the albumin-binding domain expressed by gram-positive bacteria as a part of scaffold. Not only, albumin-binding domain is important for bacteria, but also can be used for protein purification and immobilization  and olfactory ensheathing cells (OECs). ications . Also, Jst cells . Ferreroet al developed an albumin-derived peptide scaffold which was from the sequence of the IIA binding site of HSA. They revealed that the scaffold could bind to various molecules and had catalytic properties . Furthermore, Weszl feration . In thisferation . TCP leaferation . Al-MunaLickorisha et al synthesized a polyester/TCP scaffold for bone tissue repair. They presented the third generation of scaffold with enhanced in vitro and in vivo performances  and BSA. Based on their results, controllable dual-release was seen by the PLGA/TCP scaffold (et al synthesized TCP porous scaffold by albumin as a foam generating agent. They indicated that a variety of scaffold geometries has been prepared by the heating of HSA (et al used CNTs as a scaffold. They showed the cells maintained their shape and function on the scaffold for at least 21 days (ormances . Yang etscaffold . Ribeirog of HSA . In case 21 days . et al showed the effect of poly L-lactic acid scaffold on the mouse spermatogonial stem cell. They revealed the scaffold increased formation of cell cluster and differentiation of spermatogonial stem cells (Shakeriet al investigated electrospun nanofibrillar scaffolds. They showed increase number of colonies, the number of cells in each colony, and the average area of colonies after 7 days (et al designed a novel scaffold based on HSA and hydroxyapatite NPs. They showed the scaffold had no cytotoxic effects on mouse SCL (Eslahi em cells . Shakerir 7 days . Jebali ouse SCL .Taken together, this study demonstrated that HSA/TCP NPs scaffold could be easily synthesized, and has no remarkably cytotoxic effects on mouse SCL. It can be concluded that the increase of TCP concentration in HSA/TCP NPs scaffold does not affect its cytotoxicity. Moreover, the increase of incubation time led to increase of cell death. Here, it was found that TCP NPs and HSA could not release from the scaffolds. HSA/TCP NPs scaffold must be used in vivo, in order to evaluate its efficacy at actual conditions. Certainly, the proliferation and differentiation of human spermatogonial cells must be checked over different incubation times. In the future, the scaffold may be beneficial to treat male infertility.